Effect of Biofilm Formation on the Color Properties of CAD/CAM Materials

Effect of Biofilm Formation on the Color Properties of CAD/CAM Materials | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Effect of Biofilm Formation on the Color Properties of CAD/CAM Materials Sümeyra Topçu, Eda Yazıcı Özcelik, Dilan Kopuz, Fetiye Kolaylı, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3221496/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Objectives To investigate the effect of biofilm formation on the coloration properties of CAD/CAM materials. Materials and Methods 106 samples of 2-mm thickness were prepared from two different CAD/CAM materials (IPS e.max CAD; Cerasmart) and a total of 53 samples were prepared from extracted caries-free human-teeth. Five samples from each sample group were used to measure the amount of alive biomass in the biofilm. The remaining 48 samples in each group were divided into four subgroups: group-1, kept in water without the formation of dental biofilm; group-2, kept in tea without the formation of dental biofilm; group-3, kept in water after the formation of dental biofilm; group-4: kept in tea after the formation of dental biofilm (n=12). After finishing and polishing, color and surface roughness measurements were made. After baseline measurements, a biofilm layer was formed in groups-3 and 4, and the measurements were repeated. Afterward, all samples were brushed, and a third measurement was performed. Data were statistically analyzed (p<0.05). Results The lowest roughness value was observed in Cerasmart. Tooth-IPS e.max CAD gave similar results. The Cerasmart material had the most viable biomass, whereas the IPS e.max CAD material had the least. Group-4 had the highest ΔE1 value for all materials and group-1 had the lowest. The presence of biofilm on CAD/CAM materials immersed in water caused an unacceptable degree of coloration (ΔE>1.8), and immersion in tea caused much more color change. Conclusions The brushing process allowed CAD/CAM materials to return to their original color. Clinical Relevance The adhesion of biofilm to restorative dental materials plays an important role in the coloring of dental treatment. CAD/CAM Color Biofilm Formation Biofilm Surface Roughness Figures Figure 1 Figure 2 Introduction Biofilms that are in equilibrium with the host under healthy conditions are complex ecosystems. As a result of changes in this balance, bacterial colonization and biofilm formation on teeth may cause dental plaque, then this process causes enamel demineralization, dental caries, periodontitis, or many diseases [ 1 ]. Adhesion is related to the material's surface structure and its surface roughness, which is the most important component in determining the quantity of plaque accumulation [ 2 – 4 ]. It is very important to finish restorations with a smooth surface to achieve an esthetically and functionally successful restoration [ 5 , 6 ]. Cracks, grooves, and worn surfaces often lead to greater surface roughness, creating areas suitable for bacterial adhesion [ 7 ]. These surface irregularities protect bacteria from shear forces in the oral cavity and cause bacteria to bind more strongly to the substrate [ 8 ]. For these reasons, the choice of material for biofilm formation is important in restorative material selection. The preservation of color stability of the material is also a very important factor in evaluating the success of a restoration. Teeth and restoration materials are exposed to many factors such as biofilm, acidic by-products formed in the biofilm, and coloring foods and beverages [ 8 ]. The results of in vitro testing of materials exposed to these conditions can be used to predict the clinical performance of restorations regarding longevity and esthetic results. Our study aimed to investigate the color changes that occurred in CAD/CAM materials and human teeth by forming a biofilm layer on the CAD/CAM materials used in the restoration of teeth and to what extent this color change was reversed. The null hypotheses created in the study: Increasing the degree of roughness of CAD/CAM materials does not increase the amount of live biomass formed on the material. Increasing the degree of roughness of CAD/CAM materials does not cause more coloration of the material. The