摘要
Macrophages(MQ)are major constituents of chronically inflamed periapical tissues in apical periodontitis.This study aimed to investigate the immunomodulatory effect of engineered bioactive chitosan-based nanoparticles(CSnp)antibiofilm medication on MQ cocultured with periodontal ligament fibroblasts(PdLF).Cells viability,spreading,PdLF migration,and intracellular CSnp uptake were characterized.Tandem Mass Tag-based proteomics was applied to analyze MQ global protein expression profiles after interaction with Enterococcus faecalis biofilm,CSnp-treated biofilm,and CSnp.Secreted inflammatory mediators were analyzed.Following bioinformatics analyses,candidate proteins were validated via targeted proteomics.CSnp maintained cells viability,increased MQ spreading,and PdLF migration(p<0.05).Transmission electron micrographs demonstrated CSnp internalization via macropinocytosis,clathrin-mediated endocytosis,and phagocytosis.Proteomic analysis revealed that CSnp-treated biofilm upregulated proteins(>1.5-folds,p<0.05)showed functional enrichment in the pathway of metal sequestration by antimicrobial proteins,while downregulated proteins showed enrichment in ferroptosis.CSnp upregulated proteins exhibiting antioxidant and immunoregulatory properties.Upregulation of SERPINB1 by CSnp(>1.5-folds,p<0.05)was validated.CSnp-treated biofilm reduced pro-inflammatory IL-1β and nitric oxide but enhanced anti-inflammatory IL-10 and TGF-β1(p<0.05).Internalized engineered bioactive CSnp reprogrammed MQ proteomic and cytokine profiles to modulate biofilm-mediated inflammation,and prompted PdLF migration,emphasizing its potential to regulate healing process in the treatment of apical periodontitis.
基金
supported in part by a research grant from the American Association of Endodontists Foundation[#509641]
the Natural Sciences and Engineering Research Council of Canada:Discovery Grant-AK[RGPIN-2020-05844]
supported by a scholarship funded by the Egyptian Ministry of Higher Education.
