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Induction of tendon cell-related genes by BMP-12 in rat BM-MSC implants in calcaneal tendon in vivo. Samples were collected as described in Methods and Materials. mRNA levels were determined by qRT-PCR and normalized against samples of the non-BMP-12treated group. The gene expression of all experimental groups was determined as fold changes relative to native tendon samples. While unseeded implants expressed minimal Scx, Tnmd, Col I, and Tn-C, BMP-12-treated implants had significantly increased expression of these genes compared to non-BMP-12-treated implants. Data are expressed as mean 6 S.D. (n = 3). * represents p,0.05. doi:10.1371/journal.pone.0017531.g005
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We characterized the differentiation of rat bone marrow-derived mesenchymal stem cells (BM-MSCs) into tenocyte-like cells in response to bone morphogenetic protein-12 (BMP-12). BM-MSCs were prepared from Sprague-Dawley rats and cultured as monolayers. Recombinant BMP-12 treatment (10 ng/ml) of BM-MSCs for 12 hours in vitro markedly increased expres...
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... treated groups (Fig. 4). The expression levels of both Scx and Tnmd in BMP-12-treated scaffold implants were even higher than in the naı¨venaı¨ve tendons (Fig. 4). Consistently, RT-PCR analysis revealed that cells from the seeded/BMP-12-treated implants expressed much higher mRNA levels of Scx, Tnmd, Col I, and Tn-C than control samples (Fig. 5). These differences in vivo were even greater than those observed in 3D cultures in vitro. Taken together, these data suggest that rat BM-MSCs pretreated with BMP-12 have acquired full tenogenic capability, thus being able to form tendon-like tissues after implantation in ...
Citations
... Notavelmente, TGF-b1 foi o fator isolado mais potente que estimula a regulação positiva do gene tenogênico e a produção de matriz extracelular, os autores chegaram à conclusão que as BMSCs responderam melhor ao TGF-b1/CTGF combinados e TGF-b1. (Yin et al. 2016;Lee et al. 2011;Jiang et al. 2016;Zhang et al. 2018 Gissi, C. et al. Extracellular vesicles from rat-bone-marrow mesenchymal stromal/stem cells improve tendon repair in rat Achilles tendon injury model in dose-dependent manner: A pilot study. ...
Mesenchymal stem cells are of great interest due to their regenerative potential and capacity to transdifferentiate into various cell lineages. The present study executed a narrative review of the literature on the influence of mesenchymal stem cells on the regeneration of tendon injuries. To acomplish this, a search was carried out in the Pubmed and Scielo databases using descriptors such as: Mesenchymal stem cell regeneration and application tendon rat; Embryonic Stem Cell-Derived Mesenchymal Stem Cells for Tendon Tissue rat. Only research articles carried out on rats between 2011 and 2023 were included. 935 articles were found in the Pubmed database, and 43 articles were included based on the criteria of categorization, analysis of titles and abstracts. The articles showed benefits in relation to the use of stem cells in rat tendon injuries, but some studies showed that this repair was not 100% guaranteed to the state of the tendon originally, but associated with growth factors obtained a better induction of tenogenesis.
... MSC treatment of tendon/ligament injuries improve tissue strength, provides a more favourable type I collagen composition, indicating a beneficial therapeutic response to these cells [254]. There are several clinical studies using BM-MSCs as the therapeutic option for tendon repair, perhaps because it is the most studied tissue source of MSCs [255,256]. Still, a recent study compared them with UC-MSC, in vitro, and concluded UC-MSC surpasses other MSCs in its ability to differentiate into tendonlike lineage cells and establish a well-organized tendon-like matrix. ...
Musculoskeletal injuries such as equine osteoarthritis, osteoarticular defects, tendonitis/desmitis, and muscular disorders are prevalent among sport horses, with a fair prognosis for returning to exercise or previous performance levels. The field of equine medicine has witnessed rapid and fruitful development, resulting in a diverse range of therapeutic options for musculoskeletal problems. Staying abreast of these advancements can be challenging, prompting the need for a comprehensive review of commonly used and recent treatments. The aim is to compile current therapeutic options for managing these injuries, spanning from simple to complex physiotherapy techniques, conservative treatments including steroidal and non-steroidal anti-inflammatory drugs, hyaluronic acid, and polyacrylamides, to the promising regenerative therapies such as hemoderivatives and stem cell-based therapies. Each therapeutic modality is scrutinized for its benefits, limitations, and potential synergistic actions to facilitate their most effective application for intended healing/regeneration of the injured tissue/organ and subsequent patient recovery. While stem cell-based therapies emerge as particularly promising for equine musculoskeletal injuries, a multidisciplinary approach is underscored throughout the discussion, emphasizing the importance of considering various therapeutic modalities in tandem.
