Efeito da foto-ozonioterapia frente ao Pythium insidiosum e na 360 viabilidade de queratinócitos da linhagem HaCaT

• Ano de defesa: 2022
• Autor(a) principal: Trevisani, Leonardo de Arruda Campos
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade de Franca; Brasil; Pós-Graduação; Programa de Mestrado em Ciência Animal; UNIFRAN
• Palavras-chave em Português: Equinos; Ozonioterapia; Antimicrobiano; Cavalo; Laser; Ozônio; Pitiose; CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
• Link de acesso: https://repositorio.cruzeirodosul.edu.br/handle/123456789/4099

Cutaneous pythiosis, caused by Pythium insidiosum, is a life-threatening infectious disease. Low-level laser (LLLT) and ozone gas (O3) therapies have been used individually and successfully in the treatment of skin infections. Therefore, the objectives of the study were: a) to characterize the antimicrobial action of the LLLT-O3 association (photo-ozone therapy) against P. insidiosum, and b) to evaluate the cytotoxic and antiproliferative potentials of LLLT-O3 in epidermal keratinocytes of the HaCAT lineage. Equine P. insidiosum hyphae plugs and keratinocytes were subjected to a single treatment with LLLT (660nm, 65 mW cm2 and 80 J cm2), O3 (50 μg O3 mL- 1) or LLLT-O3 (LLLT and O3 in sequence). Additionally, untreated pathogens were used as controls. The measurement of the hyphal growth area was used to characterize the antimicrobial potential of the respective treatments (n=30 plugs/group). The cytotoxic and antiproliferative potentials in keratinocytes were evaluated, respectively, by colorimetric and clonogenic efficiency assays (n=3 samples/group). Photo-ozone therapy had a negative effect (P < 0.001) on the growth of P. insidiosum hyphae. The LLLT-O3 and O3 treatments inactivated, respectively, 93.3 and 30% of the hyphal plugs. The absence of growth of all recultured plugs confirmed the germicidal effect of LLLT-O3. Microbial growth was detected in all plugs submitted only to LLLT and from untreated samples. Keratinocyte viability was not affected by treatment with LLLT or O3 alone (P > 0.2). However, the LLLT-O3 association showed mild cytotoxicity (cell viability = 80%; P < 0.05). When compared to the control group, all treatments (LLLT, O3 and LLLT-O3) significantly reduced (P < 0.05) the survival fraction of HaCat lineage keratinocytes. The present study described, for the first time, the antimicrobial action of photo-ozone therapy against P. insidiosum isolated from horses. In addition, the LLLT-O3 association showed a slight impairment of cutaneous cell viability. Keywords: Antimicrobial, horse, laser, ozone, pythiosis.