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Biological Effect of Hydrolyzed Collagen on Bone Metabolism
Critical Reviews In Food Science and Nutrition
Abstract
Osteoporosis is a chronic and asymptomatic disease characterized by low bone mass and skeletal microarchitectural deterioration, increased risk of fracture and associated co-morbidities most prevalent in the elderly. Due to an increasingly aging population, osteoporosis has become a major health issue requiring innovative disease management. Proteins are important for bone by providing building blocks and by exerting specific regulatory function. This is why adequate protein intake plays a considerable role in both bone development and bone maintenance. More specifically, since an increase in the overall metabolism of collagen can lead to severe dysfunctions and a more fragile bone matrix and because orally administered collagen can be digested in the gut, cross the intestinal barrier, enter the circulation and become available for metabolic processes in the target tissues, one may speculate that a collagen-enriched diet provides benefits for the skeleton. Collagen-derived products such as gelatin or hydrolyzed collagen (HC) are well acknowledged for their safety from a nutritional point of view, however what is their impact on bone biology? In this manuscript, we critically review the evidence from literature for an effect of HC on bone tissues in order to determine whether HC may represent a relevant alternative in the design of future nutritional approaches to manage osteoporosis prevention.
... Bone is a highly mineralized connective tissue [13,17]. In synovial joints, the bone consists of the trabecular bone and the calcified cartilage zone. ...
... When consumed, CHs, BAPs, and any other nutraceuticals or medications taken orally must undergo digestion and absorption before exerting their bioactive effects [17,75]. It is mainly in the small intestine (SI) that proteins are broken down into peptide and AA components. ...
... Human clinical studies have shown that BAPs and AAs generated from orally ingested CHs and gelatin appear in the systemic circulation and are excreted in urine [45][46][47][48][49][50]92]. These bioavailable BAPs and AAs have also been shown to build up in joint tissues such as cartilage and bone [17,[93][94][95], which likely explains why CHs demonstrate possible clinical potential. As with bioaccessibility measurements, the assessment of CH and peptide bioavailability using human trials continues to be lengthy, costly, and with restricted experimental opportunities for sampling due to ethical considerations, as well as limited methodologies for identifying and detecting both peptides and AAs in plasma or blood. ...
Osteoarthritis (OA) is the most common joint disorder, with a social and financial burden that is expected to increase in the coming years. Currently, there are no effective medications to treat it. Due to limited treatment options, patients often resort to supplements, such as collagen hydrolysates (CHs). CHs are products with low molecular weight (MW) peptides, often between 3 and 6 kDa, and are a result of industrialized processed collagen. Collagen extraction is often a by-product of the meat industry, with the main source for collagen-based products being bovine, although it can also be obtained from porcine and piscine sources. CHs have demonstrated positive results in clinical trials related to joint health, such as decreased joint pain, increased mobility, and structural joint improvements. The bioactivity of CHs is primarily attributed to their bioactive peptide (BAP) content. However, there are significant knowledge gaps regarding the digestion, bioavailability, and bioactivity of CH-derived BAPs, and how different CH products compare in that regard. The present review discusses CHs and their BAP content as potential treatments for OA.
... The newly formed osteoblast cells result in a favorable OPG/RANKL ratio (Osteoprotegerin/ Receptor activator of nuclear factor kappa-Β ligand) which inhibits the differentiation of stem cells into osteoclasts ( Figure 2). The net effect of BCP is to increase the number of osteoblast cells and decreases the number of osteoclast cells [12] . BCP increases bones' organic component and improves vertebrae's metabolism, microarchitecture, and biomechanical resistance. ...
... Supplementation with calcium and vitamin D formulation and appropriate catabolic or anabolic treatment for osteoporosis helps improve bone mineral density and reduces the risk of fractures [6] . This process is further facilitated by using bioactive collagen peptides (Type 1), which help replenish bones' collagen backbone [12] . Type II collagen supplementation helps slow down osteoarthritic joints' wear and tear and supports collagen synthesis and the standard of care treatment [8] . ...
... It will help manage 60 plus fitness addict senior citizens, 60 plus senior citizens nonresponsive to the standard of care, and 20 plus fitness freak youngsters. The importance of "nutribiography" and geroprotection is critical in the outpatient clinic of every orthopedician [12] . Nutribiography and nutraceutical science will help every clinician/ surgeon optimize/ individualize the intervention approaches and help advance patient care, and facilitate geroprotection. ...
... While this contamination may not necessarily have biological consequences in vivo, it can act as a strong confounder in vitro (Heinrich et al. 2023). Hence, there is a need for models using digested CH, for example, by using serum obtained after oral CH ingestion, but this has not yet been tested in in vitro models in canine or human OA (Daneault et al. 2017). ...
... Lastly, further research on the biological mechanisms of CH in dog-specific in vitro models may give additional insight on the patient population that may benefit most from its supplementation. Ideally, these studies simulate changes in CH-peptides that occur during gastro-intestinal passage, digestion and absorption, for instance by using CH-enriched serum methods (Daneault et al. 2017;Wauquier et al. 2019). ...
Osteoarthritis (OA) is a common disease in dogs with severe impact on their welfare. The multimodal management of OA includes feeding therapeutic diets and nutraceuticals to slow down OA progression. Collagen hydrolysates (CH) are a nutritional supplement that may exert anabolic effects on osteoarthritic joint cartilage as well as disease-modifying effects. After oral intake, CH is absorbed, mainly as amino acids, di- and tripeptides that are transported amongst others to the joint. In addition to reducing cartilage degradation, CH metabolites may reduce synovial inflammation and subchondral bone sclerosis during OA. Preliminary evidence in dogs suffering from the consequences of OA support the clinical efficacy of CH with reported reductions in lameness. However, effects on biomarker level of cartilage metabolism and inflammation are inconclusive. Additionally, current studies show a lack of standardised dosing regimens and the use of not validated outcomes. Future work should therefore elucidate further on the bioavailability of CH in dogs in order to establish adequate dosing recommendations. Furthermore, high-quality placebo-controlled randomised controlled trials are essential to dstudies have evaluated the cetermine the clinical efficacy of CH to reduce lameness, prevent OA progression and thereby improve the level of evidence.
... These mechanisms include the anti-inflammatory and antioxidant capacities of collagen peptides, and its ability to stimulate collagen synthesis and promote bone formation [22][23][24]. In vitro and in vivo studies showed a reduction of pro-inflammatory cytokines, including IL-1β, IL-6, and TNF-α after collagen peptide administration [22,25]. Collagen peptide also displays antioxidant activities measured by oxygen radical absorbance capacity and radical scavenging assay [24]. ...
... Bovine collagen hydrolysate was shown to stimulate osteoblast differentiation and mineralized bone matrix formation through increased runt-related transcription factor 2 (Runx2) expression and activity [23], and may serve as an effective supplement for preventing bone loss by significantly enhancing the organic substance content of bone [28]. The promotion of bone formation could be further explained by the downregulation of the aforementioned pro-inflammatory molecules, because these cytokines are responsible for the upregulation of receptor activator for nuclear factor kappa-B ligand (RANKL) for osteoclast recruitment, which may lead to bone loss [25]. Current scientific research indicates that consistent consumption of collagen peptides has been associated with a reduction in joint pain and bone density loss [21,22,29]. ...
Background
The management of knee osteoarthritis involves various treatment strategies. It is important to explore alternative therapies that are both safe and effective. Collagen peptides have emerged as a potential intervention for knee osteoarthritis. This study aims to evaluate the analgesic effects and safety of collagen peptide in patients diagnosed with knee osteoarthritis.
Methods
We conducted a systematic literature search following the guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement. Multiple databases including PubMed, Scopus, EMBASE, Web of Science, Cochrane, and ClinicalTrials.gov were searched for randomized controlled trials (RCTs) published up to 27 May 2023 that focused on the analgesic outcomes and adverse events associated with collagen peptides or hydrolyzed collagen in patients with osteoarthritis. We assessed the quality of the included studies and the strength of evidence using the Cochrane ROB 2.0 tool and Grading of Recommendations, Assessment, Development, and Evaluations.
Results
Four trials involving 507 patients with knee osteoarthritis were included and analyzed using the random-effects model. All these trials were considered to have a high risk of bias. Our results revealed a significant difference in pain relief between the collagen peptide group and the placebo group in patients with knee osteoarthritis (standardized mean difference: − 0.58; 95% CI − 0.98, − 0.18, p = 0.004; I2: 68%; quality of evidence: moderate). However, there was no significant difference in the risk of adverse events between collagen peptide and placebo (odds ratio: 1.66; 95% CI 0.99, 2.78, p = 0.05; I2: 0%; quality of evidence: very low).
Conclusions
Our findings demonstrate significant pain relief in patients with knee osteoarthritis who received collagen peptides compared to those who received placebo. In addition, the risk of adverse events did not differ significantly between the collagen peptide group and the placebo group. However, due to potential biases and limitations, well-designed randomized controlled trials are needed to validate and confirm these findings.
... Possibly, the presence of proline and hydroxyproline in its structure is responsible for the formation of bioactive peptides, since both provide resistance against the action of proteases, limiting hydrolysis [19]. The bioaccessibility of amino acids and peptides in the intestine, their absorption rate and availability in the bloodstream define their bioavailability for the regulation of metabolic processes in target tissues [20]. ...
... These variations in experimental protocols (target population, sex, dose, duration of treatment, presence, or absence of orthopedic alterations) contribute to the lack of agreement on the observed results. Furthermore, a better understanding of the physiological mechanisms that promote these benefits is still required [20]. Therefore, considering the positive effect of collagen consumption on joint diseases and physiology and the wide variety of intervention protocols, future clinical studies should better address guidelines regarding the optimal dose for consumption, the type of collagen, and duration of intervention. ...
Collagen is one of the main components of the extracellular matrix of the dermis and articular cartilage and influences the body’s mechanical, organizational, and tissue formation properties. Produced from food industry by-products, it is considered a nutraceutical product widely used as an ingredient or supplement in food, pharmaceutical, and cosmetic industries. This study aimed to conduct a literature review on the scientific evidence regarding the beneficial effects of collagen consumption in the treatment of skin and orthopedic diseases. Literature data have shown that hydrolyzed collagen supplementation promotes skin changes, such as decreased wrinkle formation; increased skin elasticity; increased hydration; increased collagen content, density, and synthesis, which are factors closely associated with aging-related skin damage. Regarding orthopedic changes, collagen supplementation increases bone strength, density, and mass; improves joint stiffness/mobility, and functionality; and reduces pain. These aspects are associated with bone loss due to aging and damage caused by strenuous physical activity. Thus, this review addresses the economic and health potential of this source of amino acids and bioactive peptides extracted from food industry by-products.
... These peptides have various biological activities, such as stimulating the growth of skin fibroblasts 3,8 , promoting osteoblast differentiation 9,10 , and chemotactic activity on skin fibroblasts, tenocytes, and peripheral blood neutrophils [11][12][13] . The collagenderived bioactive oligopeptides possessing high oral bioavailability are possibly the major contributing factor to the beneficial effects of collagen hydrolysate ingestion on bone 14 , joint 15 , skin 16 , and other targets 17 , which have been reported by many groups, particularly in recent years. ...
... Similar discussions can be applied to the promotional activity of osteoblast differentiation by Gly-3Hyp-4Hyp observed here. Our in vitro data suggest the therapeutic potential of this bioactive tripeptide for damage in skin and bone, both showing beneficial effects of collagen hydrolysate ingestion in several clinical trials 14,16,31 . Although Pro-4Hyp deposited in tissue after oral administration of the peptide is reported to be further converted to amino acids and other metabolites 24,45 , Gly-3Hyp-4Hyp would more effectively function in tissue owing to the exceptionally high stability. ...
There are increasing reports demonstrating high bioavailability of 4-hydroxyproline (4Hyp)-containing oligopeptides after oral ingestion of collagen hydrolysate and their bioactivity. In contrast, no study investigates the fate of another collagen-specific but minor amino acid, 3Hyp. Here, we identified Gly-3Hyp-4Hyp tripeptide in human blood at high concentrations, comparable to other 4Hyp-containing oligopeptides, after ingesting porcine skin collagen hydrolysate. Additionally, Gly-3Hyp-4Hyp uniquely maintained the maximum concentration until 4 h after the ingestion due to its exceptionally high resistance to peptidase/protease demonstrated by incubation with mouse plasma. In mice, oral administration of collagen hydrolysate prepared from bovine tendon, which contains a higher amount of 3Hyp, further increased blood Gly-3Hyp-4Hyp levels compared to that from bovine skin. Furthermore, Gly-3Hyp-4Hyp showed chemotactic activity on skin fibroblasts and promoted osteoblast differentiation. These results highlight the specific nature of the Gly-3Hyp-4Hyp tripeptide and its potential for health promotion and disease treatment.
... The investigational product in this current trial (CalGo™, produced by HBC) is derived from offcuts of salmon filet production, and contains 60% calcium salts, primarily in the form of natural hydroxyapatite (nHAP) and 36% protein, mainly Collagen. Studies have shown that the intake of Collagen has a positive effect of increasing BMD in several in-vivo bone growth models [10]. CalGo™ has also shown an osteoinductive effect by increasing osteoblast activity in-vitro [11] and in-vivo [12]. ...
... In clinical studies, natural hydroxyapatite appears to be more effective in building bone than traditional supplementation with calcium carbonate [14,15]. Bone meal also has a high collagen content, which appears to confer osteo-inductive properties, as indicated by several studies, both in-vitro and in-vivo [10]. focused on delivering high levels of elemental calcium but this has resulted in a rather limited impact on bone density and strength. ...
... The investigational product in this current trial (CalGo™, produced by HBC) is derived from offcuts of salmon filet production, and contains 60% calcium salts, primarily in the form of natural hydroxyapatite (nHAP) and 36% protein, mainly Collagen. Studies have shown that the intake of Collagen has a positive effect of increasing BMD in several in-vivo bone growth models [10]. CalGo™ has also shown an osteoinductive effect by increasing osteoblast activity in-vitro [11] and in-vivo [12]. ...
... In clinical studies, natural hydroxyapatite appears to be more effective in building bone than traditional supplementation with calcium carbonate [14,15]. Bone meal also has a high collagen content, which appears to confer osteo-inductive properties, as indicated by several studies, both in-vitro and in-vivo [10]. focused on delivering high levels of elemental calcium but this has resulted in a rather limited impact on bone density and strength. ...