presence of biofilm on CAD/CAM materials does not cause further coloration of the materials by the effect of coloring beverages. Brushing would not influence discoloration. Materials and methods 106 CAD/CAM block sections of 2 mm thickness were prepared, Cerasmart (A2 HT, GC Corporation, Tokyo, Japan) (n = 53) and IPS e.max CAD (A2 HT, Ivoclar Vivadent, Schaan, Liechtenstein) (n = 53), and 53 extracted caries-free third-molar human teeth were collected for testing. The materials used in our study are given in Table 1 . Table 1 Materials used in the study Material Contents Manufacturer LOT Cerasmart CAD/CAM Bloc 71% silica and barium glass nanoparticles, 20 nm silica, 300 nm barium glass nanoparticles, 29% Bis-MEPP, UDMA and DMA polymers GC Corporation, Tokyo, Japan 2012151 IPS e.max CAD CAD/CAM Bloc (57–80%) SiO 2 , (11–19%) Li 2 O, (0–13%) K 2 O, (0–11%) P 2 O 5 , (0–8%) ZrO 2 , (0–8%) ZnO, (% 0–12) other oxides Ivoclar Vivadent, Schaan, Liechtenstein Z00FTN Abbreviations : Bis-MEPP: 2,2-Bis (4-methacryloxy polyethoxy phenyl) propane, UDMA: Urethane Dimethacrylate, DMA: Dimethacrylate, SiO 2 : Silicon Dioxide, Li 2 O: Lithium oxide, K 2 O: Potassium oxide, P 2 O 5 : Diphosphorus penta oxide, ZrO 2 : Zirconium oxide, ZnO: Zinc oxide Five samples from each group (IPS e.max CAD, Cerasmart, teeth) were separated for the measurement of live biomass in the biofilm. The remaining 48 samples from each group were divided into 4 experimental subgroups (n = 12 in each): group 1, kept in water without forming a dental biofilm; group 2, kept in tea without forming a dental biofilm; group 3, kept in water after the dental biofilm was formed; group 4, kept in the tea after the dental biofilm was formed (Fig. 1 ). The surfaces of the samples were sanded for 60 seconds with 600, 800, and 1000-grit silicon carbide (SIC) sandpaper to provide standardization before the polishing process. According to the manufacturer's instructions, the IPS e.max CAD blocks were fired to crystallize and were glazed in the Ivoclar Programat P300 oven. Cerasmart blocks were finished and polished using a two-stage Sof-Lex Diamond Polishing system with a slow-speed micromotor running at 15,000–20,000 rpm. A beige-colored disc was used for pre-polishing, and a pink-colored disc was used for polishing. These procedures were performed underwater for 15 seconds. The samples were cleaned using deionized water in an ultrasonic cleaner for 10 minutes following the completion of the surface treatments. After surface finishing applications were completed, roughness measurements of all samples were performed using a Nanovea 3D Non-Contact Profilometer PS50 (Nanovea, 6 Morgan Ste 156, Irvine, CA, USA). The purpose of this measurement was to compare the effect of surface roughness on surface discoloration. Color measurements of the samples were performed using a VITA Easyshade Compact device (VITA Zahnfabrik, Bad Säckingen, Germany). Each measurement began with the calibration of the device. The measurements were taken on a standard white background and under constant light for the elimination of the background effect [ 9 ]. During the measurements, the fiber optic end of the device was held perpendicular to the samples and parallel to the ground. Each measurement was repeated three times and the average of L, a, b, C, and H values ​​were recorded. A CIEDE2000 system (ΔE 00 ) was used to calculate the color change. Then all samples were sterilized using an ethylene oxide gas sterilization system (Steris Amsco Eagle, Mentor, AND). To ensure bacterial attachment and to create a pellicle layer on the samples, artificial saliva was created. For 2 liters of artificial saliva, 8.4 mg NaF, 2560 mg NaCl, 332.97 mg CaCl 2 , 250.00 mg MgCl 2 (6H 2 O), 189.48 mg KCl, 0.1 mL H 3 PO 4 (85%), and 0.1 mmol NaOH were used. All ingredients were mixed and the pH of the resulting mixture was measured using a pH meter. To adjust the pH of the mixture between 6.5-7, 30 mL of 0.1 M NaOH was added [ 10 ]. The mixture was sterilized by passing a 0.22 µm syringe tip through a PES filter (BIOSORFA, China) and then 140 mg of Type II mucin (Sigma-Aldrich Chemie GmbH, Deisenhofen, Germany) was added under aseptic conditions at a rate of 140 mg per 100 mL. A biofilm culture medium was made for the bacteria