... BMP-12 and surgically implanted around experimentally transected AT in a rat model improved and accelerated tendon healing and influenced early tissue regeneration, leading to quicker recovery and improved biomechanical properties of the AT (Majewski et al., 2008). Lee et al. showed that BMMSCs implanted into an animal model after loading onto a three-dimensional collagen sponge scaffold and induction by BMP-12 promoted tendon-like tissue formation (Lee et al., 2011). In addition, the number of tenocytes increased, which were arranged regularly along the tension axis. ...
Achilles tendon (AT) injury is one of the most common tendon injuries, especially in athletes, the elderly, and working-age people. In AT injury, the biomechanical properties of the tendon are severely affected, leading to abnormal function. In recent years, many efforts have been underway to develop effective treatments for AT injuries to enable patients to return to sports faster. For instance, several new techniques for tissue-engineered biological augmentation for tendon healing, growth factors (GFs), gene therapy, and mesenchymal stem cells were introduced. Increasing evidence has suggested that GFs can reduce inflammation, promote extracellular matrix production, and accelerate AT repair. In this review, we highlighted some recent investigations regarding the role of GFs, such as transforming GF-β(TGF-β), bone morphogenetic proteins (BMP), fibroblast GF (FGF), vascular endothelial GF (VEGF), platelet-derived GF (PDGF), and insulin-like GF (IGF), in tendon healing. In addition, we summarized the clinical trials and animal experiments on the efficacy of GFs in AT repair. We also highlighted the advantages and disadvantages of the different isoforms of TGF-β and BMPs, including GFs combined with stem cells, scaffolds, or other GFs. The strategies discussed in this review are currently in the early stages of development. It is noteworthy that although these emerging technologies may potentially develop into substantial clinical treatment options for AT injury, definitive conclusions on the use of these techniques for routine management of tendon ailments could not be drawn due to the lack of data.
... Moreover, the results of Azan staining indicated that the sham group was strongly stained with azocarmine red, whereas at 4 weeks after the operation, the injured area was strongly stained with aniline blue (Figure 2I,L). It is commonly assumed that Achilles tendons, which are normally composed of collagen fibers, are stained blue; however, Lee et al. reported that the original Achilles tendon stained red [33]. Therefore, we suspect that the blue staining observed at 4 weeks after the operation may represent collagen fibers that have formed scar tissue. ...
Tendons help transmit forces from the skeletal muscles and bones. However, tendons have inferior regenerative ability compared to muscles. Despite studies on the regeneration of muscles and bone tissue, only a few have focused on tendinous tissue regeneration, especially tendon regeneration. Sex-determining region Y-box transcription factor 9 (Sox9) is an SRY-related transcription factor with a DNA-binding domain and is an important control factor for cartilage formation. Sox9 is critical to the early-to-middle stages of tendon development. However, how Sox9 participates in the healing process after tendon injury is unclear. We hypothesized that Sox9 is expressed in damaged tendons and is crucially involved in restoring tendon functions. We constructed a mouse model of an Achilles tendon injury by performing a 0.3 mm wide partial excision in the Achilles tendon of mice, and chronologically evaluated the function restoration and localization of the Sox9 expressed in the damaged sites. The results reveal that Sox9 was expressed simultaneously with the formation of the pre-structure of the epitenon, an essential part of the tendinous tissue, indicating that its expression is linked to the functional restoration of tendons. Lineage tracing for Sox9 expressed during tendon restoration revealed the tendon restoration involvement of cells that switched into Sox9-expressing cells after tendon injury. The stem cells involved in tendon regeneration may begin to express Sox9 after injury.
