This Article Abstract Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Sawa, Y. Articles by Yoshida, S. Search for Related Content PubMed PubMed Citation Articles by Sawa, Y. Articles by Yoshida, S. Social Bookmarking                         What's this?

Production of Colony-stimulating Factor in Human Dental Pulp Fibroblasts

Department of Oral Functional Science, Graduate School of Dental Medicine, Hokkaido University, N13 W7, Kita-ku, Sapporo 060-8586, Japan;

View larger version (121K): [in this window] [in a new window]   Figure 1. Immunostaining of dental pulp sections for CSFs. The reaction products are visualized by green fluorescence. (a,c,e,g) Serial sections from a normal tooth. (b,d,f,h) Serial sections from a tooth with dentinal caries. (a,b) H-E staining. (c,d) G-CSF. (e,f) M-CSF. (g,h) GM-CSF. In sections from the normal tooth, dental pulp fibroblasts (arrowheads) are stained by anti-G-CSF, but not by anti-M-CSF and anti-GM-CSF. In sections from the tooth with dentinal caries, dental pulp fibroblasts (arrowheads) are stained by anti-G-CSF and anti-M-CSF, especially strongly in the odontoblast layer (arrow), and by anti-GM-CSF. Bar = 100 µm.

View larger version (90K): [in this window] [in a new window]   Figure 2. Immunostaining of cultured dental pulp fibroblasts. The reaction products of anti-CSFs are visualized by green fluorescence, and of anti-Ki-67 by red fluorescence in the nuclei. (a1) G-CSF. (a2) G-CSF in IMR-90. (b) M-CSF. (c) M-CSF, with TNF-. (d) GM-CSF, with TNF-. Cells are stained by anti-G-CSF, while IMR-90 as a control was not stained. Cells are not stained by anti-M-CSF. Cells cultured with TNF- are stained by anti-M-CSF and anti-GM-CSF to a weaker extent than anti-G-CSF. Both cells in the cell-division cycle expressing Ki-67 in the nuclei and dormant cells not expressing Ki-67 are stained by anti-CSFs. There are accumulated proliferative cells (d, arrowhead). Bar = 100 µm.

View larger version (30K): [in this window] [in a new window]   Figure 3. (A) Immunoblot analysis of immunoprecipitated products with anti-CSFs. In whole-cell proteins of dental pulp fibroblasts cultured without TNF-, G-CSF (19 kDa) was detected, while M-CSF (85 kDa) and GM-CSF (22 kDa) were not detected. In whole-cell proteins of dental pulp fibroblasts cultured with TNF-, G-CSF and M-CSF were detected, while GM-CSF was not detected. M1, M2 = molecular-weight markers of 19.3, 86 kDa. (B) Expressions of G-CSF, M-CSF, and GM-CSF genes. (a) Dental pulp fibroblasts. (b) VEC. PCR products for G-CSF and M-CSF mRNAs were detected in VEC at an intensity similar to that in a control. GM-CSF mRNA was detected at an intensity lower than that in VEC, and also lower than in G-CSF and M-CSF mRNAs. PCR products for β-actin showed similar intensities in all samples.

View larger version (22K): [in this window] [in a new window]   Figure 4. Effect of TNF- on CSF production, examined by anti-CSFs binding activity to lysate of cells cultured in the absence (open bar) or presence (filled bar) of TNF-. Each bar, expressing absorbance change at 415 nm per min, represents the mean value (labeled) and standard deviation of 5 determinations. Controls are shown in the left margin (cont), indicating the non-specific IgG binding activity of second antibody. *Significantly different from control (p < 0.05). **Significantly different from control, and from cells without TNF- (p < 0.005).

Journal of Dental Research, Vol. 82, No. 2, 96-100 (2003)
DOI: 10.1177/154405910308200204


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?