Análise da ação do peptídeo antimicrobiano Lrot3 nanoestruturado contra as bactérias Escherichia coli e Staphylococcus epidermidis

Abstract

Pathogenic microorganisms are responsible for the development of various infections, particularly in hospital environment, leading to death in many patients. It is therefore necessary to change this scenario, it is difficult, however, because the bacteria have very distinct resistance mechanisms after mutations in their DNA. Furthermore, the indiscriminate use of antibiotics continuously generates the selection and propagation of more resistant strains, causing loss of effectiveness of antibiotics over the years. In this case, the development of a nanocarrier containing Antimicrobial Peptides (AMPs) becomes an interesting alternative against pathogenic bacteria. This paper aims to synthesize the delivery of AMPs to bacterial cells of Escherichia coli 25922 and Staphylococcus epidermidis 12228 through the use of nanospheres of sodium alginate by ionotropic gelation method. Alginate stands out for being a polyanionic, biodegradable and biocompatible polymer, which show interaction with Lrot3 cationic AMP. The antimicrobial activity of free and nanostructured peptides (128, 64, 32, 16, 8 e 4 µg/mL) was evaluated by the minimum inhibitory concentration (MIC) and the cytotoxic potential investigated in human HEK 293 cells using the MTT Assay (Tetrazolium thiazolyl blue). The data were evaluated by ANOVA and the means compared by Tukey test. For Gramnegative E. coli 25922 bacteria, the free peptide at concentration of 64 µg/mL stood out for being the only concentration with significant antimicrobial activity (P˂0,01), showing similar action to the chloramphenicol, at concentrations of 128 µg/mL and 64 µg/mL, respectively. NPs, in turn, showed inhibitory activity at all concentrations (P˂0,01). Unlike occurred in S. epidermidis 12228, which was observed more bacterial resistance to all treatments. The MTT data suggest that alginate nanoparticles do not exhibit in vitro cytotoxicity, encouraging the continuation of this study. In conclusion, the nanostructured AMPs has the potential to be used to fight infections caused by E. coli.