... Orally administered gelatin is digested in the gut, crosses the intestinal barrier, becoming available for the metabolic process in the tissues. It has been proposed that HC peptides are only digested reaching the blood by passing through the enterocyte (transcytosis) at a level of approximately 10% [20]. ...
... In a first study, Adam et al. showed the effects of calcitonin, alone or in combination with a hydrolyzed collagen, on bone metabolism in postmenopausal women. The results underlined that a daily ingestion of 10 g of hydrolyzed collagen, in association with intramuscular injection of calcitonin (100 UI) twice a week for 24 weeks, enhanced and prolonged the effect of the drug as shown by a fall in urinary pyridinoline cross-link levels [20]. ...
This chapter discusses the physiologic, metabolic, and clinical aspects of collagen, including the role of nutritional factors in a new nosographic entity, called “extended collagen carential disease.” Except water and possibly fats, carbohydrates, and other structural proteins, perhaps there is more collagen in the mammalian body than anything else. Moreover, collagen participates in almost all of the body functions, adjusting its structure constantly in response to changes in environment, development, growth, and external clues. Collagens found in bones and nails are different from collagens found in body fluids and other biological structures, such as basement membrane, skin, tendons, muscles, and hair. The ubiquity of collagen functions accounts for its phylogenetic ubiquity, involving any tissue, organ, and apparatus. This is shown by the so-called “collagen carential disease,” involving nails, hair, osteoarticular and gastrointestinal systems. For instance, the Ehlers-Danlos syndrome describes another group of genetic collagen disorders, affecting the collagen processing and structure. Some of them are inherited in an autosomal dominant manner, while others emerge in the absence of essential nutritional factors. It is the case of Vitamin C, which plays a critical role in the maintenance of a normal mature collagen network. Hence, the idea of an “extended collagen carential disease,” applicable to the absence of essential nutritional factors.
... The terms "hydrolyzed gelatin," "collagen hydrolysate," "hydrolyzed collagen," or sometimes "collagen peptides" used in publications describe the same product. Gelatin is obtained by partial thermal hydrolysis of collagen, which partially separates the chains by destroying the cross-links (Daneault et al. 2015). Upon hydrolysis, the breaking of peptide bonds implies the fragmentation of collagen molecules leading to the disruption of hydrogen bonds within collagen helices. ...
This review provides a general overview of collagen structure, biosynthesis, and biological properties, with a particular focus on marine collagen sources, especially fisheries discards and by-catches. Additionally, well-documented applications of collagen are presented, with special emphasis not only on its final use but also on the processes enabling sustainable and safe recovery from materials that would otherwise go to waste. Particular attention is given to the extraction process, highlighting key aspects essential for the industrialization of fish sidestreams, such as hygiene standards, adherence to good manufacturing practices, and ensuring minimal environmental impact. In this context, the EcoeFISHent projects have provided valuable insights, aiming to create replicable, systemic, and sustainable territorial clusters based on a multi-circular economy and industrial symbiosis. The main goal of this project is to increase the monetary income of certain categories, such as fishery and aquaculture activities, through the valorization of underutilized biomass.
... HC also shows a near-significant increase in protein intake despite not reaching a daily protein intake of 60 g per day. However, current research shows no benefits of collagen supplementation on muscle protein synthesis or the loss of lean mass [3,17,29], but collagen could benefit other post-MBS issues like bone health and wound healing [6,8]. Therefore, HC may need to be combined with another protein source that stimulates muscle protein synthesis to prevent FFM loss. ...
Purpose
Disproportional fat-free mass loss often occurs post-bariatric surgery, partly due to insufficient protein intake during the post-surgery recovery phase. We compared five protein-enhancing strategies (PES) on patient tolerability, satisfaction and protein intake.
Materials and Methods
Ninety-four participants, scheduled for bariatric surgery, were enrolled and allocated to either of the following: (1) whey powder, (2) hydrolysed collagen powder, (3) plant-based powder, (4) protein-rich products, (5) protein gel, or control. PES groups were instructed to add 30 g of powder or 2 gels or protein products to their diet. Patient satisfaction and tolerability were evaluated with questionnaires. Dietary intake was assessed prior to and during PES use.
Results
Seven patients dropped out (i.e. loss of contact, personal reasons or post-surgery complications) yielding an analytical cohort of 87 participants. The majority of patients (61%) did not experience dietary complaints from PES and could use PES ≥ 5 days of the week. PES non-usage was mainly related to taste dislike (58%). Hydrolysed collagen scored highest on tolerability and satisfaction: 86% of the participants could use HC ≥ 5 days and 71% were satisfied with the product. PES increased protein intake from 54.7 ± 21.5 g/day to 64.7 ± 23.4 g/day during the intervention (p = 0.002), which differed from the control group (+ 10.1 ± 24.5 g/day vs. − 6.3 ± 23.8 g/day for controls, p = 0.019). Whey showed the highest increase, namely + 18.3 ± 16.3 g/day (p = 0.009).
Conclusion
PES were tolerated by the majority of participants, and an improved protein intake with PES use was seen. However, the taste of the products could be improved to further enhance satisfaction and tolerability.
Graphical Abstract
... Therefore, there is an urgent need to discover new effective natural substances to play an anti-osteoporosis role with less side effects. There is growing evidence that bioactive peptides can promote bone health [6][7][8][9] . Hydrolyzed egg yolk peptide (YPEP) is a bioactive peptide extracted from egg yolk. ...
Hydrolyzed egg yolk peptide (YPEP) was shown to increase bone mineral density in ovariectomized rats. However, the underlying mechanism of YPEP on osteoporosis has not been explored. Recent studies have shown that Wnt/β-catenin signaling pathway and gut microbiota may be involved in the regulation of bone metabolism and the progression of osteoporosis. The present study aimed to explore the preventive effect of the YPEP supplementation on osteoporosis in ovariectomized (OVX) rats and to verify whether YPEP can improve osteoporosis by regulating Wnt/β-catenin signaling pathway and gut microbiota. The experiment included five groups: sham surgery group (SHAM), ovariectomy group (OVX), 17-β estradiol group (E2: 25 µg /kg/d 17β-estradiol), OVX with low-dose YPEP group (LYPEP: 10 mg /kg/d YPEP) and OVX with high-dose YPEP group (HYPEP: 40 mg /kg/d YPEP). In this study, all the bone samples used were femurs. Micro-CT analysis revealed improvements in both bone mineral density (BMD) and microstructure by YPEP treatment. The three-point mechanical bending test indicated an enhancement in the biomechanical properties of the YPEP groups. The serum levels of bone alkaline phosphatase (BALP), bone gla protein (BGP), calcium (Ca), and phosphorus (P) were markedly higher in the YPEP groups than in the OVX group. The LYPEP group had markedly lower levels of alkaline phosphatase (ALP), tartrate-resistant acid phosphatase (TRAP) and C-terminal telopeptide of type I collagen (CTX-I) than the OVX group. The YPEP groups had significantly higher protein levels of the Wnt3a, β-catenin, LRP5, RUNX2 and OPG of the Wnt/β-catenin signaling pathway compared with the OVX group. Compared to the OVX group, the ratio of OPG/RANKL was markedly higher in the LYPEP group. At the genus level, there was a significantly increase in relative abundance of Lachnospiraceae_NK4A136_group and a decrease in Escherichia_Shigella in YPEP groups, compared with the OVX group. However, in the correlation analysis, there was no correlation between these two bacteria and bone metabolism and microstructure indexes. These findings demonstrate that YPEP has the potential to improve osteoporosis, and the mechanism may be associated with its modulating effect on Wnt/β-catenin signaling pathway.
... SCP could affect osteoclasts by increasing the OPG/RANKL ratio, potentially reducing bone resorption. Further research is necessary to understand the exact mechanism [26] . ...
... Collagen constitutes the principal protein in animal bones and connective tissues, accounting for approximately 30% of the total protein in animals and serving as the most abundant protein in mammals [5,6]. At least 29 types of collagen have been identified, which have different amino acid compositions and sequences to perform different biological functions in the organism [7,8]. ...
Antler ossified tissue has been widely used for the extraction of bioactive peptides. In this study, collagen was prepared from antler ossified tissue via acetic acid and pepsin. Five different proteases were used to hydrolyze the collagen and the hydrolysate treated by neutrase (collagen peptide named ACP) showed the highest DPPH radical clearance rate. The extraction process of ACP was optimized by response surface methodology, and the optimal conditions were as follows: a temperature of 52 °C, a pH of 6.1, and an enzyme concentration of 3200 U/g, which resulted in the maximum DPPH clearance rate of 74.41 ± 0.48%. The peptides (ACP-3) with the strongest antioxidant activity were obtained after isolation and purification, and its DPPH free radical clearance rate was 90.58 ± 1.27%; at the same time, it exhibited good scavenging activity for ABTS, hydroxyl radical, and superoxide anion radical. The study investigated the protective effect of ACP-3 on oxidative damage in HaCaT cells. The findings revealed that all groups that received ACP-3 pretreatment exhibited increased activities of SOD, GSH-Px, and CAT compared to the model group. Furthermore, ACP-3 pretreatment reduced the levels of ROS and MDA in HaCaT cells subjected to H2O2-induced oxidative damage. These results suggest that collagen peptides derived from deer antler ossified tissue can effectively mitigate the oxidative damage caused by H2O2 in HaCaT cells, thereby providing a foundation for the utilization of collagen peptides in pharmaceuticals and cosmetics.
... For this reason, attempts to find novel therapies or solutions to facilitate skin regeneration are crucial from a socio-economic point of view. In recent years, in order to accelerate and improve wound healing, different skin substitutes have been designed [28,30,31] that are mainly based on the most abundant components of the ECM [32][33][34], such as collagen [7][8][9]. This latter is mainly of mammalian origin, a feature that may present medical and ethical concerns; therefore, an economically sustainable and disease-transmission safe collagen source is still needed and marine collagens are a promising alternative [35]. ...
The mutable collagenous tissue (MCT) of echinoderms possesses biological peculiarities that facilitate native collagen extraction and employment for biomedical applications such as regenerative purposes for the treatment of skin wounds. Strategies for skin regeneration have been developed and dermal substitutes have been used to cover the lesion to facilitate cell proliferation, although very little is known about the application of novel matrix obtained from marine collagen. From food waste we isolated eco-friendly collagen, naturally enriched with glycosaminoglycans, to produce an innovative marine-derived biomaterial assembled as a novel bi-layered skin substitute (Marine Collagen Dermal Template or MCDT). The present work carried out a preliminary experimental in vivo comparative analysis between the MCDT and Integra, one of the most widely used dermal templates for wound management, in a rat model of full-thickness skin wounds. Clinical, histological, and molecular evaluations showed that the MCDT might be a valuable tool in promoting and supporting skin wound healing: it is biocompatible, as no adverse reactions were observed, along with stimulating angiogenesis and the deposition of mature collagen. Therefore, the two dermal templates used in this study displayed similar biocompatibility and outcome with focus on full-thickness skin wounds, although a peculiar cellular behavior involving the angiogenesis process was observed for the MCDT.
... As expected, the most abundant protein functional domain in the Phylobone dataset is collagen, which is present in 12% of human proteins, covering 90% of the organic fraction of the bone ECM. 2 Collagen is essential for homeostasis maintenance, and it serves as a scaffold to many other macromolecules and hydroxyapatite, enabling cell attachment and bone resistance to mechanical forces. 1,42,43 Furthermore, 214 non-collagenous functional domains are present in human sequences of the dataset ( Fig. 2 and Table S4). They are mostly common in bone formation, resorption, cell attachment, or as intermediaries in a variety of metabolic pathways. ...
The bone extracellular matrix (ECM) contains minerals deposited on highly crosslinked collagen fibrils and hundreds of non-collagenous proteins. Some of these proteins are key to the regulation of bone formation and regeneration via signaling pathways, and play important regulatory and structural roles. However, the complete list of bone extracellular matrix proteins, their roles, and the extent of individual and cross-species variations have not been fully captured in both humans and model organisms. Here, we introduce the most comprehensive resource of bone extracellular matrix (ECM) proteins that can be used in research fields such as bone regeneration, osteoporosis, and mechanobiology. The Phylobone database (available at https://phylobone.com ) includes 255 proteins potentially expressed in the bone extracellular matrix (ECM) of humans and 30 species of vertebrates. A bioinformatics pipeline was used to identify the evolutionary relationships of bone ECM proteins. The analysis facilitated the identification of potential model organisms to study the molecular mechanisms of bone regeneration. A network analysis showed high connectivity of bone ECM proteins. A total of 214 functional protein domains were identified, including collagen and the domains involved in bone formation and resorption. Information from public drug repositories was used to identify potential repurposing of existing drugs. The Phylobone database provides a platform to study bone regeneration and osteoporosis in light of (biological) evolution, and will substantially contribute to the identification of molecular mechanisms and drug targets.
... Bovine collagen hydrolysate was shown to stimulate osteoblast differentiation and mineralized bone matrix formation through increased runt-related transcription factor 2 (Runx2) expression and activity [23], and may serve as an effective supplement for preventing bone loss by signi cantly enhancing the organic substance content of bone [28]. The promotion of bone formation could be further explained by the downregulation of the aforementioned pro-in ammatory molecules, because these cytokines are responsible for the upregulation of receptor activator for nuclear factor kappa-B ligand (RANKL) for osteoclast recruitment, which may lead to bone loss [25]. Current scienti c research indicates that consistent consumption of collagen peptides has been associated with a reduction in joint pain and bone density loss [21,22,29]. ...
Background:
The management of knee osteoarthritis involves various treatment strategies. It is important to explore alternative therapies that are both safe and effective. Collagen peptides have emerged as a potential intervention for knee osteoarthritis. This study aims to evaluate the analgesic effects and safety of collagen peptide in patients diagnosed with knee osteoarthritis.
Methods:
We conducted a systematic literature search following the guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement. Multiple databases including PubMed, Scopus, EMBASE, Web of Science, Cochrane, and ClinicalTrials.gov were searched for randomized controlled trials (RCTs) published up to 27 May, 2023 that focused on the analgesic outcomes and adverse events associated with collagen peptides or hydrolyzed collagen in patients with osteoarthritis. We assessed the quality of the included studies and the strength of evidence using the Cochrane ROB 2.0 tool and Grading of Recommendations, Assessment, Development, and Evaluations.
