Amniotic membrane extract provides significant improvement in in vitro wound healing model while preserving the basic...

Although many preclinical and clinical studies are ongoing on amniotic membrane extract (AME), an amniotic membrane-derived product developed to support ocular surface healing, the effect of AME on the basic cellular functions and properties of human corneal epithelial cells (hCECs) has not been clearly defined. In this study, we aimed to evaluate the effect of AME supplementation to the culture media, on basic cellular functions of hCECs and on expression of specific cell markers of hCECs, as well as to determine its effectiveness in an experimental in vitro wound model. hCECs were seeded with the constant cell density in 6, 24 and 48 well plates. The next day, the media was refreshed with 0 mg/ml, 0.75 mg/ml, 1.5 mg/ml and 3 mg/ml final concentration of AME supplemented complete growth medium. Cellular morphology, viability, metabolic activity, proliferation assessments and immunocytochemistry were conducted on the time points at day 3, 6 and 9. Then, in vitro wound healing assay was performed on hCECs under the effect of AME. AME did not affect cellular morphology, viability, metabolic activity and proliferation. AME supplementation induced CK3 and CK12 expression of hCECs significantly (p < 0.05) higher than without AME group. In vitro wound healing assay revealed that while control and AME-treated cultures both exhibited healing, AME-treated mechanical abrasions closed at a greater rate compared to control. Our study shows that AME promotes in vitro wound healing and cell characteristics in terms of CK3 and CK12 protein expressions, while preserving basic cellular functions of corneal epithelial cells.