Study Of Allogenic Demineralized Bone Matrix Of Mongrel Combined With Cell-sheet And Endothelial Progenitor Cells

PURPOSE:To investigate the growth situation of BMSCs cell-sheet and endothelial progenitor cells (EPC) on demineralized bone matrix (DBM) and explore the effect of DBM/BMSCs cell-sheet and DBM/EPC complex on the construction of functional and vascularized tissue-engineered bone.METHODS:BMSC cell-sheet was prepared with cell-sheet engineering approach, EPC were isolated from canine bone marrow and DBM was prepared from homologous bone. BMSCs cell-sheet and EPC were inoculated on DBM respectively and observed by inverted phase contrast microscope and scanning electron microscope. The porosity of DBM was measured. The back of dog was invided into four areas. The DBM/BMSC/EPC/BMSC cell-sheet was implanted in the area of A, the DBM/BMSC/EPC was implanted in the area B and C, and the DBM/BMSC was implanted in the area of D. The implantes in the areas A and B were prepared into cylindrical to wrap the thoracodorsal artery and vein of the dog. The implantes in the areas C and D were prepared into flake and were wraped by the myofascial of Latissimus dorsi. The plantes were taken out 12 weeks later for examination. RESULTS:The DBM/BMSCs cell-sheet and DBM/EPC complex were successfully constructed. The BMSCs cell-sheet and EPC adhered to DBM well and grew rapidly. The degree of osteogenesis formation of the implantes was A>B and the blood vessel formation was B>C>D.CONCLUSION:The BMSCs cell-sheet and EPC have good biocompatibility with DBM. The complex of DBM/BMSCs cell-sheet, DBM/EPC could provide good conditions to acquire functional and vascularized tissue-engineered bone.