Zhimning Zhang, Xue Lü, Jian Hu, Jian Wang, En Luo
Hua Xi Kou Qiang Yi Xue Za Zhi. 2014 Oct;32(5):488-92. doi: 10.7518/hxkq.2014.05.015.
We aimed to construct a recombinant adenovirus Ad-APN-EGFP and to evaluate the transfection efficiency and effect of the adenovirus promoting osteogenesis around implants.
Recombinant adenovirus Ad-APN-EGFP containing adiponectin (APN) was constructed by DNA sequencing and restriction enzyme digestion. Rat bone marrow stem cells (BMSCs) were transfected in vitro. Transfection efficiency and APN mRNA expression were tested. Animal models of rat femoral epiphysis and hydroxyapatite (HA)-coated implants were established. The Ad-APN-EGFP adenovirus at 10 μL was injected into the defect around HA-coated implants in the experimental group, whereas the same volume of phosphate-buffered saline was injected into the defect in the control group. Osteogenesis around implants was observed by hematoxylin-eosin staining four weeks after implantation.
Ad-APN-EGFP was successfully constructed by DNA sequencing and restriction enzyme digestion. The transfection efficiency of Ad-APN-EGFP to BMSCs was up to ≥ 90%. APN mRNA expression of BMSCs transfected with Ad-APN-EGFP was higher than that of the control group. Osteogenesis in the experimental group was more evident than that in the control group in vivo.
Ad-APN-EGFP could be transfected into BMSCs and express APN mRNA at a high level. Ad-APN-EGFP could improve osteogenesis around implants in vivo.
构建重组腺病毒Ad-APN-EGFP,并评估该腺病毒对种植体周围成骨的转染效率及促进作用。
通过DNA测序和限制性内切酶消化构建含脂联素(APN)的重组腺病毒Ad-APN-EGFP。体外转染大鼠骨髓间充质干细胞(BMSCs),检测转染效率及APN mRNA表达。建立大鼠股骨骨骺和羟基磷灰石(HA)涂层种植体的动物模型。实验组将10 μL Ad-APN-EGFP腺病毒注入HA涂层种植体周围的缺损处,而对照组在缺损处注入相同体积的磷酸盐缓冲盐水。植入4周后通过苏木精-伊红染色观察种植体周围的成骨情况。
通过DNA测序和限制性内切酶消化成功构建Ad-APN-EGFP。Ad-APN-EGFP对BMSCs的转染效率高达≥90%。转染Ad-APN-EGFP的BMSCs的APN mRNA表达高于对照组。实验组在体内的成骨比对照组更明显。
Ad-APN-EGFP可转染至BMSCs并高水平表达APN mRNA。Ad-APN-EGFP可改善体内种植体周围的成骨情况。