The Autogenous Bone Marrow Derived MSCs Being Induced To Myoblast And Hhk Scaffold Material For Defect Skeletal Muscle Repair

The damage and defect of skeletal muscle is common surgery. In order to investigate the repair of defect skeletal muscle, the autogenous bone marrow derived mesenchymal stem cells (MSCs) were induced to myoblasts at first. Then the compound of the myoblasts and human hair keratin (HHK) was used for the repair of the defect skeletal muscle.The bone marrow derived MSCs possessed of the ability of proliferation in high rate and could grow in stable, in the lower concentration of serum, through the observation of morphology and the assay of proliferation capability and cell cycle. In this condition, we could obtain sufficient cells for the next inducing experiments.In the process of inducing the bone marrow derived MSCs to myoblasts in vitro, l0μmol/L 5-azacytidin was effective, through the detection of the activation of myoblast determination genes with RT-PCR, and the expression of skeletal muscle desmin with flow cytometry and immiinohistochemistry. The cell population containing higher proportion of myoblasts, therefore, it could be used for the next implantation.The degradation progress of HHK scaffold material was observed through the routine morphological and ubiquitin immunohistochemical study, after the implantation of the compound of the induced myoblasts and HHK scaffold material. The results made clear that the degradation was in the peak after implantation 3-6^th week, and accomplished in about 9th week. The HHK scaffold material was degraded into small particles by ubiquitin system at first, and then those particles phagocytosed by macrophages. At the same time, because of directivity of HHK scaffold material, from our works, it could lead the cells to grow in parallel with the muscle fiber.There were three groups for the comparison of the repair results: HHKscaffold material implantation group (1st group), the compound of the autogenous bone marrow derived MSCs and HHK scaffold material implantation group (2nd group), and the compound of the autogenous induced myoblasts and HHK scaffold material implantation group (3rd group). Among the three groups, the repair effectiveness of the 3rd group was the best than the others, on the basis of the time of regeneration myofibers formation and it density, through the morphological and myosin immunohistochemical study...