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Effect of transforming growth factor‐beta1 on expression of the connective tissue growth factor ( CCN2/CTGF ) gene in normal human gingival fibroblasts and periodontal ligament cells
Author(s) -
Takeuchi H.,
Kubota S.,
Murakashi E.,
Fukada T.,
Hashimoto S.,
Takigawa M.,
Numabe Y.
Publication year - 2009
Publication title -
journal of periodontal research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.31
H-Index - 83
eISSN - 1600-0765
pISSN - 0022-3484
DOI - 10.1111/j.1600-0765.2008.01093.x
Subject(s) - ctgf , periodontal fiber , connective tissue , growth factor , fetal bovine serum , extracellular matrix , chemistry , transforming growth factor , biology , microbiology and biotechnology , pathology , medicine , cell , dentistry , biochemistry , receptor
Background and Objective: Connective tissue growth factor (CCN2/CTGF) plays an important role in wound healing and regulation of the extracellular matrix in periodontal tissue. However, the functional relationship between altered transforming growth factor‐beta1 levels and CCN2/CTGF has not been extensively investigated in human gingival fibroblasts and periodontal ligament cells. This study investigated the effects of transforming growth factor‐beta1 on the expression of the CCN2/CTGF gene in human gingival fibroblasts and periodontal ligament cells in vitro . Material and Methods: Cells were isolated from normal periodontal tissues and cultured in Dulbecco’s modified Eagle’s minimal essential medium/F12 containing 10% fetal bovine serum. Subconfluent cells were maintained under serum deprivation for 24 h then treated with Dulbecco’s modified Eagle’s minimal essential medium/F12 containing 0.5% fetal bovine serum (control) and 0.1, 1, 5 or 10 ng/mL of transforming growth factor‐beta1 for 24, 48 or 72 h. The effects of transforming growth factor‐beta1 on CCN2/CTGF mRNA expression were measured by reverse transcription–polymerase chain reaction. CCN2/CTGF protein was quantitatively analyzed using enzyme‐liked immunosorbent assay. Subcellular distribution of CCN2/CTGF protein in both human gingival fibroblasts and periodontal ligament cells was observed using immunofluorescence microscopy. Results: In both human gingival fibroblasts and periodontal ligament cells, the expression of CCN2/CTGF mRNA and CCN2/CTGF protein was significantly increased, in a dose‐ and time‐dependent manner, in the presence of transforming growth factor‐beta1. Moreover, immunofluorescence analysis indicated that immunoreactivity to CCN2/CTGF showed a granular pattern of protein localization. Conclusion: The expression of CCN2/CTGF mRNA and protein was induced by transforming growth factor‐beta1 in human gingival fibroblasts and periodontal ligament cells. These results suggest that CCN2/CTGF plays an important role in wound healing and in the regeneration of periodontal tissue.