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

• Published in: International endodontic journal Vol. 52; no. 5; pp. 639 - 645
• Main Authors: Muramatsu, T., Kashiwagi, S., Ishizuka, H., Matsuura, Y., Furusawa, M., Kimura, M., Shibukawa, Y.
• Format: Journal Article
• Language: English
• Published: England Wiley Subscription Services, Inc 01.05.2019
• Subjects: Alkalies; alkaline conditions; Alkaline phosphatase; Ankyrins; calcium; Carbon dioxide; cementoblast; Dentistry; Endodontics; Mineralization; pH effects; Proliferation; Transient receptor potential proteins; TRPA1; Variance analysis; cementoblast; mineralization; calcium; Proliferation; TRPA1; alkaline conditions
• ISSN: 0143-2885; 1365-2591
• DOI: 10.1111/iej.13044

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 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 upregulated expression of phospho‐p44/42 under alkaline conditions. ALP activity was also increased at pH 8.0 and 8.4. Alizarin red staining revealed 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.