Department of Operative Dentistry, Cariology and Pulp Biology, Tokyo Dental College, Tokyo, Japan.
Department of Endodontics, Tokyo Dental College, Tokyo, Japan.
Int Endod J. 2019 May;52(5):639-645. doi: 10.1111/iej.13044. Epub 2018 Dec 11.
To investigate the proliferation and mineralization of a human cementoblast cell line under alkaline conditions.
A human cementoblast cell line was cultured in alkaline media with several pHs (pH 7.6, 8.0 and 8.4) without CO . 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).
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.
Extracellular alkaline conditions promoted the proliferation and mineralization of human cementoblasts in vitro via TRPA1.
研究碱性条件下人成牙骨质细胞系的增殖和矿化情况。
将人成牙骨质细胞系在无 CO 的碱性培养基中于不同 pH 值(7.6、8.0 和 8.4)下培养。评估细胞数量、磷酸化 p44/42 表达、碱性磷酸酶(ALP)活性和矿化情况。使用双因素方差分析 15(ANOVA),随后进行 Bonferroni 多重比较检验(α=0.01)评估组间差异的显著性。
高 pH 培养基组的细胞数量随时间呈依赖性增加。Western blot 分析显示碱性条件下磷酸化 p44/42 的表达上调。ALP 活性在 pH 8.0 和 8.4 时也增加。茜素红染色显示高 pH 培养基组的矿化增加。瞬时受体电位阳离子通道亚家族成员 1(TRPA1)拮抗剂 HC030031 的加入显著否定了对增殖和矿化的影响。
细胞外碱性条件通过 TRPA1 促进人成牙骨质细胞的增殖和矿化。