The Effect of Irradiated Riboflavin in Human Tenon’s Fibroblast – A Study on Cellular Viability

ABSTRACT

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.

KEYWORDS:

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

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the supplementary material. https://doi.org/10.1080/02713683.2021.2011326

Acknowledgments

The authors would like to thank Puan Anis Syamimi Mohamed, the laboratory assistant and Miss Lim Chien Joo for the statistical expertise.

Declarations

This manuscript has not been published elsewhere and that it has not been submitted simultaneously for publication elsewhere.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Supplementary material

Supplemental data for this article can be accessed on the publisher’s website

Additional information

Funding

This thesis is funded by High Impact Research, University of Malaya, Ministry of Higher Education (HIR grant: H-20001-00-E000058) and Fundamental Research Grant Scheme (FRGS), Ministry of Higher Education, Malaysia (Grant number FP037-2015A).