... Hence, this is a unique study of fabricating and characterizing phytoextract incorporated tendon-engineered construct, an unmet clinical need in skeletal reconstructive surgeries.Mesenchymal stem cells (MSCs) can differentiate in to mesenchymal lineages such as adipocytes, chondrocytes and osteocytes.They were potentially transdifferentiate into non-mesenchymal cell types such as pancreatic cells and cardiomyocytes.[46][47][48][49] MSCs are studied as the ideal source of cellular therapeutic agents for tendon repair.[50][51][52][53][54][55] A number of growth factors, particularly members of the bone morphogenetic proteins BMP family have been shown to encourage tenocytic differentiation from the multipotent MSC. ...
Achilles tendon, which connects the calf muscles to heel, is the strongest tendon in the body. Despite its strength, it is more prone to injury due to its limited blood supply. Tendon‐related injuries are more common in sportspersons, people with labor‐intensive work and the aged community. The currently available treatment mode is surgery which is expensive with chances of re‐injury. Present study made an attempt to fabricate a tissue‐engineered tendon product using decellularized tendon (DT) seeded with stem cells and bioactive components of Tinospora cordifolia extract (TCE). The bare DT tissue scaffold/substitute may also serve as a drug delivery platform for growth factors and cells with a new approach to promote tissue regeneration in clinical applications. DT construct showed good regenerative potential and easily promoted new tissue formation. Decellularization of the tendon was carried out by chemical method using tri (n‐butyl) phosphate (TnBP). DT was physicochemically characterized by contact angle measurement, thermal gravimetric analysis (TGA), and mechanical testing. Rabbit adipose derived mesenchymal stem cells (RADMSCs) were isolated and phenotypically characterized by flow cytometry analysis, tri lineage differentiation, and so forth. Further, stem cell seeded DT scaffolds were prepared and found to be non‐toxic by cytotoxicity, cell adhesion by scanning electron microscope (SEM) analysis, cell viability by live dead assays, and so forth. The findings of this study yield valid proof for the employability of cell‐seeded DT construct as a natural scaffold in repairing injured tendons—the toughest chords of the skeleton. This is a cost effective method for the replacement of injured/damaged tendons for athletes, people in labor‐intensive occupations, the elderly population, and so forth—a boon towards the repair of the tendon in damage/injury.
... [79] BMP Improves molecular, organizational, and mechanical properties of healing tendon. [181][182][183][184] Epigenetic regulation miRNA and lncRNA Guide tendon stem cell differentiation and tissue regeneration, promote tendon healing, and reduce adhesion. [161][162][163] EGR1: early growth response 1; Scx: scleraxis; TGF-β: transformational growth factor-beta; VEGF: vascular endothelial growth factor; FGF: fibroblast growth factor; PDGF: platelet-derived growth factor; ECM: extracellular matrix; IGF: insulin-like growth factor; BMP: bone morphogenetic protein. ...
Tendons are an important part of the musculoskeletal system. Connecting muscles to bones, tendons convert force into movement. Tendon injury can be acute or chronic. Noticeably, tendon healing requires a long time span and includes inflammation, proliferation, and remodeling processes. The mismatch between endogenous and exogenous healing may lead to adhesion causing further negative effects. Management of tendon injuries and complications such as subsequent adhesion formation are still challenges for clinicians. Due to numerous factors, tendon healing is a complex process. This review introduces the role of various biological and mechanical factors and epigenetic regulation processes involved in tendon healing.
... The present study and other previous studies found that BMPs play a crucial role in tendon development and regeneration. For instance, our study and others have demonstrated that BMP12 can promote tenogenic differentiation [15,27] by stimulating SCX expression [20] via the Smad1/5/8 pathway [31]. Overexpression of BMP12 can promote tendon regeneration and formation [23]. ...
The Wingless and Int-1 (WNT) and bone morphogenic protein/growth differentiation factor (BMP/GDF) signalling pathways contribute significantly to the development of the musculoskeletal system. The mechanism by which they contribute is as follows: BMP/ GDF signalling usually promotes tendon differentiation, whereas WNT signalling inhibits it. We hypothesised that inhibiting WNT and subsequently stimulating BMP signalling may enhance the tenogenic differentiation of stem cells. The objective of this study was to determine whether a combination of WNT inhibitor (KY02111) and BMP12/GDF7 protein could enhance the differentiation of bone marrow-derived equine mesenchymal stromal cells (BM-eMSCs) into tenocytes. Cells were cultured in five treatments: control, BMP12, and three different combinations of BMP12 and KY02111. The results indicated that a 1-day treatment with KY02111 followed by a 13-day treatment with BMP12 resulted in the highest tenogenic differentiation score in this experiment. The effect of KY02111 is dependent on the incubation time, with 1 day being better than 3 or 5 days. This combination increased tenogenic gene marker expression, including SCX, TNMD, DCN, and TNC, as well as COL1 protein expression. In conclusion, we propose that a combination of BMP12 and KY02111 can enhance the in vitro tenogenic differentiation of BM-eMSCs more than BMP12 alone. The findings of this study might be useful for improving tendon differentiation protocols for stem cell transplantation and application to tendon regeneration.
