Basic Research| Volume 47, ISSUE 9, P1435-1444, September 2021

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Engineered Chitosan-based Nanoparticles Modulate Macrophage–Periodontal Ligament Fibroblast Interactions in Biofilm-mediated Inflammation

  • Hebatullah Hussein
    The Kishen Lab, Dental Research Institute, University of Toronto, Toronto, Canada

    Faculty of Dentistry, University of Toronto, Toronto, Canada

    Faculty of Dentistry, Endodontics Department, Ain Shams University, Cairo, Egypt
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  • Anil Kishen
    Address requests for reprints to Dr Anil Kishen, Dental Research Institute, Faculty of Dentistry, University of Toronto, 124 Edward Street, Toronto, Ontario, Canada M5G 1G6.
    The Kishen Lab, Dental Research Institute, University of Toronto, Toronto, Canada

    Faculty of Dentistry, University of Toronto, Toronto, Canada

    Department of Dentistry, Mount Sinai Health System, Mount Sinai Hospital, Toronto, Ontario, Canada
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      Crosstalk between immune cells and tissue-resident cells regulates the pathophysiology and posttreatment healing of apical periodontitis. This investigation aimed to understand the influence of residual root canal biofilm on macrophage (MQ)–periodontal ligament fibroblast (PdLF) interaction and evaluate the effect of engineered chitosan-based nanoparticles (CSnp) on MQ-PdLF interactions in residual biofilm-mediated inflammation.


      Six-week-old Enterococcus faecalis biofilms in root canal models were disinfected conventionally using sodium hypochlorite alone or followed by calcium hydroxide medication or CSnp dispersed in carboxymethylated chitosan (CMCS). The effect of the treated biofilms (n = 25/group) on the inflammatory response of THP-1–differentiated MQ monoculture versus coculture with PdLF was evaluated for cell viability, MQ morphometric characterization, inflammatory mediators (nitric oxide, tumor necrosis factor alpha, interleukin [IL]-1 beta, IL-1RA, IL-6, transforming growth factor beta 1 [TGF-β1], and IL-10), and the expression of transcription factors (pSTAT1/pSTAT6)/cluster of differentiation markers (CD80/206) after 24, 48, and 72 hours of interaction. PdLF transwell migration was evaluated after 8 and 24 hours. Unstimulated cells served as the negative control, whereas untreated biofilm was the positive control.


      Biofilm increased nitric oxide and IL-1β but suppressed IL-10, IL-1RA, and PdLF migration with significant cytotoxic effects. CSnp/CMCS reduced nitric oxide and IL-1β (P < .01) while maintaining ≥90% cell survival up to 72 hours with evident M2-like MQ phenotypic changes in coculture. CSnp/CMCS also increased the IL-1RA/IL-1β ratio and enhanced TGF-β1 production over time (P < .05, 72 hours). In coculture, CSnp/CMCS showed the highest IL-10 level at 72 hours (P < .01), reduced the pSTAT1/pSTAT6 ratio, and enhanced PdLF migration (P < .01, 24 hours).


      CSnp/CMCS medication facilitated MQ switch toward M2 (regulatory/anti-inflammatory) phenotype and PdLF migration via paracrine signaling.

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