Department and Research Institute of Dental Biomaterials and Bioengineering, BK21 PLUS Project, College of Dentistry, Yonsei University, Seoul, Republic of Korea; Department of Biomaterials Science, College of Dentistry, Dankook University, Cheonan, Republic of Korea; Institute of Tissue Regeneration Engineering (ITREN), Dankook University, Cheonan, Republic of Korea.
Department of Biomaterials Science, College of Dentistry, Dankook University, Cheonan, Republic of Korea; Institute of Tissue Regeneration Engineering (ITREN), Dankook University, Cheonan, Republic of Korea.
Dent Mater. 2016 May;32(5):e93-104. doi: 10.1016/j.dental.2016.01.003. Epub 2016 Mar 5.
The objective of this study is to assess the cytotoxic and anti-inflammatory effects of ZOE cement during setting in two-dimensional (2D) or three-dimensional (3D) cultures of immortalized human oral keratinocytes (IHOKs) with determining the extract components responsible for these effects.
Extracts of mixed ZOE at different stages of setting were analyzed by a digital pH meter, ICP-MS, and GC-MS. Serial concentrations of extract and their mixture of ZnCl2, ZnSO4·H2O, and eugenol liquid were added to the 2D and 3D IHOK cultures to determine the half maximal effective concentration in investigating the cause of cytotoxicity by means of WST assay and to investigate mRNA expression levels of inflammatory cytokines by RT-PCR.
Zn(2+) and eugenol (4-19 ppm) were detected in the extracts. In the early setting stage, significant cytotoxicity was observed in the 2D and 3D IHOK cultures (P<0.05). The EC50 of Zn(2+) from ZnCl2 was 5-44 ppm in both cultures, whereas the EC50 of eugenol was not detectable under 100 ppm. Along with the lower levels of inflammatory cytokine gene expressions in the extract, treatment of the 2D IHOKs with Zn(2+) alone and treatment of the 3D IHOKs with Zn(2+) plus eugenol resulted in significantly lower expression levels of IL-1β, IL-6, and IL-8 (P<0.05).
The cytotoxic effect of ZOE on IHOKs was greater during the setting stage owing to the presence of Zn(2+). The anti-inflammatory response to ZOE was induced by a combination of Zn(2+) and eugenol. Cytotoxic and anti-inflammatory effects differed between the 2D and 3D IHOK cultures.
本研究旨在评估在二维（2D）或三维（3D）培养的永生化人口腔角质细胞（IHOKs）中，混合 ZOE 水泥在凝固过程中的细胞毒性和抗炎作用，并确定导致这些作用的提取成分。
使用数字 pH 计、ICP-MS 和 GC-MS 分析不同凝固阶段的 ZOE 提取物。将不同浓度的提取物及其 ZnCl2、ZnSO4·H2O 和丁香酚液体混合物添加到 2D 和 3D IHOK 培养物中，通过 WST 测定法确定半最大有效浓度，以调查细胞毒性的原因，并通过 RT-PCR 研究炎症细胞因子的 mRNA 表达水平。
在提取物中检测到 Zn(2+)和丁香酚（4-19ppm）。在早期凝固阶段，2D 和 3D IHOK 培养物中观察到明显的细胞毒性（P<0.05）。两种培养物中，来自 ZnCl2 的 Zn(2+)的 EC50 为 5-44ppm，而丁香酚的 EC50 在 100ppm 以下不可检测。随着提取物中炎症细胞因子基因表达水平的降低，单独用 Zn(2+)处理 2D IHOKs 以及用 Zn(2+)和丁香酚处理 3D IHOKs 导致 IL-1β、IL-6 和 IL-8 的表达水平显著降低（P<0.05）。
由于存在 Zn(2+)，ZOE 对 IHOKs 的细胞毒性作用在凝固阶段更大。ZOE 的抗炎反应是由 Zn(2+)和丁香酚的组合引起的。2D 和 3D IHOK 培养物的细胞毒性和抗炎作用不同。
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