... The treatment of tendon injuries using MSCs and scaffolds have been assured by progress in tissue engineering (Yang et al. 2013). MSCs can be obtained from various tissues, such as bone marrow, adipose tissue, and tendons (Bi et al. 2007;Cheng et al. 2009;Yin et al. 2010) and they have the ability to make tissues of musculoskeletal lineages, including tenocytes (Lee et al. 2011). Among these sources, harvesting ASCs is more easy and yields to a high number of ASCs per volume of tissue (Naderi et al. 2017). ...
... A recent research has presented the key role of the ECM in tendon differentiation (Bi et al. 2007;Marturano et al. 2013) and the role of several growth factors in the healing process for tendons and ligaments has been documented (Molloy et al. 2003). Bone morphogenic proteins 12 and 13 (BMP12 / 13) as members of transforming growth factor beta (TGF-b) superfamily, additionally referred to as growth differentiation factor (GDF7 / 6), are shown to play an very important role in proliferation, cell differentiation, and matrix synthesis (Wang et al. 1988;Wozney et al. 1988;Wozney 1992;Sieber et al. 2009;Lee et al. 2011;Javanshir et al. 2020). They also play an important role in regulating evolution and morphogenesis of multiple organs and tissues (Wang et al. 1988;Wozney et al. 1988;Wozney 1992;Sieber et al. 2009). ...
Mesenchymal Stem Cells (MSCs) are important in regenerative medicine and tissue engineering and will be a very sensible choice for repair and regeneration of tendon. New biological practices, such as cellular therapy using stem cells, are promising for facilitating or expediting tendon therapy. Before using these cells clinically, it is best to check and confirm the optimal conditions for differentiation of these cells in the laboratory. Hence, in the present study, the impacts of PDGF-BB and GDF-6 supplementation on adipose-derived MSCs (ASCs) culture were studied. The frozen ASC were recovered and expanded in basic culture medium (DMEM with 10%FBS). The cells after passage five (P5) were treated with basic medium containing L-Prolin, Ascorbic Acid and only PDGF-BB or GDF-6 (20 ng/ml) or both of them (mix) as 3 groups for 14 days to investigate efficiency of ASCs differentiation towards tenocytes. The cells culturing in basic medium were used as control group. To validate tenogenic differentiation, H&E and Sirius Red staining were used to assess cell morphology and collagen production, respectively. In addition, mRNA levels of collagen I and III, Scleraxis and Tenomodulin as tenogenic markers were analyzed using qPCR. In all test groups, cells appeared slenderer, elongated cytoplasmic attributes compared to the control cells. The intensity of Sirius Red staining was significantly higher in GDF-6, PDGF-BB alone, than in group without supplements. The optical density was higher in the GDF-6 than PDGF-BB and mix-group. QPCR results showed that Col I and III gene expression was increased in all groups compared to the control. SCX expression was significantly increased only in the PDGF-BB group. TNMD mRNA expression was not significant among groups. In this study, we have corroborated that human ASCs are reactionary to tenogenic induction by GDF-6 and PDGF-BB alone or in combination. These outcomes will help greater insight into GDF-6 and PDGF-BB driven tenogenesis of ASCs and new directions of discovery in the design of ASC-based treatments for tendon healing.
... Studies have indicated that in vitro-induced differentiation of bone marrow MSCs into tendon cells can be achieved by a combination of biophysical, soluble, and extracellular matrix factor modulation [29][30][31][32] . A number of in vivo studies have also indicated that treatment with transplanted MSCs can improve tendon healing for various parts in animal models, including Achilles tendon defects [33][34][35][36][37] , rotator cuff defects 35,38 , and patellar tendinitis 39 . ...
