Avaliação in vitro do potencial antimicrobiano e da viabilidade celular de diferentes concentrações de Ozonioterapia

Abstract

Cutaneous wound is a clinical condition with high prevalence and represents a public health problem. Although ozone therapy (O3), ozone therapy, is widely used in clinical practice due to its antimicrobial and restorative effects, evidence on ideal dosimetries is still scarce. Thus, the aim of the present study was to evaluate and compare, in vitro, the cytotoxicity of different concentrations of ozone therapy in keratinocyte (NOK) and fibroblast (L929) lines, as well as the antimicrobial potential for Staphylococcus aureus (S.aureus) and Escherichia coli (E. coli). NOK and L929 cells were seeded at a concentration of 1x104 and strains of gram-positive bacteria S.aureus and gram negative bacteria E. coli at a concentration of 1x108 and divided into 7 experimental groups treated with ozone at doses of 0 (G0), 5 (G5), 10 (G10), 20 (G20), 30 (G30), 40 (G40) and 50 μg/mL (G50). The effect of ozone on cell viability/cytotoxicity was determined using the alamarBlue® assay, the Live/Dead® assay, and morphometric analysis of the percentage of live and dead cells. The effect of the gas on the microorganisms was evaluated by counting the colony forming unit (CFU). The results obtained show that ozone at concentrations of 5, 10 and 20 μg/mL presented the highest viability values of NOK (keratinocytes) and L929 (fibroblasts) cells, as well as a higher density and percentage of live cells in the experimental period of 72 hours when compared to other concentrations. However, it was not possible to observe the difference between the percentage of dead cells between the experimental groups. Furthermore, ozone promoted a reduction in CFUs at all concentrations tested. With the results obtained in the present study, it is possible to confirm the reparative potential of ozone therapy in a dose-dependent response, as well as its antimicrobial potential. However, studies are needed to elucidate the molecular mechanisms by which it exerts such biological effects.