Wang Hong-Jun, Cai Bo, Zhao Xing-Yu, Li Shu-Qiang, Feng Wei, Liu Jian-Guo, Li Dong-Song
Department of Bone and Joint, No.1 Hospital of Jilin University, Changchun 130021, Jilin, China;
Zhongguo Gu Shang. 2017 Jun 25;30(6):545-551. doi: 10.3969/j.issn.1003-0034.2017.06.012.
To explore repairing results of VEGF165 gene modified adipose-derived stem cells for diabetic rats with bone defect.
Seventy-eight male Wistar rats weighted 180 to 220 g were selected, 72 rats were established diabetic animal models by streptozotocin inducement method, blood glucose level was more than 16.7 mmol/L. Experimental animals were randomly divided into 5 groups, 6 rats in normal group and each 18 rats in other groups. VEGF165 gene modified adipose-derived stem cells were implanted into normal group with bone defect; single diabetic rats with bone defect were named as diabetic group;vascular endothelial growth factor implanted into single diabetic rats with bone defect named as growth factor group; adipose-derived stem cells implanted into diabetic rats with bone defect names as stem cell group; VEGF165 gene modified adipose-derived stem cells implanted diabetic rats with bone defect named as experimental group. After combination of VEGF165-ADSCs (5×106) cells combined with gel sponge, implanted into diabetic rats with bone defect. On the forth week, general form of defect repairing tissue were observed by optical microscopy;local density of micro-vessel were detected by immunohistochemistry method; content of Ca, P and ALP of repairing callus were detected by IRIS Intrepid XSP inductively coupled plasma emission spectrometer. Efficacy of the VEGF165-ADSCs repairing function was evaluated by SPSS statistic software.
Fluorescent staining results showed that expression of VEGF165 located on cytoplasm of ADSCs, expression percentage was more than 87%; general histology results showed that callus formation and quality was near to normal group, repairing results in diabetes group, growth factor group and stem cell group were poor. On the Forth week after implantation, content of Ca, P and ALP of repairing callus in experimental group were higher than those in growth group and stem cell group, and without significant differences compared with normal group; blood vessel density in experimental group was lower than normal group, but higher than other groups.
VEGF165 gene modified adipose-derived stem cells for repairing diabetic rats with bone defect has advantages of osteogenesis and angiogenesis, and should be one of the effective method for repairing diabetic rats with bone defect.
探讨血管内皮生长因子165（VEGF165）基因修饰的脂肪干细胞对糖尿病大鼠骨缺损的修复效果。
选取体重180～220 g的雄性Wistar大鼠78只，采用链脲佐菌素诱导法将72只大鼠制成糖尿病动物模型，血糖水平高于16.7 mmol/L。将实验动物随机分为5组，正常组6只，其余各组18只。将VEGF165基因修饰的脂肪干细胞植入有骨缺损的正常组大鼠；将单纯有骨缺损的糖尿病大鼠设为糖尿病组；将血管内皮生长因子植入单纯有骨缺损的糖尿病大鼠设为生长因子组；将脂肪干细胞植入有骨缺损的糖尿病大鼠设为干细胞组；将VEGF165基因修饰的脂肪干细胞植入有骨缺损的糖尿病大鼠设为实验组。将VEGF165 - ADSCs（5×10⁶）细胞与明胶海绵混合后，植入有骨缺损的糖尿病大鼠。第4周时，通过光学显微镜观察缺损修复组织的大体形态；采用免疫组织化学方法检测局部微血管密度；用IRIS Intrepid XSP电感耦合等离子体发射光谱仪检测修复骨痂中钙、磷和碱性磷酸酶的含量。采用SPSS统计软件评估VEGF165 - ADSCs的修复功能疗效。
荧光染色结果显示，VEGF165在脂肪干细胞的细胞质中表达，表达率大于87%；大体组织学结果显示，实验组骨痂形成及质量接近正常组，糖尿病组、生长因子组和干细胞组的修复效果较差。植入后第4周，实验组修复骨痂中钙、磷和碱性磷酸酶的含量高于生长因子组和干细胞组，与正常组相比无显著差异；实验组血管密度低于正常组，但高于其他组。
VEGF165基因修饰的脂肪干细胞修复糖尿病大鼠骨缺损具有成骨和血管生成优势，应是修复糖尿病大鼠骨缺损的有效方法之一。