A rotator cuff tear is an age-related common cause of pain and disability. Studies including our previously published ones have demonstrated that mesenchymal stem cells cultured under hypoxic conditions [hypoxic multipotent stromal cells (MSCs)] facilitate the retention of transplanted cells and promote wound healing. However, there are very few, if any, reports targeting the punctured supraspinatus tendons to create more or equally serous wounds as age-related tears of rotator cuff. It remains to be determined whether transplantation of bone-marrow-derived hypoxic MSCs into the punctured supraspinatus tendon improves tendon repair and, when combined with ultrasound-guided delivery, could be used for future clinical applications. In this study, we used a total of 33 Sprague-Dawley rats in different groups for normal no-punched control, hypoxic MSC treatment, nontreated vehicle control, and MSC preparation, and then evaluated treatment outcomes by biomechanical testing and histological analysis. We found that the ultimate failure load of the hypoxic MSC-treated group was close to that of the normal tendon and significantly greater than that of the nontreated vehicle control group. In vivo tracking of cells labeled with superparamagnetic iron oxide (SPIO) nanoparticles revealed an enhanced retention of transplanted cells at the tear site. Our study demonstrates that hypoxic MSCs improve rotator cuff tear repair in a rat model.
... BMP is a multifunctional acidic hydrophobic glycoprotein. Previously, it was indicated that these BMPs, particularly BMP-12, can initiate the tenogenic pathway in rat and human derived MSCs (Dai et al. 2015;Lee et al. 2011;Shen et al. 2013). It was shown that transient stimulation with BMP-12 in vitro was sufficient to induce the differentiation of rat bone marrow mesenchymal stem cells BM-MSCs into tendon cells, and this phenotype was maintained in vivo (Lee et al. 2011). ...
... Previously, it was indicated that these BMPs, particularly BMP-12, can initiate the tenogenic pathway in rat and human derived MSCs (Dai et al. 2015;Lee et al. 2011;Shen et al. 2013). It was shown that transient stimulation with BMP-12 in vitro was sufficient to induce the differentiation of rat bone marrow mesenchymal stem cells BM-MSCs into tendon cells, and this phenotype was maintained in vivo (Lee et al. 2011). In addition, other researchers used a rat Achilles tendon healing model to demonstrate that sutures that release BMP-12 can accelerate tendon healing, and that the delivery method determines the response of the healing tissue to BMP-12 (Chamberlain et al. 2015). ...
... Different concentrations of BMP-12 were added to α-MEM medium containing 10% serum and induced for different durations; the relative expression levels of tendon proteins Scleraxis, Tenascin C and Tenomodulin were measured, and it was found that the tendon cell proteins Scleraxis, Tenascin C, and Tenomodulin were significantly increased after induction by 10 ng/ml BMP-12 for 48 h. This is consistent with previous research (Lee et al. 2011), which shows that the bone morphogenetic protein BMP-12 can induce the tendon differentiation of C3H10T1/2 cells. In addition, some studies also suggested that activation YAP can enhance tendon regeneration (Tao et al. 2021;Huang et al. 2020). ...
The Yes-associated protein (YAP) transcription co-activator is recognized as a key mediator and has been implicated in the regulation of stem cell fate; however, the role of YAP in the tenogenic differentiation of mesenchymal stem cells (MSCs) is not well understood. In the present study, we characterized tenogenic differentiation of C3H10T1/2 cells induced by BMP-12 and evaluated the function of YAP in the regulation of the BMP-12-induced tenogenesis. We found that the tendon-related proteins Scleraxis, Tenascin C, and Tenomodulin were significantly increased after induction by 10 ng/ml BMP-12 for 48 h in C3H10T1/2 cells. In addition, the expression of YAP was significantly enhanced and transferred from the cytoplasm to the nucleus during tenogenic differentiation. In contrast, downregulation of YAP suppressed the cells’ tenogenic differentiation, and the expression of YAP transferred from the nucleus to the cytoplasm. These results indicate that YAP is required for the expression of tenogenic markers in the tenocytic differentiation process of C3H10T1/2 cells. Collectively, we demonstrate that YAP transcription co-activator is a novel regulator in the process of BMP-12-induced MSCs tenogenesis and has a correlation with the other tenogenic regulators and markers. These results shed new light on the function of YAP in tendon healing and regeneration.
