Regulating Osteogenic Differentiation Of Bmscs By Cell-secreted Decellularized Extracellular Matrixes From Different Cell Origins

The development of tissue engineering is undoubtedly a key part of tissue engineering scaffold materials.In vivo,various biophysics and biochemical stimuli control the growth and development of tissues and organs,maintain their physiological structure and function,and use these factors to improve the functional expression of tissue engineering implants,which is one of the important ideas in the research of tissue engineering.Extracellular matrix(ECM)is secreted by specific cells from organisms,and there is a reversible dynamic balance between cells and cells.It plays an important role in cell attachment,differentiation,and the connection between cells and biomaterials.Decellularized extracellular matrix(D-ECM)scaffolds obtained by acellular technology have been widely used,and are favored because of their good biocompatibility,biodegradability and induction of differentiation.However,the D-ECM scaffolds of different tissue sources have different effects on cell proliferation and differentiation,especially the regularities of the directional differentiation of pluripotent stem cells,which still need to be studied.To better retain the growth ability and function of cells,and to obtain a complete lamellar extracellular matrix material,a new idea of obtaining seed cells by cell sheet technology is provided in tissue engineering research.In this paper,three kinds of cells,fibroblasts(L929),osteoblasts(MC3T3)and bone marrow mesenchymal stem cells(BMSCs)were selected from different origins.The cell sheet was constructed by culture in vitro,and then three kinds of D-ECM were obtained by deactivating the cell sheet by physical and the biological enzymes.Materials were cultured in vitro and subcutaneously implanted in rats,and their effects on proliferation and osteogenic differentiation of BMSCs were compared.The proliferation and osteogenic differentiation of BMSCs were characterized by three kinds of D-ECM.The results showed that all D-ECM were beneficial to the attachment and proliferation of BMSCs,and BMSCs derived D-ECM promoted cell proliferation and osteogenic differentiation were the best,followed by osteoblast D-ECM,and finally,fibroblast D-ECM.The reason for the analysis was that D-ECM of BMSCs origin contains the most abundant COL-1 and GAG,and VEGF and BMP-2 expression.Finally,the polylactic acid/gelatin fiber membrane obtained by electrostatic spinning was used as the support to incubate the three kinds of cell sheets.The composite structure of the D-ECM and the electrospun membrane was obtained by acellular treatment.They were implanted into the back of the mice and cultured for 3 months to show that the electrospun membrane can promote the formation of the cell sheet and endowed the D-ECM with the operation.Compared with the simple fibrous membrane group,the fibrous membrane of the D-ECM loaded didn't cause large immune rejection and inflammatory reaction,which showed good biocompatibility.Therefore,D-ECM/fiber membrane complex is expected to be a new kind of regenerative repair material for guided tissue.It has a better prospect in the field of periodontium and bone tissue regeneration.