In Vitro Study Of Bio-gide Collagen Membrane As Bone Mesenchymal Stem Cells Scaffolds

Abstract:Background:Osteonecrosis of the femoral head (ONFH) is a kind of common disease in Department of orthopedics. This type of tumor is characterized by high incidence and high disability rate. ONFH can cause destruction of the bone structure, and result in loss of bone structure collapse, joint function and joint pain. How to repair cartilage lesions and prevent further damage of the cartilage is an important project for the treatment of head-preserving procedures with ONFH. It was new theory that combined culture of bone marrow stromal stem cells in vitro and Bio-gide collagen membrane to repair cartilage defects.Objective:This study was to research canine bone marrow stromal stem cells isolation, culture and identification methods of ONFH as a cartilage defect repair tissue engineering seed cells is feasible. And in vitro composited cultured of BMSCs with Bio-gide,to explored the advantages of the Bio-gide collagen membrane carrier as a bone marrow stromal stem cells.Methods:In vitro cultured BMSCs, inverted microscope/transmission electron microscope morphological observation,HE staining, immunofluorescenceexamination,,and the cell surface antigen test; In vitro by BMSCs induced to differentiation into chondrocytes culture and toluidine blue, type â…¡ collagen immunohistochemical staining, and other biological identification; will BMSCs with Bio-gide membrane collagen in vitro co-culture by inverted phase contrast microscope/transmission electron microscopy to observation.Results:The combination of bone marrow adherent blood centrifugation screening method to get high purity active BMSCs.MTT test result the3rd-passage cells showed a high growth rate and conspicuous syntropic characteristics. Immunofluorescence detected high expression of CD44and CD90positive, CD34negative, indication that the cells are not fibroblast cells,hematopoietic cells and endothelial cells; BMSCs under certain conditions can be transformed into chondrocytes, toluidine blue staining to see the chondrocytes were coloring, staining faded after the passage. Immunohistochemical staining of collagen type â…¡ see the brown-yellow precipitatein less than five generations of cartilage cells; BMSCs in the Bio-gide membrane attached to collagen proliferation, and multi-layer growth.Conclusion:BMSCs easily obtained and expanded differentiation ability. Potential conversion to chondrocytes in vitro certain conditions.BMSCs can be used as ONFH in tissue engineering of cartilage defects source of seed cells. Bio-gide membrane collagen as a carrier of cell growth.