RESUMEN
The antimicrobial photodynamic therapy (aPDT) has stood out as an alternative and promising method of disinfection and has been exploited for the treatment of oral bacteria. In this study, we evaluate in vitro the action of aPDT, mediated by methylene blue, chlorin-e6, and curcumin against clinical subgingival plaques that were resistant to metronidazole. The sensitivity profile of the samples to metronidazole was analyzed by the agar dilution method. Cell viability in the planktonic and biofilm phase was assessed by CFU / mL. The composition of the biofilm was evaluated by the checkboard DNA-DNA Hibrydization technique. Photosensitizers internalization was qualitatively assessed by confocal fluorescence microscopy (CLSM). The aPDT mediated by the three photosensitizers tested was able to reduce the totality of the planktonic microbial load and partially reduce the biofilm samples. The analysis performed by CLSM showed that the photosensitizers used in the application of aPDT were able to permeate the interior of the biofilm. The aPDT has been shown to be useful in a supportive and effective approach to the treatment of periodontal disease.
Asunto(s)
Antibacterianos/farmacología , Placa Dental/microbiología , Farmacorresistencia Bacteriana/efectos de los fármacos , Metronidazol/farmacología , Fotoquimioterapia/métodos , Biopelículas/efectos de los fármacos , Clorofilidas , Curcumina/farmacología , Humanos , Azul de Metileno/farmacología , Pruebas de Sensibilidad Microbiana , Enfermedades Periodontales/tratamiento farmacológico , Fármacos Fotosensibilizantes/farmacología , Fármacos Fotosensibilizantes/uso terapéutico , Porfirinas/farmacologíaRESUMEN
In the past few years, the World Health Organization has been warning that the post-antibiotic era is an increasingly real threat. The rising and disseminated resistance to antibiotics made mandatory the search for new drugs and/or alternative therapies that are able to eliminate resistant microorganisms and impair the development of new forms of resistance. In this context, antimicrobial photodynamic therapy (aPDT) and helical cationic antimicrobial peptides (AMP) are highlighted for the treatment of localized infections. This study aimed to combine the AMP aurein 1.2 to aPDT using Enterococcus faecalis as a model strain. Our results demonstrate that the combination of aPDT with aurein 1.2 proved to be a feasible alternative capable of completely eliminating E. faecalis employing low concentrations of both PS and AMP, in comparison with the individual therapies. Aurein 1.2 is capable of enhancing the aPDT activity whenever mediated by methylene blue or chlorin-e6, but not by curcumin, revealing a PS-dependent mechanism. The combined treatment was also effective against different strains; noteworthy, it completely eliminated a vancomycin-resistant strain of Enterococcus faecium. Our results suggest that this combined protocol must be exploited for clinical applications in localized infections as an alternative to antibiotics.
Asunto(s)
Péptidos Catiónicos Antimicrobianos/farmacología , Fotoquimioterapia , Fármacos Fotosensibilizantes/farmacología , Péptidos Catiónicos Antimicrobianos/metabolismo , Transporte Biológico , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Membrana Celular/efectos de la radiación , Farmacorresistencia Bacteriana/efectos de los fármacos , Farmacorresistencia Bacteriana/efectos de la radiación , Sinergismo Farmacológico , Enterococcus faecalis/citología , Enterococcus faecalis/efectos de los fármacos , Enterococcus faecalis/metabolismo , Enterococcus faecalis/efectos de la radiación , Humanos , Especies Reactivas de Oxígeno/metabolismoRESUMEN
The action of diflubenzuron (DFB) was evaluated in a freshwater fish, Prochilodus lineatus, after exposure to 0.06, 0.12, 0.25, or 0.50 mg L(-1) DFB for 14 days. Erythrocyte nuclear abnormalities (ENA), the gill activity of Na(+)/K(+)-ATPase, H(+)-ATPase and carbonic anhydrase (CA), and lipid peroxidation (LPO) and histopathological changes in the gills and liver were determined. The number of micronuclei increased in fish exposed to 0.25 and 0.50 mg L(-1) DFB. Plasma Cl(-) and the CA activity decreased, while the activity of Na(+)/K(+)-ATPase and of H(+)-ATPase increased in fish exposed to 0.25 and 0.50 mg L(-1) DFB. LPO did not change in the gills but increased in the liver of fish exposed to 0.25 and 0.50 mg L(-1) DFB. In the gills, histopathological changes indicated disperse lesions and slight to moderate damage in fish exposed to 0.50 mg L(-1) DFB, whereas in the liver, these changes were significantly greater in fish exposed to 0.25 and 0.50 mg L(-1) DFB, indicating moderate to severe damage. Continuous exposure to DFB is potentially toxic to P. lineatus, causing heath disorders when the fish is exposed to the two highest DFB concentrations, which are applied to control parasites in aquaculture and to control mosquito populations in the environment.