West China Journal of Stomatology

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Recombinant adenovirus Ad-APN-EGFP constructing and promoting osteogenesis around implants

Zhang Zhiming1, Lü Xue2, Hu Jian3, Wang Jian2, Luo En2.   

  1. 1. Dept. of Prosthodontics, Stomatology Hospital of Shaoxing, Shaoxing 312000, China; 2. State Key Laboratory of Oral Diseases, Dept. of Oral and Maxillofacial Surgery, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China; 3. Dept. of Prosthodontics, Hospital of Stomatology, Wuhan University, Wuhan 430072, China
  • Online:2014-10-01 Published:2014-10-01

Abstract:

Objective 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. Methods Recombinant adenovirus Ad-APNEGFP 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. Results 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. Conclusion 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.

Key words: adiponectin, bone marrow stem cells, osteogenesis, gene transfection, implant