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Abstract

Stem cell technology has been a great hope for the treatment of many common problems such as Parkinson's disease, Alzheimer's disease, diabetes, cancer, and tissue regeneration. Therefore, the main challenge in hard tissue engineering is to make a successful combination of stem cells and efficient inductors in the concept of stem cell differentiation into odontogenic and osteogenic cell types. Although some boron derivatives have been reported to promote bone and teeth growth in vivo, the molecular mechanism of bone formation has not been elucidated yet. Different concentrations of sodium pentaborate pentahydrate (NaB) were prepared for the analysis of cell toxicity and differentiation evaluations. The odontogenic, osteogenic differentiation and biomineralization of human tooth germ stem cells (hTGSCs) were evaluated by analyzing the mRNA expression levels, odontogenic and osteogenic protein expressions, alkaline phosphatase (ALP) activity, mineralization, and calcium deposits. The NaB-treated group displayed the highest ALP activity and expression of osteo- and odontogenic-related genes and proteins compared to the other groups and baseline. In the current study, increased in vitro odontogenic and osteogenic differentiation capacity of hTGSCs by NaB application has been shown for the first time. The study offers considerable promise for the development of new scaffold systems combined with NaB in both functional bone and tooth tissue engineering.

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... Boron supplementation increased mineralization and osteogenic differentiation of osteoblasts and bone marrow stromal cells in vitro (Hakki et al., 2010;Ying et al., 2011). In a recent work, we also showed the increase in osteogenic and odontogenic differentiation capacity of human tooth germ stem cells after they were treated with boron (Taşlı et al., 2013). ...
... Experimental composites (4 mm in depth and 6 mm in diameter) were placed on the upper surface of the Transwell culture plate to provide indirect contract between cells and dental materials. hDPSCs were induced to differentiate into osteogenic and odontogenic cell lineages according to the protocol previously used by our group (Taşlı et al., 2013). Briefly, cells were treated with an osteogenic medium composed of growth medium supplemented with 0.1 mmol/L dexamethasone, 10 mmol/L β-glycerolphosphate, and 50 mmol/L ascorbate as well as an odontogenic medium composed of growth medium supplemented with 10 nmol/L dexamethasone, 10 mmol/L β-glycerol-phosphate, and 50 mmol/L ascorbate (Sigma Chemical Co., USA). ...
... Boron has previously been shown to have a role in healing the alveolar bone after tooth extraction (Gorustovich et al., 2008a(Gorustovich et al., , 2008b. Moreover, we have previously reported that boron increases the osteogenic and odontogenic differentiation of dental stem cells (Taşlı et al., 2013;Demirci et al., 2014). Consistent with these findings, the results revealed that the osteogenic and odontogenic potential of hDPSCs was found to be higher in the presence of boron-containing dental composites. ...
Article
Secondary dental caries are one of the major reasons for restoration replacements. Incorporating antimicrobial properties into dental materials would limit the initiation and progression of dental caries. In the current study, dental composites having 1%, 5%, and 10% (w/w) sodium pentaborate pentahydrate were prepared and analyzed for their mechanical properties, degree of monomer conversion (DC) rate, antibacterial effects against Streptococcus mutans, and biocompatibility with human dental pulp stem cells (hDPSCs). Incorporation of boron into the composites significantly decreased flexural strength and DC in a dose-dependent manner, but the value for 1% boron-containing composite still remained within acceptable levels. Compressive strength and diametral tensile strength were not found to be different from those of controls. Although no inhibition zone was detected in an agar-well diffusion assay for any materials tested, significant bacterial growth inhibition was obtained in a direct contact test for boron-containing composites. Immunocytochemical and lineage-specific gene expression analysis revealed that composites with boron content increased the osteogenic and odontogenic capacity of hDPSCs. Boron-containing dental composites showed promising results for future clinical applications, displaying nontoxic, osteogenic, and odontogenic-inducing characteristics with remarkable antibacterial activity against S. mutans, and are hence potentially able to prevent secondary caries.
... The cells were seeded on a six-well plate for RNA isolation and a 48-well plate (BIOFIL, TCP) for immunocytochemistry at a density of 150,000 cells/well and 10,000 cells/well respectively for each differentiation. For osteogenic differentiation the cells were seeded on six-well plates followed by the addition of osteogenic differentiation medium, which consisted of DMEM, 10% (v/v) FBS, 0.1 µM dexamethasone (AppliChem, Germany), 10 mM β-glycerol phosphate (Sigma, USA), and 50 µg/mL ascorbic acid (Sigma) (Taşlı et al., 2013a), with or without 100 nM melatonin. The medium was changed every other day and the cells were treated with the osteogenic differentiation medium for 14 days. ...
... The chondrogenic differentiation lasted for 3 weeks, changing the medium every 2-3 days. The odontogenic differentiation was conducted by treating the cells with differentiation medium containing DMEM, 10% (v/v) FBS, 10 -8 M dexamethasone, 5 mmol/L KH 2 PO 4 , and 50 µg/mL ascorbic acid, with or without melatonin (Taşlı et al., 2013a). Eventually, myogenic differentiation was done with DMEM medium supplemented with 5% (v/v) horse serum, 0.1 µM dexamethasone, and 50 µM hydrocortisone (Sigma) (Taşlı et al., 2013b). ...
Article
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Stem cell-based applications have become a popular and promising approach for therapy for a number of disorders including neurodegenerative diseases, degenerative muscle diseases, and osteoporosis, as well as trauma, inflammations, burns, and injuries. Human tooth germ stem cells are an adult stem cell source; they have mesenchymal stem cell properties and show high proliferative and differentiation capacity. Melatonin has been demonstrated to regulate differentiation of human and mouse mesenchymal stem cells into various cell lineages in addition to its other functions in the body. In the current study, the effects of melatonin on osteogenic, neurogenic, adipogenic, chondrogenic, myogenic, and odontogenic differentiation of human tooth germ stem cells were investigated. The results showed that melatonin increases the viability of cells. It significantly augments osteogenic, neurogenic, chondrogenic, myogenic, and odontogenic differentiation of the cells, whereas it reduces adipogenic differentiation capability. These results suggest that melatonin has a great potential to increase differentiation capacity of human tooth germ stem cells and might be useful in regenerative therapy applications involving stem cell differentiations in addition to defining potential treatments for obesity because of its suppressor effects on adipogenesis.
... Human newborn foreskin stem cells (hnFSSCs) were characterized according to the protocol described previously by our group [27]. Cells were trypsinized and incubated with the primary antibodies which were prepared in PBS. ...
... Immunocytochemistry analyses were completed according to the previously described protocol [27]. At the end of the differentiation procedures of hnFSSCs, the cells were incubated with 2 % (w/v) paraformaldehyde for 30 min at 4°C for fixation and permeabilized with 0.1 % (v/v) Triton X-100 for 5 min and prepared in PBS. ...
Article
Circumcision is described as a cultural, medical, and religious process which states surgical removal of the foreskin either partly or fully. Cells isolated from the circumcised tissues are referred as foreskin cells. They have been thought as feeder cell lines for embryonic stem cells. Their fibroblastic properties were also utilized for several experiments. The waste tissues that remain after the circumcision thought to have stem cell properties. Therefore, there have been very few attempts to expose their stem cell properties without turning them into induced pluripotent stem cells. Although stem cell isolation from prepuce and their mesenchymal multilineage differentiation potential have been presented many times in the literature, the current study explored hematopoietical phenotype of newborn foreskin stem cells for the first time. According to the results, human newborn foreskin stem cells (hnFSSCs) were identified by their capability to turn into all three germ layer cell types under in vitro conditions. In addition, these cells have exhibited a stable phenotype and have remained as a monolayer in vitro. hnFSSCs suggested to carry different treatment potentials for bone damages, cartilage problems, nerve damages, lesion formations, and other diseases that are derive from mesodermal, endodermal, and ectodermal origins. Owing to the location of the tissue in the body and differentiation capabilities of hnFSSCs, these cells can be considered as easily obtainable and utilizable even better than the other stem cell sources. In addition, hnFSSCs offers a great potential for tissue engineering approaches due to exhibiting embryonic stem cell-like characteristics, not having any ethical issues, and teratoma induction as in embryonic stem cell applications.
... B at a physiological amounts (1.0 mg/d) has a beneficial effect on bone health [19,20], while B deficiency (0.07 mg B/kg diet) is detrimental to bone strength, shape, and mineral element compositions [21,22]. B-related materials, such as boric acid, sodium tetraborate, and borate bioactive glasses have exhibited distinctive biological properties like stimulation of bone healing and angiogenesis in vitro and in vivo [23][24][25][26]. In vivo studies indicated that B-modified bioactive glasses induced a higher area of neoformed bone tissue and showed better osseointegration compared with the glasses without B [27,28]. ...
... In the osteogenesis process, ALP is generally used as a typical early marker for osteoblastic differentiation [43]. The cells incubated on the B-CS coating showed evidently greater levels of ALP secretion than that on the CS coating at day 7. Ying et al. [44] reported that the BMSCs exhibited higher levels of ALP activity and the mRNA expression after being exposed to 10 and 100 ng/ml B. Pakize et al. also found the NaB-treated sample presented the higher ALP activity, as well as expression of osteogenic-related genes of human tooth germ stem cells [24]. In addition, the MSCs cultured on the B nitride nanotubes layer (1-10 μg mL −1 ), show obviously increased ALP activity in comparison with the control [40]. ...
Article
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In recent years, CaSiO3 bio-ceramic coatings have attracted great attention because of their good bioactivity. However, their high degradation rates in physiological environment restrict their practical applications. In this work, boron-modified CaSiO3 ceramic (Ca11Si4B2O22, B-CS) coating was developed on Ti substrates by plasma-spraying technique attempting to obtain enhanced chemical stability and osteogenic activity. The B-CS coating possessed significantly increased chemical stability due to the introduction of boron and consequently the modified crystal structure, while maintaining good bioactivity. Scanning electron microscope and immunofluorescence studies showed that better cellular adhesion and extinctive filopodia-like processes were observed on the B-CS coating. Compared with the pure CaSiO3 (CS) coating, the B-CS coating promoted MC3T3-E1 cells attachment and proliferation. In addition, enhanced collagen I (COL-I) secretion, alkaline phosphatase activity, and extracellular matrix mineralization levels were detected from the B-CS coating. According to RT-PCR results, notable up-regulation expressions of mineralized tissue-related genes, such as runt-related transcription factor 2 (Runx2), bone sialoprotein and osteocalcin, and bone morphogenetic protein 7 (BMP-7) were observed on the B-CS coating compared with the CS coating. The above results suggested that Ca11Si4B2O22 coatings possess excellent osteogenic activity and might be a promising candidate for orthopedic applications.
... Sodium pentaborate pentahydrate (NaB; NaB 5 O 8 .5H 2 O) is one of the basic refined boron compound containing 5 moles of water (7). NaB has been found to be successful in a research for odontogenic and osteogenic differentiation capacity (14). NaB added dental composite has been evaluated on S. mutans in an aim of avoiding secondary caries and the physical changes of composite material (15). ...
... However, these results were encouraging for further research on increased concentration of NaB. The present study also confirms previous findings and contributes additional evidence that suggests NaB as an irrigation solution due to the osteoinductive properties of NaB (14). ...
... Moreover, shRNA-mediated knockdown of β-catenin rescued the differentiation potential of boron-treated cells, verifying that the Wnt/β-catenin pathway was at least partially involved in the boron-induced inhibition of adipogenesis. These results were in line with our previous findings about boron's stimulatory role on osteogenic differentiation [38,39] as Wnt signaling stimulates osteogenesis and represses adipogenesis. ...
Article
Obesity is a worldwide medical problem resulting in serious morbidity and mortality involving differentiation of pre-adipocytes into mature adipocytes (adipogenesis). Boron treatment has been reported to be associated with weight reduction in experimental animals; however, its effects on pre-adipocyte differentiation and anti-adipogenic molecular mechanisms are unknown. In this study, we demonstrate the inhibitory activities of boric acid (BA) and sodium pentaborate pentahydrate (NaB) on adipogenesis using common cellular models. Boron treatment repressed the expression of adipogenesis-related genes and proteins, including CCAAT-enhancer-binding protein α and peroxisome proliferator-activated receptor γ, by regulating critical growth factors and the β-catenin, AKT, and extracellular signal-regulated kinase signaling pathways. In addition, although boron treatment did not induce apoptosis in pre-adipocytes, it depressed mitotic clonal expansion by regulation of cell cycle genes. Overall, these data offer promising insights into the prevention/treatment of obesity and associated diseases.
