Research on promoting periodontal regeneration with human basic fibroblast growth factor-modified bone marrow mesenchymal stromal cell gene therapy

被引:44
作者
Tan, Zhen [1 ]
Zhao, Qing [2 ]
Gong, Ping [1 ,3 ]
Wu, Yang [4 ]
Wei, Na [1 ]
Yuan, Quan [1 ]
Wang, Chuan [5 ]
Liao, Dapeng [1 ]
Tang, Hua [1 ]
机构
[1] Sichuan Univ, Oral Implant Ctr, W China Coll Stomatol, Chengdu 610041, Sichuan, Peoples R China
[2] Sichuan Univ, W China Coll Stomatol, Dept Orthodont, Chengdu 610041, Sichuan, Peoples R China
[3] Sichuan Univ, State Key Lab Oral Dis, Chengdu 610041, Sichuan, Peoples R China
[4] Sichuan Univ, State Key Lab Biotherapy Human Dis, Chengdu 610041, Sichuan, Peoples R China
[5] Sichuan Univ, W China Sch Publ Hlth, Dept Med Technol, Chengdu 610041, Sichuan, Peoples R China
基金
美国国家科学基金会;
关键词
Basic fibroblast growth factor; bone marrow mesenchymal stromal cells; gene therapy; periodontal regeneration; STEM-CELLS; IN-VITRO; TISSUE REGENERATION; REPAIR; DIFFERENTIATION; DEFECTS; DOGS; ENHANCEMENT; DELIVERY; TENDON;
D O I
10.1080/14653240902824757
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Background aims Recently, it has been found that effective periodontal regeneration can be induced by bone marrow mesenchymal stromal cell (BMSC) transplantation or local application of basic fibroblast growth factor (bFGF). The aim of the present study was to assess, in dogs, the efficacy of periodontal regeneration via the delivery of BMSC transfected with bFGF to repair destruction of periodontal tissue. Methods BMSC from dogs were isolated, cultured and purified via density-gradient centrifugation. Polymerase chain reaction (PCR) was employed to clone bFGF cDNA from human periodontal cells, and the product was then ligated into the eukaryotic expression vector pDC316-IREs-EGFP. BMSC transfected with pDC316bFGF-IREs-EGFP were transplanted into root furcation defects of beagle dogs. After 6 weeks, regeneration in defects was assessed via clinical examination, X-ray, histologic observation and micro-CT analysis. Results DNA sequence analysis showed that the bFGF sequence of recombinant plasmid pDC316bFGF-IREs-EGFP was consistent with that reported by GeneBank. bFGF expression was detected with Western blotting, and active bFGF in supernatant was also observed. Our animal experiment proved that the regenerating speed of periodontal bone tissue in groups transplanted with BMSC containing the modified bFGF gene was higher than in those transplanted with BMSC alone. Conclusions A successfully constructed eukaryotic expression vector containing human bFGF in pDC316bFGF-IREs-EGFP could produce bioactive bFGF in vitro. bFGF overexpression mediated by the recombinant plasmid pDC316bFGF- IREs-EGFP accelerated periodontal regeneration.
引用
收藏
页码:317 / 325
页数:9
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