Research on promoting periodontal regeneration with human basic fibroblast growth factor-modified bone marrow mesenchymal stromal cell gene therapy.
OBJECTIVE
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.
Tan
Zhen
Z
Oral Implant Center, West China College of Stomatology, Sichuan University, Chengdu, China.
Zhao
Qing
Q
Gong
Ping
P
Wu
Yang
Y
Wei
Na
N
Yuan
Quan
Q
Wang
Chuan
C
Liao
Dapeng
D
Tang
Hua
H
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism*; D017690:Cell Transplantation; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004285:Dogs; D016222:Fibroblast Growth Factor 2 / Q000235:genetics / Q000378:metabolism*; D017823:Furcation Defects / Q000473:pathology / Q000503:physiopathology / Q000628:therapy*; D015316:Genetic Therapy; D016557:Guided Tissue Regeneration, Periodontal; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism*; D014162:Transfection; D019076:Transgenes
D016222:Fibroblast Growth Factor 2