Study On Constructing Of Tissue-Engineering Skull With Gene Thansfering Technology

Skull defect caused of varios reasons is one of the most common diseases in neurosurgery. The traditional strategies of healing the disease are to repair the defect. At present, there are many mending materials. All these materials undergo some disadvantages such as scarce source, immunologic rejection and disease transmission. In recent years, rapidly developed gene engineering and tissue engineering technology give a promising method to deal with skull defect. It has become a hot research spot. According to the primary method of bone tissue engineering, we used porous CPC as extracellular matrix, VEGF gene thansferred marrow stromal cells (MSCs) and osteogenic induced MSCs as seed cells to make artificial skull of tissue engineering in this study. In order to determine whether the artificial bone have osteogenic activity, we have observed the capabilities of the vascularization and bone defect repairing in vivo. The osteogenic induced MSCs contructed skull was used as comparison. I.The revascularization and osteogenesis activity of the VEGF gene transferred tissue engineering skull in vivo. Objective: To observe osteogenesis and vascularization of tissue engineering skull compounded with osteogenic induced MSCs and MSCs transfected with VEGF in vivo. Methods: Autogenous marrow stromal cells were obtained from femurs and tibias of 10 adult rabbits under general anesthesia and sterile condition and cultured in DMEM supplemented with 10%FBS. VEGF165 gene was transfected into stroma cells by means of LipofectAMINETM2000 reagent transfection. The stably gene expressive cells were screened with G-418 for fourteen days. After osteogenic induced of MSCs, we mixed osteogenic induced MSCs and MSCs transfected with VEGF at the rate of 2:1. The cell mixture were seeded in porous calcium phosphate bioceramic and cultured in vitro for another 7 days. After inoculating the cells onto the surface of porous calcium phosphate ceramic, we surveyed the characteristic of proliferation by cell using scanning electronic microscope. The cell-ceramic compound was implanted in the muscle pouches of in the corresponding rabbit. Implanted substances were taken out of 2, 4 and 8 weeks later. Slices were observed to study the osteogenesis activity of implanted substance. Results: Rat bone marrow stroma cell could be attached to and extended on the surface of porous calcium phosphate ceramic, and normally grown, proliferated. Porous calcium phosphate ceramic could promote cell proliferation. In the group of tissue engineering bone compounded with MSCs transfected VEGF, more osteoblasts and capillary vessels appeared, compared with those in the group of tissue engineering bone compounded with osteogenic induced MSCs. Conclusion: The results show that porous calcium phosphate ceramic has good biocompatibility with rabbit MSCs transfected VEGF, they can be used as biomaterials in bone tissue engeering. Tissue engineering bone compounded with MSCs transfected VEGF had good osteogenetic activity. II.The repair of skull defect with the VEGF gene transferred tissue engineering skull in vivo. Objective: To evaluate the effect of repairing of skull defect with the VEGF gene transferred tissue engineering bone. Methods: To establish large segment...