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Chlorhexidine Inhibits the Proteolytic Activity of Root and Coronal Carious Dentin in vitro

Garcia M.B.a · Carrilho M.R.b, c · Nör J.E.d · Anauate-Netto C.b · Anido-Anido A.b · Amore R.b · Tjäderhane L.f · Bretz W.A.e
aDepartment of Biochemistry, Universidade São Francisco, School of Dentistry and School of Pharmacy, Bragança Paulista, bBandeirante University of São Paulo (UNIBAN), School of Dentistry, São Paulo, cDepartment of Restorative Dentistry, University of Campinas, School of Dentistry, Piracicaba, Brazil; dDepartment of Cariology, Restorative Sciences, and Endodontics, University of Michigan, School of Dentistry, Ann Arbor, Mich., eDepartment of Cariology and Comprehensive Care, New York University, College of Dentistry, New York, N.Y., USA; fInstitute of Dentistry, University of Oulu and Oulu University Hospital, Oulu, Finland Caries Res 2009;43:92–96 (DOI:10.1159/000209340)


The purpose of this study was to evaluate the effect of chlorhexidine on the proteolytic activity of carious coronal and root dentin collected from patients. Sound dentin from freshly extracted human teeth was used as a control. Dentin fragments were mixed with a synthetic substrate for proteolytic enzymes (N-benzoyl-DL-arginine-naphthylamide – BANA) and the suspensions mixed with either 0.12% chlorhexidine digluconate or distilled water. These mixtures were incubated for 18 h at 37°C, color was developed by the addition of 0.1% Fast Garnet and their optical density was recorded spectrophotometrically. BANA hydrolysis measured by the optical density of incubated specimens was detected in all tested groups, but was significantly higher for carious than for sound dentin (p < 0.05). The proteolytic activity was reduced for carious coronal and root dentin by chlorhexidine (p < 0.05; 50 and 30%, respectively). Chlorhexidine also reduced the proteolytic activity in sound root dentin (p < 0.05; 20%). Conversely, changes in the proteolytic activity of sound coronal dentin were not observed in the presence of chlorhexidine. The reduction in proteolytic activity by chlorhexidine was significantly higher in carious coronal dentin than in carious root dentin (p < 0.05). In conclusion, part of the effect of chlorhexidine in controlling caries progression in humans may be due to a decrease in the proteolytic activity of carious coronal and root dentin. Because of the prolonged incubation time in the present study, similar results may be obtained clinically with prolonged dentin exposure to chlorhexidine, e.g. chlorhexidine-containing varnishes.


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