Streptococcus mutans (S. mutans) HF76, Streptococcus sanguinis (S. sanguinis) ATCC 10556, and Candida albicans (C. albicans) ATCC 90028 strains. S. mutans and S. sanguinis strains were removed from the freezer and inoculated on 5% sheep blood agar (SBA) medium and incubated at 37°C in 5% CO 2 . C. albicans were seeded on Sabouraud dextrose agar (SDA) medium and incubated at 37°C for 24 hours. After incubation, S. mutans, S. sangiunis, and C. albicans were subcultured. The subcultured tubes were centrifuged at 3000 g at 19°C for 5 minutes and the supernatant was discarded. Then, sterile phosphate-buffered saline (PBS) (pH 7.0) was added and washed, and this process was repeated twice. To form a biofilm, a bacterial density of 0.5 McFarland (1.5x108 cfu/mL) S. mutans and S. sangiunis was prepared by suspending brain heart infusion (BHI) broth with 3% sucrose, 0.5 McFarland C. albicans SDB [ 11 ]. In the biofilm formation phase of the study, e.max CAD (n = 29) and Cerasmart (n = 29) CAD/CAM blocks, and caries-free third-molar human teeth (n = 29) which previously sterilized using ethylene oxide were used. Sterile CAD/CAM samples were placed in 24-well plates and dental samples were placed in tubes and incubated at 37°C for 1 hour by adding artificial saliva until the material was completely covered to ensure pellicle formation. After incubation, artificial saliva was removed from the environment. For 48 hours, the CAD/CAM samples and dental samples were incubated at 37°C in an environment containing 5% CO 2 by inoculating equal amounts of the prepared bacterial suspension and yeast suspension on plates [ 12 – 14 ]. Measurement of biofilm mass through Live Biomass Assessment (MTT method) was performed with reference to previous studies [ 15 ]. CAD/CAM block samples on which biofilm was formed were placed in new 24-well plates and the teeth were placed in new tubes and washed 3 times with PBS (VWR USA) to remove microorganisms that were not in the biofilm structure. Two different solutions, distilled water and tea, were prepared for coloring the samples. Two tea bags (Yellow Label Tea, Lipton, Rize, Turkey) (2 x 2 g) were dissolved in 300 mL of boiling distilled water for 10 minutes to create the tea solution. Tea was added to half (n = 12) of the biofilm-formed CAD/CAM samples and teeth until they were completely covered, and water was added to the other half. The control group CAD/CAM samples, on which no biofilm was formed, were placed in 24-well plates, and teeth were placed in tubes; tea was added to half of the samples (n = 12) until they were completely covered, and distilled water was added to the other half, and all samples were kept at 37℃ for 24 hours [ 16 ]. After the samples were kept in solutions for 24 hours, second color measurements were performed. To remove the plaque on the samples, a rechargeable Braun Oral B Ultra Plaque Remover toothbrush (Braun D9, Braun AG, Kronberg, Germany) was used and fixed to a stand to simulate brushing under 2 N load, according to ISO 14569-1 [ 17 , 18 ]. The experiment samples were brushed under a 2 N load for 10 seconds, using toothpaste (ProNamel, Sensodyne) containing 1450 ppm sodium fluoride with a relative dentin abrasivity (RDA) value of 34, and a slurry prepared by mixing homogeneously with distilled water in a 1:1 ratio [ 19 ]. After brushing, the samples were washed in distilled water and cleaned, then color measurements were made again. The IBM SPSS 20.0 (IBM Corp., Armonk, NY, USA) package program was used for statistical evaluation. Differences between groups were examined using the independent sample t-test and one-way analysis of variance (ANOVA) for normally distributed variables, and the Mann-Whitney U and Kruskal-Wallis tests for non-normally distributed data. Tukey and Dunn tests were used for multiple comparisons (p < 0.05). Results After applying polishing procedures, the initial surface roughness of the tooth, Cerasmart, IPS e.max CAD materials were measured, and Ra, Rz, and Sa values were compared. The values are shown in Table 2 . Three-dimensional images of the samples are shown in Fig. 2 . Table 2 Surface roughness values (Ra, Rz, Sa µm) (median values, 25–75% percentile, mean ± standard deviation) Material Ra Rz Sa Cerasmart Median 0,132 A Median 0,729 D Median 