MSCs And MGF Combined With CPC/FG To Construct Tissue Engineered Bone For The Treatment Of Osteoporotic Bone Defect In Rats

Objective The calcium phosphate cement/fibrin glue was used as the composite carrier to prepare tissue engineered bone carrying bone marrow mesenchymal stem cells and force growth factor.The situation of repairing osteoporotic bone defect in rats by implanting this material was observed,and the feasibility and effectiveness of this method were observed,so as to find an ideal material for repairing osteoporotic bone injury.Methods(1)BMSCs were isolated and cultured from the bone marrow of rats using the whole bone marrow method,and their morphology and growth were observed,and the differentiation into osteoblasts was attempted in vitro.(2)Using absorbable calcium phosphate cement / fibrin glue as a carrier,bone marrow mesenchymal stem cells and force growth factors were combined to construct tissue engineering bone scaffold in vitro,and the growth of cells was observed.(3)The osteoporotic bone defect model of rats was established.The composite bone scaffold material(Group C: CPC/FG/MGF/BMSCs)constructed in vitro was implanted into the osteoporotic bone defect area.At the same time,group A(CPC/FG)and group B(CPC/FG/BMSCs)were set for comparative experiments.Samples were taken at 4 and 8 weeks after operation.The repair of bone defect was observed by gross morphology,X-ray photography,histological staining,and immunohistochemistry,and the feasibility and effectiveness of the prepared tissue engineered bone were preliminarily evaluated.Results(1)Rat bone marrow mesenchymal stem cells were isolated and cultured from rat bone marrow by whole bone marrow method,which conformed to the characteristics of stem cells and had strong self-proliferation ability.Osteoblasts can be induced to differentiate into osteoblasts under osteogenic induction.(2)Rat bone marrow mesenchymal stem cells can grow and proliferate in calcium phosphate cement / fibrin glue composited with force growth factors constructed in vitro.(3)The osteoporosis model of ovariectomized female SD rats was evaluated by bone mineral density detection method,and it was confirmed that the osteoporosis model could be successfully established 12 weeks after castration.(4)The preliminary evaluation results of the tissue-engineered bone scaffold prepared in this experiment for the repair of radial defects in rats showed that the bone repair process of rats in group C was faster than that in group A and group B at 4 and 8 weeks after operation.The results of immunohistochemistry showed that group C had higher ALP positive expression than group B and group A at 4 weeks after operation.The positive expression of ALP in group C and group B was significantly higher than that in group A at 8 weeks after operation,and there was no significant difference between group C and group B.Conclusions 1.It is feasible to isolate and culture bone marrow mesenchymal stem cells in vitro by whole bone marrow method.The isolated cells show the characteristics of stem cells in vitro,and can differentiate into osteoblasts under osteogenic induction environment.2.Rat BMSCs cultured in vitro can adhere and grow on CPC / FG / MGF composite scaffolds and can be used to construct tissue engineered bone in vitro.3.The osteoporosis model of ovariectomized female SD rats was evaluated by bone mineral density detection,and it was confirmed that the osteoporosis model could be successfully established 12 weeks after castration.4.Preliminary evaluation of in vivo repair experiment of osteoporotic rat radial defect model showed that calcium phosphate cement / fibrin glue composite scaffold loaded with growth factor and bone marrow mesenchymal stem cells had good ability to repair osteoporotic bone defect,which was worthy of further study of its mechanism and effectiveness.