Construction Of Tissue Engineered Periosteum Of Two Different Scaffold Materials And In Vitro Determination Of Osteogenic Activity

ObjectiveBased on rabbit bone marrow mesenchymal stem cells,tissue engineered periosteum was constructed in vitro using human acellular amniotic membrane and porcine small intestine submucosa as scaffolds.The growth and differentiation of rabbit mesenchymal stem cells were observed and the osteogenic activity of the two tissue engineered periostees was determined in vitro.We can provide experimental data support for finding scaffold materials that can construct ideal tissue engineered periosteum.MethodThe test can be divided into three groups.The human acellular amniotic membrane is loaded with rabbit bone marrow mesenchymal stem cells,which is recorded as HAAM group,and the rabbit small intestinal submucosa is loaded with rabbit bone marrow mesenchymal stem cells,which are recorded as SIS group,and rabbit bone marrow without stent material.Mesenchymal stem cells.Recorded as a blank control group.The extraction of bone marrow mesenchymal stem cells from the long bones of rabbits was the first step of the experiment.After osteogenic induction culture,histological observation and growth curve were drawn.Secondly,human amniotic membrane and submucosal layer of pig small intestine were decellularized and disinfected,and the morphology was observed under electron microscope.The tissue-engineered periosteum was constructed in vitro by loading the third generation rabbit bone marrow mesenchymal stem cells,and then the morphology of the two tissues was observed under electron microscope.The MTT assay was used to detect the rabbit bone marrow mesenchymal stem cells in human acellular amniotic membrane and porcine small intestinal mucosa.The proliferation of the lower layer.Finally,the amount of alkaline phosphatase and osteocalcin secretion was measured in vitro.Result1.After in vitro osteogenic induction culture,rabbit bone marrow mesenchymal stem cells can survive and proliferate,and have the characteristics of osteoblasts.2.The human acellular amniotic membrane and the submucosal layer of the small intestine of the pig are treated with disinfection and sterilization.There are no residual cells on both surfaces,and the spatial structure is basically intact,and clear fibrous tissue can be seen.3.The human acellular amniotic membrane has more cell proliferation than the porcine small intestine submucosa and the blank group,and more alkaline phosphate and osteocalcin are secreted than the other two groups.Conclusion1.Rabbit bone marrow mesenchymal stem cells have the same morphology and effect as osteoblasts after in vitro osteogenic induction.2.The cell proliferation carried on the scaffold material is more 3.The human amniotic membrane and the porcine small intestine submucosa are treated without carrying cells,only collagen fibers are present,no cytotoxicity,and there is room for growth and differentiation of bone marrow mesenchymal stem cells.structure.4.The specific reticular fiber structure of human acellular amniotic membrane is more suitable for rabbit bone marrow mesenchymal stem cell proliferation and expression of osteoblast phenotype than the parallel fibrous structure of porcine small intestine submucosa.More alkaline phosphatase and bone calcium are secreted.Prime.More suitable for use as a scaffold material to construct tissue engineered periosteum.