Results:
Four trials involving 507 patients with knee osteoarthritis were included and analyzed using the random-effects model. All of these trials were considered to have a high risk of bias. Our results revealed a significant difference in pain relief between the collagen peptide group and the placebo group in patients with knee osteoarthritis (standardized mean difference of the mean difference: -0.63; 95% CI: -0.86, -0.39, p < 0.00001; I²: 52%; quality of evidence: moderate). However, there was no significant difference in the risk of adverse events between collagen peptide and placebo (odds ratio: 1.66; 95% CI: 0.99, 2.78, p = 0.05; I2: 0%; quality of evidence: very low).
Conclusions:
Our findings demonstrate significant pain relief in patients with knee osteoarthritis who received collagen peptides compared to those who received placebo. In addition, the risk of adverse events did not differ significantly between the collagen peptide group and the placebo group. However, due to potential biases and limitations, well-designed randomized controlled trials are needed to validate and confirm these findings.
... As expected, the most abundant protein functional domain in the Phylobone dataset is collagen, which is present in 12% of human proteins, covering 90% of the organic fraction of the bone ECM 2 . Collagen is essential for homeostasis maintenance, and it serves as a scaffold to many other macromolecules and hydroxyapatite, enabling cell attachment and bone resistance to mechanical forces 1,42,43 . Furthermore, 214 non-collagenous functional domains are present in human sequences of the dataset ( Figure 2 and Supplementary Table S4). ...
The bone extracellular matrix (ECM) contains minerals deposited on highly crosslinked collagen fibrils and hundreds of non-collagenous proteins. Some of these proteins are key to the regulation of bone formation and regeneration via signaling pathways, and play important regulatory and structural roles. However, the complete list of bone extracellular matrix proteins, their roles, and the extent of individual and cross-species variations have not been fully captured in both humans and model organisms. Here, we introduce the most comprehensive resource of bone extracellular matrix (ECM) proteins that can be used in research fields such as bone regeneration, osteoporosis, and mechanobiology. The Phylobone database (available at https://phylobone.com) includes 255 proteins potentially expressed in the bone extracellular matrix (ECM) of humans and 30 species of vertebrates. A bioinformatics pipeline was used to identify the evolutionary relationships of bone ECM proteins. The analysis facilitated the identification of potential model organisms to study the molecular mechanisms of bone regeneration. A network analysis showed high connectivity of bone ECM proteins. A total of 214 functional protein domains were identified, including collagen and the domains involved in bone formation and resorption. Information from public drug repositories was used to identify potential repurposing of existing drugs. The Phylobone database provides a platform to study bone regeneration and osteoporosis in light of (biological) evolution, and will substantially contribute to the identification of molecular mechanisms and drug targets.
... Bone tissue is composed of numerous collagen, of which glycine, proline, and hydroxyproline are the most common amino acids in collagen [44]. Studies conducted in the past have revealed that dietary AKG supplementation can result in a heightened concentration of glycine and proline in the serum of animals after eating [45,46]. ...
Simple Summary
Intensive farming in the modern pig industry has resulted in an increase in phosphorus pollution, which is a major environmental concern. The dietary intake of phosphorus is primarily employed for the growth and metabolism of pig bones. Dietary strategies to improve bone growth can be utilized to optimize the utilization of dietary phosphorus and reduce its release into the environment. Interestingly, recent studies have shown that alpha-ketoglutarate, an intermediate metabolite in the tricarboxylic acid cycle, improves osteogenesis in vitro. Therefore, we hypothesized that dietary alpha-ketoglutarate supplementation would have a positive effect on bone growth, and thereby improve the utilization of dietary phosphorus and calcium in piglets. In the present study, we found that dietary alpha-ketoglutarate supplementation improves bone growth, such as bone density, length, and weight in piglets. Of note, our study further demonstrated that alpha-ketoglutarate supplementation improves the apparent ileal and total tract digestibility of phosphorus and calcium in piglets’ diets. Our findings may provide a nutritional strategy for diminishing phosphorus pollution originating from the pig industry.
Abstract
Phosphorus (P) pollution from modern swine production is a major environmental problem. Dietary interventions to promote bone growth can improve the utilization of dietary P, and thereby reduce its emission. Recent in vitro studies have shown that alpha-ketoglutarate (AKG) exerts a pro-osteogenic effect on osteoblast cells. This study aimed to evaluate the effects of AKG supplementation on bone growth, P and Ca digestion, and the gut microbial profile in piglets. Thirty-two piglets were randomly assigned into two dietary groups. The piglets were fed a basic diet containing 10 g/kg AKG or 10 g/kg maize starch (control) for 28 days. On days 21–28, titanium dioxide was used as an indicator to determine the apparent digestibility of P. AKG supplementation improved the bone mineral density, length, weight, and geometrical and strength properties of the femur and tibia. Furthermore, AKG supplementation increased apparent ileal and total tract digestibility of P. Colonic microbiota analysis results showed that AKG supplementation increased α-diversity and beneficial bacteria, including Lactobacillus and Clostridium butyricum, and decreased nitrogen fixation and chemoheterotrophy. Together, AKG supplementation improves bone growth, the utilization of dietary P, and the colonic microbial profile, which may provide a nutritional strategy for diminishing P pollution originating from the pig industry.
... Gelatin may be used as a supplement to reduce the risk of osteoporosis, or a thinning of the bones. [159,160] 10 Purification of water A gelatinous yeast biosorbent is used to improve the extraction and biosorption of Cr(VI) from water. Removal of metal ions, phenols, pesticides, dyes, PCBs, DTT, amino acids, proteins, oil, and greases. ...
Meat by-products such as bones, skin, horns, hooves, feet, skull, etc., are produced from slaughtered mammals. Innovative solutions are very important to achieving sustainability and obtaining the added value of meat by-products with the least impact on the environment. Gelatin, which is obtained from products high in collagen, such as dried skin and bones, is used in food processing, and pharmaceuticals. Chitosan is derived from chitin and is well recognized as an edible polymer. It is a natural product that is non-toxic and environmentally friendly. Recently, chitosan has attracted researchers' interests due to its biological activities, including antimicrobial, anti-tumor, and antioxidant properties. In this review, article, we highlighted the recent available information on the application of gelatin and chitosan as antioxidants, antimicrobials, food edible coating , enzyme immobilization, biologically active compound encapsulation, water treatment, and cancer diagnosis.
... Bio-peptides derived from the hydrolysis of collagen are acknowledged as active ingredients and thus naturally stand as relevant candidates for nutricosmetics. To date, oral intake of collagen peptides has already been linked to a beneficial effect on the osteoarticular system, either on bone or joint tissues [15][16][17]. In this context, the potential interest of this same FCH in the treatment of joint discomfort associated with osteoarthritis in rodents was recently demonstrated [18]. ...
Due to its significant exposure to stressful environmental factors, the skin undergoes a high remodeling rate over time, which alters not only its appearance but also its functionality. This alteration of the skin, namely photoaging, is characterized by dryness and a loss of elasticity that mainly originates from the dysregulation of dermal fibroblast activities. In order to overcome such tissue outcome, cosmetic products have evolved toward nutricosmetics, thus promoting beauty from within. Among bio-actives of interest, bio-peptides deriving from plant or animal sources may exert various biological activities beyond their nutritional value. However, studies remain mostly descriptive and the mode of action at the cellular level in clinic remains a concern. In a recent clinical trial, it was showed that supplementation with a fish cartilage hydrolysate (FCH) improved signs of chronological and photoaging-induced skin changes in healthy women. Here, using an original ex vivo clinical approach adapted to nutricosmetic purpose, we further demonstrated that this fish cartilage hydrolysate was absorbed and that the circulating metabolites produced in humans following FCH intake stimulate human dermal fibroblast growth, promote specific hyaluronan production, up-regulate elastin synthesis and inhibit MMP-1 and 3 expression along with the enhancement of TGF-β release. Altogether, these data provide clues on the mechanisms likely contributing to the beneficial impact of FCH on human skin functionality by supporting hydration, elasticity and limiting the expression of catabolic factors involved in photoaging onset.
... First studies on hydrolysed collagen date back to the 80s, but during the last two decades, its number and diversity have grown notably. Clinical and in vivo studies have reported improvement in most prevalent degenerative diseases in joints and bones (Daneault et al., 2017;Porfírio & Fanaro, 2016), skin properties (de Miranda et al., 2021, pressure ulcer and wound healing (Nomoto & Iizaka, 2020), sarcopenia (Zdzieblik et al., 2015) and fibromialgia (Olson et al., 2016). Other studies have reported physiological effects as an antioxidant (Hong et al., 2019), antihypertensive (Kouguchi et al., 2013), and antidiabetic (Huang et al., 2014). ...
Immunonutrition or modulation of immune capacity through food and supplements has been gaining significant importance. Hydrolysed collagen has long been used as a functional ingredient, showing multiple physiological activities, including enhancement of immune functions. However, how collagen peptides may affect the immune system still needs further research. This study investigates bone collagen peptides (BCP) immunomodulatory activity on human monocytic THP-1 and human Jurkat T lymphocyte cell lines, using cytokine mRNA expressions as biomarkers. In vitro, gastrointestinal digestion and Caco-2 cell absorption allow obtaining digested and absorbed BCP fractions, respectively, which are tested on immune cells. Results show: (1) Immunostimulatory effect on M0 macrophages, but not on M1 macrophages (lipopolysaccharide (LPS)-activated), (2) Significant T lymphocyte proliferation after incubation with absorbed BCP fraction. (3) Significant increase of anti-inflammatory IL-10 cytokine biomarker. These results suggest that BCP could act as an immunonutrient, modulating the immune response and inflammatory processes.
Abbreviations: ATCC: American Type Culture Collection BCP: bone collagen peptide DMSO: dimethyl sulphoxide FBS: fetal bovine serum ECM: extracellular matrix GRAS: Generally recognized as safe; IFN: interferon IL: interleukin, LPS: lipopolysaccharide SEM: standard error of the mean TEER: transepithelial electrical resistance
... Evidence has consistently shown that gelatin hydrolysates play an active role in the treatment of OP, joint disease, and osteoarthritis [8,9]. Gelatin hydrolysates could also stimulate the proliferation and differentiation of osteoblasts, inhibit the formation of osteoclasts, and improve calcium (Ca) absorption in vitro [10]. ...
Tilapia skin gelatin hydrolysates (TSGH) were obtained by complex protease hydrolysis. The amino acid sequences of 50 peptides in TSGH were identified, and most of these peptides were found to contain the -Gly-Pro-sequence. The osteoporosis (OP) rat model induced by retinoic acid was prepared, and the effects of different doses of TSGH on OP in vivo were evaluated. Serum calcium (Ca) and phosphate (P), alkaline phosphatase activity, and osteocalcin levels in OP rats were regulated by TSGH. The bone length, dry weight index, maximum load, and Ca content of OP rats significantly increased by treatment TSGH in a dose-dependent manner. Micro-CT images of the femurs and tibias of the rats indicated that the bone mineral density, cortical bone thickness, and cortical/trabecular bone area ratios were recovered and that OP symptoms were improved. Tartrate-resistant acid phosphate and hematoxylin-eosin staining showed that osteoclast numbers and histomorphological changes in the femurs in OP rats could be recovered by TSGH.
... Similar to isoflavone, fsCH containing collagen peptides of GPH, GP, and PH was effective in counteracting estrogen loss-induced uterus atrophy and osteoclastic bone loss. This study revealed that fsCH enhanced the formation of trabecular bones and collagenous matrix in the metaphysis and diaphysis of OVX mice through reducing expression of the osteoclastic biomarkers of CAII, V-ATPase, and cathepsin K. Other investigations have shown that collagen hydrolysates have a beneficial effect on osteoarthritis and osteoporosis [31,32]. However, the action mechanisms of fsCH on bone health in the postmenopausal state have not yet been defined. ...
Osteoporosis manifest in postmenopausal women is an osteolytic disease characterized by bone loss, leading to increased susceptibility to bone fractures and frailty. The use of complementary therapies to alleviate postmenopausal osteoporosis is fairly widespread among women. The current study examined that Pangasius hypophthalmus fish skin collagen hydrolysates (fsCH) inhibited ovariectomy (OVX)-induced bone loss by conducting inter-comparative experiments for anti-osteoporotic activity among 206-618 mg/kg fsCH, 2 mg/kg isoflavone, 15 mg/kg glycine-proline-hydroxyproline (GPH) tripeptide, and calcium lactate. Surgical estrogen loss of mice for 8 weeks reduced serum 17β-estradiol levels with uterus atrophy, which was ameliorated by orally administering fsCH or isoflavone to mice. Similar to isoflavone, fsCH containing GPH-enhanced bone mineral density reduced levels of cathepsin K and proton-handling proteins, and elevated collagen 1 level in OVX bones. The treatment with fsCH and isoflavone enhanced the serum levels of collagen synthesis-related procollagen type 1 carboxy/amino-terminal propeptides reduced by OVX, whereas serum levels of osteocalcin and alkaline phosphatase, as well as collagen breakdown-related carboxy/amino-terminal telopeptides of type 1 collagen were reduced in OVX mice treated with fsCH, isoflavone, and calcium lactate. The trabecular bones were newly formed in OVX bones treated with isoflavone and fsCH, but not with calcium lactate. However, a low-dose combination of fsCH and calcium lactate had a beneficial synergy effect on postmenopausal osteoporosis. Furthermore, similar to isoflavone, 15-70 μg/mL fsCH, with its constituents of GPH and dipeptides of glycine-proline and proline-hydroxyproline, enhanced osteogenesis through stimulating differentiation, matrix mineralization, and calcium deposition of MC3T3-E1 osteoblasts. Accordingly, the presence of fsCH may encumber estrogen deficiency-induced bone loss through enhancing osteoclastogenic differentiation and matrix collagen synthesis. Therefore, fsCH may be a natural compound retarding postmenopausal osteoporosis and pathological osteoresorptive disorders.
... Adequate protein intake contributes to bone development and bone maintenance (7). As key constituents of the bone mineral matrix, proteins regulate bone metabolism by providing building blocks and performing specific regulatory functions (8). In recent years, the geriatric nutritional risk index (GNRI) has been used as a significant tool to access the nutritional status of the elderly. ...
Background
The geriatric nutritional risk index (GNRI) has been used as a significant tool to access the nutritional status of the elderly. However, the relationship between the GNRI and femur bone mineral density (BMD) and the risk of osteoporosis remains unclear in American postmenopausal women.
