Understanding the protective role of Antioxidant in preventing epithelial ultraviolet radiation damage.

Abstract

Background: UVR has a definitive impact on the DNA of the keratinocyte which could lead to more complicated skin diseases. However, antioxidants play a role in the prevention of ATP depletion and metabolic perturbations induced by UV radiation and can increase cellular energy and enhance DNA repair. Therefore, the need to reveal the optimal efficacy that can be provided by antioxidants is important to minimize the UVR effect. hence, the project aimed to understand and compare the protective effect of three antioxidants ( NA, TA, and HA ). Materials and Methods: The project was done within sterile cell culture techniques involving HDF and NTERT1 lines. From these cells monolayers and 3D models were created. Both monolayer and 3D models were subjected to UV radiation, followed by treatment with antioxidants (NA, TA, HA). Viability was assessed using LDH and MTT assays. real-time qPCR conducted for gene expression evaluation. The histological starts with formalin fixation, paraffin embedding, and sectioning of 3D models, followed by H&E staining and light microscope imaging. Result: NTERT1 cell monolayers, observing changes in cell architecture post-UVR exposure and viability decreased with longer UVR exposure times. Based on the MTT assay, TA had fluctuating toxicity, while HA exhibited a concentration-dependent positive impact. Based on the LDH assay of monolayer cells, HA was more effective in preventing damage than TA against UVR. However, the LDH assay did not show a markable difference in the cell viability between the antioxidants with 3D model cells. HA increased IL-6 and CCL2 expression levels, while TA and NA showed minimal effects in promoting inflammatory response gene expression. 3D models demonstrated that the expression of IL-6 and CCL2 is highest with HA treated models. Conclusion: Within the study limitation, it was concluded that the UVR had a significant effect on the keratinocytes. Moreover, it was found that TA had notable toxicity at 31.25 µM and enhanced proliferation at 15.63 µM while HA exhibited a proportional viability increase. QPCR analysis in 2D, HA triggered immune responses and elevated CCL2 expression, suggesting a potential role in mitigating UVR damage. 3D models showed HA's protective role against UVR-induced LDH effects, altering IL-6 and CCL2 levels.