Dudok David V, Nagdee Imraan, Cheung Kevin, Liu Hong, Vedovelli Luca, Ghinelli Emiliano, Kenyon Kenneth, Parapuram Sunil, Hutnik Cindy M
Ivey Eye Institute, Western University, London, Ontario, Canada.
Division of Nephrology, Cell Biology, Human Biology and Physiology, Massachusetts General Hospital, Boston, Massachusetts, USA.
Clin Exp Ophthalmol. 2015 Jul;43(5):443-8. doi: 10.1111/ceo.12480. Epub 2015 Jan 15.
To assess the effects of amniotic membrane extract (AMX) on cellular activity of primary human corneal epithelial (HCE) cells under mechanical and oxidative stress, and on human limbal cells under oxidative stress.
Corneal mechanical stress was simulated with a linear scratch in confluent HCE cell plates, then incubated with 0.1% AMX for 48 and 72 h. Subjecting HCE cultures to 0.5 mmol/L tertiary-butylhydroperoxide for 1 h simulated an oxidative stress. 0.1% AMX-treated cultures were compared with controls at 24 and 48 h using cellular viability assay, along with 12-h AMX pretreatment and human limbal cell comparisons.
Mechanical stress on HCE cultures revealed a statistically significant distance ratio at 48 and 72 h in favour of 0.1% AMX-treated cultures (P = 0.021 and 0.035, respectively). Oxidative stress did not reveal any significant difference in cellular viability of AMX-treated versus control cultures. Twelve hour AMX pre-treatment prior to oxidative stress revealed a significant difference after 24 h from oxidative injury (73.3% AMX vs. 66.0% control, P = 0.035), but not after 48 h. Human limbal cells demonstrated significantly improved oxidative viability compared with HCE cells, with (91.0% vs. 82.0% control, P = 0.017) and without 0.1% AMX pre-treatment (91.2% vs. 83.7% control, P = 0.019).
HCE cells treated with AMX healed faster after mechanical insult, suggesting a potential benefit in acute corneal injuries. Under oxidative stress, human limbal cells, a more proliferative cell type, showed superior viability compared with HCE cells.
评估羊膜提取物（AMX）对机械和氧化应激下原代人角膜上皮（HCE）细胞以及氧化应激下人角膜缘细胞的细胞活性的影响。
在融合的HCE细胞平板上进行线性划痕模拟角膜机械应激，然后用0.1% AMX孵育48和72小时。用0.5 mmol/L叔丁基过氧化氢处理HCE培养物1小时模拟氧化应激。在24和48小时使用细胞活力测定法将0.1% AMX处理的培养物与对照进行比较，同时进行12小时AMX预处理和人角膜缘细胞比较。
对HCE培养物的机械应激显示，在48和72小时时，支持0.1% AMX处理培养物的距离比具有统计学意义（分别为P = 0.021和0.035）。氧化应激未显示AMX处理培养物与对照培养物在细胞活力上有任何显著差异。氧化应激前12小时的AMX预处理在氧化损伤后24小时显示出显著差异（AMX为73.3%，对照为66.0%，P = 0.035），但在48小时后无差异。与人角膜上皮细胞相比，人角膜缘细胞在有（91.0%对82.0%对照，P = 0.017）和无0.1% AMX预处理（91.2%对83.7%对照，P = 0.019）的情况下，氧化活力均显著提高。
用AMX处理的HCE细胞在机械损伤后愈合更快，提示在急性角膜损伤中可能有益。在氧化应激下，人角膜缘细胞这种增殖性更强的细胞类型与人角膜上皮细胞相比显示出更高的活力。