Objectives
We aimed to explore associations between the GNRI with femur BMD and the risk of osteoporosis in American postmenopausal women.
Methods
We merged the continuous National Health and Nutrition Examination Survey (NHANES) 2005–2006, 2007–2008, 2009–2010, 2013–2014, and 2017–2018 to ensure a large and representative sample, including 3,152 participants. The linear relationship between the GNRI and femur BMD was assessed via a weighted multivariate linear regression model. The odds ratios (ORs) and 95% confidence intervals (95% CIs) for the association between the GNRI and the risk of osteoporosis were assessed by a weighted logistic regression model. Moreover, the nonlinear relationship was also characterized by smooth curve fitting (SCF) and a weighted generalized additive model (GAM).
Results
After adjusting for all covariates, the weighted multivariable linear regression models demonstrated that the GNRI was positively correlated with femur BMD. The weighted logistic regression models demonstrated that each unit of increased GNRI value was associated with a decreased risk of osteoporosis of 4.13%. When categorizing GNRI based on quartiles, ORs between the risk of osteoporosis and the GNRI across quintiles 2, 3, and 4 compared with quintile 1 were 0.5565 (95% CI: 0.4791, 0.6463; P < 0.000001), 0.5580 (95% CI: 0.4600, 0.6769; P < 0.000001), and 0.3475 (95% CI: 0.2681, 0.4505; P < 0.000001). The trends similar to the above were also observed in SCF and GAM.
Conclusion
This study indicated that nutritional status, represented by the GNRI, was positively associated with femur BMD and negatively associated with the risk of osteoporosis in American postmenopausal women. The GNRI may be a good tool to identify American postmenopausal women who need further bone health nutritional support.
... Hydrolysis has been shown to improve the functional and nutritional properties of proteins, and collagen peptides have exhibited several physiological activities (15). Among these, it is worth noting its osteogenic capacity, whose mechanism revealed in in vitro studies (16,17) supports the results of in vivo studies (18,19). Furthermore, bone beneficial effects of calcium and vitamin D supplementation increase remarkably if it is accompained by oral hydrolyzed collagen intake (20)(21)(22). ...
Introduction:
osteoporosis is the most prevalent bone disease and one of the main causes of chronic disability in middle and advanced ages. Conventional pharmacological treatments are still limited, and their prolonged use can cause adverse effects that motivate poor adherence to treatment. Nutritional strategies are traditionally based on supplementing the diet with calcium and vitamin D. Recent studies confirm that the results of this supplementation are significantly improved if it is accompanied by the intake of oral hydrolyzed collagen.
Objective:
to evaluate the possible in vitro osteogenic activity of a peptide-mineral complex formed by bovine hydrolyzed collagen and bovine hydroxyapatite (Phoscollagen®, PHC®).
Methods:
the digestion and absorption of PHC® were simulated using the dynamic gastrointestinal digester of AINIA and Caco-2 cell model, respectively. Primary cultures of human osteoblasts were treated with the resulting fraction of PHC® and changes were evaluated in the proliferation of preosteoblasts and in the mRNA expression of osteogenic biomarkers at different stages of osteoblast maturation: Runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), osteocalcin (OC) and type I collagen (ColA1).
Results:
an increase in preosteoblastic proliferation was observed (p ≤ 0,05). No changes were detected in the biomarkers of osteoblasts with 5 days of differentiation, but were with 14 days, registering an increase in Runx2 (p = 0.0008), ColA1 (p = 0.035), OC (p = 0.027) and ALP (without significance).
Conclusion:
these results show that the PHC® peptide-mineral complex stimulates the activity of mature osteoblasts, being capable of promoting bone formation.
... Collagen peptides have other bioactivities such as antibacterial, antiaging, stimulating wound healing and cell proliferation, treating bone and joint diseases, and improving skin health (Ahmed, Verma, & Patel, 2020;Daneault, Prawitt, Fabien Soulé, Coxam, & Wittrant, 2017;Fu et al., 2019;Pal & Suresh, 2016;Zhao et al., 2021). ...
Collagen is the most abundant extracellular matrix protein in food-producing animals. Gelatin is partially degraded collagen. Collagen peptides refer to the peptides with specific properties identified from collagen hydrolysate who produced by hydrolysis of collagen/gelatin. Due to the specific structural and bio- and physical-chemical properties, collagen and its derivatives are used in the field of food industry. In this review, the structure of the collagen molecule and its biosynthetic process in vivo are introduced, and the production methods and structures of gelatin and collagen peptides described. Then the inherent self-assembly property of collagen, the mechanical properties of collagen and gelatin gels, functional properties of collagen and gelatin, and bioactive properties of collagen peptides are reviewed. Finally, the applications of collagen and its derivatives that are correlated with their properties in food industry are summarized. The mechanisms and advantages of the applications of collagen and its derivatives in food industry are raised, and the limitations and challenges of these applications are also discussed. And possible studies to address the challenges of the applications in different areas are indicated.
... Osteoporosis is mostly seen in the elderly and is characterized as low bone mineral density, loss of mechanical ability, architectural decay, and increased risk of fracture (Daneault et al., 2017). To support bone health, a number of studies focused on the supplementation of FPH to the individual have been carried out. ...
Fish processing industries result in an ample number of protein‐rich byproducts, which have been used to produce protein hydrolysate (PH) for human consumption. Chemical, microbial, and enzymatic hydrolysis processes have been implemented for the production of fish PH (FPH) from diverse types of fish processing byproducts. FPH has been reported to possess bioactive active peptides known to exhibit various biological activities such as antioxidant, antimicrobial, angiotensin‐I converting enzyme inhibition, calcium‐binding ability, dipeptidyl peptidase‐IV inhibition, immunomodulation, and antiproliferative activity, which are discussed comprehensively in this review. Appropriate conditions for the hydrolysis process (e.g., type and concentration of enzymes, time, and temperature) play an important role in achieving the desired level of hydrolysis, thus affecting the functional and bioactive properties and stability of FPH. This review provides an in‐depth and comprehensive discussion on the sources, process parameters, purification as well as functional and bioactive properties of FPHs. The most recent research findings on the impact of production parameters, bitterness of peptide, storage, and food processing conditions on functional properties and stability of FPH were also reported. More importantly, the recent studies on biological activities of FPH and in vivo health benefits were discussed with the possible mechanism of action. Furthermore, FPH‐polyphenol conjugate, encapsulation, and digestive stability of FPH were discussed in terms of their potential to be utilized as a nutraceutical ingredient. Last but not the least, various industrial applications of FPH and the fate of FPH in terms of limitations, hurdles, future research directions, and challenges have been addressed.
Surface structure plays a crucial role in determining cell behavior on biomaterials, influencing cell adhesion, proliferation, differentiation, as well as immune cells and macrophage polarization. While grooves and ridges stimulate M2 polarization and pits and bumps promote M1 polarization, these structures do not accurately mimic the real bone surface. Consequently, the impact of mimicking bone surface topography on macrophage polarization remains unknown. Understanding the synergistic sequential roles of M1 and M2 macrophages in osteoimmunomodulation is crucial for effective bone tissue engineering. Thus, exploring the impact of bone surface microstructure mimicking biomaterials on macrophage polarization is critical. In this study, we aimed to sequentially activate M1 and M2 macrophages using Poly-l-Lactic acid (PLA) membranes with bone surface topographical features mimicked through the soft lithography technique. To mimic the bone surface topography, a bovine femur was used as a model surface, and the membranes were further modified with collagen type-I and hydroxyapatite to mimic the bone surface microenvironment. To determine the effect of these biomaterials on macrophage polarization, we conducted experimental analysis that contained estimating cytokine release profiles and characterizing cell morphology. Our results demonstrated the potential of the hydroxyapatite-deposited bone surface-mimicked PLA membranes to trigger sequential and synergistic M1 and M2 macrophage polarizations, suggesting their ability to achieve osteoimmunomodulatory macrophage polarization for bone tissue engineering applications. Although further experimental studies are required to completely investigate the osteoimmunomodulatory effects of these biomaterials, our results provide valuable insights into the potential advantages of biomaterials that mimic the complex microenvironment of bone surfaces.
In this study, the chemical substances of Heiguteng Zhuifeng Huoluo Capsule (HZFC) and its potential active ingredients for the treatment of rheumatoid arthritis (RA) were characterized and analyzed by medicinal chemistry combined with bioinformatics methods. Also, the potential active ingredients of HZFC against RA were verified by lipopolysaccharide (LPS)-induced macrophage activation model. The results showed that 79 chemical constituents were successfully identified, mainly including phenylpropanoids, flavonoids, and alkaloids. Among them, 13 active components were closely related to the nine core targets (FASN, ALOX5, EGFR, MMP1, CYP2D6, CNR1, AR, MAOA, and FKBP5) of HZFC in the treatment of RA. Molecular docking further proved that 13 active components had strong docking activity with 9 core targets. In the verification experiment of the LPS-induced RAW 264.7 macrophage model, the verified components (magnoflorine, N-feruloyltyramine, canadine, rutin, quercetin-3-O-glucoside, and pseudocolumbamine) all showed a clear inhibitory effect on the secretion of inflammatory factors in model cells. The above research results suggest that 13 components such as stepharanine, rutin, quercetin-3-O-glucoside, corydine methyl ether, canadine, 8-oxoepiberberine, disinomenine, deosinomenine glucoside, tuduranine, magnoflorine, isosinomenine, pseudocolumbamine, and N-feruloyltyramine may be the main active substances of HZFC in the treatment of RA.
There are scarce published data suggesting, that collagen extracted from fish skin may be an attractive alternative to mammalian-derived collagen for the in vitro cell cultures. In this study, we investigated proliferation potential and differentiation capability into osteogenic and adipogenic lineages of rat adipose-derived mesenchymal stem cells (rASCs) and human adipose-derived mesenchymal stem cells (hASCs) cultured on collagen extracted from silver carp and African sharptooth catfish skins, compared with commercially available mammalian collagen and collagen-free culture dishes. Our results revealed no significant differences between fish collagen and mammalian collagen in supporting cell viability and proliferation capacity. Fish-derived collagen is a cheap material derived from production waste, does not contain transmissible pathogens of mammalian origin, supports human cell cultures at comparable level to conventional collagen sources, and may be considered as the product of choice for the in vitro cell cultures.
This study reveals that low-molecular-weight collagen peptide (LMWCP) can stimulate the differentiation and the mineralization of MC3T3-E1 cells in vitro and attenuate the bone remodeling process in ovariectomized (OVX) Sprague-Dawley rats in vivo. Moreover, the assessed LMWCP increased the activity of alkaline phosphatase (ALP), synthesis of collagen, and mineralization in MC3T3-E1 cells. Additionally, mRNA levels of bone metabolism-related factors such as the collagen type I alpha 1 chain, osteocalcin (OCN), osterix, bone sialoprotein, and the Runt family-associated transcription factor 2 were increased in cells treated with 1,000 μg/ml of LMWCP. Furthermore, we demonstrated that critical bone morphometric parameters exhibited significant differences between the LMWCP (400 mg/kg)-receiving and vehicle-treated rat groups. Moreover, the expression of type I collagen and the activity of ALP were found to be higher in both the femur and lumbar vertebrae of OVX rats treated with LMWCP. Finally, the administration of LMWCP managed to alleviate osteogenic parameters such as the ALP activity and the levels of the bone alkaline phosphatase, the OCN, and the procollagen type 1 N-terminal propeptide in OVX rats. Thus, our findings suggest that LMWCP is a promising candidate for the development of food-based prevention strategies against osteoporosis.
Movement and nutrition play a very important role in maintaining health throughout life and preventing chronic diseases. Obesity and physical inactivity are among the leading causes of death at an early age, and factors that define an individual’s lifestyle, such as exercise and healthy nutrition, can reduce the risk of developing many chronic diseases. Regular physical activity and a balanced diet not only prolong life but also improve the quality of life and lead to a healthy life. In this chapter, the effects of nutrition on musculoskeletal health will be considered.KeywordsMovementNutritionHealthy lifestyleExerciseChronic diseasesQuality of life
Caulerpa lentillifera is a potential cash crop, the current promotion and cultivation in China is in the development stage. Take Caulerpa lentillifera as a raw material to extract the active peptides. Then, investigate the effects of active peptides on the proliferation and differentiation of osteoblasts. Simultaneously, the metabolomics strategies was used to find the different metabolites and metabolic pathways, combining with other molecular biology methods and metabolomics results, to discuss the molecular mechanism of active peptide extracts. This filed of research is to provide scientific basis for the exploitation of marine resources and the development of new anti-osteoporosis drugs.KeywordsProgressOsteogenic activityPeptide
Caulerpa Lentillifera
Collagen is considered a nutraceutical, and its consumption has been expanding due to the increased life expectancy, rising per capita income, and increased consumer awareness of health care. This study aimed to evaluate consumers' perceptions, knowledge, attitudes, and practices about the consumption of collagen-based products by using an online questionnaire and to correlate them with socio-economic data. A market survey (pharmacy stores and online) was also conducted to evaluate the available products. In total, 275 participants answered the survey, 73.3% from the Southeast region, mostly female (84.0%). Most participants reported three months as the period of collagen intake (31.6%), and the consumption period was associated with the perception of the health benefits (p < 0.001). Furthermore, the participants' knowledge and perceptions regarding collagen intake are frequently associated with dermatological and orthopedic changes. Collagen-based products supplementation is a growing market with a broad target audience (genders, age groups, and socio-economic levels). The commercial presentation of collagen has been diversified over the years, and powder collagen is the most consumed (52.7%) and cheapest compared with capsules, pills, or gummies. The results of the present study demonstrate that most consumers of this type of supplement associate its benefits with aesthetic care such as skin, hair, and nails, although the scientific literature has shown its effects in treating osteoarticular diseases, for example. Undoubtedly, the correct dose prescription, treatment time, and choice of product presentation must be analyzed carefully, as they significantly impact treatment results.
