Department of Clinical Sciences - Restorative, Faculty of Dentistry, University of Toronto, Toronto, ON M5G 1G6, Canada.
Biomaterials and Biotechnology and Innovation in Health Programs, Anhanguera University of Sao Paulo, Brazil.
Dent Mater. 2018 Jun;34(6):e107-e114. doi: 10.1016/j.dental.2018.01.025. Epub 2018 Feb 7.
To screen the effect of two compounds, chlorhexidine diacetate (CHX) and epigallocatechin-gallate (EGCG), on the levels of cytokines produced by odontoblast-like cells (MDPC-23).
Cells were seeded at 24h and 48h with serial dilution of the compounds to determine cell metabolic activity by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (n=3). Cells with no compound treatment were used as control (Ctr). For the highest equal non-cytotoxic compound dilution tested at 48h cell treatment, total protein concentration was measured using a Pierce bicinchoninic acid (BCA) assay (n=3), and expression of 23 cytokines was analyzed using the Bio-Plex cytokine assay (n=2). Data were analyzed by one-way ANOVA and Tukey's test (α=5%).
The MTT assay revealed that at 24h and 48h, CHX and EGCG did not reduce cell metabolic activity at concentrations of 2.5-20μM (CHX) and 2.5-160μM (EGCG), respectively (p>0.05). At 48h, total protein levels were consistent across all groups for 20μM compound dilution (Ctr: 1.04mg/mL; CHX: 0.98mg/mL; and EGCG: 1.06mg/mL). At 20μM dilution, both CHX and EGCG significantly increased the secretion of IL-1β, IL-10, IL-12, KC, MIP-1α, IFN-γ and IL-6 (p<0.05). Treatment with CHX significantly increased secretion of IL-4 and RANTES (p<0.05).
with EGCG significantly increased Eotaxin secretion (p<0.05). Both CHX and EGCG significantly decreased secretion of IL-17 (p<0.05). GM-CSF and TNF-α did not present significant change in secretion after treatment with either CHX or EGCG (p>0.05).
Both CHX and EGCG modulate secretion of various inflammatory and anti-inflammatory mediators in odontoblastic cells.
筛选两种化合物——醋酸洗必泰（CHX）和表没食子儿茶素没食子酸酯（EGCG）对成牙本质样细胞（MDPC-23）产生细胞因子水平的影响。
将细胞在 24 小时和 48 小时分别用化合物的系列稀释液播种，通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴化物（MTT）测定法（n=3）测定细胞代谢活性。无化合物处理的细胞用作对照（Ctr）。在 48 小时细胞处理时，用最高等非细胞毒性化合物稀释度进行测试，使用 Pierce 双缩脲（BCA）测定法（n=3）测量总蛋白浓度，并使用 Bio-Plex 细胞因子测定法（n=2）分析 23 种细胞因子的表达。通过单因素方差分析和 Tukey 检验（α=5%）进行数据分析。
MTT 测定法显示，在 24 小时和 48 小时时，CHX 和 EGCG 在 2.5-20μM（CHX）和 2.5-160μM（EGCG）浓度下均未降低细胞代谢活性（p>0.05）。在 48 小时时，对于 20μM 化合物稀释度，所有组的总蛋白水平均一致（Ctr：1.04mg/mL；CHX：0.98mg/mL；和 EGCG：1.06mg/mL）。在 20μM 稀释度下，CHX 和 EGCG 均显著增加了 IL-1β、IL-10、IL-12、KC、MIP-1α、IFN-γ 和 IL-6 的分泌（p<0.05）。CHX 处理显著增加了 IL-4 和 RANTES 的分泌（p<0.05）。EGCG 处理显著增加了 Eotaxin 的分泌（p<0.05）。CHX 和 EGCG 均显著降低了 IL-17 的分泌（p<0.05）。GM-CSF 和 TNF-α 在用 CHX 或 EGCG 处理后其分泌均未发生显著变化（p>0.05）。
CHX 和 EGCG 均可调节成牙本质细胞中各种炎症和抗炎介质的分泌。
