The Effect of Irradiated Riboflavin in Human Tenon's Fibroblast - A Study on Cellular Viability

• Published in: Current eye research Vol. 47; no. 4; pp. 525 - 530
• Main Authors: See, Wendy Yen Nee, Ismail, Fazliana, Sheikh Abdul Kadir, Siti Hamimah, Subrayan, Visvaraja
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 03.04.2022
• Subjects: cellular viability; human tenon fibroblast; irradiated riboflavin; MTT assay; Riboflavin; Riboflavin; irradiated riboflavin; cellular viability; human tenon fibroblast; MTT assay
• ISSN: 0271-3683; 1460-2202
• DOI: 10.1080/02713683.2021.2011326

The main purpose of this work is to study the cellular viability effect of irradiated riboflavin in cultured human tenon fibroblasts. The tenon tissue was harvested from a patient undergoing strabismus surgery. The human tenon fibroblast cell culture and isolation were performed according to the standard laboratory cell culturing protocol. The cells were divided into three groups: control, treatment with irradiated and non-irradiated riboflavin. There were five different concentrations (0.00156%, 0.003125%, 0.00625%, 0.0125%, 0.025%) in each group of riboflavin. The fibroblasts were treated with riboflavin and the cellular viability was assessed at 24-hour and 48-hour post treatment with MTT 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyl tetrazolium bromide colorimetric assay. The absorbance values were analysed using Magellan microplate reader data analysis. A triplicate of readings was taken. The data were presented as mean ± standard deviation of the triplicates. Statistical analysis was performed with Statistical Package for Social Sciences (SPSS) analysis version 23. Irradiated riboflavin caused a concentration-dependent cell death in human tenon fibroblast cell culture (p < .05). The antiproliferative difference between irradiated and non-irradiated riboflavin was significant up to 48 hours (p < .05). Post hoc multiple comparisons showed higher concentrations of irradiated riboflavin (0.0125% and 0.025%) caused more reduction in cellular viability in human tenon fibroblast cells (p < .05). The duration of treatment is not a causative factor in this study. This pilot experiment demonstrated that irradiated riboflavin induced cell death in human tenon fibroblast culture in a concentration-dependent manner, but is not time-dependent. Further exploratory investigations should be performed to determine the mechanism of cell death. We postulate that apoptosis occurred in these irradiated riboflavin-treated cells.