0,353 F Percentile %25 0,120 Percentile %25 0,635 Percentile %25 0,319 %75 0,145 %75 0,790 %75 0,497 Mean 0,181 Mean 0,700 Mean 0,402 Standard deviation ± 0,186 Standard deviation ± 0,099 Standard deviation ± 0,108 IPS e.max CAD Median 0,410 B Median 2,102 E Median 4,122 G Percentile %25 0,302 Percentile %25 1,637 Percentile %25 2,960 %75 0,592 %75 2,918 %75 5,286 Mean 0,427 Mean 2,209 Mean 4,827 Standard deviation ± 0,141 Standard deviation ± 0,732 Standard deviation ± 2,623 Tooth Median 0,233 AB Median 0,767 D Median 4,441 G Percentile %25 0,166 Percentile %25 0,595 Percentile %25 3,792 %75 0,298 %75 1,116 %75 6,181 Mean 0,238 Mean 0,838 Mean 4,811 Standard deviation ± 0,080 Standard deviation ± 0,261 Standard deviation ± 1,553 While the different capital letters indicated indicate a statistically significant difference within the same column, the same capital letters indicate that there is no statistically significant difference within the same column. When the Ra values ​​of the samples were examined, the median values ​​of Cerasmart, IPS e.max CAD, and tooth groups were 0.132, 0.410, and 0.233, respectively. Accordingly, the lowest roughness was observed in Cerasmart samples, and the highest roughness values ​​were observed in IPS e.max CAD samples. In the pairwise comparison of the groups, there was a significant difference between Cerasmart-IPS e.max CAD (p = 0.001). No statistically significant difference was found between the Cerasmart blocks and teeth (p = 0.067) and the teeth and IPS e.max CAD blocks (p = 0.078). When the Rz values ​​of the groups were examined, the median values ​​of Cerasmart, IPS e.max CAD, and the teeth were 0.729, 2.102, and 0.767, respectively. Accordingly, the lowest roughness was observed in Cerasmart samples, and the highest roughness values ​​were observed in IPS e.max CAD samples. In the pairwise comparison of the groups, there was a significant difference between Cerasmart-IPS e.max CAD (p = 0.001) and dental-IPS e.max CAD (p = 0.001). No statistically significant difference was found between the Cerasmart samples and teeth (p = 0.737). When the Sa values ​​of the groups were examined, the median values ​​of Cerasmart, IPS e.max CAD, and tooth groups were 0.353, 4.122, and 4.441, respectively. Accordingly, the lowest roughness was observed in Cerasmart samples, and the highest roughness values ​​were observed in the teeth. In the pairwise comparison of the groups, there was a significant difference between Cerasmart and IPS e.max CAD (p = 0.001), and Cerasmart and teeth (p = 0.001). No statistically significant difference was found between the teeth and IPS e.max CAD (p = 0.999). The mean (± standard deviation) and median (mean value, 25th percentile-75th percentile) color change values of the CAD/CAM materials and tooth samples after dental biofilm was formed on them are shown in Table 3 . Table 3 ΔE values mean (mean, ± standard deviation) and median (mean value, 25th percentile-75th percentile) ΔE1 ΔE2 ΔE3 Mean ± Std dev Median Per(%25–75) Mean ± Std dev Median Per(%25–75) Mean ± Std dev Mean ± Std dev CERASMART G1 0,893 ± 0,634 0,641 (0,306-1,668) Aa 0,832 ± 0,332 0,749 (0,559-1,111) Aab 0,696 ± 0,509 0,566 (0,375-0,822) Aa G2 2,265 ± 0,780 2,132 (1,681-2,791) Ba 2,244 ± 0,843 1,990 (1,659-2,834) Ba 0,496 ± 0,231 0,538 (0,335-0,592) Aa G3 2,917 ± 0,438 2,928 (2,600-3,222) Ca 2,897 ± 0,412 2,953 (2,544-3,154) Ca 0,553 ± 0,218 0,559 (0,342-0,721) Aa G4 6,153 ± 1,365 6,185 (4,972-7,217) Da 6,214 ± 1,325 6,447 (4,922-7,368) Dab 0,479 ± 0,322 0,334 (0,227-0,729) Aa IPS e-max CAD G1 0,444 ± 0,103 0,427 (0,379-0,480) Aa 0,652 ± 0,167 0,648 (0,569-0,694) Ab 0,636 ± 0,165 0,606 (0,546-0,720) Aa G2 2,118 ± 1,019 2,050 (1,303-2,406) Ba 1,629 ± 0,599 1,619 (1,049 − 2,256) Ba 0,948 ± 1,282 0,335 (0,256-0,947) Aa G 3 2,722 ± 0,883 2,583 (2,279-2,893) Ca 3,009 ± 0,940 2,886 (2,410-3,321) Ca 0,432 ± 0,120 0,416 (0,326-0,498) Ba G 4 7,787 ± 4,563 7,716 (3,596 − 10,510) Da 7,863 ± 4,508 7,759 (3,767 − 10,667) Db 0,347 ± 0,217 0,284 (0,217-0,422) Aa TOOTH G1 3,575 ± 1,228 3,510 (2,407-4,926) Ab 1,567 ± 1,033 1,205 (0,703-2,246) Aa 3,725 ± 1,003 3,959 (2,631-4,618) Ab G 2 6,373 ± 5,492 4,015 (2,634-9,306) ABb 3,731 ± 5,076 1,543 (1,032 − 3,776) ABa 3,996 ± 2,594 3,058 (2,568-5,215) ACb G 3 2,683 ± 