Effect of LPS treatment on the viability and chemokine synthesis by epithelial cells and gingival fibroblasts
Effect of LPS treatment on the viability and chemokine synthesis by epithelial cells and gingival fibroblasts
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DOI：
10.1016/j.archoralbio.2015.04.010
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发表时间：
2015-08-01
影响因子：
3
通讯作者：
Hebling, J.
中科院分区：
文献类型：
--
作者：
Basso, F. G.;Soares, D. G.;Hebling, J.
目的：几种局部因素可影响创面愈合过程，延缓创面愈合进程，延缓组织动态平衡。众所周知，局部炎症与伤口愈合有关；然而，炎症反应的维持会损害口腔粘膜细胞的增殖和迁移。设计：将人牙龈成纤维细胞和上皮细胞(1×10(5)个/孔)接种于24孔板中，培养24小时，以模拟细胞对局部慢性口腔黏膜炎症的反应。我们在Dulbecco‘s改良的Eagle’s培养基中加入内毒素(10mU g/ml)，分别与成纤维细胞和上皮细胞接触24、48、48、24、48、24小时检测细胞活性(AlamarBlue法)、细胞数量(台盼蓝法)、CCL2和CCL5的表达(酶联免疫吸附试验(ELISA))。数据经非参数Kruskal-Wallis和Mann-Whitney检验在5%的显着性水平上进行统计学分析。结果：两种细胞经脂多糖处理后，细胞存活率均显著降低。上皮细胞未观察到时间依赖性效应。两组细胞CCL2和CCL5的合成均显著增加，且随时间延长而降低。结论：内毒素维持炎症细胞的刺激使培养的口腔黏膜细胞的数量和活性降低，这可能与创伤愈合延迟有关。(C)2015爱思唯尔有限公司。保留所有权利。
Objective: Several local factors can affect the wound-healing process, delaying its progression and postponing tissue homeostasis. It is known that local inflammation is related to wound healing; however, the maintenance of the inflammatory reaction can impair the proliferation and migration of oral mucosal cells. The aim of this study was to evaluate the viability and chemokine expression of epithelial cells and gingival fibroblasts exposed to long-term lipopolysaccharide (LPS) treatment.Design: Epithelial cells (HaCaT, Cell Lines Service, 300493) and human gingival fibroblasts (HGFs) were seeded (1 x 10(5) cells/well) in 24-well plates and incubated for 24 h. To simulate the responses of cells to a local chronic oral mucosal inflammation, we added LPS of Escherichia coli (10 mu g/ml) to Dulbecco's modified Eagle's medium (DMEM), kept in contact with fibroblasts and epithelial cells for 24, 48, and 72 h. Then the cells were assessed for viability (alamarBlue assay), number (trypan blue assay), and expression of CCL2 and CCL5 inflammatory chemokines (enzyme-linked immunosorbent assay (ELISA)). Data were statistically analyzed by nonparametric Kruskal-Wallis and Mann-Whitney tests at a significance level of 5%.Results: Cell treatment with LPS caused significant decrease in viability for both cell lines. No time-dependent effect was observed for epithelial cells. However, reduction in fibroblast viability was greater at 48 and 72 h. CCL2 and CCL5 synthesis was significantly increased for both LPS-treated cells, and this expression decreased with time.Conclusion: The maintenance of an inflammatory cell stimulus by LPS decreases the number and viability of cultured oral mucosal cells, which may be related to delayed wound healing. (C) 2015 Elsevier Ltd. All rights reserved.