Effects of adenoviral-mediated coexpression of bone morphogenetic protein-7 and insulin-like growth factor-1 on human periodontal ligament cells.
OBJECTIVE
Bone morphogenetic protein-7 (BMP-7) and insulin-like growth factor-1 (IGF-1) are important in periodontal reconstruction. However, their synergistic effect in periodontal regeneration by gene delivery has not been reported. In this study, gene delivery of these two growth factors to human periodontal ligament cells (hPDLCs) was examined for its effects on cell proliferation and differentiation.


METHODS
Recombinant adenoviruses containing both human BMP-7 and IGF-1 cDNA created by introducing the internal ribosome entry site (IRES) sequence were used to transfer the genes into hPDLCs. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and cell cycle analysis were used to observe their effects on cell proliferation, while alkaline phosphatase activity measurement, RT-PCR and in vivo tests were conducted to investigate their effects on cell differentiation.


RESULTS
The proliferation of hPDLCs transduced by adenoviruses coexpressing BMP-7 and IGF-1 was suppressed while their differentiation ability was enhanced. There was a synergism of BMP-7 and IGF-1 in up-regulating alkaline phosphatase activity and mRNA levels of collagen type I and Runx2. Implantation in vivo with scaffolds illustrated that the transduced cells exhibited osteogenic differentiation and formed bone-like structures.


CONCLUSIONS
The combined delivery of BMP-7 and IGF-1 genes using an IRES-based strategy synergistically enhanced differentiation of hPDLCs. It is suggested that this could be a new potential method in gene therapy for periodontal reconstruction.
Yang
L
L
Key Laboratory for Oral Biomedical Engineering, Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, PR China.
Zhang
Y
Y
Dong
R
R
Peng
L
L
Liu
X
X
Wang
Y
Y
Cheng
X
X
D000256:Adenoviridae / Q000235:genetics*; D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D055419:Bone Morphogenetic Protein 7 / Q000235:genetics*; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics; D049109:Cell Proliferation; D018920:Coculture Techniques; D024042:Collagen Type I / Q000032:analysis; D004396:Coloring Agents; D050659:Core Binding Factor Alpha 1 Subunit / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics; D018014:Gene Transfer Techniques; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000235:genetics*; D051379:Mice; D010012:Osteogenesis / Q000502:physiology; D010513:Periodontal Ligament / Q000166:cytology*; D010957:Plasmids / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D040521:Subcutaneous Tissue / Q000473:pathology; D013778:Tetrazolium Salts; D013844:Thiazoles; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D014162:Transfection; D019076:Transgenes / Q000235:genetics; D015854:Up-Regulation
C525819:BMP7 protein, human; D015415:Biomarkers; D055419:Bone Morphogenetic Protein 7; D024042:Collagen Type I; D004396:Coloring Agents; D050659:Core Binding Factor Alpha 1 Subunit; C493709:RUNX2 protein, human; D013778:Tetrazolium Salts; D013844:Thiazoles; D007334:Insulin-Like Growth Factor I; D000469:Alkaline Phosphatase; C022616:thiazolyl blue