In vitro studies of virulence suppression on P. aeruginosa by phosphate / polyphosphate-loaded nanoparticles.

摘要

Critically ill patients harbor multi-drug resistant pathogens that can activate their virulence in the response to low nutrient conditions and host stress derived factors. It was recently shown that the oversupply of inorganic phosphate to bacterial environment can profoundly suppress the virulence of pathogens. Here we hypothesized that phosphate- and/or polyphosphate-loaded nanoparticles can present a tool to deliver and slowly release phosphate in pathogen-rich niche, thereby suppressing bacterial virulence. In this work, a designed study on effect of different phosphate levels (including the phosphate released from hydrogel nanoparticles) on virulence of P. aeruginosa is addressed.;In this work, we developed formulations for preparing hexametaphosphate-loaded nanoparticles on the basis of that for phosphate loaded nanoparticles. We utilized inverse miniemulsion polymerization in the synthesis of these nanoparticles. Polyethylene glycol diacrylate (PEGDA, moleculat weight of 575 Da) and N-vinyl pyrrolidone (molecular weight: 111.14 Da ) were chosen to be the initial monomers because the main crosslinker, polyethylene glycol is a kind of biocompatible material that has been approved by the U.S. Food and Drug Administration (FDA). Several parameter could be adjusted among the experiment. We selected the monomer mole fraction of PEGDA-575 as our parameter. After the synthesis, a nanoparticle size distribution between 110 nm and 150 nm was obtained. And these nanoparticles were proved to be able to release phosphate and hexametaphosphate as drug molecules. Although there were release bursts in the test of release kinetics, the crosslink density could be adjusted in following researches.;The second part of this study is to test the virulence suppression effect of the nanoparticles in in vitro experiment on a kind of opportunistic pathogen, P. aeruginosa. This kind of gram-negative bacteria is one of the common intestinal microbial communities. We presented the strategy of suppressing virulence while containing rather than killing the bacteria. As a result, polyphosphate-loaded nanoparticles showed to be the most effective one among several experiment groups. This result gave this study a promising future in further research in several aspects, such as in vivo test in biomedical and biomedical engineering.