Virginia Via Research Day Book 2026

Undergraduate Student Research Biomedical

03 DUAL ASSAYS TO EVALUATE COPPER TOXICITY ON HUMAN KERATINOCYTES

Claire Cho, Alyssa Morrison, Claudia Ramos Lopez, Austin Mason, Allyson Meador, Dr. William Ducker, Dr. Marion Ehrich, and Dr. Jia-Qiang He Corresponding author: clairec28@vt.edu

Department of Biomedical Sciences and Pathobiology in Virginia-Maryland College of Veterinary Science, and Department of Chemical Engineering, College of Science, Virginia Tech

The antimicrobial properties of copper could lead to integrating copper in high-touch surfaces, such as door handles, to decrease bacterial transmission. However, the effects on copper with keratinocytes (skin cells) raises a concern to whether or not copper could have a toxic effect on human skin cells when coming into contact. This study aimed to investigate the impacts of copper on the viability of human keratinocytes. To this end, human keratinocytes were cultured in 12-well petri dishes until reaching around 80% confluency,and then uncoated block (UCB, negative control), copper coated block (CCB), or 0.1% Triton X- 100 (TX-100, positive control) were added in the wells. Specifically, glass blocks were used, and the CCB was coated with Cu2O particles. Both lactate dehydrogenase (LDH) and a resazurin-based toxicology assay were performed according to the manufacturer's protocols. LDH was used to measure the activity of LDH,which is released

the TX-100 wells compared to both UCB and CCB. Additionally, for the toxicity assay, the wells containing UCB and CCB were dyed in red while the wells with TX-100 were dyed blue. This shows that there was a high cell death rate found in wells with TX-100 and a low cell death rate in wells with UCB and CCB. Comparing the UCB and CCB wells, there was no significant difference for either assay, suggesting that the copper particles did not induce significant cell death within 24 hours incubation.

into the culture medium when cell membrane is damaged. In this assay, a plate reader was used to read the optical density (OD) of culture medium at 565 nm at 0 minutes and 25 minutes, while the resazurin-based test measures cell viability and metabolic activity following incubation of the resazurin solution in the culture. Since the dye is a reduction-oxidation indicator, the wells that are dyed in red indicate metabolic healthy cells, while ones that are dyed in blue are unhealthy. The absorbance at 690 nm and 600 nm were measured for each well to quantify the surviving cell population relative to the controls as well as the incubation time. For each assay, there were three plates comparing the time effects of copper on cell viability: 4, 8, and 24 hours. The data that has been collected so far has shown no significant differences between the cells with UCB and cells with CCB in both LDH and toxicity assays. However, there was a significant difference in

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