... hADSCs were from our prior study were also used as the in vitro model in the current study Preservation of stem cell markers by these cells that were at late passages was verified according to the protocol described previously [22]. For characterization, cells were trypsinized and incubated with primary antibodies against CD14 (ab82434), CD31 (ab28364), CD34 (ab18227), CD44 (ab157107), CD45 (ab134202), CD73 (ab157335), CD90 (ab95700), CD105 (ab53321), Integrin (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), and CD29 (Zymed, San Francisco, CA, USA) that were prepared in phosphate-buffered saline (PBS). ...
Article
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Over the past years, adipose tissue has become an invaluable source of mesenchymal stem cells (MSCs) due to development of improved isolation methodologies. In a recent work, our group established a primary culture of human adipose-derived stem cells (hADSCs), which were characterized for their stem cell characteristics in detail and studied their myogenic differentiation potential in presence of boron. In the current study, we focused on the effects of a boron-containing compound, sodium pentaborate pentahydrate (NaB), on the adipogenic differentiation of hADSCs. Incorporation of boron in various chemical derivates has been a novel interest in drug-discovery attempts due to increasing number of reports on their anticancer, antibacterial, antiviral, and antifungal activities. In this report, a striking suppressive activity of boron on adipogenic differentiation of hADSCs is observed in a dose-dependent manner. Higher concentrations of NaB (20, 50, and 100 μg/mL (68, 170 and 340 μM)) resulted in a progressive decrease of lipid deposition, suppressed master regulators of adipogenesis transcriptional programming at the mRNA and protein levels, while having no evident cytotoxicity on the cells. The findings of this study are encouraging to undertake further investigations on potential beneficial effects boron in terms of its impact on normal and dysfunctional adipose biology. In that respect, these results pave the path to evaluate boron-based compounds in prevention and treatment of obesity which is a modern age pandemic that is predominant worldwide and found in strong association with comorbidities, including type 2 diabetes, hypertension, cardiovascular disease, cancers, and others."
... NOTE: All experimental procedures should be carried out under sterile conditions in laminar flow hood. This non-enzymatic technique has been previously used 21,22,23 . 3. Place small pulp tissues inside the tissue culture treated 6-well plates and add 200 µL of complete DMEM media to cover each small pulp tissue pieces. ...
Article
Cancer as a multistep process and complicated disease is not only regulated by individual cell proliferation and growth but also controlled by tumor environment and cell-cell interactions. Identification of cancer and stem cell interactions, including changes in extracellular environment, physical interactions, and secreted factors, might enable the discovery of new therapy options. We combine known co-culture techniques to create a model system for mesenchymal stem cells (MSCs) and cancer cell interactions. In the current study, dental pulp stem cells (DPSCs) and PC-3 prostate cancer cell interactions were examined by direct and indirect co-culture techniques. Condition medium (CM) obtained from DPSCs and 0.4 µm pore sized trans-well membranes were used to study paracrine activity. Co-culture of different cell types together was performed to study direct cell-cell interaction. The results revealed that CM increased cell proliferation and decreased apoptosis in prostate cancer cell cultures. Both CM and trans-well system increased cell migration capacity of PC-3 cells. Cells stained with different membrane dyes were seeded into the same culture vessels, and DPSCs participated in a self-organized structure with PC-3 cells under this direct co-culture condition. Overall, the results indicated that co-culture techniques could be useful for cancer and MSC interactions as a model system.
... Так, бор повышает одонтогенную и остеогенную дифференцировку клеток ростка стволовых клеток зубов. Прием пентабората натрия оказывал дозозависимый эффект на активность щелочной фосфатазы и экспрессию генов, связанных с одонтогенезом [34]. Поэтому дефицит бора во время беременности, наряду с дефицитами кальция и других микронутриентов, также будет способствовать нарушениям развития зубов и у беременной, и у ребенка. ...
Article
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In this paper the results of experimental and clinical studies indicating the importance of compensation micronutrient in the prevention and treatment of osteoporosis, osteopenia and rachitis are discussed.
... This filler releases multiple ions including F − , Sr 2+ , Na + , BO 3 3− , Al 3+ , and SiO 3 2− [20,22] and the ion release may have an effect on various biological phenomena. For example, fluoride ions are reported to have an antimicrobial effect [23,24] and induce remineralization of enamel [25], strontium ions are reported to enhance bone formation [26,27], silicate ions are known to have antioxidant effects and enhance bone formation [28], and boron ions are reported to promote osteogenic and odontogenic differentiation [29]. These findings suggest that S-PRG filler could have a positive effect on tertiary dentin formation after dental pulp injury, but there is only one report about its use as a pulp capping material containing in a resinous material [30]. ...
Article
Full-text available
Objectives To evaluate new pulp capping cements containing surface pre-reacted glass ionomer (S-PRG) filler and to investigate ion release kinetics and pH shift of eluates from the cement. Materials and methods Molars of Wistar rats were directly pulp capped using three kinds of cement containing S-PRG filler and mineral tri-oxide aggregate (MTA) was used as a control. After 1, 2, or 4 weeks, histological evaluation was performed and differences of tertiary dentin formation were analyzed. Release of Sr²⁺, BO3³⁻, SiO3²⁻, Na⁺, and Al³⁺ ions was determined by inductively coupled plasma-atomic emission spectrometry, and F⁻ ion release was measured using a fluoride ion selective electrode. The pH of the eluate from each cement after mixing was measured with a pH electrode. Results One of S-PRG cements promoted tertiary dentin formation to the same extent as the control (p > 0.05) and it showed a tendency of less inflammatory response. This cement released more BO3³⁻ and SiO3²⁻, but less Sr²⁺, Na⁺, and F⁻ than other S-PRG specimens. Each cement recovered nearly neutral compared with glass ionomer cement. Conclusions S-PRG cement induced tertiary dentin formation based on multiple ion releases, suggesting that it is suitable as a pulp capping material. Clinical relevance This new material can be an alternative pulp capping agent to MTA.
... In addition, ions of boron, silicon, and strontium have the ability to promote bone formation by inducing osteoblastic differentiation of undifferentiated mesenchymal cells as follows: i) Boron ions promoted extracellular matrix (ECM) mineralization by inducing osteoblastic differentiation of rat bone marrow-derived mesenchymal stem cells (MSCs) (8). Boron ions also induced odontogenic and osteogenic differentiation of human tooth germ stem cells (9). In addition, Doğan et al demonstrated that boron-containing poly-(lactide-co-glycolide-acid) (PLGA) scaffolds promoted the in vivo healing of a bone defect in a femur. ...
Article
Full-text available
Surface pre-reacted glass‑ionomer (S‑PRG)-containing dental materials, including composite and coating resins have been used for the restoration and/or prevention of dental cavities. S‑PRG is known to have the ability to release aluminum, boron, fluorine, silicon, and strontium ions. Aluminum ions are known to be inhibitors whereas boron, fluorine, silicon, and strontium ions are known to be promoters of mineralization, via osteoblasts. However, it remains to be clarified how an aqueous eluate obtained from S‑PRG containing these ions affects the ability of mesenchymal stem cells (MSCs), which are known to be present in dental pulp and bone marrow, to differentiate into osteogenic cell types. The present study demonstrated that 200‑ to 1,000‑fold‑diluted aqueous eluates obtained from S‑PRG significantly upregulated the mRNA expression level of the osteogenic differentiation marker alkaline phosphatase in human MSCs (hMSCs) without exhibiting the cytotoxic effect. In addition, the 500‑ to 1,000‑fold‑diluted aqueous eluates obtained from S‑PRG significantly and clearly promoted mineralization of the extracellular matrix of hMSCs. It was additionally demonstrated that hMSCs cultured on the cured resin composites containing S‑PRG fillers exhibited osteogenic differentiation in direct correlation with the weight percent of S‑PRG fillers. These results strongly suggested that aqueous eluates of S‑PRG fillers promoted hard tissue formation by hMSCs, implicating that resins containing S‑PRG may act as a useful biomaterial to cover accidental exposure of dental pulp.
... Some experimental data support these effects [8,9]; however, there is not enough evidence to support its regular employment [10]. For instance, boron deprivation disrupts bone developmental stages in mice [11], and its administration induces bone mineralization and tooth mineral composition [12,13]. Additionally, boric acid administration induces many physiological changes (some linked to boron-induced toxicity [2,14]), and among these changes are changes in bone, muscle, inflammatory responses and hormone plasma levels [1,7]. ...
... Moreover, shRNA-mediated knockdown of β-catenin rescued the differentiation potential of boron-treated cells, verifying that the Wnt/β-catenin pathway was at least partially involved in the boron-induced inhibition of adipogenesis. These results were in line with our previous findings about boron's stimulatory role on osteogenic differentiation [38,39] as Wnt signaling stimulates osteogenesis and represses adipogenesis. ...
Chapter
Dental Stem Cells (DSCs) are emerging adult stem cells (ASCs) as a good cell source for stem cell-based therapies and tissue engineering approaches. MicroRNAs (miRNAs) are approximately 20 nucleotide long small RNA molecules which can coordinate many genes and pathways. miRNAs effectively regulate stem cell types ranging from embryonic stem cells (ESCs) to mesenchymal stem cells (MSCs), and DSCs are no exception. Numerous studies in which miRNA profiling of DSCs from different sources of the mature and immature teeth including pulp, periodontal ligament, follicle and apical papilla tissues have been made in order to determine the role of specific miRNAs in various functions, which may be important for DSC biology. Moreover, the miRNA expression levels are being monitored both between different steps of differentiation and in cells that are differentiated in vitro. Such data will help to better retain stemness of cells, and manipulate the differentiation process into desired lineages. In this book chapter, we seek to compile data resulting from miRNA studies in DSCs.
... The degree of mineralization and bone-assisted protein expression in osteoblasts (Hakki et al., 2010), in dental stem cells (Tasli et al., 2013) and bone marrow stromal cells (Ying et al., 2011) was found to be increased with higher concentrations of boron. For this reason, the use of nontoxic doses of boron compounds are of fundamental importance for clinical applications, providing calcium mineralization in tissue engineering. ...
Article
Full-text available
Calcium phosphate derivatives have been widely employed in medical and dental applications for hard tissue repair, as they are the main inorganic constitution of hard tissue; such as bones and teeth. Owing to their excellent osteoconductive and bioactive properties, hydroxyapatite- (HA) based ceramics are the best candidates of this group for medical, bioscience, and dental applications. However, when replacing a bone or tooth, HA is not able to sustain similar mechanical properties. In this study, to improve the mechanical properties, nanoscale hexagonal boron nitride with different compositional percentages was added to the nano HA to form composites. The effect of compositional changes and sintering parameters on microstructural and morphological properties of the ceramic composites was comparatively investigated. Detailed chemical characterization of the composite materials was carried out using X-ray diffraction, Fourier transform infrared spectroscopy, Raman spectroscopy, and energy-dispersive X-ray spectroscopy, whereas scanning electron microscopy and atomic force microscopy investigations were employed to monitor morphological and surface features. Additional transmission electron microscopy investigations were carried out to reveal the nanostructure and crystal structure of the composites.
... Human newborn foreskin stem cells were isolated and characterized, according to the protocol defined formerly by the authors (20). They were trypsinized and raised with primary antibodies that were arranged in PBS. ...
Article
Full-text available
A previous study performed by the authors of the current study revealed the characterization and differentiation of newly defined stem cells known as human newborn foreskin stem cells (hnFSSCs). According to their stem cell properties, this study aimed at investigating myogenic differentiation and related tissue engineering. Human newborn foreskin stem cells were characterized by flow cytometry. The results showed that hnFSSCs carries a noble prospective for myogenic differentiation and can be used as a beneficial method for muscle related diseases, including muscular dystrophy, neuromuscular disorders, muscle damages, muscle weakness, lesion formations, and other problems associated with tissue obtainability and multi-potency; these cells may be accepted as effortlessly accessible and functional, and even superior to other stem cell origins. Furthermore, hnFFSCs were also seeded onto 3D micro-wells and Polycaprolactone (PCL) scaffolds in order to examine tissue development. Human newborn foreskin stem cells on PCL scaffolds showed good cell-cell integration, so that they may be thought as a stem cell basis for tissue engineering.
... Recent studies support boron's dose-dependent role as an anti-osteoporosis factor and a biological factor also influencing other related functions [20][21][22][23][24][25][26]. Animal studies suggest boron salts are easily absorbed. ...
Article
We will present 2 cases of patients suffering from osteoarthrosis, characterized by joint swelling and pain, treated with an organic boron salt, calcium fructoborate containing 6 mg of elemental Boron daily, resulting in a clinical significant decrease of pain and swelling. Boron salts have been implied to be essential for plants since the 20s of last century, and suggestions for such essential element status started to emerge for animals and humans since the 80s of last century, although the role of boron salts in a number of physiological functions have remained somewhat controversial and did not penetrate mainstream thinking. However, since the last few years’ new data emerged supporting the role
... Boron is a trace mineral that proven essential throughout in the lifecycle for organizations across a phylogenetic kingdom. This mineral found in the form of boric acid (H 3 BO 3 ) at physiological pH, studies show that boron is required for a good bone health, playing a vital role in embryogenesis, bone growth, and immune functions psychomotor [3][4][5]. ...
