Abstract
BackgroundAcacia honey is a natural product which has proven to have therapeutic effects on skin wound healing, but its potential healing effects in corneal wound healing have not been studied. This study aimed to explore the effects of Acacia honey (AH) on corneal keratocytes morphology, proliferative capacity, cell cycle, gene and protein analyses. Keratocytes from the corneal stroma of six New Zealand white rabbits were isolated and cultured until passage 1. The optimal dose of AH in the basal medium (FD) and medium containing serum (FDS) for keratocytes proliferation was identified using MTT assay. The morphological changes, gene and protein expressions of aldehyde dehydrogenase (ALDH), marker for quiescent keratocytes and vimentin, marker for fibroblasts were detected using q-RTPCR and immunocytochemistry respectively. Flowcytometry was performed to evaluate the cell cycle analysis of corneal keratocytes.ResultsCultured keratocytes supplemented with AH showed no morphological changes compared to control. Keratocytes cultured in FD and FDS media supplemented with 0.025% AH showed optimal proliferative potential compared with FD and FDS media, respectively. Gene expressions of ADLH and vimentin were increased in keratocytes cultured with AH enriched media. All proteins were expressed in keratocytes cultured in all media in accordance to the gene expression findings. No chromosomal changes were detected in keratocytes in AH enriched media.ConclusionCorneal keratocytes cultured in media supplemented with 0.025% AH showed an increase in proliferative capacity while retaining their morphology, gene and protein expressions with normal cell cycle. The results of the present study show promising role of AH role in accelerating the initial stage of corneal wound healing.
Highlights
Acacia honey is a natural product which has proven to have therapeutic effects on skin wound healing, but its potential healing effects in corneal wound healing have not been studied
Cell viability and proliferation assay Corneal keratocytes cultured in the free medium (FD) medium supplemented with low concentrations of Acacia honey (AH) from 0.00038% to 0.025% exhibited increase in proliferative capacity compared to the FD medium alone
Corneal keratocytes cultured in the FD medium supplemented by 0.012% AH (p = 0.024) and 0.025% AH (p = 0.003) showed a significant increase in the proliferative capacity (p < 0.05) when compared to the FD medium alone (Figure 1A)
Summary
Acacia honey is a natural product which has proven to have therapeutic effects on skin wound healing, but its potential healing effects in corneal wound healing have not been studied. This study aimed to explore the effects of Acacia honey (AH) on corneal keratocytes morphology, proliferative capacity, cell cycle, gene and protein analyses. Cornea is an avascular and transparent tissue located at the anterior aspect of the eye It consists of five layers, i.e. the epithelium, Bowman’s membrane, stroma, Descemet’s membrane and endothelium. Eye injuries frequently caused by chemical, thermal, infection, trauma or surgical procedure such as keratectomy [3,4]. These injuries may involve only the epithelium, for example, in corneal abrasion [5] or may extend deep into the stroma such as corneal ulceration [6,7]. Even at very low concentrations, appeared to be cytotoxic for ocular cells and leads to apoptosis and free radical production [14,15]
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