Objective

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).

Methods

Cells were seeded at 24 h and 48 h 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 48 h 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%).

Results

The MTT assay revealed that at 24 h and 48 h, CHX and EGCG did not reduce cell metabolic activity at concentrations of 2.5–20 μM (CHX) and 2.5–160 μM (EGCG), respectively (p > 0.05). At 48 h, total protein levels were consistent across all groups for 20 μM compound dilution (Ctr: 1.04 mg/mL; CHX: 0.98 mg/mL; and EGCG: 1.06 mg/mL). At 20 μM 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). Treatment: 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).

Significance

Both CHX and EGCG modulate secretion of various inflammatory and anti-inflammatory mediators in odontoblastic cells.

中文翻译：

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洗必泰和EGCG处理的成牙本质细胞样细胞中的细胞因子水平分析

客观的

为了筛选两种化合物（双乙酸洗必泰（CHX）和表没食子儿茶素没食子酸酯（EGCG））对成牙本质细胞样细胞（MDPC-23）产生的细胞因子水平的影响。

方法

用化合物的连续稀释液在24小时和48小时接种细胞，以通过3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四唑溴化物（MTT）测定法测定细胞代谢活性（n  = 3） 。未经化合物处理的细胞用作对照（Ctr）。对于在48 h细胞处理后测试的最高相等的非细胞毒性化合物稀释度，使用皮尔斯二辛可宁酸（BCA）分析法（n  = 3）测量总蛋白浓度，并使用Bio-Plex细胞因子分析法分析23种细胞因子的表达。 （n  = 2）。数据通过单因素方差分析和Tukey检验（α  = 5％）进行分析。

结果

MTT分析表明，在浓度分别为2.5–20μM（CHX）和2.5–160μM（EGCG）的浓度下，CHX和EGCG在24 h和48 h不会降低细胞代谢活性（p> 0.05）。在第48小时，对于20μM化合物稀释液（Ctr：1.04 mg / mL； CHX：0.98 mg / mL； EGCG：1.06 mg / mL），所有组的总蛋白水平均保持一致。在稀释20μM时，CHX和EGCG均显着增加IL-1β，IL-10，IL-12，KC，MIP-1α，IFN-γ和IL-6的分泌（p <0.05）。CHX处理可显着增加IL-4和RANTES的分泌（p <0.05）。治疗：用EGCG明显增加了趋化因子分泌（p <0.05）。CHX和EGCG均显着降低IL-17的分泌（p <0.05）。用CHX或EGCG治疗后，GM-CSF和TNF-α的分泌没有显着变化（p> 0.05）。

意义

CHX和EGCG都可以调节牙本质细胞中各种炎症和抗炎介质的分泌。