... Only at the 28th day, a significant increase was seen in B-HAp/Ch group when compared with the 14th day results (Fig. 5b). Taşlı et al. (2013) noted that On expression of hTGSCs treated with different concentrations of sodium borate was significantly increased compared with control group. ...
Article
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Boron (B), which is a beneficial bioactive element for human, has an increasing interest in tissue engineering for the last 5 years. However, the effective B concentration in cell culture is still unknown. The aim of the present study is to investigate in vitro osteogenic potential of mesenchymal stem cells, isolated from adipose tissue (AdMSCs), on boron containing 2D and 3D cell cultures. At first, the effects of B concentrations between 1 and 20 lg/mL were evaluated on the survival and osteogenic differentiation of AdMSCs cultured on 2D cell cultures. The 3D cultures were established by using chitosan (Ch) scaffolds prepared by freeze-drying and Ch scaffolds combined with hydroxyapatite (HAp) and B containing hydroxyapatite (B-HAp) that are produced by microwave-induced biomimetic method. The proliferation and osteogenic differentiation of AdMSCs on Ch, HAp/Ch and B-HAp/Ch scaffolds were investigated by in vitro cell culture studies. The results were evaluated with respect to cell viability, bone related ECM gene expressions, and cellular morphology. It was demonstrated that cellular functions of AdMSCs were enhanced by boron in both 2D and 3D cultures. Especially, B-HAp/Ch scaffolds, which have both osteoinductive and osteoconductive properties based on presence of B and HAp in its structure, promoted adhesion, proliferation and osteogenic differentiation of AdMSCs.
... Bone growth retardation is correlated with dietary B deprivation in mice model, and therefore, B is essential to bone formation via altering periodontal alveolar bone modelling and remodelling [22]. Moreover, there are cellular and molecular evidences that B can significantly increase the level of mineralization and the expression of bone associated proteins in osteoblasts [23], bone marrow stromal cells [24] and tooth germ stem cells [25]. Recently, Wu et al. reported that B-containing mesoporous bioactive glass could lead to controllable release of B ions and significantly improve osteogenic differentiation of osteoblasts [26]. ...
... Therefore, Sr ions released from the adhesive may also have contributed to dentin bridge formation in group 2. B ion is associated with brain function, immune regulation, and bone and hormone metabolism 38) . Taşlı et al. investigated the cell viability of hTGSCs (human tooth germ stem cell) incubated with different concentrations of NAB (Na pentaborate pentahydrate) using the MTS assay 48) . Their results showed that the cell viability of hTGSCs was increased at low concentrations (10 and 20 μg/mL) of NAB, but cytotoxicity for hTGSCs was recognized at high concentrations of NAB. ...
Article
The purpose of this study was to evaluate pulpal healing and reparative dentin formation after 14 and 28 days in exposed rat pulp directly capped with an experimentally developed all-in-one adhesive containing surface reaction-type pre-reacted glass-ionomer (S-PRG) filler. The four experimental groups and the control group were compared using the Kruskal-Wallis test, followed by the Steel-Dwass post-hoc test to compare the histopathological score. The Mann-Whitney U test was used to compare the histopathological score at 14 and 28 days for each observation item. All experimental adhesives containing S-PRG fillers developed for direct pulp capping showed no pulpal inflammation. After 14 days, the experimental adhesives containing S-PRG fillers and the control group formed tertiary dentin around the exposed pulp. After 28 days, the experimental adhesives containing 13 and 27 wt% of S-PRG fillers formed dentin bridge equal to the control.
... Although N-BPs adsorb strongly to bone surfaces via noncovalent bonds, it is possible for extrinsic ions to dissociate bound N-BPs from mineralized bone by competitive desorption [13,14]. Boron plays a crucial role in osteogenesis and bone homeostasis [15] by enhancing odontogenic and osteogenic differentiation in human stem cells [16,17]. Water-soluble microfibrous borate glass dressings (MBG) are capable of rejuvenating bone tissues via ionexchange, speedy dissolution and rapid conversion into carbonated apatite [18]. ...
Article
Statement of significance: Long-term oral consumption and injections of nitrogen-containing bisphosphonates (N-BPs) may result in death of the jaw bone when there is traumatic injury to the bone tissues. To date, there is no effective treatment for such a condition. This work reported the use of an ionic cocktail derived from water-soluble borate glass microfibers to displace the most potent type of N-BPs that are bound strongly to the mineral component on bone surfaces. The mechanism responsible for such an effect has been identified to be cation-mediated complexation of borate anions with negatively-charged N-BPs, allowing them to be released from the mineral surface. This borate-containing cocktail may be developed into a novel topical rinse for removing mineral-bound N-BPs from exposed dead bone.
... In an in vitro study by Taşl et al., the odontogenic and osteogenic differentiation capacity of mesenchymal cells isolated from dental germ stem cells (hTGSCs) cultured with different concentrations of sodium pentaborate pentahydrate (borax) was evaluated. The results obtained showed that the cells treated with B showed an increase in alkaline phosphatase activity and the expression of genes related to odontogenesis and osteogenesis compared to the control group without treatment [16]. ...
Article
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Cobalt–chromium (Co-Cr) alloys have been used in a wide variety of biomedical applications, including dental, cardiovascular, and orthopedic devices. In vitro studies have shown that the mineralization of cells involved in osteogenesis is regulated by boron. The development of a new cobalt-chromium-boron (Co-Cr-B) alloy improves the mechanical properties of the metal, such as wear resistance, and meets biocompatibility requirements. Therefore, the objective of this study was to evaluate the osteogenic differentiation and biocompatibility in in vitro assays. Human dental pulp mesenchymal cells (hDPSCs) were isolated from volunteers and then co-cultured with the Co-Cr plus boron alloy from 0.3% to 1% for 15 days, while the formation of calcium deposits was quantified by Alizarin red staining and the expression of genes was related to osteodifferentiation by RT-qPCR. Simultaneously, the cytotoxicity of our alloy was evaluated by MTT assay and the change in the gene expression of cytokines commonly associated with inflammatory processes. The results showed low cytotoxicity when cells were treated with the Co-Cr-B alloy, and no change in the gene expression of IL-1β, TNF-α, IL-6, and IL-8 was observed compared to the untreated control (p > 0.05). The osteoinduction results shown an increase in mineralization in hDPSCs treated with Co-Cr-B alloy with 1.0% B. In addition, a significant increase in mRNA levels for collagen type 1 in with 0.3% boron and alkaline phosphatase and Runx2 with 0.6% boron were observed. The addition of Boron to the ASTM F75 Co-Cr base alloy improves the biocompatible characteristics. No cytotoxicity and any change of the expression of the pro-inflammatory cytokines IL-1β, TNF-α, IL-6, and IL-8 in human peripheral blood mononuclear cells treated with the cobalt-chromium-boron alloy was observed in vitro assays. Furthermore, our alloy acts as an osteoinductive in osteogenic differentiation in vitro. Therefore, our results could set the standard for the development of in vivo trials and in the future, it could be considered as an alternative for regenerative therapy.
... Boron is a natural product and found in many human tissues. Up to now, developmental benefits of the B have been shown in various studies [19,20]. B compounds have been used in prostate, lung, cervical, and breast cancer and caused positive effects against cancer [11,13,[21][22][23]. ...
Article
Neuroblastoma occurs in childhood with high aggressiveness and is one of the most common solid tumors. Although there are many different strategies to fight neuroblastoma including surgical treatment, chemotherapy, radiotherapy, and immunotherapy, ultimately successful treatment has not been evaluated yet. Effective, safe, and less toxic options must be investigated. Zoledronic acid (ZOL) is a type of amino-bisphosphonates and has been used in bone-related diseases for more than 20 years and the anti-tumor ability of the ZOL is known. Boron is a natural product and many regenerative properties of boron compounds such as myogenic, osteogenic, and odontogenic induction potential have been discovered. Besides, the boron compound also displayed anti-cancer characteristics in different studies. In the current study, we evaluated the possible synergistic effects of the ZOL and Sodium pentaborat tetrahydrate (SPT) on the neuroblastoma cells, SH-SY5Y. As a result, ZOL and SPT combination exhibited the most favorable anti-proliferative, pro-apoptotic and anti-migratory effects compared to the ZOL and SPT alone and control groups. Moreover, molecular evidence has indicated that while expression of the proliferative gene, NFκB was significantly decreased in the combination group compared to all other groups, pro-apoptotic genes, were overexpressed. To sum up, obtained results from the recent study lead it necessary to carry out more detailed studies.
... hBMSCs were seeded on 24-well plates (BIOFIL) at a density of 20,000 cells/well. Osteogenic differentiation medium consisted of Dulbecco modified Eagle medium, 10% (v/v) fetal bovine serum, 0.1 mmol/L dexamethasone (AppliChem, Darmstadt, Germany), 10 mmol/L b-glycerol phosphate (Sigma-Aldrich, St Louis, MO), and 50 mg/mL ascorbic acid (Sigma-Aldrich) (29) with or without mate- rials. The medium was changed every other day, and the cells were treated with osteogenic differentiation medium for 14 days. ...
Article
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Introduction: Stem cell technology has been a great hope for the regeneration of cells of pulp-dentin complex and dental structures together with surrounding bone and periodontium. The main challenge in the regeneration process is a successful combination of stem cells and efficient inductors such as inductive biomaterials. In this regard, today, manufacturers propose novel tooth filling materials. The current study was aimed to compare the effect of ProRoot MTA (Dentsply Tulsa Dental, Tulsa, OK), Biodentine (Septodont, Saint Maur des Fossés, France), and MM-MTA (Micro-Mega, Besançon Cedex, France) on the cell viability, hard tissue deposition capacity, and osteogenic differentiation of human bone marrow stem cells (hBMSCs) derived from mandibular bone. Methods: Dental materials were packed into Teflon rings (Grover Corp, Milwaukee, WI) and placed on Transwell inserts (Corning, Corning, NY) to determine the toxicity of tooth filling materials by the 3-(4,5-dimethyl-thiazol-2-yl)-5-(3-carboxy-methoxy-phenyl)-2-(4-sulfo-phenyl)-2H tetrazolium assay on days 1, 3, 7, and 14; 20% dimethyl sulfoxide (DMSO) was used as a positive control for the toxicity assay. hBMSCs were characterized by their surface markers with mesenchymal stem cell antibodies. Teflon rings were cocultured with hBMSCs followed by the induction of osteogenic differentiation. The osteogenic differentiation of hBMSCs and hard tissue formation of the materials were evaluated by analyzing the messenger RNA expression levels of osteonectin, Runt-related transcription factor 2, and collagen type 1A by real-time polymerase chain reaction expression analysis, measurement of alkaline phosphatase activity, and visualization of calcium deposits by alizarin red staining. Results: MTA, Biodentine, and MM-MTA did not exhibit a cytotoxic effect on hBMSCs after 14 days in culture. Even though all the materials significantly stimulate (P < .05) osteogenic differentiation of hBMSCs compared with the negative control, ProRoot MTA showed greater osteoinductivity than Biodentine or MM-MTA according to the messenger RNA expression, alkaline phosphatase, immunocytochemistry, and alizarin red staining data. Conclusions: All of the dental materials used in this study show the osteogenic differentiation potential of hBMSCs. Therefore, newly introduced MM-MTA can also be used as a material of choice in routine dental treatment.
... However, most of these studies relate the positive biological effect of S-PRG filler to the release of different types of bioactive ions, such as Sr, B, Si, F, and Al. The effect of each ion has been previously reported: Sr and B ions are postulated to downregulate inflammatory responses [27], the former ion also promotes bone formation [28,29], and the latter has been reported to induce osteogenesis and odontogenic differentiation [30]. Si ions exhibit antioxidant activity and enhance bone formation [31]. ...
Article
The effects of surface pre‐reacted glass‐ionomer (S‐PRG) filler on pulpal cells and on the composition of dentinal deposits were investigated. Proliferation (CCK‐8), cytotoxicity (LDH), and differentiation activity (ALP) tests, along with cell morphology observations, were conducted at 6 and 24 h after treatment of pulpal cells with different S‐PRG filler eluate concentrations. Dentinal surfaces were immersed in deionized water or S‐PRG filler eluate followed by immersion in deionized water or simulated body fluid and observed under scanning electron microscope and elemental analysis using energy dispersive x‐ray spectrometer. At 24 h, there were significant differences in CCK‐8 and ALP activity values between the groups in a concentration‐dependent manner. LDH test data were not significantly different among the groups. Cell morphology was not altered at either exposure time. However, decreased cellular density was observed with the highest eluate concentration. Crystalline deposits and occluded dentinal tubules were observed in samples immersed in S‐PRG filler with a later immersion in simulated body fluid, which also showed higher concentrations of certain ions compared to surfaces that were not initially treated with S‐PRG filler. The lowest two eluate concentrations did not show significant toxicity. S‐PRG enhanced the effect of simulated body fluid in the formation of mineral deposits.
