Comparison of cytotoxicity and wound healing effect of carboxymethylcellulose and hyaluronic acid on human corneal epithelial cells
Comparison of cytotoxicity and wound healing effect of carboxymethylcellulose and hyaluronic acid on human corneal epithelial cells
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DOI：
10.3980/j.issn.2222-3959.2015.02.01
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发表时间：
2015-04-18
影响因子：
1.4
通讯作者：
Lee, Ji-Eun
中科院分区：
文献类型：
--
作者：
Lee, Long Soo;Lee, Scans Uk;Lee, Ji-Eun
目的：研究羧甲基纤维素（CMC）和透明质酸（HA）对人角膜上皮细胞（HCEC）的细胞毒性作用以及促进角膜上皮伤口愈合的能力。方法：将HCEC暴露于0.5％CMC（Refresh plus（R），Allergan，尔湾，加利福尼亚州，美国）和0.1％和0.3％HA（Kynex（R） Alcon（韩国首尔）和 Hyalein mini（右），Santen（日本大阪））持续 30 分钟、4、12 和 24 小时。进行基于甲基噻唑基四氮杂环 (MU) 的量热测定来评估细胞增殖的代谢活性，并进行乳酸脱氢酶 (LDH) 渗漏测定来评估细胞毒性。通过流式细胞分析和膜联蛋白 V 和碘化丙啶荧光染色评估细胞凋亡反应。通过倒置相差光学显微镜和电子显微镜评估细胞形态。汇合的HCECs划伤24h后测量伤口宽度。结果：对人角膜上皮增殖的抑制作用和细胞毒性呈时间依赖性反应，但无明显效果。在流式细胞术中发生细胞凋亡，并且在荧光显微照片中证实了凋亡细胞。受损的 HCEC 从培养皿底部脱离，并显示出发育良好的液泡结构。 CMC和HA均能刺激划伤的HCECs上皮化，这种现象在CMC中更为明显。结论：CMC和HA用于人工泪液制剂中，对HCECs没有任何明显的毒性作用。两者均显着刺激角膜伤口的 HCEC 再上皮化。
AIM: To investigate the cytotoxic effect on human corneal epithelial cells (HCECs) and the ability to faciliate corneal epithelial wound healing of carboxymethylcellulose (CMC) and hyaluronic acid (HA).METHODS: HCECs were exposed to 0.5% CMC (Refresh plus (R), Allergan, Irvine, California, USA) and 0.1% and 0.3% HA (Kynex (R) Alcon, Seoul, Korea, and Hyalein mini (R), Santen, Osaka, Japan) for the period of 30min, and 4, 12, and 24h. Methyl thiazolyl tetrazoiun (MU) -based calorimetric assay was performed to assess the metabolic activity of cellular proliferation and lactate dehydrogenase (LDH) leakage assay to assess the cytotoxicity. apoptotic response was evaluated with flow cytometric analysis and fluorescence staining with Annexin V and propiodium iodide. Cellular morphology was evaluated by Inverted phase-contrast light microscopy and electron microscopy. The wound widths were measured 24h after confluent HCECs were scratch wounded.RESULTS: The inhibitory effect of human corneal epithelial proliferation and cytotoxicity showed the time-dependent response but no significant effect. Apoptosis developed in flow cytometry and apoptotic cells were demonstrated in fluorescent micrograph. The damaged HCECs were detached from the bottom of the dish and showed the well -developed vacuole formations. Both CMC and HA stimulated reepithehlialization of HCECs scratched, which were more observed In CMC.CONCLUSION: CMC and HA, used in artificial tear formulation, could be utilized without any significant toxic effect on HCECs. Both significantly stimulated HCEC reepithelialization of corneal wounds.