Effect Of BMP-2 Transfection Mediated By Adenovirus On The Bone Marrow Mesenchymal Stem Cells

Bone is an active tissue, which maintains its integrity and mineralization balance through bone remodeling, that is bone formation by osteoblasts and bone resorption by osteoclasts. Oral bone defects and jaw bone resorption, defects and atrophy, caused by tumor, inflammation, damage or infection, is one of the most common manifestations in oral clinics. It not only influences functions such as chewing, pronouncing, beautiful outlook and eating, but also significantly influences patients'living quality and physical and mental health. How to find a way to prevent bone resorption and promote bone formation efficiently is one of the problems oral medicine meet.Osteoblast is the main cell that participates in bone formation. Its precursor, that is bone marrow mesenchymal stem cells (BMMSCs), can differentiate into multiple cells under certain conditions, such as chondroblast, osteoblast, lipoblast, myoblast, etc. Therefore, studying the regulation of the differentiation and function of BMMSCs into osteoblast not only has significance in treatment of oral and maxillofacial bone resorption diseases, but also provides new references.Bone morphogenetic protein (BMP) is an acidic protein, which is in bone matrix. It belongs to TGF-βsuper family and is a kind of multi-function factor. There are altogether at least 40 BMP members, among which BMP-2 is widely investigated in bone tissue engineering and is now known to be one of the strongest factors in inducing bone formation.BMP-2 has a significant role during bone formation and restoration, and is one of the most important growth factors in ectopia osteogenesis. We tried to introduce BMP-2 gene into BMMSCs by gene transfer and examine the effect of BMP-2 on the proliferation, differentiation and function of BMMSCs, which can further provide experiment evidence in clinical use.In this experiment, BMMSCs were cultured in vitro, and vectors of adenovirus carrying BMP-2 gene were constructed. Then BMMSCs were transferred and examined. We tried to investigate the feasibility of transferring BMMSCs and examine the effect of BMP-2 on BMMSCs, which provides theoretical base and experiment evidence in treating bone defect diseases by gene transfer.Firstly, BMMSCs were isolated and cultured using whole bone marrow adherence method. BMMSCs were identified using ALP staining and lipo-formation induction. Then, BMMSCs were transferred by adenovirus carrying BMP-2 gene. Finally, proliferation and differentiation were tested by cellular biology and molecular biology.Conclusions:1. BMMSCs were isolated, amplified, cultured and identified using whole bone marrow adherence method; BMMSCs obtained have a high purity and a strong ability of proliferation;2. It was confirmed the feasibility of gene transfer in BMMSCs, which made the target gene expression persistently, stably and efficiently;3. It was demonstrated that BMP-2 can significantly promote the osteogenic and function of BMMSCs;4. It provided theoretical base and experimental evidence in bone tissue engineering using BMMSCs as seed cells.