... Boron has been shown to be essential for the completion of the life cycle in all organisms [1,2]. In the literature, the effects of boron and boron deprivation have been mainly investigated on osteoblast proliferation, osteogenic differentiation of stem cells and osteoporosis [1][2][3][4][5][6][7][8]. Experimental studies on boron nitride (BN) have reported promising results in respect of biocompatibility and biosafety for orthopedic applications [9][10][11][12][13]. ...
... They suggested that continuous media flow could help avoid the immense ion accumulation. On the other hand, Taşlı et al. employed 20 mg/L sodium borate (NaB) to induce differentiation of DPSCs and determined that B was not cytotoxic and increased ALP production [54]. Najafabi et al. demonstrated that 6 mg/L of H 3 BO 3 was optimal to achieve the highest ALP activity of cells [55]. ...
Article
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Boron-doped hydroxyapatite/tricalcium phosphates (BHTs) were synthesized to study boron uptake and correlate structural alterations of incremental boron addition (0 to 10 mol%). BHTs with a Ca/P ratio of 1.6 were prepared by a wet precipitation/microwave reflux method, sieved (< 70 μm) and characterized. XRD and FTIR analyses revealed that boron slightly distorted apatite crystal, increased crystallinity (95.78 ± 2.08% for 5BHT) and crystallite size (103.39 ± 23.47 nm for 5BHT) and still, boron addition did not show any further detrimental effects. Total surface area (4.05 ± 0.82 m2/g for 10BHT) and mesoporosity (23.90 ± 7.92 μL/g for 10BHT) were expanded as boron content was increased. Moreover, boron addition made grains become smaller (0.21 ± 0.06 μm for 5BHT) and ordered while hardness (10.51 ± 0.86 GPa for 10BHT) increased. Boron incorporation enhanced bioactivity with significantly highest calcium phosphate deposition and protein adsorption (135.29 ± 29.58 μg on 10BHT). In return, boron favored highest alkaline phosphatase activity (4.80 ± 0.40 MALP/ngDNA.min), intracellular calcium (23.61 ± 0.68 g/gDNA), phosphate (31.84 ± 4.68 g/gDNA), and protein (23.70 ± 3.46 g/gDNA) storage in 5BHT without cytotoxicity (128 ± 18% viability compared to pure HT). Compared to literature, it can be pointed out that we successfully employed an optimal procedure for production of BHTs and incorporated significantly higher boron content in HT (5.23 mol%). Additionally, results tended to conclude that 5BHT samples (5 mol% boron in HT) demonstrated a very high potential to be used in composite bone tissue constructs.
... Bunun yanında borik asidin kemik yapımını stimüle etmesi ve mineralize doku oluşumunu teşvik etmesiyle ilgili çalışmalar da mevcuttur (32)(33)(34)(35). Ayrıca diş çekim sorası alveoller kemikte iyileşmeyi stimüle etmesi ile birlikte dental kök hücrelerde borun osteojenik ve odontojenik farklılaşma oluşturduğu tespit edilmiştir (36,37 (20). Çalışmamızda %5'lik borik asit solüsyonu, sınıf V kavitelerde restorasyon öncesi kavite dezenfektanı olarak uygulanmıştır. ...
... B-MBG scaffolds illustrated potent promoting effects of boron on osteogenesis and bone remodeling in vitro and in vivo [4,12,41]. Our previous study showed that B-MBG scaffolds slowly released boron to activate Wnt/β-catenin pathway to promote osteogenic genetic transcription to enhance bone regeneration [13]. ...
... The molecular level of boric acid could increase the level of mineralization and levels of bone growth-related mRNA such as collagen I and osteocalcin in mouse osteoblast cell line, MC3T3-E1 [61]. Additionally, the sodium pentaborate pentahydrate (NaB) was reported to have ability to stimulate osteogenic and odontogenic differentiation of human tooth germ stem cells [62]. Therefore, it is reasonable to speculate that TCP-PDLLA-5LB scaffolds simultaneously released bioactive La 3+ and BO 3 3− ions which exerted synergetic effects on the osteogenesis of rBMSCs. ...
Article
Bone tumors may lead to osteoporosis. It is of great significance to fabricate functional scaffolds for treating bone tumor, osteoporosis and repairing bone defects. In this study, LaB6 micro-nanoparticles/poly(d,l-lactide)-modified β-tricalcium phosphate scaffolds (TCP-PDLLA-LB) were successfully prepared. The LaB6 surface chemistry-reinforced TCP-PDLLA-LB scaffolds were endowed with the following excellent properties: (1) the enhanced mechanical strength. The TCP-PDLLA-5LB scaffolds possessed higher compression resistant capacity than TCP scaffolds owing to the strong adhesive property of PDLLA that bound the LaB6 micro-nanoparticles and scaffolds tightly; (2) the excellent photothermal performance. TCP-PDLLA-LB scaffolds presented distinct photothermal property because of localized surface plasmon resonance effect of LaB6 micro-nanoparticles when exposed to the near-infrared (NIR) light (wavelength: 808 nm) and it was flexibly controllable through changing the quantity of LaB6 micro-nanoparticles, power density of NIR light and environmental humidity; (3) the excellent biological properties in vitro. When culturing with saos-2 bone tumor cells, the TCP-PDLLA-5LB scaffolds effectively killed the saos-2 cells through adjusting power density of NIR light, irradiation duration time of NIR light and irradiation times of NIR light. Additionally, the TCP-PDLLA-5LB scaffolds were able to release bioactive ions which were beneficial to the in vitro osteogenesis of rabbit bone marrow stromal cells (rBMSCs) by supporting the attachment and proliferation of rBMSCs as well as promoting the expressions of osteogenic genes BMP2, RUNX2 and COL 1; and (4) the abilities of effectively ablating bone tumors and supporting bone tissue regeneration in vivo. The bone tumor tissues were significantly ablated and suppressed through hyperthermia due to the existence of LaB6 micro-nanoparticles on the scaffolds. Simultaneously, TCP-PDLLA-5LB scaffolds implanted in bone defects effectively helped new bone formation regardless of NIR light irradiation. In conclusion, LaB6 surface chemistry-reinforced scaffolds possess the dual functions of ablating bone tumor and repairing bone defects, which offers a promising therapy strategy for tumor/disease-related bone tissue engineering.
... 11 The addition of MTS to the cell culture medium results in the formation of colored formazan crystals, indicating cellular viability. 14,15 The alteration in color was measured as the absorbance using an ELISA plate reader (BioTek Instruments Inc, Winooski, VT). ...
Article
Objective: The aim of the present study was to evaluate the viability and proliferative capacity of adipose-derived stem cells obtained by laser-assisted liposuction (LAL). Methods: Fat tissue was obtained from 7 male patients treated surgically for gynecomastia. On one side, harvesting was made before LAL, while it was implemented after LAL on the contralateral side. Viability, cell surface antigens, pluripotency, and apoptosis were assessed and compared in these samples. Results: Cells harvested before and after LAL did not exhibit any significant difference in terms of surface cell markers. Number of viable stem cells was lower initially after exposure to laser, while this difference was reversed at the end of 72 hours. Genetic indicators of cellular differentiation were similar in both groups. Apoptosis indicators were increased remarkably after laser exposure in the first 24 hours, but this increase was absent 72 hours after LAL procedure. Conclusion: The authors' results have promising clinical relevance since mesenchymal stem cells harvested during LAL have maintained appropriate cellular features to be used for autologous fat transfer and fat grafting.
... Бор оказывает выраженное воздействие на процессы роста клеток костной ткани и хряща, повышение уровня белков остеогенеза -остеокальцина, коллагена 1-го типа, белков морфогенеза костей 4, 6 и 7, а также остеопонтина, сиалопротеина кости (ген BSP), белка Runx2 и др. [40][41][42]. ...
... Therefore, we sought for an alternative to boric acid that is potentially less toxic to mammalian system. Recent study conducted by our group confirmed that sodium pentaborate pentahydrate (NaB) is a good alternative source for boron in the regeneration studies [22]. Apart from the studies related to bone regeneration, boron was also investigated in studies of lipid metabolism and suggested that sodium borate could be worth pursuing as a drug candidate for lowering plasma lipid levels in humans and other animals based on the observations of lipid profiles of dogs fed a fatty diet [23] as well as dairy cows where a decrease in fatty liver was recorded [24]. ...
Article
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Obesity is a major public health problem worldwide and a risk factor for certain diseases, including cardiovascular disease, diabetes, cancer, and depression. Unfortunately, currently available anti-obesity drugs have failed in the long-term maintenance of weight control. It has been a challenge to design novel drugs that could potentially treat obesity or prevent uncontrolled weight-gain which lies underneath the pathology of obesity. Since obesity in a way is a consequence of the accumulating new mature adipocytes from undifferentiated precursors which is a process also termed as adipogenesis, drugs that might control adipogenesis could be beneficial for the treatment of obesity. In the current study, combined effect of sodium pentaborate pentahydrate (NaB) and pluronic F68 on adipogenic differentiation was examined by administering various combinations of the two agents to human adipose-derived stem cells (hADSCs) in in vitro. Immunocytochemistry and quantitative RT-PCR were performed to evaluate the levels of adipogenesis-promoting genes such as peroxisome proliferator-activated receptor-γ (PPARγ), fatty acid binding protein (FABP4), and adiponectin. Results indicated that expressions of all these three genes were restrained. Furthermore, Oil Red O staining revealed that lipid vesicle formation was reduced in hADSCs treated with differentiation medium containing NaB/F68 combination. Finally, expression levels of Hippo pathway kinases Lats2, MST1, and scaffold protein Sav1 were reduced in these cells, suggesting a possible link between Hippo pathway-dependent downregulation of PPARγ and the NaB/F68 treatment. Herein, we showed that combination of NaB and F68 curtails adipocyte differentiation by inhibiting the adipogenic transcriptional program leading to a decrease in lipid accumulation in adipocytes even at very low doses, thereby uncovered a striking opportunity to use this combination in obesity treatment.
... Boron at 10 and 100 ng/mL also increased mRNA expression of alkaline phosphatase, osteocalcin, collagen type 1, and bone morphogenetic protein-7. Boron also was found to increase alkaline phosphatase activity and enhance the expression of osteogenic markers collagen type 1 and osteocalcin in human tooth germ stem cells in vitro (Tașh et al., 2013). ...
Chapter
Since 1970, numerous elements have been suggested to be essential based on an apparently beneficial action in experimental animals. These elements include aluminum, arsenic, boron, bromine, cadmium, chromium, fluoride, germanium, lead, lithium, nickel, rubidium, silicon, strontium, tin, and vanadium. The lack of any of these elements has not been shown to interrupt the life cycle or have a defined biochemical function in humans; thus they are generally considered nonessential. The benefits of supranutritional intakes of chromium and fluoride are presented in other chapters. Two of the other elements, boron and silicon, in nutritional or physiological amounts, have beneficial effects in humans. An intake of boron of 1 mg/day or more has been found to be beneficial for bone growth and maintenance, central nervous system function, and hormone action. Silicon intakes greater than 15 mg/day have been associated with bone health. Plausible mechanisms of actions for these effects are described herein.
Article
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Here we present the integration of Boron (NaB) with Graphene Oxide (GO) to develop a new class of membranes which are bio-compatible and cost-effective for cell and tissue culture studies. Ethanol (EtOH) assisted the uniform dispersion of GO flakes on top a glass substrate. We investigated the effect of GO+NaB membrane on growth and proliferation of hASCs. hASC’s showed better cell viability on NaB integrated GO membranes compared to their respective controls. The concentrations of NaB and GO are 0.02% and 1/20 of stock (0.024%) respectively. To our knowledge this is the first time that enhanced cell proliferation and attachment ability of hASCs with NaB integrated GO membranes has been observed. Our study provides a platform for the development of 3D-GO scaffold systems combined with NaB in tissue engineering.
