Teeth were hemi-sectioned in a mesial-distal direction through the center of the marked lesion area with a http://www.selleckchem.com/products/CAL-101.html high-speed drill and fine diamond bur. Speed was set at 800 rpm and a moderate weight (100 g) was chosen to guide the diamond-coated saw blade during the cutting procedure. One half of each tooth was processed for histological evaluations. Subsequently, specimens were dried in alcohol solvents of increasing concentration and embedded in Technovit 9100 New (Heraeus Kulzer, Wehrheim, Germany). Serial sections of 200 ��m were prepared and polished to a final thickness of approximately 150 ��m. Wet sections were viewed under a polarized light microscope (Axiovert 135, Carl Zeiss, Jena, Germany) at ��10 magnification.
Specimens were immersed in water, and the test sites were viewed under a polarized light microscope for the presence or absence of residual caries.15 To prevent examiner bias, the reproducibility of the histological validation was determined by re-examining the sections of teeth after a minimum break of 1 week. The examiner was blind to the previous results. Intra-examiner correlation (Pearson) was rp = 0.91�C0.98. SEM evaluation One half of each tooth was prepared for scanning electron microscope (SEM) investigation of the treated dentinal surface morphology. The samples were rinsed and were ultrasonically cleaned during 12 minutes. Then, the specimens were mounted separately on aluminum stubs, coated with gold/palladium by means of an ion coating equipment (SEM Coating Unit E 500, POLARON Equipment Limited, Barcelona, Spain), then evaluated under SEM (JSM-6400, JEOL, Tokyo, Japan) and micrographs were obtained.
Statistical analysis Statistical analysis were performed by means of SPSS 13.0 for Windows software (SPSS Inc., Chicago IL, USA). Cohen��s Kappa was calculated for intra-examiner calibration of detecting residual dentinal caries. The Kappa values were determined using the established cut-off levels for caries status. A two-way ANOVA with repeated measures were applied to do the comparisons among the groups and between the initial and final DIAGNOdent measures. In each group, whether there is a difference between methods used for determining residual caries is analyzed by McNemar test, and differences in the comparisons between all the groups are analyzed by Fischer`s exact test. Statistical significance were set at P=.
05. RESULTS Our initial Cohen��s Kappa evaluation of intra-examiner reproducibility agreement produced Kappa values from 0.49 to 0.93. There was not GSK-3 difference among the groups in the initial DIAGNOdent readings. There were statistical differences among initial and final DIAGNOdent values within the same groups (P<.05) (Table 1). In addition, the mean DIAGNOdent readings between all groups were stated significant (P<05) and the mean DIAGNOdent values in the groups 2 and 3 were significantly higher than those obtained from the group 1 (Table 1).
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