title: In vitro load-induced dentin collagen-stabilization against MMPs degradation.
abstract: BACKGROUND
  Teeth are continuously subjected to stresses during mastication, swallowing and parafunctional habits, producing a significant reduction of the bonding efficacy in adhesive restorations. The purpose of this study was to evaluate the metalloproteinases (MMPs)-mediated dentin collagen degradation of hybrid layers created by using different demineralization processes, previous resin infiltration, and in vitro mechanical loading.
  
  
  METHODS
  Human dentin beams (0.75×0.75×5.0mm) were subjected to different treatments: (1) untreated dentin; (2) demineralization by 37% phosphoric acid (PA) or by 0.5% M ethylenediaminetetraacetic acid (EDTA); (3) demineralization by PA, followed by application of Adper(™) Single Bond (SB); (4) demineralization by EDTA, followed by application of SB. In half of the specimens, mechanical loadings (100,000 cycles, 2Hz, 49N) were applied to dentin beams. Specimens were stored in artificial saliva. C-terminal telopeptide (ICTP), determinations (which indicates the amount of collagen degradation) (radioimmunoassay) were performed after 24h, 1 week and 4 weeks.
  
  
  RESULTS
  Load cycling decreased collagen degradation when dentin was untreated or PA-demineralized and EDTA-treated. ICTP values increased when both PA-demineralized and EDTA-treated and infiltrated with SB dentin beams were loaded, except in samples that were subjected to EDTA treatment and SB infiltration after 4w of storage, which showed similar values of collagenolytic activity than the non loaded specimens. Load cycling preserved the initial (24h) ICTP determination at any time point, in all groups of the study, except in PA-demineralized and SB infiltrated dentin which showed an increased of collagen degradation values, over time. This same trend was observed in all groups without loading.
  
  
  CONCLUSIONS
  Mechanical loading enhances collagen's resistance to enzymatic degradation in natural and demineralized dentin. Mild acids (EDTA) lead to a lower volume of demineralized/unprotected collagen to be cleaved by MMPs. Load cycling produced an increase of collagen degradation when PA-demineralized dentin and EDTA-treated dentin were infiltrated with resin, but EDTA-treated dentin showed a constant collagenolytic degradation, over time.
authors.affiliation: Department of Dental Materials, School of Dentistry, University of Granada, E-18071 Granada, Spain. Electronic address: toledano@ugr.es.
authors.forename: Manuel
authors.initials: M
authors.lastname: Toledano
authors.forename: Fátima S
authors.initials: FS
authors.lastname: Aguilera
authors.forename: Monica
authors.initials: M
authors.lastname: Yamauti
authors.forename: María Estrella
authors.initials: ME
authors.lastname: Ruiz-Requena
authors.forename: Raquel
authors.initials: R
authors.lastname: Osorio
mesh_terms: D003094:Collagen / Q000737:chemistry / Q000378:metabolism*; D003804:Dentin / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D008422:Materials Testing; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D008963:Molar / Q000378:metabolism / Q000502:physiology; D055550:Protein Stability; D059748:Proteolysis; D016474:Weight-Bearing
chemical_list: D003094:Collagen; D020782:Matrix Metalloproteinases
keywords: Adhesion; Collagen degradation; Dentin; Load cycling; Longevity; Resin infiltration; Restoration