title: Impact of protease inhibitors on dentin matrix degradation by collagenase.
abstract: This proof-of-concept study assessed whether the reduction of the degradation of the demineralized organic matrix (DOM) by pre-treatment with protease inhibitors (PI) is effective against dentin matrix loss. Bovine dentin slices were demineralized with 0.87 M citric acid, pH 2.3, for 36 hrs. In sequence, specimens were treated or not (UT, untreated) for 1 min with gels containing epigallocatechin 3-gallate (EGCG, 400 µM), chlorhexidine (CHX, 0.012%), FeSO(4) (1 mM), NaF (1.23%), or no active compound (P, placebo). Specimens were then stored in artificial saliva (5 days, 37°C) with the addition of collagenase (Clostridium histolyticum, 100 U/mL). We analyzed collagen degradation by assaying hydroxyproline (HYP) in the incubation solutions (n = 5) and evaluated the dentin matrix loss by profilometry (n = 12). Data were analyzed by ANOVA and Tukey's test (p < 0.05). Treatment with gels containing EGCG, CHX, or FeSO(4) led to significantly lower HYP concentrations in solution and dentin matrix loss when compared with the other treatments. These results strongly suggest that the preventive effects of the PI tested against dentin erosion are due to their ability to reduce the degradation of the DOM.
authors.affiliation: Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, SP, Al. Octávio Pinheiro Brisolla, 9-75, Bauru-SP, 17012-901, Brazil.
authors.forename: M T
authors.initials: MT
authors.lastname: Kato
authors.forename: A L
authors.initials: AL
authors.lastname: Leite
authors.forename: A R
authors.initials: AR
authors.lastname: Hannas
authors.forename: M P
authors.initials: MP
authors.lastname: Calabria
authors.forename: A C
authors.initials: AC
authors.lastname: Magalhães
authors.forename: J C
authors.initials: JC
authors.lastname: Pereira
authors.forename: M A R
authors.initials: MA
authors.lastname: Buzalaf
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D002327:Cariostatic Agents / Q000627:therapeutic use*; D002392:Catechin / Q000031:analogs & derivatives / Q000627:therapeutic use; D002417:Cattle; D002710:Chlorhexidine / Q000627:therapeutic use; D003094:Collagen / Q000378:metabolism*; D017364:Collagenases / Q000378:metabolism*; D003804:Dentin / Q000378:metabolism* / Q000648:ultrastructure; D005109:Extracellular Matrix / Q000378:metabolism; D005296:Ferrous Compounds / Q000627:therapeutic use; D011480:Protease Inhibitors / Q000627:therapeutic use*; D012969:Sodium Fluoride / Q000627:therapeutic use; D017001:Tooth Demineralization / Q000201:enzymology / Q000517:prevention & control; D014077:Tooth Erosion / Q000201:enzymology / Q000517:prevention & control*
chemical_list: D001426:Bacterial Proteins; D002327:Cariostatic Agents; D005296:Ferrous Compounds; D011480:Protease Inhibitors; D002392:Catechin; D012969:Sodium Fluoride; D003094:Collagen; C045651:epigallocatechin gallate; D017364:Collagenases; D002710:Chlorhexidine