Differential effects of various growth factors and cytokines on the syntheses of DNA, type I collagen, laminin, fibronectin, osteonectin/secreted protein, acidic and rich in cysteine (SPARC), and alkaline phosphatase by human pulp cells in culture.
The purpose of this study is to differentiate roles of several growth factors and cytokines in proliferation and differentiation of pulp cells during development and repair. In human pulp cell cultures, laminin and type I collagen levels per cell remained almost constant during the whole culture period (22 days). On the other hand, secreted protein, acidic and rich in cysteine (SPARC/osteonectin) and alkaline phosphatase (ALPase) levels markedly increased after the cultures reached confluence. Laminin and type I collagen, as well as fibronectin, stimulated the spreading of pulp cells within 1 h. Adding transforming growth factor-beta (TGF-beta) decreased laminin and ALPase levels, whereas it increased SPARC and fibronectin levels 3- to 10-fold. Western and Northern blots showed that TGF-beta enhanced SPARC synthesis at the protein and mRNA levels. Basic fibroblast growth factor (bFGF) decreased type I collagen, laminin, SPARC, and ALPase levels without changing the fibronectin level. Platelet-derived growth factor (PDGF) selectively decreased laminin, SPARC, and ALPase levels. Epidermal growth factor (EGF) also decreased SPARC and ALPase levels. Tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) decreased type I collagen and laminin levels, and abolished SPARC and ALPase syntheses. Of these peptides, bFGF and PDGF showed the greatest stimulation of [3H]thymidine incorporation into DNA. TGF-beta, EGF, and TNF-alpha had less effect on DNA synthesis, whereas IL-1beta inhibited DNA synthesis. These findings demonstrated that TGF-beta, bFGF, EGF, PDGF, TNF-alpha, and IL-1beta have characteristically different patterns of actions on DNA, laminin, type I collagen, fibronectin, ALPase, and SPARC syntheses by pulp cells.
Shiba
H
H
Department of Endodontology and Periodontology, Hiroshima University School of Dentistry, Japan.
Fujita
T
T
Doi
N
N
Nakamura
S
S
Nakanishi
K
K
Takemoto
T
T
Hino
T
T
Noshiro
M
M
Kawamoto
T
T
Kurihara
H
H
Kato
Y
Y
D000469:Alkaline Phosphatase / Q000096:biosynthesis*; D015152:Blotting, Northern; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis*; D016207:Cytokines / Q000494:pharmacology*; D004247:DNA / Q000096:biosynthesis*; D003782:Dental Pulp / Q000378:metabolism*; D005353:Fibronectins / Q000096:biosynthesis*; D006133:Growth Substances / Q000494:pharmacology*; D006801:Humans; D007797:Laminin / Q000096:biosynthesis*; D015676:Osteonectin / Q000096:biosynthesis*
D016207:Cytokines; D005353:Fibronectins; D006133:Growth Substances; D007797:Laminin; D015676:Osteonectin; D003094:Collagen; D004247:DNA; D000469:Alkaline Phosphatase