种植转染VEGF165基因骨髓基质细胞的脱蛋白异体骨在裸鼠体内的成骨

目的:探讨脱蛋白骨支架种植转染血管内皮生长因子(vascular endothelial growth factor,VEGF)165基因的骨髓基质细胞体内的成骨作用.方法:从狗的髂骨穿刺骨髓分离骨髓基质细胞,应用脂质体转染VEGF165基因,以RT-PCR检测转染的基因表达.随机选取4周龄BALB/C裸鼠30只,随机分为3组,各组10只.实验组A为脱蛋白骨 转染pcDNA3-hVEGF165基因的骨髓基质细胞;实验组B为脱蛋白骨 骨髓基质细胞;对照组C为单纯植入脱蛋白骨 DMEM.在脊柱两侧分开肌间隙,每只裸鼠植入2块脱蛋白骨材料至肌腹内.术后4、8周处死取材,经大体观察、组织形态学与免疫组化方法检查细胞材料复合物植入后的成骨作用及VEGF表达.结果:组织学检查A、B组均有成骨,并随时间延长而新骨增多,但其中转基因的A组其成骨作用明显,并在植入4周后免疫组化可见VEGF表达呈阳性;对照组C无成骨现象.结论:转染VEGF165基因骨髓基质细胞复合脱蛋白骨在体内具有较好的成骨作用.

Osteogenic potentiality of deproteinized bone composited with dog marrow stromal cells transfected with VEGF165 gene after implantation into nude mice

Objective: To study the osteogenic potentiality of deproteinized bone composited with dog marrow stromal cells(MSCs)transfected with VEGF_(165) gene implantation in vivo.Methods:Marrow stromal cells were obtained from the dual ilium of three adult dogs.The recombinant pcDNA_3-hVEGF_(165) plasmid was transfected into the MSCs with lipofectin and expression of VEGF_(165) in MSC s was detected by RT-PCR.Deproteinized bone was composed with transfected MSCs or untransfected MSCs cultured for one week before being implanted into nude mice.30 nude mices were divided into three groups at random as follows:the group A was implanted with deproteinized bone composed with transfected MSCs;group B used deproteinized bone composed with untransfected MSCs;the control group C used deproteinized bone composed with culture medium DMEM.The osteogenesis of material with cells was studied through gross observation,hematoxylin and eosin staining,immunochemsitry and quantitative analysis.Results:Deproteinized bone composed with transfected or untransfected MSCs could produce cartilage tissue and bone in vivo,and more new bone tissue produced gradually as the time after implantation went by.The transfected MSCs material group had more contents of ossification and angiogenesis than those in the untransfected group.Expression of VEGF_(165) could be detected 4 weeks after implantation and many capillaries were observed in the transfected MSCs material group.However,there was no bone production in the control group.Conclusion:MSCs transfceted with VEGF_(165) gene has good osteogenesis and angiogenesis potentiality composed with deprotienized bone implantation.