Article
Background Boron (B) is an element involved in many physiological processes in humans and accelerates wound healing and increases angiogenesis. This study aimed to evaluate the possible effects of sodium pentaborate pentahydrate (NaB) on hair growth and reveal its effects on Wnt-1, β-catenin, vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), and transforming growth factor-β1 (TGF-β1) signaling pathways, which are important molecular mechanisms involved in hair growth. Methods Thirty-five Sprague-Dawley/Wistar albino rats were randomly divided into five groups: non-shaved control, shaved control, NaB 1 mg (shaved + NaB 1 mg elemental B/kg CA), NaB 2 mg (shaved + NaB 2 mg elemental B/kg CA), and NaB 4 mg (shaved + NaB 4 mg elemental B/kg CA). Hair density was measured using the trichoscopy method. Dorsal skin samples were examined histopathologically at the end of the 42nd day, and follicle count, follicle diameter, and subcutaneous tissue thickness were recorded. Wnt-1, β-catenin, PDGF, VEGF, TGF-β1, and collagen I levels were analyzed with the Western blot method. Results In trichoscopy measurements, hair density increased in the NaB 4 mg group (90.9%). In histopathological examination, anagen follicles were observed to increase in the NaB 1 mg and 2 mg groups (p<0.05). Follicle diameter increased in all NaB groups (p<0.05). The Wnt-1, β-catenin, PDGF, VEGF, TGF-β1, and collagen I level increased in the NaB 1 mg and 2 mg groups (p<0.05), but they were similar in the NaB 4 mg group compared to the control groups (p>0.05). Conclusion NaB 1 and 2 mg B/kg supplementation induces the anagen phase in rats via Wnt-1, β-catenin, VEGF, PDGF, and TGF-β1 signaling pathways. NaB 4 mg B/kg suppresses these pathways and adversely affects hair growth.
Article
Every year, many dental restoration methods are carried out in the world and most of them do not succeed. High cost of these restorations and rejection possibility of the implants are main drawbacks. For this reason, a regenerative approach for repairing the damaged dentin-pulp complex or generating a new tissue is needed. In this study, the potential of three-dimensional cellulose acetate/oxidized pullulan/gelatin-based dentin-like constructs containing 10 or 20% bioactive glass nanoparticles was studied to explore their potential for dentin regeneration. Three-dimensional nano biocomposite structures were prepared by freeze-drying/metal mold pressing methods and characterized by in vitro degradation analysis, water absorption capacity and porosity measurements, scanning electron microscopy, in vitro biomineralization analysis. During one-month incubation in phosphate buffered saline solution at 37°C, scaffolds lost about 25-30% of their weight and water absorption capacity gradually decreased with time. Scanning electron microscopy examinations showed that mean diameter of the tubular structures was about 420 µm and the distance between walls of the tubules was around 560 µm. Calcium phosphate precipitates were formed on scaffolds surfaces treated with simulated body fluid, which was enhanced by boron-modified bioactive glass addition. For cell culture studies human dental pulp stem cells were isolated from patient teeth. An improvement in cellular viability was observed for different groups over the incubation period with the highest human dental pulp stem cells viability on B7-20 scaffolds. ICP-OES analysis revealed that concentration of boron ion released from the scaffolds was between 0.2 and 1.1 mM, which was below toxic levels. Alkaline phosphatase activity and intracellular calcium amounts significantly increased 14 days after incubation with highest values in B14-10 group. Von Kossa staining revealed higher levels of mineral deposition in these groups. In this work, results indicated that developed dentin-like constructs are promising for dentin regeneration owing to presence of boron-modified bioactive glass nanoparticles.
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Implant-associated infections and aseptic loosening are some of the main reasons for implant failure. Therefore, there is an urgent need to improve the osseointegration and antibacterial capabilities of implant materials. In recent years, a large number of breakthroughs in the biological application of tantalum and its derivatives have been achieved. Owing to their corrosion resistance, biocompatibility, osseointegration ability, and antibacterial properties, they have shown considerable potential in orthopedic and dental implant applications. In this review, we provide the latest progress and achievements in the research on osseointegration and antibacterial properties of tantalum as well as its derivatives, and summarize the surface modification methods to enhance their osseointegration and antibacterial properties.
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Dental caries is a dental disease affecting public health, which results in many socio-economic consequences. This disease causes loss of tooth hard tissue and subsequent inflammation and loss of the dental pulp. In this study, it was aimed to develop and characterize boron (B) modified bioactive glass nanoparticles (BG-NPs) containing cellulose acetate/oxidized pullulan/gelatin (CA/ox-PULL/GEL) three dimensional scaffolds with tubular morphology for dentin regeneration. 3D nanobiocomposite structures were prepared by thermally induced phase separation and porogen leaching methods and characterized by in vitro degradation analysis, water absorption (WA) capacity measurement, SEM, in vitro biomineralization analysis, porosity measurement and mechanical tests. Scaffolds lost about (30–40)% of their weight during one month and WA capacity decreased with increase in immersion time in phosphate buffer solution (PBS) during one month. According to SEM, aligned and tubular structures were formed with mean diameter of about 11 μm, and BG-NPS were distributed evenly in all parts of the scaffolds. Scaffolds (without BG-NPs) possessed the highest porosity percentage. Addition of 10% BG-NPs improved the mechanical properties of scaffolds. Scaffolds surfaces were fully covered by calcium phosphate (Ca-P) deposits after conditioning in simulated body fluid for 14 days with higher quantity of deposition in groups with inclusion of B-BG-NPs. Human dental pulp stem cells (hDPSCs) were isolated from third molar teeth and used in cell culture studies. In all groups, cells adhered well 1 day after culture. Group B14–10 showed a slight increase of proliferation than group (without BG-NPs) after 7 days of incubation. Alkaline phosphatase activity (ALP) and intracellular calcium amounts increased significantly 14 days after incubation with highest values in B14–10 and B14–20 groups. Confocal laser scanning microscopy (CSLM) analysis, showed that cells on B14–10 and B21–10 scaffold groups, spread more 14 days after culture, and they also possessed extended processes specific to odontoblasts. Alizarin Red quantification showed that the highest calcium deposition was observed on B14–10 scaffolds. Immunohistochemical and Von Kossa stainings showed that scaffolds positively affected the odontoblastic differentiation of the hDPSCs. In this work, results showed that boron modified BG-NPs (B-BG-NPs) incorporated dentin-like constructs bring a new approach for dental tissue engineering applications.
Poster
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Secreted from all types of cells, exosomes are nanosized vesicles with their sizes ranging between 30-100 nm. Although it is a well-known fact that the communication between eukaryotic cells is carried out through exosome secretion, the role of plants exosomes remains unexplored. Here, we examined for the first time the effects of pineapple exosomes on human adipose stem cells. In order to achieve that, first the characterization of pineapple exosomes are performed and then followed with the uptake analysis of the exosomes inside the stem cells. The effect of varying doses of pineapple exosomes is also evaluated through MTS assay, cell cycle analysis and doubling time.
Presentation
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Exosomes are lipid bilayer nano vesicular messengers secreted by virtually all cell types, adapting to and conveying the physiological conditions of the cell. Exosomes carry out essential roles in cellular communications and transfer of biological material -such as proteins, RNA and DNA- from one cell to another. They carry membrane proteins characteristic to their cell-of-origin which are capable of eliciting responses to cells they interact with -such as antigen presenting exosomes from dendritic cells-, meaning that exosomes are capable of acting as mini-cells. Their diverse range of biological functions and the rich biomolecular content has attracted considerable interest over the last decade, and researchers from many different fields of biology are now contributing to our understanding of exosomes. Exosomes prove useful in a wide range of applications. They are capable drug carriers, able to pass through the blood-brain barrier and last for longer periods of time than most other conventional drug vehicles, and circulating exosomes are being investigated for diagnostic purposes against many diseases including cancer. Furthermore, as our understanding with exosomes grows, so does our understanding of their importance for the physiology of our cells and pathophysiology of diseases. Unfortunately, the influx of new papers must be scrutinized closely - the mutable nature of extracellular vesicle (EV) content and the difficulty in purifying exosomes (especially true with specific populations of EVs, such as exosomes or small EVs) means that great care must be taken while preparing samples for exosome studies. Experimental results of exosome studies may be easily skewed due to small details in experimental procedure affecting exosomes content, while using isolation methods unsuitable for a given study may result in co-isolation of biologically active soluble factors, reduced integrity of exosomes, and insufficient exosome concentrations. Furthermore, many of the biological mechanisms behind EV secretion, uptake, and determination of EV cargo are still unknown. Most commonly used exosomes isolation methods are currently incapable of producing exosomes with the quantity and quality necessary for exosomes research. Recent years has seen the development of modern exosomes isolation methods that address both issues of traditional EV isolation - however, these methods are yet to see widespread use in the exosomes field. Efficient isolation of exosomes remains to be a great obstacle for both exosome research and their potential applications. Discoveries in exosome and EV research will surely continue and lead to more discoveries in both their roles in biology and clinical applications as we learn more about these tiny, but essential elements of life.
Conference Paper
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Today, many studies are performed to explore novel wound, healing agent. Extracellular vesicles (EVs) isolated from various plants are used as wound healing agents. Human keratinocyte HACAT cells can be used as a model cell line for wound healing studies. In the current study, EVs obtained from grapefruit was selected because it has been previously proven that grapefruit increases cell proliferation of some cells. Firstly, EVs were isolated from the grapefruit and the effects of these vesicles on the cell viability of HACAT, the cell migration ability of HACAT and expression of related genes were investigated. For cell viability, EVs isolated from grapefruit were applied for 72 hours at concentrations of 1.25, 2.5, 5, 10 and 20 μg/mL, and at the end of 72 hours, cell viability was increased in a dose-dependent manner except for the lowest dose. According to the cell viability experiments; the cell migration assay was performed with 2.5 μg/mL (low dose) and 20 μg/mL (high dose). At the end of this essay, the cell migration ability of HACAT was increased in a dose-dependent manner. Then, the cells were collected from culture and total RNA was isolated for a real-time polymerase chain reaction. In gene expression analysis marker genes including collagen, laminin and fibronectin were selected. As a result, EVs isolated from grapefruit significantly increased in laminin and fibronectin gene expressions; it did not affect collagen expression. The results show that EVs isolated from grapefruit are a promising agent for wound healing studies.
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Barrier membranes are used in periodontal tissue engineering for successful neo‐bone tissue formation and prevention of bacterial colonization. We aimed to prepare and characterize novel 7% boron modified bioactive glass (7B‐BG) containing bilayered membrane for this end. We hypothesized that presence of 7B‐BG could promote structural and biological properties of GBR membrane. Cellulose acetate (CA) layer was prepared by solvent casting and functionally graded layer of CA/gelatin/BG nanoparticles was prepared by electrospinning. 0B‐BG, and 7B‐BG were synthesized by quick alkali‐mediated sol‐gel method and were characterized by SEM and FT‐Raman spectroscopy. Membranes were cross‐linked with glutaraldehyde to preserve their stability. SEM analysis showed the asymmetric nature of membranes consisting of a smooth membrane layer and a rough surface composed of (0B‐BG, 7B‐BG) containing nanofibers. 7B‐BG addition increased surface wettability (from 110.5° ± 0.8 to 73.46° ± 7.6) and biodegradability of the membranes. Additionally, a significant increase in Ca‐P layer formation was observed in 7B‐BG containing group after 1‐week incubation in stimulated body fluid. 7B‐BG incorporation resulted in a decrease in tensile strength and Young's modulus values. Human Dental Pulp Stem Cells (hDPSCs) showed better attachment, spreading and proliferation on 7B‐BG containing bilayered membranes. Osteogenic differentiation analysis revealed higher ALP enzyme activity of cells (~1.5 fold), higher intracellular calcium deposition (~2 fold), and higher calcium deposition revealed by Alizarin red staining on 7B‐BG containing bilayered membranes. Overall, results suggested that functionally graded bilayered membranes hold potential for GBR applications in regenerative dentistry.
Conference Paper
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Wound healing is a complex process where the skin and the sub-tissues repair themselves after damage. Various external substances are usually unfamiliar to the body, so they may trigger the immune system. To overcome this issue, exosomes are good candidates for the wound healing process. Due to the fact that they have the same cell surface antigens, exosomes can easily penetrate through the cell without any immune response. Human foreskin stem cells (hFSCs) are isolated from circumcised tissue normally discarded after surgery. Through embryonic development, the layer of hFSCs provides nutrients to the cells. Their potential can promote the wound healing process through proliferation, migration, and angiogenesis. In this study, it was aimed to evaluate the effect of exosomes derived from hFSCs on the wound healing.
