title: A model for clinical evaluation of the effect of antimicrobial agents on carious dentin.
abstract: OBJECTIVE
  To evaluate, longitudinally, the effect of a chlorhexidine varnish on the proteolytic activity of dentin caries in vivo.
  
  
  METHODS
  20 permanent molars and 8 primary molars with carious lesions in dentin were studied in subjects 18-35 yrs old (n=20), and 5-6 yrs old (n=8) respectively. These lesions were clinically evaluated according to texture and color. Carious dentin specimens were obtained by means of biopsies performed with a #4 carbide bur at the initial visit (TO) before application of a 10% chlorhexidine varnish and 2, 4, 8, and 12 wks thereafter. The dentin biopsies were immersed in Sorensen's buffer, vortexed for 30 s, and mixed with a 1.67 mM solution of n-benzoyl-DL-arginine-naphthylamide (BANA), a substrate for proteolytic enzymes. Samples were incubated overnight at 37 degrees C and color was developed with 0.1% fast garnet. The optical density (OD) of reaction mixtures was recorded photometrically. All teeth were grouped for analysis, as Mann-Whitney tests revealed no statistically significant differences between median values for OD for both age groups. ANOVA was used to compare progressive inhibition of proteolytic activity in dentin caries samples over time.
  
  
  RESULTS
  The average proteolytic activity at the dentin substrates (OD) at TO and 2, 4, 8 and 12 wks thereafter were 0.794+/-0.089, 0.741+/-0.071, 0.676+/-0.087, 0.600+/-0.094, and 0.508+/-0.108 respectively. The chlorhexidine varnish mediated a significant inhibition of the proteolytic activity present in dentin caries after 12 wks (P<0.0001). At T0, 100% of the carious lesions examined were characterized as soft upon exploration. After 12 wks, 54% (15/28) of the lesions were partially hardened and 46% (13/28) hardened/nonprogressing. The dentin color was yellow/light brown in 100% of the lesions at baseline, and dark brown/black in 86% (24/28) after 12 wks.
  
  
  CONCLUSIONS
  This study demonstrated that chlorhexidine varnishes arrested active caries in vivo and inhibited the proteolytic activity present in these lesions. These findings strengthen the rationale for including chlorhexidine in the overall treatment strategy for patients with high caries activity.
authors.affiliation: Department of Biochemistry, School of Dentistry, Universidade São Francisco, Bragança, Paulista, SP, Brazil.
authors.forename: M B
authors.initials: MB
authors.lastname: Garcia
authors.forename: J E
authors.initials: JE
authors.lastname: Nör
authors.forename: L G
authors.initials: LG
authors.lastname: Schneider
authors.forename: W A
authors.initials: WA
authors.lastname: Bretz
mesh_terms: D000293:Adolescent; D000328:Adult; D000367:Age Factors; D000704:Analysis of Variance; D000891:Anti-Infective Agents, Local / Q000008:administration & dosage / Q000627:therapeutic use*; D001587:Benzoylarginine-2-Naphthylamide; D002648:Child; D002675:Child, Preschool; D002710:Chlorhexidine / Q000008:administration & dosage / Q000627:therapeutic use*; D003116:Color; D004396:Coloring Agents; D003731:Dental Caries / Q000201:enzymology / Q000517:prevention & control*; D003804:Dentin / Q000187:drug effects* / Q000201:enzymology; D010450:Endopeptidases / Q000187:drug effects; D005500:Follow-Up Studies; D006801:Humans; D008137:Longitudinal Studies; D008963:Molar; D055095:Optics and Photonics; D010150:Paint; D010783:Photometry; D011480:Protease Inhibitors / Q000008:administration & dosage / Q000627:therapeutic use; D018709:Statistics, Nonparametric; D014089:Tooth Remineralization; D014094:Tooth, Deciduous
chemical_list: D000891:Anti-Infective Agents, Local; D004396:Coloring Agents; D011480:Protease Inhibitors; D001587:Benzoylarginine-2-Naphthylamide; D010450:Endopeptidases; D002710:Chlorhexidine