Alkaline extracellular conditions promote the proliferation and mineralization of a human cementoblast cell line

Abstract

Aim

To investigate the proliferation and mineralization of a human cementoblast cell line under alkaline conditions.

Methodology

A human cementoblast cell line was cultured in alkaline media with several pHs (pH 7.6, 8.0 and 8.4) without CO₂. Cell numbers, phospho‐p44/42 expression, alkaline phosphatase (ALP) activity, and mineralization were evaluated. The significance of differences between groups was assessed using a two‐way analysis of variance 15 (ANOVA) followed by Bonferroni's multiple comparison test (α=0.01).

Results

Cell numbers increased in a time‐dependent manner in the high pH medium groups. Western blot analysis revealed the up‐regulated expression of phospho‐p44/42 under alkaline conditions. ALP activity was also increased at pH 8.0 and 8.4. Alizarin red staining showed increased mineralization in the high pH medium groups. The incorporation of the transient receptor potential ankyrin subfamily member 1 (TRPA1) antagonist HC030031 markedly negated the effect on proliferation and mineralization.

Conclusions

Extracellular alkaline conditions promoted the proliferation and mineralization of human cementoblasts in vitro via TRPA1.

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