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A proper biological microenvironment conducive to tissue repair and regeneration, while the bio‐implant interface directly affects the local microenvironment. In this study, to improve the biological microenvironment, a nano‐sized tantalum boride (Ta‐B) was coated on a titanium alloy substrate (Ti6Al4V, TC4) using magnetron co‐sputtering. The sample surface was characterized by X‐ray diffraction (XRD) and transmission electron microscopy (TEM). To investigate the effects of tantalum boride coating on the microenvironment, rabbit bone marrow stromal cells (BMSCs) and RAW 264.7 cells were respectively seeded on the sample surface and relevant experiments were conducted in vitro. The pure tantalum coating (Ta) and naked TC4 were prepared as controls. Our results showed that the Ta‐B coating enhanced cell proliferation and adhesion and inhibited the inflammatory response. Findings of alkaline phosphatase (ALP) staining, alizarin red staining and real‐time PCR for osteoblastic gene expression indicated that Ta‐B and Ta coating improve the osteogenesis, in which Ta‐B coating showed higher osteogenesis than Ta coating. Thus, this study suggests that Ta‐B coating with excellent biocompatibility could have new applications for wound healing in bone tissue engineering.
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Objectives Bisphosphonate‐related osteonecrosis of the jaw (BRONJ) is a severe complication of systemic nitrogen‐containing bisphosphonates (N‐BPs) administration, which lead to osteonecrosis, pain and infection. Despite much effort, effective remedies are yet to be established. This study aimed to investigate potential recovery effect of borate bioactive glass (BBG) in vitro and in vivo . Methods The effect of BBG on zoledronate treated bone marrow mesenchymal cells (BMSCs) and human umbilical vein endothelial cells (HUVECs) was explored by CCK‐8, EdU, flow cytometry, alkaline phosphatase staining, alizarin red staining, angiogenesis experiment and real time‐quantitative polymerase chain reaction. The preventive effect of BBG on zoledronate‐induced osteonecrosis of the jaw in rat model was examined by micro‐CT, HE staining and immunohistochemistry. Results Exposure of BBG to BMSCs and HUVECs increased cell proliferation and restored their osteogenesis and angiogenesis potential in vitro . The BRONJ lesions were satisfactorily repaired and bone mineral density, bone volume/tissue volume, trabecula number, OCN positive cells and CD31 positive cells were increased in the BBG‐treated groups compared with saline‐treated groups. Conclusions Exposure of BMSCs and HUVECs to BBG restores osteogenesis and angiogenesis inhibited by zoledronate. BBG successfully restores extraction socket healing of BRONJ in rat model.
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Objective This study aimed to evaluate the effect of direct pulp capping using an experimental self-adhesive resin for direct pulp capping (SRD) containing silica and surface pre-reacted glass-ionomer (S-PRG) filler on pulpal healing and to monitor the dentin bridge formation in rat pulp 2–4 weeks after operation. Methods Five types of SRDs (SRD-0: S-PRG fillers 0 wt%; SRD-1: S-PRG fillers 9.1 wt%; SRD-2: S-PRG fillers 18.4 wt%; SRD-3: S-PRG fillers 27.8 wt%; and SRD-6: S-PRG fillers 57.4 wt%) were prepared, and mineral trioxide aggregate (MTA) was used as control (n = 8). Direct pulp capping was performed on rats that were sacrificed for further evaluation 2 or 4 weeks after the operation. The pulp tissue disorganization (PTD), inflammatory cell infiltration (ICI), and reparative dentin formation were histopathologically evaluated; the data were statistically analyzed using the Kruskal–Wallis and the Mann–Whitney U tests. Results The histopathological evaluation of SRD-1-treated test animals 2 weeks post-operation revealed inferior PTD and ICI when compared with that of MTA. Even 4 weeks after the operation in SRD-1- and SRD-2-treated rats, the PTD and ICI were inferior when compared with those of MTA. The dental specimens of SRD-0 and MTA showed orthodentin formation, whereas SRD-treated test animals showed osteodentin formation at a position slightly deeper than the site of the pulpal exposure. Significance The reparative dentin formed by SRD-0 and MTA was genuine, whereas that formed by SRD-3 and SRD-6 was ossified and ectopic. SRD may have the potential to be utilized clinically as a direct pulp capping material.
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Dental stem cells have been isolated from the medical waste of various dental tissues. They have been characterized by numerous markers, which are evaluated herein and differentiated into multiple cell types. They can also be used to generate cell lines and iPSCs for long-term in vitro research. Methods for utilizing these stem cells including cellular systems such as organoids or cell sheets, cell-free systems such as exosomes, and scaffold-based approaches with and without drug release concepts are reported in this review and presented with new pictures for clarification. These in vitro applications can be deployed in disease modeling and subsequent pharmaceutical research and also pave the way for tissue regeneration. The main focus herein is on the potential of dental stem cells for hard tissue regeneration, especially bone, by evaluating their potential for osteogenesis and angiogenesis, and the regulation of these two processes by growth factors and environmental stimulators. Current in vitro and in vivo publications show numerous benefits of using dental stem cells for research purposes and hard tissue regeneration. However, only a few clinical trials currently exist. The goal of this review is to pinpoint this imbalance and encourage scientists to pick up this research and proceed one step further to translation.
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Numerous studies have shown that the sustained release of ions from dental restorative materials have acid buffering capacity, prevents tooth enamel demineralization, and inhibits bacterial adhesion. Herein, the release behavior and bioresponsiveness of ions released from surface pre-reacted glass-ionomer (S-PRG) fillers were investigated in different types of media based on human dental pulp-derived stem cell (hDPSC) responses. The hDPSCs were cultured for 1–7 days in S-PRG eluates diluted with varying amounts of cell culture media. S-PRG released several types of ions, such as F⁻, Sr²⁺, Na⁺, Al³⁺, BO3³⁻, and SiO3²⁻. The balance of eluted ions differed depending on the dilution and solvent, which in turn affected the cytotoxicity, cell morphology, cell proliferation, and alkane phosphatase activity of hDPSCs, among other properties. The results suggest that tailored S-PRG filler eluates could be designed and prepared for application in dental practice.
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Background and Objective. Dental stem cell-based tissue engineered constructs are emerging as a promising alternative to autologous bone transfer for treating bone defects. The purpose of this review is to systematically assess the preclinical in vivo and in vitro studies which have evaluated the efficacy of dental stem cells on bone regeneration. Methods. A literature search was conducted in Ovid Medline, Embase, PubMed, and Web of Science up to October 2014. Implantation of dental stem cells in animal models for evaluating bone regeneration and/or in vitro studies demonstrating osteogenic potential of dental stem cells were included. The preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines were used to ensure the quality of the search. Modified ARRIVE (Animal research: reporting in invivo experiments) and CONSORT (Consolidated reporting of trials) were used to critically analyze the selected studies. Results. From 1914 citations, 207 full-text articles were screened and 137 studies were included in this review. Because of the heterogeneity observed in the studies selected, meta-analysis was not possible. Conclusion. Both in vivo and in vitro studies indicate the potential use of dental stem cells in bone regeneration. However well-designed randomized animal trials are needed before moving into clinical trials.
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Stem cell usage provides novel avenues of tissue regeneration and therapeutics across disciplines. Apart from ethical considerations, the selection and amplification of donor stem cells remain a challenge. Various biopolymers with a wide range of properties have been used extensively to deliver biomolecules such as drugs, growth factors and nucleic acids, as well as to provide biomimetic surface for cellular adhesion. Using human tooth germ stem cells with high proliferation and transformation capacity, we have investigated a range of biopolymers to assess their potential for tissue engineering. Tolerability, toxicity, and their ability to direct differentiation were evaluated. The majority of pluronics, consisting of both hydrophilic and hydrophobic poly(ethylene oxide) chains, either exerted cytotoxicity or had no significant effect on human tooth germ stem cells; whereas F68 increased the multi-potency of stem cells, and efficiently transformed them into osteogenic, chondrogenic, and adipogenic tissues. The data suggest that differentiation and maturation of stem cells can be promoted by selecting the appropriate mechanical and chemical properties of polymers. It has been shown for the first time that F68, with its unique molecular characteristics, has a great potential to increase the differentiation of cells, which may lead to the development of new tissue engineering strategies in regenerative medicine.
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Objectives: The clinical translation of stem cell-based Regenerative Endodontics demands further development of suitable injectable scaffolds. Puramatrix™ is a defined, self-assembling peptide hydrogel which instantaneously polymerizes under normal physiological conditions. Here, we assessed the compatibility of Puramatrix™ with dental pulp stem cell (DPSC) growth and differentiation. Methods: DPSC cells were grown in 0.05-0.25% Puramatrix™. Cell viability was measured colorimetrically using the WST-1 assay. Cell morphology was observed in 3D modeling using confocal microscopy. In addition, we used the human tooth slice model with Puramatrix™ to verify DPSC differentiation into odontoblast-like cells, as measured by expression of DSPP and DMP-1. Results: DPSC survived and proliferated in Puramatrix™ for at least three weeks in culture. Confocal microscopy revealed that cells seeded in Puramatrix™ presented morphological features of healthy cells, and some cells exhibited cytoplasmic elongations. Notably, after 21 days in tooth slices containing Puramatrix™, DPSC cells expressed DMP-1 and DSPP, putative markers of odontoblastic differentiation. Significance: Collectively, these data suggest that self-assembling peptide hydrogels might be useful injectable scaffolds for stem cell-based Regenerative Endodontics.
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This study details the profile of 13 cell surface cluster differentiation markers on human reserve stem cells derived from connective tissues. Stem cells were isolated from the connective tissues of dermis and skeletal muscle derived from fetal, mature, and geriatric humans. An insulin/dexamethasone phenotypic bioassay was used to determine the identity of the stem cells from each population. All populations contained lineage-committed myogenic, adipogenic, chondrogenic, and osteogenic progenitor stem cells as well as lineage-uncommitted pluripotent stem cells capable of forming muscle, adipocytes, cartilage, bone, fibroblasts, and endothelial cells. Flow cytometric analysis of adult stem cell populations revealed positive staining for CD34 and CD90 and negative staining for CD3, CD4, CD8, CD11c, CD33, CD36, CD38, CD45, CD117, Glycophorin-A, and HLA DR-II. Anat Rec 264:51–62, 2001. © 2001 Wiley-Liss, Inc.
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The differentiation of odontoblasts is initiated by the organization of differentiating ameloblasts during tooth formation. However, the exact roles of ameloblast-derived factors in odontoblast differentiation have not yet been characterized. We investigated the effects of preameloblast-conditioned medium (PA-CM) on the odontogenic differentiation of human dental pulp stem cells (hDPSCs) in vitro and in vivo. Furthermore, we analyzed the PA-CM by liquid chromatography-mass spectrometry to identify novel factors that facilitate odontoblast differentiation. In the co-culture of MDPC-23 cells or hDPSCs with mouse apical bud cells (ABCs), ABCs promoted differentiation of odontoblastic MDPC-23 cells and facilitated odontoblast differentiation of hDPSCs. PA-CM, CM from ABCs after 3 days culture, was most effective in increasing the dentin sialophosphoprotein promoter activity of odontoblastic MDPC-23 cells. When PA-CM-treated hDPSCs were transplanted into immunocompromised mice, they generated pulp-like structures lined with human odontoblast-like cells showing typical odontoblast processes. However, during recombinant human bone morphogenenetic protein 2-treated hDPSCs transplantation, some of the cells were entrapped in mineralized matrix possessing osteocyte characteristics. After proteomic analyses, we identified 113 types of proteins in PA-CM, of which we characterized 23. The results show that preameloblast-derived factors induce the odontogenic differentiation of hDPSCs and promote dentin formation.
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The goal of the present study was to evaluate the histopathologic and morphometric effects of systemic boric acid in a rat periodontitis model. Twenty-four Wistar rats were divided into three groups of eight animals each: non-ligated (NL), ligature only (LO), and ligature and treated with boric acid (BA) (3mg/kg per day for 11 days). A 4/0 silk suture was placed in a subgingival position around the mandibular first molars; after 11 days the rats were sacrificed, and changes in alveolar bone levels were measured clinically and tissues were histopathologically examined to assess the differences amongst the study groups. The ratio of presence of inflammatory cell infiltration (ICI) and osteoclast number in the LO group was significantly higher than that of the NL and BA groups (p<0.05). The ratio of presence of osteoblastic activity in the LO group was significantly lower than that of the NL and BA groups (p<0.05). Alveolar bone loss was also significantly higher in the LO group compared to the BA and NL groups (p<005). This study has demonstrated that systemic administration of boric acid reduced periodontal inflammation and alveolar bone loss in periodontal disease in rats.