Bone substitutes are widely used in maxillofacial and oral surgeries. However, in clinical practice, bone substitutes with various forms, including separated particulates, powders, and blocks, have exhibited poor handling properties and space maintenance characteristics, resulting in long surgery procedures and unstable volume of the newly formed bone. Movable separated particulates with high stiffness have induced local inflammatory responses that hinder bone regeneration. The present study aimed to develop a new method to enhance the stability and operability of bone substitutes commonly used in dentistry by premixing with photocurable hydrogel GelMA. The GelMA-encapsulated particulate had a strong capacity to aggregate separated particulates and firmly attach to the host bone defect after photocuring compared to particulates alone. Additionally, macrophages at the surface of the GelMA-stabilized particulates tended to present a more M2-like phenotype than those at the surface of Bio-Oss®, leading to more MMR⁺ multinucleated giant cell formation and the induction of blood vessel invasion and new bone formation. In conclusion, this hydrogel-coated bone substitute strategy facilitates bone regeneration with increased operability, a stable volume of osteogenic space, and a favorable osteogenic microenvironment, indicating its potential value in the field of maxillofacial and oral surgeries when bone substitutes are needed.
Aging biomarkers are a combination of biological parameters to (i) assess age-related changes, (ii) track the physiological aging process, and (iii) predict the transition into a pathological status. Although a broad spectrum of aging biomarkers has been developed, their potential uses and limitations remain poorly characterized. An immediate goal of biomarkers is to help us answer the following three fundamental questions in aging research: How old are we? Why do we get old? And how can we age slower? This review aims to address this need. Here, we summarize our current knowledge of biomarkers developed for cellular, organ, and organismal levels of aging, comprising six pillars: physiological characteristics, medical imaging, histological features, cellular alterations, molecular changes, and secretory factors. To fulfill all these requisites, we propose that aging biomarkers should qualify for being specific, systemic, and clinically relevant.
Osteoporosis commonly occurs in the older people and severe patients, with the main reason of the imbalance of bone metabolism (the rate of bone resorption exceeding the rate of bone formation), resulting in a decrease in bone mineral density and destruction of bone microstructure and further leading to the increased risk of fragility fracture. Recent studies indicate that protein nutritional support is beneficial for attenuating osteoporosis and improving bone health. This review summarized the classical mechanisms of protein intervention for alleviating osteoporosis on both suppressing bone resorption and regulating bone formation related pathways (promoting osteoblasts generation and proliferation, enhancing calcium absorption, and increasing collagen and mineral deposition), as well as the potential novel mechanisms via activating autophagy of osteoblasts, altering bone related miRNA profiles, regulating muscle-bone axis, and modulating gut microbiota abundance. Protein nutritional intervention is expected to provide novel approaches for the prevention and adjuvant therapy of osteoporosis.
Despite several attempts over decades, process scalability and sustainability remains a challenge to produce an environmental-friendly enzyme to gain industrial attention. In the present study, microbial degradation of chrome shavings (chromium-collagen leather waste) and the resulting collagen hydrolysate for producing the dehairing protease by Bacillus cereus VITSN04 were investigated in a lab-scale fermentor. Scale-up degradation of shavings resulted in higher recovery of collagen hydrolysate (76%) within 72 h compared to shake flasks (68% in 120 h). Earlier achieved medium composition of collagen hydrolysate (12 g L-1) and molasses (15 g L-1) appeared to induce amylase in high rate, despite maximal production of protease (203.8±0.18 U mL-1), which was analysed by ANS fluorescence spectroscopy. Optimization of the media containing collagen hydrolysate (12 g L-1) and molasses (5 g L-1) was effective in producing protease (170.6±0.1 U mL-1) and reduce the co-synthesis of amylase (48.2±0.09 U mL-1). The controlled fermentation process, by feeding molasses during exponential growth phase had enhanced dehairing protease production (∼2.96 fold). The produced protease then partitioned through biphasic system showed significant dehairing of goat skins in pilot scale. Thus, scalability of the process to produce dehairing enzymes using waste, generated at the site of its use offers hope for sustainable greener production of leathers.
This work investigated hydrolysed collagen (HC) powder as carrier material for particle formation via supercritical CO2 impregnation. HC solubility and volumetric expansion in supercritical CO2 (ρ: 605.8-834.8 g/L) were measured. Formation of HC particles loaded with lycopene dispersed in sunflower oil (LSO) (HC-LSO) using supercritical CO2 at 100-140 bar and 40-50 °C and HC/LSO ratios of 4:1-10:1 (w/w) was studied. HC had constant solubility (0.5-0.6 g/kg CO2) and no volumetric expansion was visibly observed. HC-LSO had similar spherical morphology, size distribution, and loading capacity (0.025 g of LSO/g of HC) between treatments. HC-LSO formed with HC/LSO ratio of 10:1 (w/w) stand out for their low agglomeration. Since HC showed low interaction with supercritical CO2 under different conditions and preserved its powder appearance after impregnation with LSO, it is suggested to be used as carrier of lipophilic bioactive compounds in particle formation via supercritical CO2 impregnation at moderate temperature (40-50°C).
Collagen extracted from fishes has been appearing as an alternative for commercial porcine and bovine collagen and it has been considered interesting especially for membrane manufacturing in tissue engineering. Despite the positive in vitro effects of fish collagen membranes, there is still no understanding of all the benefits that this natural biomaterial plays in the wound healing process, due to the lack of compilation of the results obtained in animal studies. In this sense, the purpose of this study was to perform a systematic review of the literature to examine the effects of fish collagen membranes for skin wound healing in experimental models of skin wound. The search was carried out according to the orientations of Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA), and the descriptors of the Medical Subject Headings (MeSH) were defined: “fish,” collagen,” “skin,” and “in vivo”. A total of 10 articles were retrieved from the databases PubMed and Scopus. After the elegibility analyses, this review covers the different origins of fish collagen reported in the different papers from the beginning of 2015 through the middle of 2021. The results were based mainly on histological analysis and macroscopic evaluation, and fish skin collagen was responsible for improving the wound healing rate and the process of reepithelization and collagen deposition. In conclusion, fish skin collagen has shown positive results in in vivo studies and may be a potential biomaterial in tissue engineering.
Bone fragility fractures remain an important worldwide health and economic problem due to increased morbidity and mortality. The current methods for predicting fractures are largely based on the measurement of bone mineral density and the utilization of mathematical risk calculators based on clinical risk factors for bone fragility. Despite these approaches, many bone fractures remain undiagnosed. Therefore, current research is focused on the identification of new factors such as bone turnover markers (BTM) for risk calculation. BTM are a group of proteins and peptides released during bone remodeling that can be found in serum or urine. They derive from bone resorptive and formative processes mediated by osteoclasts and osteoblasts, respectively. Potential use of BTM in monitoring these phenomenon and therefore bone fracture risk is limited by physiologic and pathophysiologic factors that influence BTM. These limitations in predicting fractures explain why their inclusion in clinical guidelines remains limited despite the large number of studies examining BTM.
Dietary habits and lifestyle-related diseases indicate that food has a direct impact on individual health. Hence, a diet containing essential nutrients is important for healthy living. Fish and fish products are important in diets worldwide because of their nutritional value, especially their easily digestible proteins with essential amino acids. Similarly, fish protein hydrolysate (FPH) obtained from fish muscle and by-products has been reported to exhibit various biological activities and to have functional properties, which make FPH a suitable nutraceutical candidate. This review focuses on the health-promoting ability of FPH in terms of skin health, bone and cartilage health, blood lipid profile, and body-weight management studied in rats and human model systems. The absorption and bioavailability of FPH in humans is discussed, and challenges and obstacles of FPH as a functional food ingredient are outlined.
Menopause drastically increases the risk of osteoporosis, and although drug therapies are available, having an efficacious dietary supplement as an adjuvant therapy or alternative is desirable. Recent findings suggest that a calcium-collagen chelate (CC) in the form of a dietary supplement is highly effective in improving bone mass in osteopenic rats. Therefore, we hypothesized that the consumption of CC reverses bone loss in postmenopausal women with osteopenia as early as three months. Women 1 to 5 years postmenopausal, not on hormone replacement therapy or any other prescribed medication known to influence bone metabolism were randomized to one of two treatment groups to receive as a dietary supplement intervention daily for three months of either of the following: 500 mg of calcium carbonate and 5 µg vitamin D (control), or 5 g of CC containing 500 mg of calcium carbonate and 5 µg vitamin D. Bone mineral density of lumbar spine and total body were assessed at baseline and at three months using dual-energy X-ray absorptiometry. Blood was collected at baseline and three months to assess bone biomarkers of bone metabolism. CC significantly increased total body bone mineral density when compared to the control group (P<0.05). A significant increase (P<0.05) in the BAP/TRAP5b ratio percent change was indicated for the CC group. Collectively, these preliminary data suggested that CC enhances bone mass potentially by increasing the rate of bone formation more than bone resorption in the process of bone turnover.
Menopause leads to an increased risk for osteoporosis in women. Although drug therapies exist, increasing numbers of people prefer alternative therapies such as dietary supplements, for example, calcium, vitamin D, and collagen hydrolysates for the prevention and treatment of osteoporosis. We have previously shown that a 3-month intervention using a calcium-collagen chelate (CC) dietary supplement was efficacious in improving bone mineral density (BMD) and blood biomarkers of bone turnover in osteopenic postmenopausal women. This study reports the long-term efficacy of CC in reducing bone loss in postmenopausal women with osteopenia. Thirty-nine women were randomly assigned to one of two groups: 5 g of CC containing 500 mg of elemental calcium and 200 IU vitamin D (1,25-dihydroxyvitamin D3) or control (500 mg of calcium and 200 IU vitamin D) daily for 12 months. Total body, lumbar, and hip BMD were evaluated at baseline, 6 and 12 months using dual-energy X-ray absorptiometry. Blood was collected at baseline, 6 and 12 months to assess levels of blood biomarkers of bone turnover. Intent-to-treat (ITT) analysis was performed using repeated measures analysis of variance pairwise comparisons and multivariate analysis to assess time and group interactions. The loss of whole body BMD in women taking CC was substantially lower than that of the control group at 12 months in those who completed the study and the ITT analysis, respectively (CC: -1.33% and -0.33% vs. control: -3.75% and -2.17%; P = .026, P = .035). The CC group had significantly reduced levels of sclerostin and tartrate-resistant acid phosphatase isoform 5b (TRAP5b) (P < .05), and higher bone-specific alkaline phosphatase/TRAP5b ratio (P < .05) than control at 6 months. These results support the use of CC in reducing bone loss in osteopenic postmenopausal women. KEY WORDS: collagen hydrolysate estrogen menopause osteoporosis sclerostin INTRODUCTION
The present study investigated the effects of egg yolk-derived peptide (YPEP) on osteogenic activities and MAPK-regulation of osteogenic gene expressions. The effects of YPEP on cell proliferation, alkaline phosphatase activity, collagen synthesis, and mineralization were measured in human osteoblastic MG-63 cells. Activation of MAPKs and downstream transcription factors such as extracellular-signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase 1/2 (JNK1/2), p38, ELK1, and cJUN were examined using western blot analysis. YPEP dose-dependently increased MG-63 cell proliferation, ALP activity, collagen synthesis, and calcium deposition. YPEP activated ERK1/2, p38, and ELK1 phosphorylation whereas JNK and cJUN were not affected by YPEP. The COL1A1 (collagen, type I, alpha 1), ALPL (alkaline phosphatase), and SPP1 (secreted phosphoprotein 1, osteopontin) gene expressions were increased while BGLAP (osteocalcin) was not affected by YPEP. The ERK1/2 inhibitor (PD98509) blocked the YPEP-induced COL1A1 and ALPL gene expressions as well as ELK1 phosphorylation. The p38 inhibitor (SB203580) blocked YPEP-induced COL1A1 and ALPL gene expressions. SPP1 gene expression was not affected by these MAPK inhibitors. In conclusion, YPEP treatment stimulates the osteogenic differentiation via the MAPK/ELK1 signaling pathway. These results could provide a mechanistic explanation for the bone-strengthening effects of YPEP.
To utilize the oyster shell containing a lot of calcium, we investigated the bioavailability of calcium compounds from oyster shell. First, calcium oxide was prepared by burning of oyster shell at . Its purity was approximately . Calcium compounds, , and , from the calcium oxide were prepared by chemical reaction. Effect on calcium absorption by the calcium compounds from oyster shell was improved using fish skin gelatin peptides.(FSGP), which was prepared by enzymatic hydrolysis of fish skin gelatin for 4hr with tuna pyloric caeca crude enzyme (TPCCE). in vitro experiment, calcium absorption by addition of FSGP in a mixture solution of calcium and phosphate was higher approximately than that by control. in vivo using calcium deficient rats, a group taken the diets with FSGP and was significantly improved amount of calcium and ash in femur and strength of femur. These results suggest that calcium compounds from oyster shell and FSGP could be used as an effective dietary calcium source.
Objective
Collagen peptides (CP) compounds, as bone health supplements, are known to play a role in the treatment of osteoporosis. However, the molecular mechanisms of this process remain unclear. This study aimed to investigate the effects of bovine CP compounds on the proliferation and differentiation of MC3T3-E1 cells.
Methods
Mouse pre-osteoblast cell line MC3T3-E1 subclone 4 cells were treated with bovine CP compounds. Cell proliferation was analyzed by MTT assays and the cell cycle was evaluated by flow cytometry scanning. Furthermore, MC3T3-E1 cell differentiation was analyzed at the RNA level by real-time PCR and at the protein level by western blot analysis for runt-related transcription factor 2 (Runx2), a colorimetric p-nitrophenyl phosphate assay for alkaline phosphatase (ALP), and ELISA for osteocalcin (OC). Finally, alizarin red staining for mineralization was measured using Image Software Pro Plus 6.0.
Results
Cell proliferation was very efficient after treatment with different concentrations of bovine CP compounds, and the best concentration was 3 mg/mL. Bovine CP compounds significantly increased the percentage of MC3T3-E1 cells in G2/S phase. Runx2 expression, ALP activity, and OC production were significantly increased after treatment with bovine CP compounds for 7 or 14 days. Quantitative analyses with alizarin red staining showed significantly increased mineralization of MC3T3-E1 cells after treatment with bovine CP compounds for 14 or 21 days.
Conclusions
Bovine CP compounds increased osteoblast proliferation, and played positive roles in osteoblast differentiation and mineralized bone matrix formation. Taking all the experiments together, our study indicates a molecular mechanism for the potential treatment of osteoarthritis and osteoporosis.