1,204 2,272 (1,710-3,682) Aa 1,496 ± 0,957 1,351 (0,636-2,591) Ab 2,680 ± 1,290 3,029 (1,627-3,547) Bb G 4 6,581 ± 4,634 5,580 (2,297-9,829) Ba 3,892 ± 2,026 3,213 (2,340-4,894) Ba 5,436 ± 2,080 5,212 (3,724-7,446) Cb While the different capital letters indicated indicate a statistically significant difference between the groups of the same material, the same capital letters indicate that there is no statistically significant difference between the groups. While the different lowercase letters indicated show a statistically significant difference between the same groups of materials, the same lowercase letters indicate that there is no statistically significant difference. The color values of the materials were recorded at the beginning, after the biofilm test, and after the brushing process. The color change value obtained after the initial biofilm test was ΔE1, the color change value obtained after the biofilm-brushing process was ΔE2, and the color change value obtained after the initial brushing process is expressed as ΔE3. In the biofilm formed on the teeth and Cerasmart and IPS e.max CAD/CAM materials, live biomass evaluation was made using the MTT method and the obtained data are shown in Table 4 . Table 4 Live Biomass Evaluation Material Live Biomass Evaluation Cerasmart Mean 0,883 Standard deviation ± 0,235 A Median 0,779 Percentile %25 0,748 Percentile %75 1,069 IPS e.max CAD Mean 0,352 Standard deviation ± 0,065 B Median 0,316 Percentile %25 0,306 Percentile %75 0,418 Tooth Mean 0,496 Standard deviation ± 0,054 AB Median 0,491 Percentile %25 0,447 Percentile %75 0,548 While the different capital letters indicated indicate a statistically significant difference within the same column, the same capital letters indicate that there is no statistically significant difference within the same column. Cerasmart had the lowest roughness measurement. Similar results were obtained in the teeth and IPS e.max CAD groups. The highest amount of viable biomass was observed in Cerasmart, and the lowest was in IPS e.max CAD. Group 4 had the highest ΔE1 value for all materials and group 1 had the lowest ΔE1 value. Brushing enabled the materials to return to their initial color or to reduce the color change in most groups. In the group 1 ΔE1 comparison between materials, the greatest color change was observed in the teeth, and the least in the IPS e.max CAD material. In the group 2 ΔE1 comparison between materials, the greatest color change was observed in the teeth, and the least was in the IPS e.max CAD material. Group 3 ΔE1 and group 4 ΔE1 comparisons between materials showed similar results. Discussion To obtain restorative materials with reduced secondary caries formation, materials should be evaluated from a microbiologic perspective [ 20 ]. (i) Our hypothesis that increasing the degree of roughness of CAD/CAM materials would not increase the amount of live biomass formed on the material was accepted because although Cerasmart material had the lowest roughness values, it showed the highest live biomass value; (ii) increasing the roughness degree of CAD/CAM materials would not cause more coloration of the material was accepted; (iii) the presence of biofilm on CAD/CAM materials would not cause further coloration of the materials by the effect of coloring beverages was rejected. In this study, CAD/CAM materials on which biofilm was formed were more colored. (iv) Brushing would not influence discoloration was rejected. To achieve an esthetically and functionally successful restoration, it is important to finish restorations with a smooth surface. Rough restoration surfaces can cause discoloration, gingival infections, plaque accumulation, and secondary caries development over time [ 21 ]. Quirynen et al. concluded that roughness took a more active role in the development of bacterial adhesion than other surface properties. It has been reported that an increase in surface roughness above a Ra value of 0.2 µm leads to a rise in in-vivo biofilm formation [ 22 ]. Bollen et al. showed that 0.2 µm Ra was a threshold value at which a decrease in bacterial adhesion was not expected [ 23 ]. Some studies also did not associate this factor with biofilm formation in dental materials [ 24 ], explaining that long-term incubation times reduce the