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Boron plays important roles in many life processes including embryogenesis, bone growth and maintenance, immune function and psychomotor skills. Thus, the delivery of boron by the degradation of borate glass is of special interest in biomedical applications. However, the cytotoxicity of borate glass which arises with the rapid release of boron has to be carefully considered. In this study, it was found that the incorporation of strontium into borate glass can not only moderate the rapid release of boron, but also induce the adhesion of osteoblast-like cells, SaOS-2, thus significantly increasing the cyto-compatibility of borate glass. The formation of multilayers of apatite with porous structure indicates that complete degradation is optimistic, and the spread of SaOS-2 covered by apatite to form a sandwich structure may induce bone-like tissue formation at earlier stages. Therefore, such novel strontium-incorporated borosilicate may act as a new generation of biomaterial for bone regeneration, which not only renders boron as a nutritious element for bone health, but also delivers strontium to stimulate formation of new bones.
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A number of studies have reported in the last decade that human tooth germs contain multipotent cells that give rise to dental and peri-odontal structures. The dental pulp, third molars in particular, have been shown to be a significant stem cell source. In this study, we isolated and characterized human tooth germ stem cells (hTGSCs) from third molars and assessed the expression of developmentally important transcription factors, such as oct4, sox2, klf4, nanog and c-myc, to determine their pluri-potency. Flow-cytometry analysis revealed that hTGSCs were positive for CD73, CD90, CD105 and CD166, but negative for CD34, CD45 and CD133, suggesting that these cells are mesenchymal-like stem cells. Under specific culture conditions, hTGSCs differentiated into osteogenic, adipogenic and neurogenic cells, as well as formed tube-like structures in Matrigel assay. hTGSCs showed significant levels of expression of sox2 and c-myc messenger RNA (mRNA), and a very high level of expression of klf4 mRNA when compared with human embryonic stem cells. This study reports for the first time that hTGSCs express developmentally important transcription factors that could render hTGSCs an attractive candidate for future somatic cell re-programming studies to differentiate germs into various tissue types, such as neurons and vascular structures. In addition, these multipotential hTGSCs could be important stem cell sources for autologous transplantation.
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Human marrow stromal cells (MSCs) were isolated from posterior illiac crest marrow aspirates obtained from 17 healthy donors, ages 19-45 years, with no apparent physical disability. First passage hMSCs exhibited growth rates in vitro that varied up to 12-fold between donors. No correlation between growth rate and the age or gender of the donor was evident (P </= 0.05). When hMSCs were cultured without passage for eight days (subconfluent cultures) or 22 days (confluent cultures) in the absence of any osteogenic agonists, levels of alkaline phosphatase enzyme activity varied 40-fold and 10-fold, respectively, between donors. When exposed to osteo-inductive media, donor populations also showed dramatic differences in levels of bone-specific gene induction. Collectively, these data demonstrate that hMSC cultures are composed of a heterogeneous mixture of cells at various stages of differentiation and with distinct osteogenic potentials. Differences in both growth rate and ALP activity were evident in hMSC cultures established from multiple aspirates obtained over a six month period from the same donors. Therefore, it appears that cellular heterogeneity produced by the method of harvest is propagated within and among different donor populations during culture expansion in vitro.
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Future cell-based therapies such as tissue engineering will benefit from a source of autologous pluripotent stem cells. For mesodermal tissue engineering, one such source of cells is the bone marrow stroma. The bone marrow compartment contains several cell populations, including mesenchymal stem cells (MSCs) that are capable of differentiating into adipogenic, osteogenic, chondrogenic, and myogenic cells. However, autologous bone marrow procurement has potential limitations. An alternate source of autologous adult stem cells that is obtainable in large quantities, under local anesthesia, with minimal discomfort would be advantageous. In this study, we determined if a population of stem cells could be isolated from human adipose tissue. Human adipose tissue, obtained by suction-assisted lipectomy (i.e., liposuction), was processed to obtain a fibroblast-like population of cells or a processed lipoaspirate (PLA). These PLA cells can be maintained in vitro for extended periods with stable population doubling and low levels of senescence. Immunofluorescence and flow cytometry show that the majority of PLA cells are of mesodermal or mesenchymal origin with low levels of contaminating pericytes, endothelial cells, and smooth muscle cells. Finally, PLA cells differentiate in vitro into adipogenic, chondrogenic, myogenic, and osteogenic cells in the presence of lineage-specific induction factors. In conclusion, the data support the hypothesis that a human lipoaspirate contains multipotent cells and may represent an alternative stem cell source to bone marrow-derived MSCs.
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We attempted the isolation and characterization of mesenchymal stem cells (MSCs) from bone marrow (BM) and umbilical cord blood (UBC) in a medium with 10% fetal bovine serum (FBS) and 10% horse serum. In the same conditions it was possible to isolate MSCs from bone marrow but not from UCB.
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We report studies of bone tissue engineering using human mesenchymal stem cells (MSCs), a protein substrate (film or scaffold; fast degrading unmodified collagen, or slowly degrading cross-linked collagen and silk), and a bioreactor (static culture, spinner flask, or perfused cartridge). MSCs were isolated from human bone marrow, characterized for the expression of cell surface markers and the ability to undergo chondrogenesis and osteogenesis in vitro, and cultured for 5 weeks. MSCs were positive for CD105/endoglin, and had a potential for chondrogenic and osteogenic differentiation. In static culture, calcium deposition was similar for MSC grown on collagen scaffolds and films. Under medium flow, MSC on collagen scaffolds deposited more calcium and had a higher alcaline phosphatase (AP) activity than MSC on collagen films. The amounts of DNA were markedly higher in constructs based on slowly degrading (modified collagen and silk) scaffolds than on fast degrading (unmodified collagen) scaffolds. In spinner flasks, medium flow around constructs resulted in the formation of bone rods within the peripheral region, that were interconnected and perpendicular to the construct surface, whereas in perfused constructs, individual bone rods oriented in the direction of fluid flow formed throughout the construct volume. These results suggest that osteogenesis in cultured MSC can be modulated by scaffold properties and flow environment.
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Tooth morphogenesis results from reciprocal interactions between oral epithelium and ectomesenchyme culminating in the formation of mineralized tissues, enamel, and dentin. During this process, epithelial cells differentiate into enamel-secreting ameloblasts. Ameloblastin, an enamel matrix protein, is expressed by differentiating ameloblasts. Here, we report the creation of ameloblastin-null mice, which developed severe enamel hypoplasia. In mutant tooth, the dental epithelium differentiated into enamel-secreting ameloblasts, but the cells were detached from the matrix and subsequently lost cell polarity, resumed proliferation, and formed multicell layers. Expression of Msx2, p27, and p75 were deregulated in mutant ameloblasts, the phenotypes of which were reversed to undifferentiated epithelium. We found that recombinant ameloblastin adhered specifically to ameloblasts and inhibited cell proliferation. The mutant mice developed an odontogenic tumor of dental epithelium origin. Thus, ameloblastin is a cell adhesion molecule essential for amelogenesis, and it plays a role in maintaining the differentiation state of secretory stage ameloblasts by binding to ameloblasts and inhibiting proliferation.
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Regenerative dental pulp strategies require the identification of precursors able to differentiate into odontoblast-like cells that secrete reparative dentin after injury. Pericytes have the ability to give rise to osteoblasts, chondrocytes, and adipocytes, a feature that has led to the suggestion that odontoblast-like cells could derive from these perivascular cells. In order to gain new insights into this hypothesis, we investigated the effects of dexamethasone (Dex), a synthetic glucocorticoid employed to induce osteogenic differentiation in vitro, in a previously reported model of human dental pulp cultures containing pericytes as identified by their expression of smooth muscle actin (SMA) and their specific ultrastructural morphology. Our data indicated that Dex (10(-8) M) significantly inhibited cell proliferation and markedly reduced the proportion of SMA-positive cells. Conversely, Dex strongly stimulated alkaline phosphatase (ALP) activity and induced the expression of the transcript encoding the major odontoblastic marker, dentin sialophosphoprotein. Nevertheless, parathyroid hormone/parathyroid hormone-related peptide receptor, core-binding factor a1/osf 2, osteonectin, and lipoprotein lipase mRNA levels were not modified by Dex treatment. Dex also increased the proportion of cells expressing STRO-1, a marker of multipotential mesenchymal progenitor cells. These observations indicate that glucocorticoids regulate the commitment of progenitors derived from dental pulp cells to form odontoblast-like cells, while reducing the proportion of SMA-positive cells. These results provide new perspectives in deciphering the cellular and molecular mechanisms leading to reparative dentinogenesis.
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Stem cells capable of differentiating to multiple lineages may be valuable for therapy. We report the isolation of human and rodent amniotic fluid-derived stem (AFS) cells that express embryonic and adult stem cell markers. Undifferentiated AFS cells expand extensively without feeders, double in 36 h and are not tumorigenic. Lines maintained for over 250 population doublings retained long telomeres and a normal karyotype. AFS cells are broadly multipotent. Clonal human lines verified by retroviral marking were induced to differentiate into cell types representing each embryonic germ layer, including cells of adipogenic, osteogenic, myogenic, endothelial, neuronal and hepatic lineages. Examples of differentiated cells derived from human AFS cells and displaying specialized functions include neuronal lineage cells secreting the neurotransmitter L-glutamate or expressing G-protein-gated inwardly rectifying potassium channels, hepatic lineage cells producing urea, and osteogenic lineage cells forming tissue-engineered bone.
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To immortalize human dental pulp (HDP) cell showing stable growth and high mineralization activities in vitro. HDP cells were obtained from a healthy third molar and immortalized by transfection with human telomerase transcriptase (hTERT) gene. To examine the characters of hTERT transfected HDP (HDP-hTERT) cells, we examined expression of mRNA for dentin sialophosphoprotein (DSSP), type I collagen (COLI), alkaline phosphatase (ALP) and bone sialoprotein (BSP) by RT-PCR. In addition, we examined ALP activity by biochemical method and nodule formation by alizarin red S (ALZ) staining. HDP-hTERT was obtained by transfection with hTERT gene. These cells bypassed the senescence and grew over 120 population doublings (PDLs) without significant growth retardation. High expression of hTERT was confirmed in HDP-hTERT by RT-PCR and showed remarkable telomerase activity. Both HDP-original (HDP-ori) and HDP-hTERT expressed DSSP, COLI, ALP and BSP mRNA and showed ALP activity and ALZ staining at the same levels. We were able to establish a cell line of immortalized human dental pulp cells with odontoblastic differentiation which will be a useful cell model for studying the mechanism of proliferation and differentiation of odontoblasts.
Article
Background— Cardiomyocytes (CMs) derived from pluripotent embryonic stem cells (ESCs) and embryonal carcinoma cells (ECCs) have some but not all characteristics of adult myocytes. ESCs have shown the ability to engraft in areas of myocardial damage, which suggests their use in cell transplantation therapy for cardiomyopathy. We studied the arrhythmogenic properties of CMs differentiated from mouse ESCs and ECCs. Methods and Results— CMs derived in vitro were studied in the whole-cell patch-clamp mode. CMs from both sources showed action potential (AP) morphology heterogeneity, with reduced maximum upstroke velocities (dV/dt) and prolonged AP durations. CMs demonstrated prolonged, spontaneous electrical activity in culture. Frequent triggered activity was observed with and without pharmacological enhancement. Phase 2 or 3 early afterdepolarizations could be induced easily by Bay K8644 plus tetraethylammonium chloride (TEA) or [TEA] o after Cs ⁺ replacement for [K ⁺ ] i , respectively. A combination of bradycardic stimulation, hypokalemia, and quinidine resulted in early afterdepolarizations. Delayed afterdepolarizations could be induced easily and reversibly by hypercalcemia or isoproterenol. Conclusions— ESCs or ECCs differentiated into at least 3 AP phenotypes. CMs showed spontaneous activity, low dV/dt, prolonged AP duration, and easily inducible triggered arrhythmias. These findings raise caution about the use of totipotent ESCs in cell transplantation therapy, because they may act as an unanticipated arrhythmogenic source from any of the 3 classic mechanisms (reentry, automaticity, or triggered activity).
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Papillon-Lefèvre syndrome (PLS) is a rare autosomal recessive disorder characterized by immune dysregulation because of a mutation in cathepsin c gene, resulting in hyperkeratosis of the palms, soles, elbows, and knees combined with premature loss of the primary and permanent dentitions. Periodontal tissue abnormalities in PLS patients were reported previously. However, less is known about dental pulp tissue derived cells of PLS patients. This study aimed to show stem cell potential of PLS dental pulp stem cells (DPSCs) and provide new evidence regarding the pathophysiology of the disease. DPSCs were characterized by using flow cytometry and immunocytochemistry. They were also induced to differentiate into adipogenic, osteogenic, chondrogenic, odontogenic, and myogenic cells. The results revealed that PLS DPSCs are stained positive for mesenchymal stem cells surface markers CD29, CD73, CD90, CD105, and CD166. PLS DPSCs were able to differentiate into adipogenic, osteogenic, chondrogenic, and odontogenic cell types properly. PLS DPSCs expressed embryonic stem cell markers Oct4, Sox2, cMYc, and Klf4 and showed similar proliferation rate compared with DPSCs isolated from healthy young controls. Interestingly, it was found that unlike the healthy DPSCs, PLS DPSCs are not able to form myotubes with correct morphology. These data are being reported for the first time; therefore, they might provide new insights to the pathology of the disease. Our results suggest that the PLS DPSCs might be an autologous stem cell source for PLS patients for cellular therapy of alveolar bone defects and other dental tissue abnormalities observed in PLS.
