Biological responses of injured human skin fibroblasts to assess the efficacy of in vitro models for cell stress studies

Abstract


Innocent Lutho Zungu, Denise Hawkins Evans, Nicolette Houreld and Heidi Abrahamse*

The classical central scratch method has been used to successfully induce a wound environment however this model has been criticized. This study aimed to compare the cellular responses of normal fibroblast cells with those of wounded, diabetic wounded, acidic and hypoxic cells to determine if the latter two models are more effective in inducing an injured environment than the central scratch method. The cellular responses that were evaluated included; cell morphology, viability, proliferation, cytotoxicity and DNA damage. The results from this study showed that the biochemical tests were sensitive enough to distinguish changes between normal and wounded, diabetic wounded, acidic and hypoxic cells. Wounded cells showed a decrease in ATP viability, increase in caspase 3/7 activity, increase in proliferation and increase in cytotoxicity and DNA damage when compared to normal cells. This study confirmed that although a central scratch method only wounds 5 - 10% of the surface cells it was sufficient to successfully induce a wound environment in vitro . The results suggest that the in vitro models may not only be used to study wound healing but also cellular responses related to other pathological conditions such as heart attack, stroke, diabetes and biological or biochemical changes in response to treatment such as laser therapy otherwise known as phototherapy.

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