David V Dudok1, Imraan Nagdee1, Kevin Cheung1, Hong Liu1, Luca Vedovelli2, Emiliano Ghinelli3, Kenneth Kenyon3,4, Sunil Parapuram1, Cindy M Hutnik1. 1. Ivey Eye Institute, Western University, London, Ontario, Canada. 2. Division of Nephrology, Cell Biology, Human Biology and Physiology, Massachusetts General Hospital, Boston, Massachusetts, USA. 3. Harvard Medical School, Schepens Eye Research Institute, Boston, Massachusetts, USA. 4. New England Medical Center, Tufts University, Boston, Massachusetts, USA.
Abstract
BACKGROUND: 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. METHODS: 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. RESULTS: 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). CONCLUSIONS: 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.
BACKGROUND: 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. METHODS: 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. RESULTS: 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). CONCLUSIONS: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.
Authors: Ghasem Yazdanpanah; Ritu Shah; Sri Raghurama R Somala; Khandaker N Anwar; Xiang Shen; Seungwon An; Meisam Omidi; Mark I Rosenblatt; Tolou Shokuhfar; Ali R Djalilian Journal: Ocul Surf Date: 2021-04-22 Impact factor: 6.268
Authors: Michael S Murri; Majid Moshirfar; Orry C Birdsong; Yasmyne C Ronquillo; Yanning Ding; Phillip C Hoopes Journal: Clin Ophthalmol Date: 2018-06-18
Authors: Nur Zahirah Binte M Yusoff; Andri K Riau; Gary H F Yam; Nuur Shahinda Humaira Binte Halim; Jodhbir S Mehta Journal: Cells Date: 2022-01-05 Impact factor: 6.600