4.3 Article

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

Journal

CURRENT EYE RESEARCH
Volume 47, Issue 4, Pages 525-530

Publisher

TAYLOR & FRANCIS INC
DOI: 10.1080/02713683.2021.2011326

Keywords

Riboflavin; irradiated riboflavin; cellular viability; human tenon fibroblast; MTT assay

Categories

Funding

  1. High Impact Research, University of Malaya, Ministry of Higher Education (HIR grant) [H-20001-00-E000058]
  2. Fundamental Research Grant Scheme (FRGS), Ministry of Higher Education, Malaysia [FP037-2015A]

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The study aimed to investigate the effect of irradiated riboflavin on cellular viability in human tenon fibroblasts. The results showed that irradiated riboflavin induced concentration-dependent cell death, with a significant impact up to 48 hours, but the duration of treatment did not significantly affect the outcome. Further investigations are needed to determine the mechanism of cell death, with a postulation of apoptosis in irradiated riboflavin-treated cells.
Purpose/ Aim The main purpose of this work is to study the cellular viability effect of irradiated riboflavin in cultured human tenon fibroblasts. Materials and Methods 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. Results 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. Conclusions 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.

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