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K.C. Shin, Y.S. In, J.–H. Ko, M.–G. Park, H.–G. Kim, W.R. Wee, J.H. Lee, M.K. Kim; The Effect of MB500 on Corneal Epithelium and Keratocyte as Hyaluronan Biosynthesis Promoter: in vitro and in vivo Study . Invest. Ophthalmol. Vis. Sci. 2005;46(13):2135.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To investigate the effect of MB500(MDbioalpha, Daejeon, Korea) on cultured human corneal epithelial cells and keratocytes and to evaluate the effect of MB500 on corneal wound healing in rabbit. Methods: Various concentrations(1–200uM) of MB 500(10ul) was applied to the cultured human epithelial cells and keratocytes. Supernatant was collected after 48 hours of incubation with MB 500. MTT assay was performed to evaluate toxicity of MB 500 to the cells. Hyaluronan(HA) synthesis assay with biotinylated hyaluronan binding protein(b–HABP), glycosaminoglycan(GAG) synthesis assay and nitric oxide(NO) assay were performed using ELISA. Epithelial defects were made using 7.0 mm trephine and application of 20% alcohol for 20 seconds in 24 rabbits. MB500(200uM) or phosphate buffer saline(PBS) as a control was topically instilled four times a day to each 12 rabbit’s eye. The ratio of the epithelial defect size to the total corneal area was calculated using Image Pro PlusTM(Media Cybernetics, MD, USA) every day until the wound was completely healed. Hyaluronan was stained with b–HABP and 3,3’–diaminobenzidine dihydrochloride at 1,2 and 5 days after injury in both treated group and control Results: Cell death was not increased in MB500–treated cells(1uM and 200uM) compared with that in control. HA increased by 1.15 folds in epithelial cell with 5uM of MB500 and by 1.35 folds in keratocytes with 5uM of MB 500. 50uM of MB500 increased the concentration of GAG by 2 folds. MB500 decrease NO concentration by 43% at 50uM compared with that in positive control with IL–1ß. Corneal wound healing is rapid in study group with MB500 compared with control group with PBS and statistically significant difference(p=0.034) is observed at 3 days after injury. Conclusions: The instillation of MB500 shows no cell toxicity. MB 500 can promote corneal wound healing possibly by increasing HA and GAG and by decreasing NO.
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