Abstract
BackgroundAcacia honey (AH) has been proven to improve skin wound healing, but its therapeutic effects on corneal epithelium has not been elucidated to date. This study aimed to investigate the effects of AH on cultured corneal epithelial cells (CEC) on in vitro corneal abrasion wound healing model. Six New Zealand white rabbits’ CEC were isolated and cultured until passage 1. Circular wound area was created onto a confluent monolayer CEC using a corneal trephine which mimicked corneal abrasion and treated with 0.025% AH supplemented in basal medium (BM) and complete cornea medium (CCM). Wound healing was measured as the percentage of wound closure by the migration of CEC on day 0, day 3 and day 6, post wound creation. The morphological changes of CEC were assessed via phase contrast microscopy. Gene and protein expressions of cytokeratin (CK3), fibronectin and cluster of differentiation 44 (CD44) in AH treated groups and control groups were determined by real-time PCR and immunocytochemistry, respectively.ResultsCultured CEC exhibited similar morphology of polygonal shaped cells in all culture media. CEC cultured in AH-supplemented media showed higher percentage of wound closure compared to the controls. Gene expression of CK3 increased in AH-supplemented groups throughout the study. Fibronectin expression was increased at the initial stage while CD44 expression was increased at day 3, post wound creation. The protein expression of CEC cultured in all media was in accordance to their respective gene expressions.ConclusionSupplementation of AH in BM and CCM media accelerates CEC wound closure of the in vitro corneal abrasion model by increasing the expression of genes and proteins associated with CEC wound healing.
Highlights
Acacia honey (AH) has been proven to improve skin wound healing, but its therapeutic effects on corneal epithelium has not been elucidated to date
Morphological evaluation corneal epithelial cells (CEC) supplemented with AH exhibited small polygonalshaped cells with distinct cell border (Figure 2A-L)
Corneal epithelial wound healing study CEC cultured in AH-supplemented media (Figure 3D-F, J-L) showed faster wound healing compared to the controls
Summary
Acacia honey (AH) has been proven to improve skin wound healing, but its therapeutic effects on corneal epithelium has not been elucidated to date. This study aimed to investigate the effects of AH on cultured corneal epithelial cells (CEC) on in vitro corneal abrasion wound healing model. Migration and proliferation of epithelial cells are essential for wound closure during initial phase of corneal epithelial wound healing. Epithelial cells express the specific corneal epithelial differentiation marker, cytokeratin 3 (CK3) and cytokeratin 12 (CK12) once the cells left the limbal basal layer during centripetal migration [4]. These cells expressed fibronectin, a prototype of cell adhesion protein required for cell attachment, migration, differentiation, wound healing and cytoskeletal organization [5,6]. The migration phase in cornea wound healing is influenced by the synthesis of cluster of differentiation
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