Osteogenic differentiation of pre-conditioned bone marrow mesenchymal stem cells with Nisin on modified poly-L-lactic-acid nanofibers.

Introduction: Human bone marrow-derived mesenchymal stem cells (hBM-MSCs) are undifferentiated cells with self-renewing ability and multi-lineage differentiation beneficial for regenerative medicine. Nano scaffolds are novel materials employed in bone repair and regeneration. Nisin is a prebiotic that can increase stem cells' lifespan and proliferation. This study attempted to provide a proper strategy for bone marrow mesenchymal stem cells differentiation into the Osteocytes on a Poly-L-lactic-acid (PLLA) scaffold after pretreating with Nisin. Methods: MSC osteogenic differentiation was evaluated by measuring Calcium, Alkaline phosphatase, and quantitative tests such as Real-Time PCR, Acridine Orange, Alizarin Red, Von Kossa, and others. Results: The result of the MTT test showed that the optimal dose of Nisin prebiotic for the MSCs' preconditioning was 200 IU/mL on the 1st, 3rd, and 5th days of culture. Real-time PCR data indicated that the expression rate of ALP, Osteonectin, Osteocalcin, and Collagen I have increased in the presence of Nisin, while the RUNX-2 gene expression has decreased. Furthermore, the results of Alizarin Red and Von Kossa tests, as well as Scanning electron microscopy (SEM), revealed that the cell proliferation in the preconditioned samples with Nisin increased significantly. Conclusions: The study concluded that the cell proliferation and differentiation increased in samples pretreated with Nisin on the PLLA Nano scaffolds.

Application of adipose-derived, muscle-derived, and co-cultured stem cells for the treatment of stress urinary incontinence in rat models.

OBJECTIVES: Based on the recent advancements in cell therapy techniques, we aimed to evaluate the efficacy of transurethral injection of autologous adipose-derived stem cells, muscle-derived stem cells, and co-cultured cells for the rehabilitation of stress urinary incontinence rat models. We hypothesized that the utilization of co-cultured stem cells could result in enhanced therapeutic outcomes attributed to their more comprehensive environment of paracrine factors and cytokines. METHODS: We performed bilateral pudendal nerve transection surgeries to simulate urinary incontinence in 25 female Wistar rats and employed urodynamic evaluations to confirm the injury. We autologously isolated and cultured adipose-derived mesenchymal stem cells, muscle-derived stem cells, and a mixed culture of the two types, which we subsequently injected into the urethral lumen of the damaged animals. Three weeks after the injection, urodynamic assays, histological staining, and immunohistochemical evaluations were performed to determine the efficacy of the implanted cell cultures in sphincter function improvements or structural modifications. RESULTS: Histological evaluations suggested a regenerative process in the muscular layer of the external sphincter 3 weeks after the injection. Also, immunohistochemical analysis revealed a thickened periurethral striated muscle layer in the co-cultured group. Postinjection urodynamic analysis indicated that the urethral pressure profile significantly increased in the co-cultured group compared with other groups. CONCLUSIONS: The outcomes of this investigation indicated that the application of co-cultured adipose-derived and muscle-derived stem cells could be associated with higher therapeutic value in stress urinary incontinence patients compared with singular-cell treatments.

Effect of dexamethasone, insulin and EGF on the myogenic potential on human endometrial stem cell.

Human endometrium contains mesenchymal stem cells (eMSC) which have the ability to differentiate into three cell lineages and the potential in therapeutic applications. We hypothesize that using environmental induction in culture media such as dexamethasone, human recombinant insulin and human epidermal growth factor (hEGF) can differentiate endometrial stem cells into myoblast. These agents have a broad range of effects in myoblast differentiation in-vitro. We used immunohystochemistry analysis and RT -PCR to evaluate the presence of skeletal muscle - specific proteins some of which are expressed in the early stage of differentiation including myoD and Desmin which expressed at later stages of differentiation. In conclusion eMSC can differentiate in culture media which contains above mentioned factors and use for therapeutic purpose in muscular degenerative disease.