Epithelial Wound Healing on Keratin Film, Amniotic Membrane and Polystyrene In Vitro. (June 2014)

• Record Type: Journal Article
• Authors: Feng, Yaqing; Borrelli, Maria; Meyer-ter-Vehn, Tobias; Reichl, Stephan; Schrader, Stefan; Geerling, Gerd
• Is Part Of: Current eye research. Volume 39:Number 6(2014:Jun.)
• Page Start: 561
• Page End: 570
• Publication Date: 2014-06
• Subjects: Ophthalmology -- Periodicals; Eye -- Diseases -- Periodicals; Ophthalmology -- Periodicals; 573.88
• DOI: 10.3109/02713683.2013.853804
• Languages: English
• ISSNs: 0271-3683

Abstract

<abstract> <title>Abstract</title> <p> <italic>Purpose</italic>: Corneal epithelial wound healing is a major issue in ocular surface (OS) reconstruction. Aim of this study was to evaluate parameters of epithelial wound healing <italic>in vitro</italic> on transparent keratin films (KFs) derived from human hair in comparison with amniotic membrane (AM) and polystyrene.</p> <p> <italic>Materials and Methods</italic>: The human corneal epithelial cell line (HCE-T) was expanded on KF, AM and commercially available 24-well polystyrene cell culture plates <italic>in vitro</italic> to compare cell proliferation, migration and attachment by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide, scratch-wound healing and adhesion assay. Cells cultured on KF and AM at an air–liquid interface for 14 d were stained with hematoxylin and eosin for histology.</p> <p> <italic>Results</italic>: The highest proliferation of HCE-T cells was observed on polystyrene at all time points (<italic>p</italic> < 0.05). At a seeding density of 5 × 10<sup>3</sup> cells/well, no difference in proliferation was found between AM and KF after 24 h and 72 h (<italic>p</italic> = 0.582 and <italic>p</italic> = 0.066), while higher proliferation was observed on AM compared to KF after 48 h (<italic>p</italic> = 0.005). At a seeding density of 1 × 10<sup>4</sup> cells/well, no difference was found between AM and KF after 24 h (<italic>p</italic> = 0.252), while higher proliferation was observed on AM compared to KF after 48 h and 72 h (<italic>p</italic> = 0.001 and <italic>p</italic> = 0.003). The significantly fastest cell migration was observed on polystyrene at all time points (<italic>p</italic> < 0.01). Cell migration was significantly higher on KF compared to AM at 48 h (<italic>p</italic> < 0.05). After 30 min, there were significantly more cells attached to AM compared to polystyrene and KF (<italic>p</italic> = 0.032 and <italic>p</italic> = 0.001). No significant difference in cell attachment was observed between KF and polystyrene (<italic>p</italic> = 0.147). Histology demonstrated that HCE-T cells cultured on KF and AM at an air–liquid interface for 14 d form a multilayered epithelium similar to normal human corneal epithelium.</p> <p> <italic>Conclusion</italic>: Transparent KFs derived from human hair support proliferation, migration, adhesion and differentiation of HCE-T cells <italic>in vitro</italic>. Therefore, it could be a promising alternative to AM for OS reconstruction.</p> </abstract>