The present study investigated the effects of CHs on osteogenic activities and MAPK-regulation on bone matrix gene expressions. The effects of CHs on cell proliferation, alkaline phosphatase (ALP) activity, collagen synthesis, and mineralization were measured in human osteoblastic MG-63 cells. Activation of MAPKs and downstream transcription factors such as extracellular-signal-regulated kinase 1/2 (ERK1/2), c-Jun N-terminal kinase 1/2 (JNK1/2), p38, ELK1, and cJUN was examined using Western blot analysis. The expressions of osteogenic genes were measured by quantitative real-time PCR. CHs dose-dependently increased MG-63 cell proliferation, ALP activity, collagen synthesis, and calcium deposition. CHs activated ERK1/2, JNK1/2, p38, and ELK1 phosphorylation except cJUN. The COL1A1 (collagen, type I, alpha 1), ALPL (alkaline phosphatase), BGLAP (osteocalcin), and SPP1 (secreted phosphoprotein 1, osteopontin) gene expressions were increased by CH treatment. The ERK1/2 inhibitor (PD98509) blocked the CH-induced COL1A1 and ALPL gene expression, as well as ELK1 phosphorylation. The JNK1/2 inhibitor (SP600125) abolished CH-induced COL1A1 expression. The p38 inhibitor (SB203580) blocked CH-induced COL1A1 and SPP1 gene expression. In conclusion, CH treatment stimulates the osteogenic activities and increases bone matrix gene expressions via the MAPK/ELK1 signaling pathway. These results could provide a mechanistic explanation for the bone-strengthening effects of CHs.
Collagen hydrolysate (CH) has been reported to exhibit a positive effect on bone. In the present study, the in vitro effects of CH (<3 kDa) were examined and the in vivo experiments confirmed the positive effects of CH in ovariectomized (OVX) rats. Bone mineral density (BMD) was examined by DXA analysis. Scanning electron microscopic analysis and quantitative 3D-color backscattered electrons imaging analysis were performed on the lumbar vertebrae. CH increased osteoblastic cell proliferation and alkaline phosphatase activity in a dose-dependent manner. Collagen synthesis and collagen, type1, alpha1 (COL1A1) gene expression were also increased by CH treatment. Furthermore, CH-induced COL1A1 gene expression was completely abolished by extracellular signal-regulated kinase (ERK) inhibitor, suggesting the involvement of ERK/MAPK signaling for transcriptional effects on COL1A1 expression. OVX rats supplemented with CH showed osteoprotective effects as the BMD levels were increased compared with control. Moreover, CH prevented the trabecular bone loss induced by OVX and improved the microarchitecture of lumbar vertebrae. CH administration dose-dependently reduced the serum procollagen type I N-terminal propeptide level, which was elevated by OVX. The present study suggests that CH isolated in this study is a promising alternative to current therapeutic agents for the management of osteoporosis.
Some studies have shown that dietary hydrolyzed collagen peptides (HC) effectively prevent age-related bone loss. However, it is not known whether the intake of HC also has positive effect on bone mass or strength when combined with exercise during growth phase.
We examined the effects of 11 weeks of HC intake and running exercise on bone mass and strength in growing rats. Rats were randomized into four groups, the 20% casein group (Casein20), the 40% casein group (Casein40), the 20% HC group (HC20), and the 40% HC group (HC40). Each group was further divided into exercise groups (Casein20 + Ex, Casein40 + Ex, HC20 + Ex, HC40 + Ex) and non-exercise group (Casein20, Casein40, HC20, HC40). In the HC intake groups, 30% of casein protein was replaced with HC. Exercise group rats were trained 6 days per week on a treadmill (25--30 m/min, 60 min) for 60 days. After being sacrificed, their bone mineral content (BMC) and bone strength were evaluated.
Exercise and dietary HC effects were observed in the adjusted BMC of lumbar spine and tibia among the 20% protein groups (p < 0.001 for exercise; p < 0.05 for dietary HC, respectively). These effects were also noted in the adjusted wet weight and dry weight of femur among the 20% protein groups (p < 0.001, p < 0.01 for exercise; p < 0.01, p < 0.001 for dietary HC, respectively). On the other hand, in adjusted bone breaking force and energy, dietary HC effect was not significant. Among the 40% protein groups, similar results were obtained in the adjusted BMC, femoral weight, bone breaking force, and energy. There were no differences between the 20% protein groups and the 40% protein groups.
The present study demonstrated that moderate HC intake (where the diet contains 20% protein, of which 30% is HC) increased bone mass during growth period and further promoted the effect of running exercise. On the other hand, a higher HC intake (where the diet contains 40% protein, of which 30% is HC) had no more beneficial effect on bone mass than the moderate HC intake.
Abstract Collagen hydrolysates (CHs) are mixtures of peptides obtained by partial hydrolysis of gelatin that are receiving scientific attention as potential oral supplements for the restoration of osteoarticular tissues. The aim of this study was to evaluate the effectiveness of CHs for promoting longitudinal bone growth in growing rats. An in vitro study was carried out in osteoblast-like MG63 cells and the most effective CH on bone formation was selected among 36 various CHs. An in vivo study confirmed the functional effects of a selected CH with molecular weight of <3 kDa on longitudinal bone growth. CHs dose-dependently promoted the longitudinal bone growth and height of the growth plate in adolescent male rats, whereas gelatin failed to affect longitudinal bone growth. Insulin-like growth factor-1 and bone morphogenetic protein-2 in the CH treated group were highly expressed in the growth plate. These results suggest that CHs isolated in this study may provide beneficial effects on bone metabolism of growing animals and humans.
To investigate the role of type X collagen in skeletal development, we have generated type X collagen-null mice. Surprisingly, mice without type X collagen were viable and fertile and had no gross abnormalities in long bone growth or development. No differences were detected between the type X collagen-null mice and controls when growth plates of both newborn and 3-week old mice were examined by histology and by immunostaining for extracellular matrix components of bone including osteopontin, osteocalcin and type II collagen. Our results suggest that type X collagen is not required for long bone development. However, mice and humans with dominant acting type X collagen mutations have bone abnormalities, suggesting that only the presence of abnormal type X collagen can modify bone growth and development.
Osteoporosis has become a major health concern, carrying a substantial burden in terms of health outcomes and costs. We constructed a model to quantify the potential effect of an additional intake of calcium from dairy foods on the risk of osteoporotic fracture, taking a health economics perspective.
Introduction
This study seeks, first, to estimate the impact of an increased dairy consumption on reducing the burden of osteoporosis in terms of health outcomes and costs, and, second, to contribute to a generic methodology for assessing the health-economic outcomes of food products.
Methods
We constructed a model that generated the number of hip fractures that potentially can be prevented with dairy foods intakes, and then calculated costs avoided, considering the healthcare costs of hip fractures and the costs of additional dairy foods, as well as the number of disability-adjusted life years (DALYs) lost due to hip fractures associated with low nutritional calcium intake. Separate analyses were done for The Netherlands, France, and Sweden, three countries with different levels of dairy products consumption.
Results
The number of hip fractures that may potentially be prevented each year with additional dairy products was highest in France (2,023), followed by Sweden (455) and The Netherlands (132). The yearly number of DALYs lost was 6,263 for France, 1,246 for Sweden, and 374 for The Netherlands. The corresponding total costs that might potentially be avoided are about 129 million, 34 million, and 6 million Euros, in these countries, respectively.
Conclusions
This study quantified the potential nutrition economic impact of increased dairy consumption on osteoporotic fractures, building connections between the fields of nutrition and health economics. Future research should further collect longitudinal population data for documenting the net benefits of increasing dairy consumption on bone health and on the related utilization of healthcare resources.
Evaluation of the efficacy and safety of a food supplement made of collagen hydrolysate 1200 mg/day versus placebo during 6 months, in subjects with joint pain at the lower or upper limbs or at the lumbar spine.
Comparative double-blind randomized multicenter study in parallel groups.
200 patients of both genders of at least 50 years old with joint pain assessed as ≥30 mm on a visual analogical scale (VAS).
Collagen hydrolysate 1200 mg/day or placebo during 6 months.
Comparison of the percentage of clinical responder between the active collagen hydrolysate group and the placebo group after 6 months of study. A responder subject was defined as a subject experiencing a clinically significant improvement (i.e. by 20% or more) in the most painful joint using the VAS score. All analyses were performed using an intent-to-treat procedure.
At 6 months, the proportion of clinical responders to the treatment, according to VAS scores, was significantly higher in the collagen hydrolysate (CH) group 51.6%, compared to the placebo group 36.5% (p<0.05). However, there was no significant difference between groups at 3 months (44.1% vs. 39.6%, p=0.53). No significant difference in terms of security and tolerability was observed between the two groups.
This study suggests that collagen hydrolysate 1200 mg/day could increase the number of clinical responders (i.e. improvement of at least 20% on the VAS) compared to placebo. More studies are needed to confirm the clinical interest of this food supplement.
Due to the increased consumption of marine collagen peptides preparation (MCP) as ingredients in functional foods and pharmaceuticals, it was necessary to carry out safety requirements in the form of an oral chronic toxicity assessment. In order to define the oral chronic toxicity of MCP, a 24-month feeding study of MCP was carried out. Sprague-Dawley (S-D) rats at the age of four-week of both sexes were treated with MCP at the diet concentrations of 0%, 2.25%, 4.5%, 9% and 18% (wt/wt). The actual food intake and bodyweight of the individual animals were recorded periodically until sacrifice. Blood and urine samples were collected for serum chemistry evaluations and urinalysis. Throughout the experimental period, there was no toxicologically significant difference between the vehicle and MCP-treated animals with respect to the survival rate, body weight, food consumption, urinalysis, clinical biochemistry parameter and relative organ weight in either sex. Moreover, incidences of non-neoplastic lesions in MCP-treated groups did not significantly increase compared with the control group. Under the present experimental conditions, no higher risk of chronic toxic effects was observed in MCP-treated rats at the diet concentrations of 2.25%, 4.5%, 9% and 18% (wt/wt) than in the rats fed with basal rodent diet.
The possibility of obtaining calcitonin and/or calcitonin-gene-related peptide (CGRP) immunorelated molecules from partly digested proteins was investigated with fish and shrimp hydrolysates. These two peptides were quantified by both radioimmunoassay and radioreceptor assay; the positive extracts were partly purified. Different hydrolysates were analysed: cod head, stomach and viscera hydrolysates, a shrimp hydrolysate and two sardine hydrolysates. Although each cod extract interacted in the CGRP radioimmunoassay, none of these extracts was able to displace the CT binding to its antibody. In contrast, shrimp and sardine hydrolysates interacted with both radioimmunoassays, Radioreceptor assays performed on the same extracts demonstrated that only three extracts contained the structural determinants that allowed them to interact in the CGRP radioreceptor assay. No interaction with the calcitonin radioreceptor assay could be demonstrated. Molecular sieving of the two sardine extracts showed that the immunoreactivity was resolved into two main fractions, The higher-molecular-mass fraction interacted only in the CG RP radioreceptor assay. The results obtained suggest the presence of a biologically related CGRP molecule in peptone hydrolysates and requires further investigation into the role of these peptide fragments in the regulation of intestinal function by partly digested proteins.
The long-term effect of ovariectomy on the loss of bone mineral density (BMD) was evaluated in rats with and without estrogen treatment; BMD was studied in the lumbar and caudal vertebrae, measured by DXA, to find how the losses of BMD occur in the axial skeleton. Seventy female Wistar rats of 3 months of age were divided into four groups as follows: group 1: control animals; group 2: ovariectomized animals; group 3: ovariectomized animals undergoing treatment with estrogen (0.25 mg/kg per week of 17-beta estradiol); group 4: ovariectomized rats undergoing estrogen treatment only during the last 3 months of the experimental period. No significant differences were found among the groups in regard to the BMD values of the caudal vertebrae at either 3 or 6 months. Likewise, in the lumbar vertebrae there were no significant differences among the groups after 3 months. However, at 6 months, a decrease in the BMDs of the ovariectomized animals with respect to the remaining groups was found: 226 +/- 11 mg/cm2 in the ovariectomized group; 262 +/- 14 mg/cm2 in the controls; 255 +/- 4 mg/cm2 in the rats receiving estrogen treatment for 6 months; and 259 +/- 5 mg/cm2 in the animals receiving estrogen for 3 months. The study also reveals the absence of differences in the bone mineral density between the ovariectomized and control rats when the former received estrogen treatment.(ABSTRACT TRUNCATED AT 250 WORDS)
In recent years, much attention has been paid to the existence of peptides with biological activities and proteins derived from foods that might have beneficial effects for humans. Fishes are rich sources of structurally diverse bioactive compounds. The importance of fish as a source of novel bioactive substances is growing rapidly. Marine fish is a major source of high-quality peptides, proteins, lipids, and a wide variety of vitamins and minerals. Marine fish-derived bioactive peptides based on their structural properties and their amino acid composition and sequences, in addition to nutrient utilization, these peptides may be involved in various biological functions and including inhibition of angiotensin-I-converting enzyme (ACE) and and antioxidant and immunomodulatory, antimicrobial and and anticoagulant activities. Development of bioactive peptides from fish proteins and their health promoting ability is increasing. Protein and bioactive peptides from marine fish may promote the production of potentially lead to promising therapeutic applications. Marine fish usually find extensive use as therapeutic and diagnostic proteins. This review paper highlights the important functions and therapeutics potentials of Marine Fish-derived Bioactive Peptides and Proteins for human therapy.
Relevant mechanical properties of bone The mechanical properties of bone material are determined by the relative amounts of its 3 major constituents: mineral, water, and organics (mainly type I collagen); by the quality of these components; and by how the resulting material is arranged in space. For our purposes, the mechanical properties of bone can be summed up as follows: modulus of elasticity, yield stress and yield strain, post-yield stress and post-yield strain, and the total area under the stress-strain curve. Also important are some fracture mechanics properties, but these are not discussed here. A typical tensile stress-strain curve for a bone specimen is shown in Fig. 1. The modulus of elasticity shows how stiff the bone material is. Indeed, stiffness is the prime property of bone, distinguishing it from tendon, which has much less tensile stiffness, almost no shear stiffness, but which is nearly as strong and is much tougher. Yield stress and strain determine how much energy can be absorbed before irreversible changes take place. Post-yield stress and strain determine mainly how much energy can be absorbed after yield but before fracture. Irreversible changes take place at yield, caused by microdamage. The total area under the stress-strain curve is equivalent to the work that must be done per unit volume on the specimen before it breaks. Fracture mechanics properties show the extent to which bone is resistant to crack initiation and to crack travel (which are different things and governed by somewhat different features). In fact, crack travel resistance is given rather well by post-yield stress and strain.
