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Figure 1. Effects of PTHrP on TRAP⁺ cell formation in co-cultures of human PDL cells with mouse spleen cells. (A) Mouse spleen cells (1 x 10⁵ cells/well) were co-cultured with PDL cells (1 x 10⁵ cells/well) in the presence or absence of PTHrP (10^–8 M), TGF-β (10 ng/mL), or EGF (10 ng/mL) in the presence of Dex (10^–7 M) for 10 days. TRAP⁺ cells were counted and expressed as number of TRAP⁺ cells per culture well (values are mean ± SEM, n = 3). Similar results were obtained in three independent experiments. *P < 0.01 vs. control cultures. (B) Mouse spleen cells (1 x 10⁵ cells/well) were co-cultured with PDL cells (1 x 10⁵ cells/well) in various concentrations of PTHrP together with Dex for 10 days. TRAP⁺ cells were counted and expressed as the number of TRAP⁺ cells per culture well (values are mean ± SEM, n = 3). Similar results were obtained in three independent experiments. *P < 0.01 vs. control cultures. (C) Microscopic view of TRAP⁺ cells. Resorption areas were stained with Mayer’s hematoxylin (lower panels). Bar = 100 µm. (D) Human PDL cells were treated with PTHrP (10^–8 M) in the presence of Dex for the indicated time. Total RNA was isolated from PDL cells, and expression levels of RANKL, OPG, and GAPDH mRNA were measured by RT-PCR analysis. Numbers below the gels represent n-fold increase in intensity of RANKL or OPG relative to corresponding GAPDH mRNA signals. Similar results were obtained in 3 independent experiments. (E) Co-cultures were pre-treated with or without either OPG (100 ng/mL) or sRANK (100 ng/mL) for 30 min, followed by incubation with PTHrP (10^–8 M) in the presence of Dex for 10 days. TRAP⁺ cells were counted and expressed as number of TRAP⁺ cells per culture well (values are mean ± SEM, n = 3). Similar results were obtained in 3 independent experiments. *P < 0.01 vs. PTHrP-treated cultures.

J DENT RES, Vol. 84, No. 4, 329-334 (2005)
DOI: 10.1177/154405910508400407

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