China Journal of Oral and Maxillofacial Surgery ›› 2017, Vol. 15 ›› Issue (3): 202-208.doi: 10.19438/j.cjoms.2017.03.003

• Original Articles • Previous Articles     Next Articles

Repair of critical sized mandibular bone defect with nHAC / PLA scaffold combing with rabbit adipose tissue derived mesenchymal stem cells transfected with BMP-9 / EPO gene

ZHANG Guang-de1, JIN Xia1, LI Rong-liang1, YANG Shi-mao2, GUO Yan-wei1, FANG Dian-ji2.   

  1. 1.Department of Oral and Maxillofacial Surgery, Jining Hospital of Stomatology. Jining 272000, Shandong Provence;
    2.Department of Oral and Maxillofacial Surgery, Jinan Hospital of Stomatology. Jinan 250000, Shandong Provence;
    3.Department of Oral Surgery,Dental Institute of Minhang District. Shanghai 201100, China
  • Received:2016-09-29 Revised:2016-12-17 Online:2017-05-20 Published:2017-06-09

Abstract: PURPOSE: To investigate the effect of recombinant adenovirus-mediated bone morphogenetic protein-9 (BMP-9) and erythropoietin (EPO) genes co-transfection combined with scaffold materials of nHAC/PLA on osteogenesis in vivo. METHODS: ADSCs were isolated with enzyme digestion and adherence method. The recombinant adenovirus-mediated bone morphogenetic protein-9 (BMP-9), erythropoietin (EPO), and BMP-9/EPO genes co-transfection were constructed respectively, and then ADSCs were transfected with recombinant adenovirus. The expression of cell fluorescence was observed under a fluorescence microscope at 14 days, and the transfection efficiency was calculated. Western blot was used to detect the expressions of BMP-9 and EPO proteins at 14 days. 50 New Zealand rabbits were selected, and bone defect was made in bilateral mandibles respectively, and then randomly divided into 5 groups. ADSCs/nHAC/PLA in each group was transplanted into the bilateral bone defects. Rabbits of each group were killed at 4, 8 and 12 weeks after operation respectively, and the healing of the defects was detected by X-ray, histological and scanning electron microscopy (SEM) examination. Statistical analysis was performed using SPSS 17.0 software package. RESULTS: Fluorescence microscope showed that BMP-9 and EPO, BMP-9/EPO recombinant adenovirus could stably transfected ADSCs, with a transfection efficiency of 80%-93%. Gene and protein of BMP-9, EPO as well as BMP-9/EPO in ADSCs could be stably expressed. Osteogenesis in BMP-9/EPO/nHAC/PLA group was significantly better than other groups. SEM indicated that the interface between materials and bone defect was combined closely in BMP-9/EPO/nHAC/PLA group. CONCLUSIONS: Recombinant adenovirus-mediated BMP-9, EPO and BMP-9/EPO genes can transfect ADSCs, which can stably express in ADSCs. ADSCs transfected with recombinant adenovirus-mediated BMP-9/EPO gene combined with nHAC/PLA could improve bone generation in mandibular defect, and provide a possible option for bone tissue engineering.

Key words: ADSCs, Adenovirus, BMP-9, EPO, Osteogenic differentiation

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