Beneficial effects of collagen peptide ingestion to reduce serum triacylglycerides have been reported, suggesting that the physiological conditions of adipocytes are modulated following collagen peptide ingestion. In this study, the effects of prolylhydroxyproline, a major digestive product of ingested collagen peptide in the blood, on the differentiation of mouse 3T3-L1 preadipocytes in vitro were investigated. 3T3-L1 preadipocytes were induced to differentiate to adipocytes and treated with prolylhydroxyproline, or with an amino acid mixture of proline and hydroxyproline as a control. The amount of lipid was not affected by these treatments. However, the size of the lipid droplet was significantly smaller when treated with prolylhydroxyproline compared to the amino acid mixture or the non-treated control. Proton-coupled oligopeptide transporters were expressed in non-differentiated and/or differentiated 3T3-L1 cells. These results suggest that prolylhydroxyproline might modulate the morphology of lipid droplets by incorporation into adipocytes through the transporters.
Fish proteins are potential sources of natural medicines and food additives. There are many studies being performed to develop underutilized fish proteins. Therefore, the aim of this study was to determine how shark protein functions as a dietary supplement for bone health.
Three groups of ovariectomized (OVX) rats were fed different diets containing 20% casein protein, 20% shark protein, or 20% cod protein for 4 wk. Bone mineral density of the right femur was measured by dual-energy x-ray absorptiometry and quantitative computed tomography. Furthermore, we prepared low-molecular-weight peptides from shark protein using protease for in vitro studies. Calcitriol was added to bone marrow cells and the receptor activator of the nuclear factor-κB ligand was added to RAW264 cells. After 7 d, the number of tartrate-resistant acid phosphatase-positive cells was counted.
In the shark protein-fed group, bone mineral density of the femur epiphysis was higher than that of the casein protein-fed group. In particular, the shark protein-fed group showed an increase in bone mineral density, represented mainly by trabecular bone. Shark protein hydrolysates inhibited osteoclast formation in bone marrow cells and RAW264 cells.
These results suggest that shark protein might suppress the bone loss caused by estrogen deficiency through the suppression of osteoclast formation.
The presence of hydroxyproline (Hyp)-containing peptides in human blood after collagen hydrolysate ingestion is believed to exert beneficial effects on human health. To estimate the effective beneficial dose of these peptides, we examined the relationship between ingested dose and food-derived Hyp levels in human plasma. Healthy volunteers (n = 4) ingested 30.8, 153.8 and 384.6 mg per kg body weight of collagen hydrolysate. The average plasma concentration of Hyp-containing peptides was dose-dependent, reaching maximum levels of 6.43, 20.17 and 32.84 nmol/ml following ingestion of 30.8, 153.8 and 384.6-mg doses of collagen hydrolysate, respectively. Ingesting over 153.8 mg of collagen hydrolysate significantly increased the average concentrations of the free and peptide forms of Hyp in plasma. The Hyp absorption limit was not reached with ingestion of as much as 384.6 mg of collagen hydrolysate. These finding suggest that ingestion of less than 30.8 mg of collagen hydrolysate is not effective for health benefits.
Peptides in the blood of subjects before and after collagen hydrolysate ingestion were fractionated by ion exchange and size-exclusion chromatographies and then derivatised with phenyl isothiocyanate. The derivatives were characterised by reserved phase (RP)-HPLC. Prolyl-hydroxyproline (Pro-Hyp), which has been identified in the previous studies, was detected as a major food-derived collagen peptide in the blood of all subjects (n=5). Another major peptide was identified as hydroxyprolyl-glycine (Hyp-Gly) in the blood of four subjects, which has not been detected in previous studies. The ratio of Hyp-Gly to Pro-Hyp depended on subjects and ranged from 0.00 to 5.04. Hyp-Gly was less susceptible to human serum peptidase than Pro-Hyp. Hyp-Gly enhanced the growth of mouse primary fibroblasts on collagen gels in a higher extent than Pro-Hyp. These findings suggest that Hyp-Gly plays a significant role in exerting the biological effects by ingestion of collagen hydrolysate.
The rising interest in the valorisation of industrial by-products is one of the main reasons why exploring different species and optimizing the extracting conditions of collagen and gelatin has attracted the attention of researchers in the last decade. The most abundant sources of gelatin are pig skin, bovine hide and, pork and cattle bones, however, the industrial use of collagen or gelatin obtained from non-mammalian species is growing in importance. The classical food, photographic, cosmetic and pharmaceutical application of gelatin is based mainly on its gel-forming properties. Recently, and especially in the food industry, an increasing number of new applications have been found for gelatin in products such as emulsifiers, foaming agents, colloid stabilizers, biodegradable film-forming materials and micro-encapsulating agents, in line with the growing trend to replace synthetic agents with more natural ones. In the last decade, a large number of studies have dealt with the enzymatic hydrolysis of collagen or gelatin for the production of bioactive peptides. Besides exploring diverse types of bioactivities, of an antimicrobial, antioxidant or antihypertensive nature, studies have also focused on the effect of oral intake in both animal and human models, revealing the excellent absorption and metabolism of Hyp-containing peptides. The present work is a compilation of recent information on collagen and gelatin extraction from new sources, as well as new processing conditions and potential novel or improved applications, many of which are largely based on induced cross-linking, blending with other biopolymers or enzymatic hydrolysis.
Collagens form critical elements of extracellular matrices that provide mechanical strength to skeletal tissues and serve as a binding platform for cells of bone and cartilage. The formation of collagen-rich extracellular matrices is a complex process that involves intracellular and extracellular steps. Mutations in genes that encode individual chains of triple-helical collagens present in bone and cartilage are associated with heritable diseases of skeletal tissues. In addition, mutations in genes encoding proteins involved in the intracellular and extracellular modifications of collagens are also responsible for developing skeletal abnormalities. In this review, we will summarize the pathomechanisms of molecular and cellular consequences of mutations that alter collagen structure and function. Moreover, we will discuss the prospects and limitations of therapeutic approaches to minimize the effects of mutations that affect collagens of skeletal tissues.
This study was designed to investigate the biological effects of fish collagen peptide (FCP) on human osteoblasts. Human osteoblasts were treated with 0.1% FCP, which was the optimal concentration confirmed by the increase in alkaline phosphatase activity. After one, three, five and seven days of culture, the number of FCP-treated cells increased significantly compared with untreated cells. In a real-time PCR analysis, the expression of osteocalcin, osteopontin, BMP-2 and integrin β3 mRNAs in FCP-treated cells showed increases compared with untreated cells after three days of culture. After seven days of culture, the expression levels of osteopontin and integrin β3 were still higher in the FCP-treated cells than in untreated cells. The production of osteocalcin, osteopontin and integrin β3 proteins in FCP-treated cells also showed increases after seven days of culture. Furthermore, FCP accelerated matrix mineralization in the cultures. The present study indicates the potential utility of FCP as a biomaterial.
The aims of this study were to report for the first time, the extraction and physico-chemical properties of
chicken skin gelatin compared to bovine gelatin. Extracted chicken skin gelatin 6.67% (w/v) had a higher
bloom value (355 � 1.48 g) than bovine gelatin (259 � 0.71 g). The dynamic viscoelastic profile of chicken
gelatin exhibited higher viscous and elastic modulus values compared to bovine gelatin for a range of
concentrations and frequencies. Thermal properties studied by differential scanning calorimetry (DSC)
showed that the melting temperature of 6.67%, chicken skin gelatin was significantly greater (p < 0.05)
than that of bovine gelatin, indicating lower stability of bovine gelatin compared to chicken skin gelatin.
Results obtained in this study showed that Gly (33.70%), Pro (13.42%), H.Pro (12.13%) and Ala (10.08%)
were the most dominant amino acids in chicken skin gelatin which contributed to the higher gel strength
and stability. Raman spectra of chicken skin and bovine gelatin were similar and displayed typical protein
spectra. Chicken gelatin showed strong hydrogen bonding compared to bovine gelatin as the tyrosine
doublet ratio (I855/I830) of chicken gelatin was significantly lower than that of bovine gelatin. Significantly,
the alpha helix and b-sheet type structures were higher for chicken skin gelatin compared with
bovine gelatin. The average molecular weight of chicken gelatin was 285,000 Da. These findings, obtained
for the first time for chicken skin gelatin, show that it has high potential for application as an
alternative to commercial gelatin.
Collagen is one of the most widely used biomaterials for tissue engineering and regenerative medicine. Fish collagen peptides (FCP) have been used as a dietary supplement, but their effects on the cellular function are still poorly understood. The objective of this study was to investigate the effects of FCP on collagen synthesis, quality and mineralization using an osteoblastic MC3T3-E1 cell culture system. Cells treated with FCP significantly upregulated the gene expression of several collagen modifying enzymes and more collagen was deposited in the cultures. Collagen in the treated group showed a greater extent of lysine hydroxylation, higher levels of hydroxylysine-aldehyde derived cross-links and accelerated cross-link maturation compared with the untreated group. Furthermore, the treated group showed accelerated matrix mineralization. These results indicate that FCP exerts a positive effect on osteoblastic cells in terms of collagen synthesis, quality and mineralization, thereby suggesting the potential utility of FCP for bone tissue engineering.
There is a need to understand the role of nutrition, beyond calcium and vitamin D, in the treatment and prevention of osteoporosis in adults. Results regarding soy compounds on bone density and bone turnover are inconclusive perhaps due to differences in dose and composition or in study population characteristics. The skeletal benefit of black cohosh and red clover are unknown. Dehydroepiandrosterone (DHEA) use may benefit elderly individuals with low serum dehydroepiandrosterone-sulfate levels, but even in this group, there are inconsistent benefits to bone density (BMD). Higher fruit and vegetable intakes may relate to higher BMD. The skeletal benefit of flavonoids, carotenoids, omega-3-fatty acids, and vitamins A, C, E and K are limited to observational data or a few clinical trials, in some cases investigating pharmacologic doses. Given limited data, it would be better to get these nutrients from fruits and vegetables. Potassium bicarbonate may improve calcium homeostasis but with little impact on bone loss. High homocysteine may relate to fracture risk, but the skeletal benefit of each B vitamin is unclear. Magnesium supplementation is likely only required in persons with low magnesium levels. Data are very limited for the role of nutritional levels of boron, strontium, silicon and phosphorus in bone health. A nutrient rich diet with adequate fruits and vegetables will generally meet skeletal needs in healthy individuals. For most healthy adults, supplementation with nutrients other than calcium and vitamin D may not be required, except in those with chronic disease and the frail elderly.
The amino acid composition and antioxidant activities of different hydrolysates from porcine collagen were analyzed. The gelatin was hydrolyzed for antioxidative peptides with various proteases, namely papain, protease from bovine pancreas, protease from Streptomyces, and cocktail mixture of protease from bovine pancreas and protease from Streptomyces. The hydrolysates were assessed using methods of DPPH radical-scavenging ability, metal-chelating ability and lipid peroxidation inhibition activity. It was found that the collagen hydrolysates by different protease treatments had different amino acid compositions and antioxidant properties. However, the contents of Hyp and Pro were improved and the content of Gly was decreased in each collagen hydrolysate compared with collagen. The hydrolysate prepared with the cocktail mixture of proteases, which exhibited the highest antioxidant activity, was separated into 6 fractions by gel filtration chromatography. Fraction 2 was further separated by ion exchange chromatography. Fraction 2b with abundant basic amino acids and Fraction 2d which was slightly acidic fractions had higher radical-scavenging and metal-chelating activities, and both Fraction 2b and 2d contained more hydrophobic amino acids. The results confirmed that the antioxidative peptides were rich in Hyp, Pro and Gly, which accounted for half of amino acid composition. This article added further support to the preparation of natural antioxidative peptides from porcine skin collagen.
Acid-solubilized collagen (ASC) and pepsin-solubilized collagen (PSC) were extracted from the skin of cobia (Rachycentron canadum). The yields of ASC and PSC were 35.5% and 12.3%, respectively. Based on the protein patterns and carboxymethyl-cellulose chromatography, ASC and PSC were composed of α1α2α3 heterotrimers and were characterised as type I collagen with no disulfide bond. Their amounts of imino acids were 203 and 191 residues per 1000 residues, respectively. LC-MS/MS analysis demonstrated the high sequences similarities of ASC and PSC. Fourier transform infrared spectroscopy spectra showed that the amide I, II and III peaks of PSC were obtained at a lower wave number compared with ASC. The thermal denaturation temperatures of ASC and PSC, as measured by viscometry, were 34.62 and 33.97°C, respectively. The transition temperatures (T(max)) were 38.17 and 36.03°C, respectively, as determined by differential scanning calorimetry (DSC). Both collagens were soluble at acidic pH and below 2% (w/v) NaCl concentration.
Quinoa protein/chitosan films were obtained by solution casting of blends of quinoa protein extract (PE) and chitosan (CH). Films from a PE/CH blend were characterized by FTIR, X-ray diffraction, thermal analysis, and SEM. The tensile mechanical, barrier, and sorption properties of the films were also evaluated. The blend of PE with CH yielded mechanically resistant films without the use of a plasticizer. The film had large elongation at break, and its water barrier properties showed that they were more hydrophilic than CH film. The thickness and water-vapor permeability of PE/CH (v/v) 1/1 blend film increased significantly compared to pure CH films. CH films are translucent in appearance and yellowish in blend with PE. By blending anionic PE with cationic CH an interaction between biopolymers was established with different physicochemical properties from those of pure CH. Drying and sorption properties show significant differences between PE/CH blend film and CH film. The structural properties determined by XRD, FTIR and TGA showed a clear interaction between quinoa proteins and CH, forming a new material with enhanced mechanical properties.Graphical abstract
The influence of oral administration of High Advanced-Collagen Tripeptide (HACP) developed by Jellice Co., Ltd (Sendai, Japan) for bone repairing process was investigated in rat. Cortical bone defects (1mm diameter) of the tibia were created through the medial cortex and medulla. After 1 day of operation, HACP/ physiological saline solution (80mg/2ml/Kg) was orally administrated as experimental group. Only physiological saline solution was orally administrated as control group. There were no significant differences in body weight, serum levels of total proteins, calcium concentration and alkaline phosphatase activity between HACP and control groups during administration periods. After 3 weeks of administration, the tibia bone was excised. Micro high-resolution microfocus x-ray computed tomography showed the formation of primary woven bone in HACP group. Histological sections stained with hematoxylin and eosin were observed under light microscope. In control group, blood clot inside the bone defect with a thin connective tissue surrounding the defect was observed. On the contrary, bone defect area was filled with granulation tissue, blood clot and a great number of osteoblasts in the HACP group. The present results suggested that oral administration of HACP may provide a beneficial effect on bone healing process.
