Characterizing the collagen stabilizing effect of crosslinked chitosan nanoparticles against collagenase degradation.
Antibacterial and chelating properties of chitosan has been widely studied for various dental applications.



To characterize the interaction between chitosan-nanoparticles (CSnp) and collagen, and understand their stabilizing effect against collagenase degradation for dentin matrix stabilization.



Phase-1: a single Type I collagen-fibril model was used to study the interaction with CSnp along with carbodiimides crosslinking treatment. Degradation of the crosslinked fibrils was studied with bacterial collagenase enzyme and monitored using Fourier Transform Infrared (FTIR) spectroscopy, turbidity measurement (400nm), ninhydrin assay and Atomic Force Microscopy (AFM). Interaction of CSnp with collagenase and Type I collagen, were evaluated using SDS-PAGE, and proteolytic cleavage potential of a synthetic peptide. Phase-2: degradation of dentin collagen crosslinked with/without CSnp was evaluated using FTIR, ninhydrin assay and Scanning Electron Microscopy (SEM). Glutaraldehyde crosslinking was used as a positive control.



Both native collagen-fibrils and dentin collagen after crosslinking showed higher resistance to collagenase degradation, as observed in turbidity measurements and FTIR spectra. AFM images showed the interaction of CSnp with single collagen-fibril and crosslinked collagen resisted collagenase degradation up to 54h. The collagen and collagenase both formed complexes with CSnp resulting in thickening of bands and reduction in collagen degradation. CSnp treated collagenase showed significantly reduced cleavage of the fluorescent peptides. Dentin collagen was coated with CSnp following crosslinking with significant increase in resistance to collagenase degradation.



Crosslinked CSnp on collagen stabilized and enhanced the resistance of dentin matrix against bacterial collagenase degradation due to non-specific interaction with both collagen and collagenase.
Discipline of Endodontics, Faculty of Dentistry, University of Toronto, 124 Edward Street, Toronto, ON M5G1G6, Canada. Electronic address: anil.kishen@dentistry.utoronto.ca.
Anil
A
Kishen
Dental Research Institute, Faculty of Dentistry, University of Toronto, 124 Edward Street, Toronto, ON M5G1G6, Canada. Electronic address: suja.shrestha@mail.utoronto.ca.
Suja
S
Shrestha
Discipline of Endodontics, Faculty of Dentistry, University of Toronto, 124 Edward Street, Toronto, ON M5G1G6, Canada. Electronic address: annie.shrestha@utoronot.ca.
Annie
A
Shrestha
Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, ON M5S 3H6, Canada. Electronic address: c.cheng@chem.utoronto.ca.
Calvin
C
Cheng
Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, ON M5S 3H6, Canada. Electronic address: cgoh@chem.utoronto.ca.
Cynthia
C
Goh
D048271:Chitosan; D003094:Collagen; D017364:Collagenases / Q000378:metabolism*; D003804:Dentin; D053758:Nanoparticles
D003094:Collagen; D048271:Chitosan; D017364:Collagenases
AFM; Chitosan-nanoparticles; Collagen; Crosslink; Dentin