BackgroundThere has been no effective treatment or agent that is available for corneal injury in promoting corneal wound healing. Previous studies on edible bird's nest extract (EBN) had reported the presence of hormone-like substance; avian epidermal growth factor that could stimulate cell division and enhance regeneration. This study aimed to investigate the effects of EBN on corneal keratocytes proliferative capacity and phenotypical changes.MethodsCorneal keratocytes from six New Zealand White Rabbits were isolated and cultured until Passage 1. The proliferative effects of EBN on corneal keratocytes were determined by MTT assay in serum-containing medium (FDS) and serum-free medium (FD). Keratocytes phenotypical changes were morphologically assessed and gene expression of aldehyde dehydrogenase (ALDH), collagen type 1 and lumican were determined through RT-PCR.ResultsThe highest cell proliferation was observed when both media were supplemented with 0.05% and 0.1% EBN. Cell proliferation was also consistently higher in FDS compared to FD. Both phase contrast micrographs and gene expression analysis confirmed the corneal keratocytes retained their phenotypes with the addition of EBN.ConclusionsThese results suggested that low concentration of EBN could synergistically induce cell proliferation, especially in serum-containing medium. This could be a novel breakthrough as both cell proliferation and functional maintenance are important during corneal wound healing. The in vitro test is considered as a crucial first step for nutri-pharmaceutical formation of EBN-based eye drops before in vivo application.
In vitro drug screening techniques provide rapid and easy to analyze data, while saving a lot of animals from being sacrificed. An important part of any in vitro drug screening platform is a biomaterial which promotes cell growth and proliferation. The potential of electrospun scaffolds made of polyhydroxybutyrate (PHB), poly (3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV), and polycaprolactone (PCL) were studied to serve as drug screening platform for corneal keratocyte tissues. The results showed that the proliferation rate was slightly higher for PCL and PHBV on day 7. Gene expression results showed that PCL was the best in maintaining keratocyte genes.
Proliferation of corneal epithelial cells (CEC) isvital in the initial stage of wound healing. This study aimed to investigate the proliferative capacity of Acacia Honey (AH) on rabbit CEC via assessment on morphology, proliferation, cell cycle, gene and protein expressions.The optimal dose of AH in basal medium (BM) and complete cornea medium (CCM) was identified via MTT assay. CEC cultured in both media supplemented with 0.025% AH showed optimal proliferative capacity compared to the control. There were no abnormal changes in morphology and cell cycle analysis. Gene and protein expression of CK3 was increased in the CEC cultured with 0.025% AH in both media. CEC cultured in media supplemented with 0.025% AH promotes proliferation while retaining its normal morphology, cell cycle, gene and protein expressions. These promising results serve as an impetus in realizing the proliferative potential of AH in promoting the initial step of corneal wound healing.
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