Abstract
Introduction
Collagen is the major structural protein of human dentin. Degradation of collagen
by bacterial enzymes can facilitate microbial penetration, compromise structural/interfacial
integrity, and lower resistance to fracture of dentin. We evaluated the ability of
photodynamic therapy (PDT), bioactive chitosan nanoparticles (CSnp), or PDT in combination
with CSnp to inhibit bacterial collagenase-mediated degradation of collagen.
Methods
Rat type 1 fibrillar collagen matrices were untreated or treated with 2.5% glutaraldehyde
(GD), 2.5% GD followed by 1% CSnp, 1% CSnp, PDT (rose bengal activated with 540 nm
light at 40 J/cm2), or 1% CSnp followed by PDT. Samples, except those used as untreated controls, were
exposed to Clostridium histolyticum collagenase (125 CDU/mL) for 24 hours. The soluble digestion products were assessed
by hydroxyproline assay, and the remaining adherent collagen was quantified by picrosirius
red staining. Fourier transform infrared spectroscopy, immunoblotting, and scanning
electron microscopy were used to study the interaction between CSnp/PDT with type
1 collagen. The data were analyzed by 1-way analysis of variance and post hoc Tukey
test.
Results
As assessed by hydroxyproline release into the medium, collagen treated with CSnp,
PDT, or a combination of CSnp and PDT exhibited less degradation than untreated controls
(3.6-fold, 1.7-fold, and 7.9-fold reduction, respectively; P < .05). Compared with all other treatments, GD-treated collagen was the most resistant
to collagenolytic degradation (239.6-fold reduction, P < .05). The abundance of post-treatment residual collagen, as measured by picrosirius
red staining, was inversely related to the extent of collagen degradation. Analysis
of collagen cross-links with Fourier transform infrared spectroscopy showed that PDT
or GD treatments enhanced collagen cross-linking. Immunoblotting of sedimented CSnp
indicated that CSnp and collagenase bound with low affinity. However, CSnp-bound collagenase
showed a significant reduction in collagenolytic activity compared with controls (P < .05).
Conclusions
Combined photochemical cross-linking of rat tail collagen by PDT and binding to CSnp
inhibit collagenolytic activity.
Key Words
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References
- On the causes of persistent apical periodontitis: a review.Int Endod J. 2006; 39: 249-281
- Current therapeutic options for endodontic biofilms.Endod Topics. 2010; 22: 79-98
- Mechanisms and risk factors for fracture predilection in endodontically treated teeth.Endod Topics. 2006; 13: 57-83
- Collagen degradation in endodontically treated teeth after clinical function.J Dent Res. 2004; 83: 414-419
- Quantitative analysis of the effect of irrigant solution sequences on dentin erosion.J Endod. 2011; 37: 1437-1441
- Effect of endodontic irrigation solutions on microhardness of root canal dentine.J Dent. 1999; 27: 43-46
- The activation of matrix metalloproteinases by a whole-cell extract from prevotella nigrescens.J Endod. 2009; 35: 55-59
- A scanning electron microscopic study of dentinal erosion by final irrigation with EDTA and NaOCl solutions.Int Endod J. 2002; 35: 934-939
- Photodynamic therapy: a new antimicrobial approach to infectious disease?.Photochem Photobiol Sci. 2004; 3: 436-450
- Photodynamically crosslinked and chitosan-incorporated dentin collagen.J Dent Res. 2011; 90: 1346-1351
- Photodynamic therapy for endodontic disinfection.J Endod. 2006; 32: 979-984
- Antimicrobial effects of photodynamic therapy on patients with necrotic pulps and periapical lesion.J Endod. 2008; 34: 138-142
- Application of crosslinkers to dentin collagen enhances the ultimate tensile strength.J Biomed Mater Res B Appl Biomater. 2007; 80: 268-272
- Chitosan as antimicrobial agent: applications and mode of action.Biomacromolecules. 2003; 4: 1457-1465
- Crosslinked collagen/chitosan matrix for artificial livers.Biomaterials. 2003; 24: 3213-3220
- An investigation on the antibacterial and antibiofilm efficacy of cationic nanoparticulates for root canal disinfection.J Endod. 2008; 34: 1515-1520
- Rapid analysis of amino acids using pre-column derivatization.J Chromatogr. 1984; 336: 93-104
- PEG-stabilized carbodiimide crosslinked collagen-chitosan hydrogels for corneal tissue engineering.Biomaterials. 2008; 29: 3960-3972
- Dentin basic structure and composition: an overview.Endod Topics. 2012; 20: 3-39
- Collagen cross-linking and ultimate tensile strength in dentin.J Dent Res. 2004; 83: 807-810
- Site comparisons of dentine collagen cross-links from extracted human teeth.Arch Oral Biol. 1993; 38: 541-546
- Human fibroblast collagenase-α-macroglobulin interactions: localization of cleavage sites in the bait regions of five mammalian α-macroglobulins.J Biol Chem. 1989; 264: 393-401
- Determining the presence and origin of collagenase in human periapical lesions.J Endod. 1987; 13: 228-232
- Polymerase chain reaction analysis of the porphyromonas gingivalis collagenase gene.Clin Infect Dis. 1995; 20: S167-S168
- Survey for collagenase gene prtC in porphyromonas gingivalis and porphyromonas endodontalis isolated from endodontic infections.J Endod. 1999; 25: 555-558
- Community as the unit of pathogenicity: an emerging concept as to the microbial pathogenesis of apical periodontitis.Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2009; 107: 870-878
- Biofilms and apical periodontitis: study of prevalence and association with clinical and histopathologic findings.J Endod. 2010; 36: 1277-1288
- Host-derived loss of dentin matrix stiffness associated with solubilization of collagen.J Biomed Mater Res B Appl Biomater. 2009; 90: 373-380
- Activation of gelatinolytic/collagenolytic activity in dentin by self-etching adhesives.Eur J Oral Sci. 2006; 114: 160-166
- Bacterial protease inhibitors.Med Res Rev. 2002; 22: 329-372
- A simple dot-blot-sirius red-based assay for collagen quantification.Anal Bioanal Chem. 2013; 405: 6863-6871
- Collagenolytic activity of black-pigmented bacteroides species.J Periodontal Res. 1987; 22: 300-306
- Evaluation of gelatin hydrogel crosslinked with various crosslinking agents as bioadhesives: in vitro study.J Biomed Mater Res. 1999; 46: 520-530
- Proanthocyanidin: a natural crosslinking reagent for stabilizing collagen matrices.J Biomed Mater Res A. 2003; 65: 118-124
- Relation between the physicochemical characteristics of collagen and its interactions with chitosan: I.Biomaterials. 1993; 14: 930-938
Article info
Publication history
Published online: December 12, 2013
Identification
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© 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.
