Evaluation of rabbit adipose derived stem cells fate in perfused multilayered silk fibroin composite scaffold for Osteochondral repair.

Development of osteochondral tissue engineering approaches using scaffolds seeded with stem cells in association with mechanical stimulations has been recently considered as a promising technique for the repair of this tissue. In this study, an integrated and biomimetic trilayered silk fibroin (SF) scaffold containing SF nanofibers in each layer was fabricated. The osteogenesis and chondrogenesis of stem cells seeded on the fabricated scaffolds were investigated under a perfusion flow. 3-Dimethylthiazol-2,5-diphenyltetrazolium bromide assay showed that the perfusion flow significantly enhanced cell viability and proliferation. Analysis of gene expression by stem cells revealed that perfusion flow had significantly upregulated the expression of osteogenic and chondrogenic genes in the bone and cartilage layers and downregulated the hypertrophic gene expression in the intermediate layer of the scaffold. In conclusion, applying flow perfusion on the prepared integrated trilayered SF-based scaffold can support osteogenic and chondrogenic differentiation for repairing osteochondral defects.

Antifungal activity of Gracilaria corticata methanol extract against Trichophyton mentagrophytes, Microsporum canis, and Microsporum gypseum on rat dermatophytosis models.

Background and Purpose: Dermatophytosis is one of the most prevalent zoonotic diseases. Increased resistance of dermatophytosis-causing pathogens against antidermatophytic agents highlights the need for alternative medicine with higher efficiency and lower side effects. In the present study, the in vitro antifungal activities of different concentrations of Gracilaria corticata methanol extract against Trichophyton mentagrophytes, Microsporum canis, and Microsporum gypseum were assessed and their efficacy was evaluated in rat dermatophytosis models. Materials and Methods: The broth microdilution and well diffusion methods were used to determine the in vitro antidermatophytic activity. The in vivo study was carried out using 40 dermatophytosis-infected adults male Wistar rats. The animals were divided into 4 groups (5% and 10% G. corticata ointment, terbinafine, and Vaseline) and treated with ointment until complete recovery. The percentage of wound closure was calculated for each group. Results: The results revealed that G. corticata methanol extract was effective to varying extents against the tested dermatophytes. The highest inhibitory activity of G. corticata was found against T. mentagrophytes with minimum inhibitory concentration and minimum fungicidal concentration values of 4 and 9 µg mL-1, respectively. The in vivo experiment revealed that 10% G. corticata ointment significantly accelerated skin lesions reduction and completely cured M. gypseum, T. mentagrophytes, and M. canis infections after 19, 25, and 38 days, respectively. Conclusion: The methanol extract of G. corticata exhibited significant antifungal activity in vitro and in vivo, suggesting that it could be used as an alternative to antidermatophytic therapy in a dose-dependent manner.

Equine tendon mechanical behaviour: Prospects for repair and regeneration applications.

Tendons are dense connective tissues that play an important role in the biomechanical function of the musculoskeletal system. The mechanical forces have been implicated in every aspect of tendon biology. Tendon injuries are frequently occurring and their response to treatments is often unsatisfactory. A better understanding of tendon biomechanics and mechanobiology can help develop treatment options to improve clinical outcomes. Recently, tendon tissue engineering has gained more attention as an alternative treatment due to its potential to overcome the limitations of current treatments. This review first provides a summary of tendon mechanical properties, focusing on recent findings of tendon mechanobiological responses. In the next step, we highlight the biomechanical parameters of equine energy-storing and positional tendons. The final section is devoted to how mechanical loading contributes to tenogenic differentiation using bioreactor systems. This study may help develop novel strategies for tendon injury prevention or accelerate and improve tendon healing.

Optimizing Tenogenic Differentiation of Equine Adipose-Derived Mesenchymal Stem Cells (eq-ASC) Using TGFB3 Along with BMP Antagonists.

OBJECTIVE: Tendon repair strategies usually are accompanied by pathological mineralization and scar tissue formation that increases the risk of re-injuries. This study aimed to establish an efficient tendon regeneration method simultaneously with a reduced risk of ectopic bone formation. MATERIALS AND METHODS: In this experimental study, tenogenic differentiation was induced through transforming growth factor- ß3 (TGFB3) treatment in combination with the inhibiting concentrations of bone morphogenetic proteins (BMP) antagonists, gremlin-2 (GREM2), and a Wnt inhibitor, namely sclerostin (SOST). The procedure's efficacy was evaluated using real-time polymerase chain reaction (qPCR) for expression analysis of tenogenic markers and osteochondrogenic marker genes. The expression level of two tenogenic markers, SCX and MKX, was also evaluated by immunocytochemistry. Sirius Red staining was performed to examine the amounts of collagen fibers. Moreover, to investigate the impact of the substrate on tenogenic differentiation, the nanofibrous scaffolds that highly resemble tendon extracellular matrix was employed. RESULTS: Aggregated features formed in spontaneous normal culture conditions followed by up-regulation of tenogenic and osteogenic marker genes, including SCX, MKX, COL1A1, RUNX2, and CTNNB1. TGFB3 treatment exaggerated morphological changes and markedly amplified tenogenic differentiation in a shorter period of time. Along with TGFB3 treatment, inhibition of BMPs by GREM2 and SOST delayed migratory events to some extent and dramatically reduced osteo-chondrogenic markers synergistically. Nanofibrous scaffolds increased tenogenic markers while declining the expression of osteo-chondrogenic genes. CONCLUSION: These findings revealed an appropriate in vitro potential of spontaneous tenogenic differentiation of eq- ASCs that can be improved by simultaneous activation of TGFB and inhibition of osteoinductive signaling pathways.

