Effects of basic fibroblast growth factor on proliferation, the expression of osteonectin (SPARC) and alkaline phosphatase, and calcification in cultures of human pulp cells.
Basic fibroblast growth factor (bFGF) may be involved in the development and repair of dentine and pulp because bFGF, its related peptides, and FGF receptors are expressed in dental mesenchymal cells. In this study, we examined the effects of bFGF on DNA synthesis, osteonectin/SPARC levels, alkaline phosphatase (ALPase) activity, their mRNA levels, and calcium levels in cultures of human pulp cells. Pulp cells were isolated from three healthy upper wisdom teeth of three patients and maintained separately. These cells produced SPARC, ALPase, and calcified nodules and there was a close correlation between the SPARC-synthetic activity of the cell lines and their levels of ALPase and calcification. The levels of SPARC, ALPase and calcium deposits in the three pulp cell cultures were 10-250 times those of human foreskin fibroblasts. Western blots showed that the pulp cells produced 38-kDa SPARC. Northern blots showed that the pulp cells expressed flg (FGF receptor type 1) transcripts throughout all culture stages, irrespective of the presence or absence of bFGF. The addition of bFGF to the pulp cultures suppressed the increases in ALPase activity, SPARC synthesis, and their mRNA levels, although it increased the incorporation of [3H]thymidine into DNA > 10-fold. The effects of bFGF on ALPase activity and SPARC synthesis were reversible. Furthermore, bFGF abolished the calcification of the extracellular matrix; the calcium content of bFGF-free cultures. These findings suggest that bFGF is a potent mitogen for human pulp cells and that it inhibits the expression of the odontoblast phenotype by the cells at least partly at pretranslational levels.
Shiba
H
H
Department of Endodontology and Periodontology, Hiroshima University School of Dentistry, Japan.
Nakamura
S
S
Shirakawa
M
M
Nakanishi
K
K
Okamoto
H
H
Satakeda
H
H
Noshiro
M
M
Kamihagi
K
K
Katayama
M
M
Kato
Y
Y
D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D004247:DNA / Q000096:biosynthesis; D018076:DNA, Complementary; D003782:Dental Pulp / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D000091344:Filaggrin Proteins; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D009804:Odontoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015676:Osteonectin / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017468:Receptors, Fibroblast Growth Factor / Q000235:genetics
D018076:DNA, Complementary; C000717249:FLG protein, human; D000091344:Filaggrin Proteins; D015676:Osteonectin; D012333:RNA, Messenger; D017468:Receptors, Fibroblast Growth Factor; D016222:Fibroblast Growth Factor 2; D004247:DNA; D000469:Alkaline Phosphatase; D002118:Calcium