Skeletal growth depends on endochondral ossification in growth plate cartilage, where proliferation of chondrocytes, matrix synthesis, and increases in chondrocyte size all contribute to the final length of a bone. To learn more about the potential role of matrix synthesis/degradation dynamics in the determination of bone growth rate, we investigated the expression of matrix collagens and collagenase 3 in tibial growth plates in three age groups of rats (21, 35, and 80 days after birth), each characterized by specific growth rates. By combining stereological and in situ hybridization techniques, it was found that the expression of matrix collagens and collagenase 3 was specifically turned on or off at specific stages of the chondrocyte-differentiation cycle, and these changes occurred as a temporal sequence that varied depending of animal growth rate. Furthermore, the expression of these matrix proteins by a growth plate chondrocyte was found to be sped up or slowed down depending of the growth rate. In addition to expression of types II and X collagen, collagenase-3 expression was found to constitute a constant event in the series of changes in gene expression that takes place during the chondrocyte-differentiation process. Collagenase-3 expression was found to show a biphasic pattern: it was intermittently expressed at the proliferative phase and uniformly expressed at the hypertrophic stage. An intimate relationship between morphological and kinetic changes associated with chondrocyte hypertrophy and changes in the expression pattern of matrix collagens and collagenase 3 was observed. Present data prove that the matrix synthesis/degradation dynamics of the growth plate cartilage varied depending on growth rate; these results support the hypothesis that changes in matrix degradation and synthesis are a critical link in the sequence of tightly regulated events that lead to chondrocytic differentiation.
Gelatin is regarded as a special and unique hydrocolloid, serving multiple functions with a wide range of applications. The main sources of gelatin include pigskin, cattle bones and cattle hide. Gelatin replacement has been a major issue in recent years due to the emerging and lucrative vegetarian, halal and kosher markets. It has recently gained increased interest, especially within Europe, with the emergence of bovine spongiform encephalopathy (BSE) in the 1980s. In this paper, we will discuss the unique properties of gelatin, the rationale for developing gelatin alternatives, the progress to date of research in development of gelatin alternatives, possible approaches for developing gelatin alternatives, and future directions for research in this area.
Squid gelatin obtained from inner and outer tunics was hydrolysed with Alcalase to isolate antioxidant peptide sequences. The ACE-inhibitory activity of the isolated peptides was also evaluated. After fractionation by ultrafiltration and size-exclusion chromatography into four fractions, the antioxidant activity of the peptide fractions was determined by radical scavenging ability and ferric reducing power. Fraction FIII showed the highest antioxidant activity, although slight differences could be expected in the antioxidant activity of the different fractions based on the amino acid composition. FIII was subjected to liquid chromatography and tandem mass spectrometry (LC–MS/MS) and two major compounds were identified: the compound with m/z 952.42, which could be mostly comprised by the carbohydrate fucose, and the peptide with m/z 1410.63. Three possible sequences were proposed and synthesised for this peptide, and the contribution of Leu or Hyp residues to the antioxidant and ACE-inhibitory activities of the resulting sequence was evaluated. The presence of Leu residues in the peptide sequence in replacement of Hyp seems to play an important role in the antioxidant and ACE-inhibitory activity.
Osteoblasts derived from Day 21 fetal rat calvaria grown on films of collagen type I exhibit an earlier and enhanced expression of the differentiated phenotype, compared to cells cultured on plastic. The temporal expression of genes characterizing three distinct periods of growth and differentiation are dramatically modified. During the initial proliferation period, expression of genes normally expressed at high levels on plastic (fibronectin, β1 integrin, and actin) was decreased from 50 to 70% in cells grown on collagen. Genes normally expressed at maximal levels in the postproliferative period (osteonectin, osteocalcin, and osteopontin) were up-regulated severalfold very early. Alkaline phosphatase enzyme activity was elevated 2- to 3-fold during the proliferation period, while mRNA levels remained low, suggesting post-transcriptional modifications. The most dramatic consequence of culture of cells on collagen is the accelerated and uniform mineralization of the matrix in contrast to the focal mineralization confined to bone nodules in cultures on plastic. Type I collagen supports maintenance of osteoblast phenotypic properties of passaged cells in the absence of glucocorticoid supplementation required for differentiation of osteoblasts subcultivated on plastic. Treatment of proliferating rat osteoblasts on plastic with 1,25(OH)2D3 blocks osteoblast differentiation and matrix mineralization. Although differentiation-related genes (alkaline phosphatase and osteocalcin) were up-regulated by vitamin D, culture on the collagen matrix could not overcome the inhibition of mineralization. Taken together, these studies define the critical role of type I collagen in mediating the signaling cascade for expression of a mature osteoblast phenotype and mineralization of the extracellular matrix in a physiological manner.
This paper relates the comparison of three methods applied to characterize the molecular weight of hydrolyzed collagen (HC). The high-pressure size-exclusion chromatography (HPSEC) performed on hydrophilic macroporous copolymer beads with an aqueous denaturing eluent allows the determination of the molecular weights distribution and of the weight—and number-average molecular weights. The viscosity measured in standard conditions and the number-average molecular weight determined by formol titration are correlated respectively with the weight-average molecular weight (log scale) and the number-average molecular weight determined by HPSEC.
In an investigation into making more effective use of under-utilized resources, type I collagen was prepared from fish skin, bone and fin, respectively. As a result, the yields of these collagens were as follows: (1) skin collagen, 51.4% (Japanese sea-bass), 49.8% (chub mackerel) and 50.1% (bullhead shark), respectively; (2) bone collagen, 42.3% (skipjack tuna), 40.7% (Japanese sea-bass), 53.6% (ayu), 40.1% (yellow sea bream) and 43.5% (horse mackerel), respectively; (3) fin collagen, 5.2% (Japanese sea-bass acid-soluble collagen) and 36.4% (Japanese sea-bass acid-insoluble collagen), on the basis of lyophilized dry weight. The denaturation temperatures of these collagens were as follows: skin collagen (25.0–26.5°C), bone collagen (29.5–30.0°C) and fin collagen (28.0–29.1°C), respectively. These values were lower about 7–12°C than that of porcine skin collagen. This report indicates that these fish waste materials have potential in supplementing the skin of land vertebrates as a source of collagen.
Food and pharmaceutical industries all over the world are witnessing an increasing demand for collagen and gelatin. Mammalian gelatins (porcine and bovine), being the most popular and widely used, are subject to major constraints and skepticism among consumers due to socio-cultural and health-related concerns. Fish gelatin (especially from warm-water fish) reportedly possesses similar characteristics to porcine gelatin and may thus be considered as an alternative to mammalian gelatin for use in food products. Production and utilization of fish gelatin not only satisfies the needs of consumers, but also serves as a means to utilize some of the byproducts of the fishing industry. This review focuses on the unique features, advantages, constraints, and challenges involved in the production and utilization of fish gelatin in order to provide a comprehensive look and deeper insight on this important food ingredient, as well as prospects for its future commercial exploitation and directions for future studies.
Prolylhydroxyproline (Pro-Hyp), which is derived from collagen hydrolysate, has been shown to be beneficial for skin and joint health. However, little is known about the distribution of Pro-Hyp in these tissues. In the present study, we investigated the biodistribution of orally administered [(14)C]Pro-Hyp in rats. Whole-body autoradiography at 30 min after administration of [(14)C]Pro-Hyp showed that radioactivity is widely distributed in tissues including skin and articular cartilage, with the highest level of radioactivity observed in the gastric and intestinal walls. Incorporation of radioactivity into cells known to respond to Pro-Hyp such as dermal fibroblasts, synovial cells, chondrocytes, osteoblasts, and osteoclasts was observed. The chemical form of [(14)C]Pro-Hyp-derived radioactivity detected in the tissues was investigated by thin layer chromatography. The radioactive constituents in cartilage extract were two proline-modified peptides (56%), intact Pro-Hyp (5%), and two nonpeptide metabolites (28%). Similar results were obtained for skin and bone marrow. Plasma analysis at 3 to 30 min post-dose suggested that the majority of Pro-Hyp is modified in its proline residue by a first-pass effect without peptide bond hydrolysis. In conclusion, we demonstrated that Pro-Hyp is partly distributed in observed tissues including skin and cartilage in its intact form, which might be responsible for its biological functions.
Bone consists of type I collagen as a major protein with minor various matrix proteins. Type VI collagen is one of bone matrix proteins but its function is not known. We therefore examined the effects of type VI collagen deficiency on bone. 3D-μCT analysis revealed that type VI collagen deficiency reduced cancellous bone mass. Cortical bone mass was not affected. Type VI collagen deficiency distorted the shape of osteoblasts both in the cancellous bone and in the cambium layer of periosteal region. Furthermore, type VI collagen deficiency disorganized collagen arrangement. These data indicate that type VI collagen contributes to maintain bone mass.
Plasma levels of prolyl-hydroxyproline (Pro-Hyp) and hydroxyprolyl-glycine (Hyp-Gly) in healthy volunteers (n=5) after ingestion of collagen hydrolysate were estimated by liquid chromatography-tandem mass spectrometry. The ratio of Hyp-Gly to Pro-Hyp was distributed in the range of 0.063-0.221. This is a first report for quantification of food-derived Hyp-Gly in human plasma.
This study evaluates the effect of hydrolyzed collagen (HC) on bone health of ovariectomized mice (OVX) at different ages. Twenty-six weeks after the OVX procedure, HC ingestion was still able to improve significantly bone mineral density (BMD) and some femur biomechanical parameters. Moreover, HC ingestion for 1 month before surgery prevented BMD decrease.
HC can play an important role in preserving BMD before osteoporosis appears. The aim of this study was to evaluate the effect of HC on bone health of ovariectomized mice at different ages.
Female C3H mice were either OVX at 3 or 6 months and fed for 6 months (first experiment) or 3 months (second experiment) with diet including 0, 10, or 25 g/kg of HC. In the second experiment, one group received HC 1 month before surgery, and two groups received the supplementation immediately after surgery, one fed ad libitum and the other by gavage. Mice treated with raloxifene were used as a positive control. BMD, femur intrinsic and extrinsic biomechanical properties, and type I collagen C-terminal telopeptide were measured after 12 and 26 weeks. Food intake and spontaneous physical activity were also recorded.
The OVX procedure increased body weight, while food intake decreased, thus suggesting that resting metabolism was decreased. Ingestion of 25 g/kg of HC for 3 or 6 months reduced bone loss significantly in, respectively, 3- and 6-month-old OVX mice. The lowest HC concentration was less efficient. HC ingestion for 3 months is as efficient as raloxifene to protect 3-month-old OVX mice from bone loss. Our results also demonstrated that HC ingestion before surgery prevented the BMD decreases.
This study confirms that dietary collagen reduces bone loss in OVX mice by increasing the diameter of the cortical areas of femurs and can have a preventive effect.
Bone collagen cross-links are now widely used to assess bone resorption levels in many metabolic bone diseases. The post-translational modifications of bone and other mineralizing collagens are significantly different from those of other type I collagen matrices, a fact that has been exploited during recent advances in the development of biochemical markers of bone resorption. The enzymatic collagen cross-linking mechanism is based upon aldehyde formation from specific telopeptide lysine or hydroxylysine residues. The immature ketoimine cross-links in bone form via the condensation of a telopeptide aldehyde with a helical lysine or hydroxylysine. Subsequent maturation to the pyridinoline and pyrrole cross-links occur by further reaction of the ketoimines with telopeptide aldehydes. In mineralizing tissues, a relatively low level of lysyl hydroxylation results in low levels of hydroxylysyl pyridinoline, and the occurrence of the largely bone specific lysyl pyridinoline and pyrrolic cross-links. The collagen post-translational modifications appear to play an integral role in matrix mineralization. The matrix of the turkey tendon only mineralizes after a remodeling of the collagen and the subsequent formation of a modified matrix more typical of bone than tendon. Further, disturbances in the post-translational modification of collagen can also affect the mineralization density and crystal structure of the tissue. In addition to their use as a convenient measure of matrix degradation, collagen cross-links are of significant importance for the biomechanical integrity of bone. Recent studies of osteoporotic bone, for example, have demonstrated that subtle perturbations in the pattern of lysine hydroxylation result in changes in the cross-link profile. These alterations, specifically changes in the level of the pyrrolic cross-link, also correlate with the strength of the bone. Further research into the biochemistry of bone collagen cross-links may expand current understanding and their clinical application in metabolic bone disease. This review also demonstrates the potential for further study into this area to provide more subtle information into the mechanisms and etiology of disease and aging of mineralizing tissues.
The aim of the study was to investigate the influence of dietary intake of commercial hydrolyzed collagen (Gelatine Royal) on bone remodeling in pre-pubertal children.
A randomized double-blind study was carried out in 60 children (9.42 +/- 1.31 years) divided into three groups according to the amount of partially hydrolyzed collagen taken daily for 4 months: placebo (G-I, n=18), collagen (G-II, n=20) and collagen+calcium (G-III, n=22) groups. Analyses of the following biochemical markers were carried out: total and bone alkaline phosphatase (tALP and bALP), osteocalcin, tartrate-resistant acid phosphatase (TRAP), type I collagen carboxy-terminal telopeptide, lipids, calcium, 25-hydroxyvitamin D, insulin-like growth factor-1 (IGF-1), thyroid-stimulating hormone, free thyroxin and intact parathormone.
There was a significantly greater increase in serum IGF-1 in G-III than in G-II (p < 0.01) or G-I (p < 0.05) during the study period, and a significantly greater increase in plasma tALP in G-III than in G-I (p < 0.05). Serum bALP behavior significantly (p < 0.05) differed between G-II (increase) and G-I (decrease). Plasma TRAP behavior significantly differed between G-II and G-I (p < 0.01) and between G-III and G-II (p < 0.05).
Daily dietary intake of hydrolyzed collagen seems to have a potential role in enhancing bone remodeling at key stages of growth and development.