Gelam honey potentiates ex vivo corneal keratocytes proliferation with desirable phenotype expression.

Alia Md Yusof,Norzana Abd Ghafar,Chua Kien Hui,Taty Anna Kamarudin,Yasmin Anum Mohd Yusof

doi:10.1186/s12906-016-1055-7

Abstract

BackgroundThis study aimed to evaluate the effects of Gelam honey on corneal keratocytes proliferative capacity and phenotypic characterization via MTT assay, gene expression and immunocytochemistry.MethodsCorneal keratocytes from New Zealand white rabbits were cultured in basal medium (BM) and serum enriched medium (BMS). Serial dilutions of Gelam honey (GH) were added to both media and cells were cultured until passage 1. MTT assay was performed on corneal keratocytes in both media to ascertain the optimal dose of GH that produced maximum proliferation.ResultsGelam honey at the concentration of 0.0015 % in both media showed the highest proliferative capacity with no morphological changes compared to their respective controls. The gene expression of aldehyde dehydrogenase (ALDH), a marker for quiescent keratocytes and vimentin, a marker for fibroblast, were higher in the GH enriched groups. The alpha smooth muscle actin (α-SMA) expression, marker for myofibroblast, was lower in GH treated groups compared to the controls. Immunocytochemistry results were in accordance to the gene expression analyses.ConclusionGelam honey at a concentration of 0.0015 % promotes ex vivo corneal keratocytes proliferation while retaining desirable phenotype expression. The results serve as a basis for the development of Gelam honey as a potential natural product in promoting corneal wound healing.

Highlights

This study aimed to evaluate the effects of Gelam honey on corneal keratocytes proliferative capacity and phenotypic characterization via MTT assay, gene expression and immunocytochemistry
Corneal keratocytes showed higher proliferation in the basal medium with serum (BMS) group compared to the basal medium (BM) group in all Gelam honey (GH) concentrations (p < 0.05)
Keratocytes cultured in BMS medium supplemented with 0.0004 to 0.0031 % GH significantly increased keratocytes proliferation compared to BM supplemented GH medium

Summary

Introduction

This study aimed to evaluate the effects of Gelam honey on corneal keratocytes proliferative capacity and phenotypic characterization via MTT assay, gene expression and immunocytochemistry. Cell proliferation is the key process in wound healing and it involves several metabolic processes which require much energy. Honey is one of the natural products that is rich in glucose and contains enzymes, amino acids, vitamins and minerals [1]. It possesses medicinal properties which include antimicrobial, antioxidant, antimutagenic, antitumour, anti-inflammatory and the capability to stimulate cell proliferation [2]. Honey proliferative effects on the in vivo study of the skin wound healing were reported earlier [3,4,5,6]. To date, there is lacunae in the existing scientific literature with regard

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