Article
Human marrow stromal cells (MSCs) were isolated from posterior illiac crest marrow aspirates obtained from 17 healthy donors, ages 19–45 years, with no apparent physical disability. First passage hMSCs exhibited growth rates in vitro that varied up to 12-fold between donors. No correlation between growth rate and the age or gender of the donor was evident (P ≤ 0.05). When hMSCs were cultured without passage for eight days (subconfluent cultures) or 22 days (confluent cultures) in the absence of any osteogenic agonists, levels of alkaline phosphatase enzyme activity varied 40-fold and 10-fold, respectively, between donors. When exposed to osteo-inductive media, donor populations also showed dramatic differences in levels of bone-specific gene induction. Collectively, these data demonstrate that hMSC cultures are composed of a heterogeneous mixture of cells at various stages of differentiation and with distinct osteogenic potentials. Differences in both growth rate and ALP activity were evident in hMSC cultures established from multiple aspirates obtained over a six month period from the same donors. Therefore, it appears that cellular heterogeneity produced by the method of harvest is propagated within and among different donor populations during culture expansion in vitro. J. Cell. Biochem. 75:424–436, 1999. © 1999 Wiley-Liss, Inc.
Article
This report documents osteoblast differentiation in vitro, as demonstrated by the 50–100× increase of proteins which are known markers of the osteoblast phenotype. Collagen type I and osteocalcin synthesis and accumulation, alkaline phosphatase activity, and matrix calcification show similar temporal relationships that are analogous to those seen during in vivo bone development. Chicken embryonic osteoblast progenitor cells were selected by initial growth at low densities in minimal medium. Upon subcultivation into nutrient-enriched medium at higher cell densities, near homogeneous populations of osteoblasts were obtained as demonstrated by the greater than 80% enrichment of cells positive for alkaline phosphatase activity. A comparison was made between cells grown in the presence or absence of 10 mM β-glycerolphosphate (β-GPO4), a chemical stimulant of matrix calcification, as a function of time. Cultures treated with β-GPO4 showed visible calcification at Day 12 when culture monolayers became confluent. By Day 30, numerous large foci of calcification were visible and a 20-fold increase in calcium (Ca) content was observed. In contrast, untreated cultures had only a 3-fold increase in Ca content with many smaller diffuse areas of calcification. DNA, RNA, and total protein levels were nearly identical between the two cultures, indicating that β-GPO4 had no marked effect on either cell proliferation or transcriptional activity. The major collagen type produced by either culture was type I, with no detectable type III as determined by CNBr peptide mapping and delayed reduction analysis. Alkaline phosphatase activity showed a rapid ∼50-fold induction by Day 18 and remained elevated in control cultures. However, cultures treated with β-GPO4 demonstrated a rapid 80% decline of enzyme activity after 18 days. In contrast, total osteocalcin levels showed a 100-fold induction by Day 18 and remained elevated in both control and β-GPO4-treated cultures throughout the time period examined. While the overall levels of osteocalcin were the same in β-GPO4-treated and untreated cultures, 2- to 5-fold more osteocalcin was associated with the more mineralized matrices of the β-GPO4-treated cultures. In order to confirm the association of osteocalcin with areas of mineralization, co-localization of mineral to osteocalcin and collagen was carried out by combining vital labeling with tetracycline and immunofluorescent staining with anti-osteocalcin and anti-collagen antibodies. Both collagen and osteocalcin showed strong localization with areas of mineralization. This culture system defined the expression of specific markers of osteoblast function during differentiation and their relationships to matrix calcification. Thus, these osteoblast cultures provide a unique model in which to study the regulation of bone-specific proteins and genes during osteoblast differentiation and matrix calcification.
Article
Stem cells are considered to be promising therapeutic options in many neuro-degenerative diseases and injuries to the central nervous system, including brain ischemia and spinal cord trauma. Apart from the gold standard embryonic and mesenchymal origin, human tooth germ stem cells (hTGSCs) have also been shown to enjoy the characteristics of mesenchymal stem cells (MSCs) and the ability to differentiate into adipo-, chondro-, osteo- and neuro-genic cells, suggesting that they might serve as potential alternatives in the cellular therapy of various maladies. Immortalization of stem cells may be useful to avoid senescence of stem cells and to increase their proliferation potential without altering their natural characteristics. This study evaluated the expression of stem cell markers, surface antigens, differentiation capacity, and karyotype of hTGSCs that have been immortalized by human telomerase reverse transcriptase (hTERT) or simian vacuolating virus 40 (SV40) large T antigen. These undying cells were also evaluated for their neuro-protective potential using an in vitro SH-SY5Y neuro-blastoma model treated with hydrogen-peroxide or doxo-rubicin. Although hTGSC-SV40 showed abnormal karyotypes, our results suggest that hTGSC-hTERT preserve their MSC characteristics, differentiation capacity and normal karyotype, and they also possess high proliferation rate and neuro-protective effects even at great passage numbers. These peculiars indicate that hTGSC-hTERT could be used as a viable model for studying adipo-, osteo-, odonto- and neuro-genesis, as well as neuro-protection of MSCs, which may serve as a springboard for potentially utilizing dental waste material in cellular therapy.
Article
Previous studies have shown the superiority of nanofibrous (NF) poly(l-lactic acid) (PLLA) scaffolds in supporting the osteogenic differentiation of a few cell types and bone regeneration. The aim of the current study was to investigate whether NF-PLLA scaffolds are advantageous for the odontogenic differentiation and mineralization of human dental pulp stem cells (DPSCs) over solid-walled (SW) PLLA scaffolds. The in vitro studies demonstrated that, compared with SW scaffolds, NF scaffolds enhanced attachment and proliferation as well as odontogenic differentiation of human DPSCs. The alkaline phosphatase (ALP) activity and the expression of odontogenic genes of human DPSCs were increased on NF scaffolds compared with that on SW scaffolds. In addition, more mineral deposition was observed on the NF scaffolds, as demonstrated by von Kossa staining, calcium content measurement and scanning electron microscopy. Consistent with the in vitro studies, NF scaffolds promoted odontogenic differentiation and hard tissue formation compared with SW scaffolds after 8 weeks of ectopic transplantation in nude mice, as confirmed by von Kossa staining, Masson's trichrome staining and immunohistochemical staining for dentin sialoprotein. In conclusion, NF-PLLA scaffolds enhanced the odontogenic differentiation of human DPSCs and mineralization both in vitro and in vivo, and are promising scaffolds for dentin regeneration.
Article
Bone marrow stromal cells (BMSCs) have been well established as an ideal source of cell-based therapy for bone tissue engineering applications. Boron (B) is a notable trace element in humans; so far, the effects of boron on the osteogenic differentiation of BMSCs have not been reported. The aim of this study was to evaluate the effects of boron (0, 1, 10,100, and 1,000 ng/ml) on osteogenic differentiation of human BMSCs. In this study, BMSCs proliferation was analyzed by cell counting kit-8 (CCK8) assay, and cell osteogenic differentiation was evaluated by alkaline phosphatase (ALP) activity assay, Von Kossa staining, and real-time PCR. The results indicated that the proliferation of BMSCs was no different from the control group when added with B at the concentration of 1, 10, and 100 ng/ml respectively (P > 0.05); in contrast, 1,000 ng/ml B inhibited the proliferation of BMSCs at days 4, 7, and 14 (P < 0.05). By ALP staining, we discovered that BMSCs treated with 10 and 100 ng/ml B presented a higher ALP activity compared with control (P < 0.05). By real-time PCR, we detected the messenger RNA expression of ALP, osteocalcin, collagen type I, and bone morphogenetic proteins 7 were also increased in 10 and 100 ng/ml B treatment groups (P < 0.05). The calcium depositions were increased in 1 and 10 ng/ml B treatment groups (P < 0.05). Taken all together, it was the first time to report that B could increase osteogenic effect by stimulating osteogenic differentiation-related marker gene synthesis during the proliferation and differentiation phase in human BMSCs and could be a promising approach for enhancing osteogenic capacity of cell-based construction in bone tissue engineering.
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Previous studies have shown that zinc chloride (ZnCl(2)) can induce metallthionein (MT) in the liver and kidney to protect tissues against toxicants and shows a better corneal wound healing than conventional drugs do. We hypothesized that ZnCl(2) can promote odontogenesis of dental pulp stem cells (DPSCs) via MT. The purpose of this study was to investigate the effects of ZnCl(2) on human DPSCs and the expression of MT. DPSCs were isolated by flow cytometry with selective surface marker CD146 and STRO-1. After they grew into confluence, DPSCs were induced into odontoblasts with or without ZnCl(2) supplemented in the culture medium for 21 days. The effect of ZnCl(2) on DPSCs differentiation was examined followed by alkaline phosphatase staining/activity and quantitative real-time polymerase chain reaction analysis. By treating DPSCs with ZnCl(2), the duration of mineralization was shortened and expressions of differentiation markers into odontoblasts were more significant than those without ZnCl(2) stimulation. Besides, the MT gene expression was increased with the increasing expressions of odontoblasts' markers after treated with ZnCl(2). This was the first report that ZnCl(2) could promote odontoblastic differentiation of DPSCs through the up-regulation of gene MT.
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Transplantation of mesenchymal stem cells (MSCs) inhibits the progression of disc degeneration in animal models. We know of no study to determine the optimal number of cells to transplant into the degenerated intervertebral disc (IVD). To determine the optimal donor cell number for maximum benefit, we conducted an in vivo study using a canine disc degeneration model. Autologous MSCs were transplanted into degenerative discs at 10(5), 10(6), or 10(7) cells per disc. The MSC-transplanted discs were evaluated for 12 weeks using plain radiography, magnetic resonance imaging, and gross and microscopic evaluation. Preservation of the disc height, annular structure was seen in MSC-transplantation groups compared to the operated control group with no MSC transplantation. Result of the number of remaining transplanted MSCs, the survival rate of NP cells, and apoptosis of NP cells in transplanted discs showed both structural microenvironment and abundant extracellular matrix maintained in 10(6) MSCs transplanted disc, while less viable cells were detected in 10(5) MSCs transplanted and more apoptotic cells in 10(7) MSCs transplanted discs. The results of this study demonstrate that the number of cells transplanted affects the regenerative capability of MSC transplants in experimentally induced degenerating canine discs. It is suggested that maintenance of extracellular matrix by its production from transplanted cells and/or resident cells is important for checking the progression of structural disruption that leads to disc degeneration.
Article
The aim of this study was to investigate the odontogenic differentiation of human dental pulp stem cells (DPSCs) on nanofibrous (NF)-poly(l-lactic acid) (PLLA) scaffolds in vitro and in vivo. Highly porous NF-PLLA scaffolds which mimic the architecture of collagen type I fibers were fabricated by the combination of a phase-separation technique and a porogen-leaching method. The human DPSCs were then seeded onto the scaffolds and cultured in different media for odontogenic differentiation: "Control" medium without supplements; "DXM" medium containing 10(-8)M dexamethasone (DXM), 50 microgml(-1) ascorbic acid and 5mM beta-glycerophosphate; "BMP-7+DXM" medium containing 10(-8)M DXM, 50 microgml(-1) ascorbic acid, 5mM beta-glycerophosphate plus 50 ngml(-1) bone morphogenetic protein 7 (BMP-7). For odontogenic differentiation study in vitro, alkaline phosphatase activity quantification, reverse transcription polymerase chain reaction, scanning electron microscopy, von Kossa staining and calcium content quantification were carried out. While both "DXM" medium and "BMP-7+DXM" medium induced the DPSCs to odontoblast-like cells, the "BMP-7+DXM" medium had greater inducing capacity than the "DXM" medium. Consistent with the in vitro studies, the "BMP-7+DXM" group presented more extracellular matrix and hard tissue formation than the "DXM" group after 8 weeks of ectopic implantation in nude mice. Differentiation of DPSCs into odontoblast-like cells was identified by the positive immunohistochemical staining for dentin sialoprotein. In conclusion, odontogenic differentiation of DPSCs can be achieved on NF-PLLA scaffolds both in vitro and in vivo; the combination of BMP-7 and DXM induced the odontogenic differentiation more effectively than DXM alone. The NF-PLLA scaffold and the combined odontogenic inductive factors provide excellent environment for DPSCs to regenerate dental pulp and dentin.