腺病毒介导骨形态发生蛋白2基因对兔软骨细胞的影响

Effect of adenovirus-mediated bone morphogenetic protein-2 genes on rabbit chondrocytes

背景:研究证实骨形态发生蛋白2具有促进软骨细胞增殖、分化的能力。目的:进一步验证由腺病毒介导的骨形态发生蛋白2对兔软骨细胞增殖的作用。设计、时间及地点:对比观察,实验于2006-01/2007-12在苏州大学附属第一医院骨科实验室完成。材料:5.0月龄健康的新西兰白兔4只,体质量3.0～4.0kg,雄雌不限。携带骨形态发生蛋白2基因的腺病毒载体,由苏州大学附属第一医院骨科实验室保存。方法:取兔耳郭的弹性软骨进行细胞培养,再以腺病毒为载体转染骨形态发生蛋白2到软骨细胞内作为实验组,未转染的作为对照组。主要观察指标:采用阿利新蓝染色、四甲基偶氮唑盐比色法和糖胺聚糖定量测定等方法观察软骨细胞增殖和分化。结果:实验组中的软骨细胞数量多,形态均一,融合率高,酸性黏多糖也较未转染组多。四甲基偶氮唑盐比色法和糖胺聚糖定量检测结果示实验组均高于未转染组。结论:腺病毒介导的骨形态发生蛋白2能促进软骨细胞增殖、保持软骨细胞的分化表型。

BACKGROUND： Previous Studies have shown that bone morphogenetic protein-2 （BMP-2） has the abilities to improve the proliferation and differentiation of chondrocytes. OBJECTIVE： To further verify the effect of adenovirus-mediated BMP-2 on proliferation of rabbit chondrocytes. DESIGN, TIME AND SETTING： A controlled observational experiment was performed at Orthopedic Laboratory, the First Affiliated Hospital of Soochow University from January 2006 to December 2007. MATERIALS： Four healthy New Zealand white rabbits of 5 months old, weighing 3-4 kg, any of gender, were selected in the experiment. Adenovirus vectors carrying human BMP-2 were preserved by Orthopedic Laboratory, the First Affiliated Hospital of Soochow University. METHODS： Elastic cartilages from rabbit auricles were used for chondrocyte culture. The chondrocytes transfected with adenovirus vectors carrying human BMP-2 gene were assigned as experimental group. The chondrocytes did not transfect were assigned as control group. MAIN OUTCOME MEASURES： Proliferation and differentiation of the chondrocytes were observed by alcian blue staining, MTT and quantitative determination of glycosaminoglycan. RESULTS： According to the alcian blue staining, the quantity of the chondrocytes in the experimental group was more than that in the control group. The chondrocytes in the experimental group had a similar morphology. Fusion rate and content of acid mucopolysaccharides in the experimental group were higher than that in the control group. The MTT and quantitative determination of glycosaminoglycan results were similar to the alcian blue staining. CONCLUSION： The adenovirus-mediated BMP-2 can improve the proliferation of chondrocytes, and maintain their differentiation phenotypes.