Biological Activity Of Rabbit BMSCs By HBMP-2 Gene-modified

Background:The mandible is a irregular bone which located in the 1/3 low face and contributes to the chewing, biting, swallowing and other important physiological activities. The mandibular defect has become more and more popular in maxillofacial surgery. With the rapid development of tissue engineering technology, the methods to treatment and reconstruction the mandibular defect have also been greatly improved. Among them, engineering technology has been widely recognized because of exhibiting significant advantages, the bone tissue. Seed cells, scaffold, and related cytokines are three elements of bone tissue engineering technology, and accessing to seed cells which have good biologically actives is the basic link to tissue engineering. In this study, by RT-PCR technology to obtain and amplify human bone morphogenetic protein-2(hBMP-2) gene fragment from the human hepatoma cell line HepG2. By enzyme digestion technology, the gene are inserted into bone marrow mesenchymal stem cells(BMSCs) genome, thereby enhancing the activity of BMP gene expression of BMSCs so that accelerate the reconstruction of bone defect area, and lay the foundation for subsequent experiments. Objective:The rabbit BMSCs were isolated and cultured, and its biological special characters were detected. The expression of the target gene was detected in BMSCs after the cells were modificated by BMP-2 gene, further more to explore the feasibility of it as seed cells for repairing bone defect. Methods:By density gradient centrifugation, the rabbit BMSCs were isolated and cultured, then the biological characteristics of BMSCs were preliminary testing. By reverse transcription polymerase chain reaction(PCR) technology, the hBMP-2 gene fragment were isolated from HepG2 cells. Target gene fragments were connected with a carrier after amplified. the recombinant plasmid was injected into the lentivirus vector. Then the target gene was transfected into BMSCs mediated by 293 T cell, and the mRNA and protein expression of BMP-2 gene were detected in BMSCs cells. Results:The rabbits BMSCs were isolated and cultured by density gradient centrifugation, and the cells had the good growth characters, and could passage stably. Through the methods of extracting, amplification and connection, the recombinant plasmid vectors were successfully constructed, as long as the genetically modified rabbit BMSCs. Genetically modified BMSCs had the ability to differentiated into bone and of adipose tissue. The target gene could expressed efficiently in the cells, and protein expression was also significantly increased BMP cells. Conclusions:The purpose of gene modified BMSCs to exhibit good biological activity, and laid the foundation for further experimental study.