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白细胞介素-6 和 -8 的过表达、2-羟乙基甲基丙烯酸酯处理后人牙髓间充质干细胞的细胞生长抑制和形态变化。

Overexpression of interleukin-6 and -8, cell growth inhibition and morphological changes in 2-hydroxyethyl methacrylate-treated human dental pulp mesenchymal stem cells.

机构信息

Department of Oral Science, Nano and Biotechnology, University G. D'Annunzio, Via Vestini 11, Chieti-Pescara, Italy.

出版信息

Int Endod J. 2012 Jan;45(1):19-25. doi: 10.1111/j.1365-2591.2011.01942.x. Epub 2011 Sep 8.

Abstract

AIM

To evaluate morphological features, cell growth and interleukin-6 (IL-6) and interleukin-8 (IL-8) secretion in expanded ex vivo human dental pulp mesenchymal stem cells (DP-MSCs) after exposure to 2-hydroxyethyl methacrylate (HEMA).

METHODOLOGY

Dental pulp mesenchymal stem cells were derived from the dental pulps of 10 young donors. After in vitro isolation, DP-MSCs were treated with 3 and 5 mmol L(-1) HEMA, and after 24, 48 and 72 h of incubation, their morphological features, cell growth, IL-6 and IL-8 secretion were analysed. Differences in the cell growth and in the interleukin secretion were analysed for statistical significance with two-way anova tests and the Holm-Sidak method for multiple comparisons.

RESULTS

Dental pulp mesenchymal stem cells revealed a decrease in cell growth with both treatments (P < 0.05), more evident at 5 mmol L(-1) . Microscopic analysis displayed extensive cytotoxic effects in treated cells, which lost their fibroblastoid features and became retracted, even roundish, with a large number of granules. An up-regulation of IL-6 and IL-8 in treated cells cytokines was evident (P < 0.05).

CONCLUSIONS

2-Hydroxyethyl methacrylate exhibited cytotoxicity, inhibited cell growth and induced morphological changes in cultured DP-MSCs. Moreover, in treated samples, an up-regulation of soluble mediators of inflammation such as IL-6 and IL-8 cytokines was found. The direct application of HEMA potentially induces an inflammation process that could be the starting point for toxic response and cell damage in DP-MSCs.

摘要

目的

评估暴露于 2-羟乙基甲基丙烯酸酯(HEMA)后体外扩增的人牙髓间充质干细胞(DP-MSCs)的形态特征、细胞生长以及白细胞介素-6(IL-6)和白细胞介素-8(IL-8)的分泌情况。

方法

从 10 位年轻供体的牙髓中分离出牙髓间充质干细胞。体外分离后,DP-MSCs 用 3 和 5 mmol/L HEMA 处理,孵育 24、48 和 72 h 后,分析其形态特征、细胞生长、IL-6 和 IL-8 的分泌情况。采用双因素方差分析和 Holm-Sidak 多重比较法对细胞生长和白细胞介素分泌的差异进行统计学分析。

结果

两种处理均导致 DP-MSCs 的细胞生长减少(P < 0.05),5 mmol/L 时更为明显。显微镜分析显示,处理后的细胞出现广泛的细胞毒性作用,失去成纤维细胞样特征,回缩,甚至呈圆形,有大量颗粒。处理后的细胞中白细胞介素-6 和白细胞介素-8 等促炎细胞因子的表达上调(P < 0.05)。

结论

2-羟乙基甲基丙烯酸酯具有细胞毒性,抑制细胞生长并诱导培养的 DP-MSCs 发生形态变化。此外,在处理样本中发现促炎介质如白细胞介素-6 和白细胞介素-8 等可溶性介质的表达上调。HEMA 的直接应用可能会引发炎症反应,从而成为 DP-MSCs 中毒性反应和细胞损伤的起点。

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