Sirenomelia: two case reports.

BACKGROUND: Sirenomelia, also called mermaid syndrome, is a rare lethal multi-system congenital deformity with an incidence of one in 60,000-70,000 pregnancies. Sirenomelia is mainly characterized by the fusion of lower limbs and is widely associated with severe urogenital and gastrointestinal malformations. The presence of a single umbilical artery derived from the vitelline artery is the main anatomical feature distinguishing sirenomelia from caudal regression syndrome. First-trimester diagnosis of this disorder and induced abortion may be the safest medical option. In this report, two cases of sirenomelia that occurred in an white family will be discussed. CASE PRESENTATION: We report two white cases of sirenomelia occurring in a 31-year-old multigravid pregnant woman. In the first pregnancy (18 weeks of gestation) abortion was performed, but in the third pregnancy (32 weeks) the stillborn baby was delivered by spontaneous vaginal birth. In the second and fourth pregnancies, however, she gave birth to normal babies. Three-dimensional ultrasound imaging showed fusion of the lower limbs. Neither she nor any member of her family had a history of diabetes. In terms of other risk factors, she had no history of exposure to teratogenic agents during her pregnancy. Also, her marriage was non-consanguineous. CONCLUSION: This report suggests the existence of a genetic background in this mother with a Mendelian inheritance pattern of 50% second-generation incidence in her offspring.

Equine adipose mesenchymal stem cells (eq-ASCs) appear to have higher potential for migration and musculoskeletal differentiation.

Equine adipose-derived mesenchymal stem cells (eq-ASCs) possess excellent regeneration potential especially for treatment of musculoskeletal disorders. Besides their common characteristics, MSCs harvested from different species reveal some species-specific and donor-dependent behaviors. Hence, the molecular analysis of MSCs may shed more light on their future clinical application of these cells. This study aimed to investigate some behavioral aspects of eq-ASCs in vitro which may influence the efficacy of stem cell therapy. For this purpose, MSCs of a donor horse were isolated, characterized and expanded under normal culture conditions. During continuous culture condition, eq-ASCs were started to formed aggregated structures that was accompanied with the up-regulation of migratory related genes including transforming growth factor beta 1 (TGFB1) and its receptor 3 (TGFBR3), and snail family transcriptional repressor 1 (SNAI1), E-cadherin (CDH1) and ß-catenin (CTNNB1). Moreover, the expression of a musculoskeletal progenitor marker, scleraxis bHLH transcription factor (SCX), was also increased after 3 days. In order to clarify the impact of TGFB signaling pathway on cultured cells, gain- and loss-of-function treatment by TGFB3 and SB431542 (TGFB inhibitor) were performed, respectively. We found that TGFB3 treatment exaggerated the aggregate formation effects, in some extend via induction of cytoskeletal actin rearrangement, while inhibition of TGFB signaling pathway by SB431542 reversed this phenomenon. Overall, our findings support the fact that eq-ASCs have an inherent capacity for migration, which was enhanced by TGFB3 treatment and, this ability may play crucial role in cell motility and wound healing of transplanted cells.

Strategies of tenogenic differentiation of equine stem cells for tendon repair: current status and challenges.

Tendon injuries, as one of the most common orthopedic disorders, are the major cause of early retirement or wastage among sport horses which mainly affect the superficial digital flexor tendon (SDFT). Tendon repair is a slow process, and tendon tissue is often replaced by scar tissue. The current treatment options are often followed by an incomplete recovery that increases the susceptibility to re-injury. Recently, cell therapy has been used in veterinary medicine to treat tendon injuries, although the risk of ectopic bone formation after cell injection is possible in some cases. In vitro tenogenic induction may overcome the mentioned risk in clinical application. Moreover, a better understanding of treatment strategies for musculoskeletal injuries in horse may have future applications for human and vice versa. This comprehensive review outlines the current strategies of stem cell therapy in equine tendon injury and in vitro tenogenic induction of equine stem cell.