importance of the roughness of the surface and bacterial adhesion on the existing biofilm instead of directly on the material surface [ 25 ]. Eick et al. showed that to investigate the adhesion of S. mutans in the biofilm formed after 48 hours on restorative materials, no correlation was found between bacterial adhesion and surface roughness [ 26 ]. In other studies, it was reported that the surface roughness threshold for plaque formation was Ra = 0.2 µm [ 27 , 28 ], the roughness value that the patient could feel with their tongue was Ra = 0.28 µm [ 29 ], and Ra < 1 µm was perceived as smooth, clinically [ 30 ]. Besides the polishing and finishing procedures, differences in the chemical composition of resin-based materials affect bacterial adhesion and biofilm formation [ 31 ]. These materials generally consist of a hydrophobic resin matrix containing hydrophilic fillers and an intermediate binding agent (silane) between the two phases. It should be considered that materials with such composition cannot form homogeneity, and topographic and chemical differences can be seen on the surface [ 32 ]. In our study, in line with the study of Wang et al. [ 32 ], it was observed that the composite-based material Cerasmart hybrid CAD/CAM material (although less roughness) caused more viable biomass formation than the IPS e.max CAD material in lithium disilicate glass-ceramic structure. Covering teeth and restorative materials with a salivary pellicle layer is the first step for bacterial colonization. Oral bacterias bind to host-derived receptors in the pellicle [ 33 ]. In our study, after forming a pellicle with artificial saliva on the samples, biofilm was formed on the pellicle layer, and biofilm biomass values ​​were obtained independent of the roughness, thus supporting the hypothesis that the surface roughness could not be transferred from the pellicle. On the other hand, it was shown that an increase in surface roughness, even in the presence of salivary pellicle, causes an increase in bacterial adhesion [ 34 ]. In light of these indicators, it can be thought that the physicochemical properties of the surfaces are partially balanced by the presence of saliva. The choice of culture medium depends on the type of inoculum, and the needs and purpose of the study. It has been noted that monocultures are more accurate for assessing specific physiologic aspects of biofilm. For monoculture biofilms, S. mutans has been the preferred microorganism [ 35 ]. S. mutans has been identified as one of the more cariogenic strains present in oral biofilms [ 36 ]. In the study, S. mutans and S. sanguinis were included because they are the main colonizers of hard surfaces in the oral cavity, functioning as a bridge between other bacteria that attach to and grow on the substrate surface. Like several other Streptococcal species, they participate in the initial colonization of tooth enamel [ 37 ]. In multi-culture-based models of maintaining clinical oral conditions, bacterial growth occurs in the presence of nutrient-limited media. In this case, bacterial growth is faster in the oral cavity and exhibits an entirely unnatural behavior [ 38 ]. This may explain why multicultural studies take a shorter time. Like some other studies using multi-culture media, a 48-hour biofilm evaluation was performed in our study [ 13 ]. To evaluate the pathogenicity of dental plaque, the amount of bacterial viability in the biofilm is important. In a study, it was determined that there was a correlation between the glass content of the materials and bacterial viability. It has been reported that the contents of ceramics affect bacterial viability during adhesion. It was determined that the number of S.oralis and S.sanguinis was low on tetragonal stabilized zircon, zirconia-reinforced glass ceramics, and glass alumina ceramics [ 39 ]. Again, in a study investigating bacterial viability, it was reported that although there was high bacterial accumulation in amalgam, the number of viable bacteria was low [ 40 ]. In our study, although the roughness rate was higher, a lower amount of live biomass was detected in the IPS e.max CAD compared with the Cerasmart material. The correct color selection is important for a successful restoration, restorative materials in the oral cavity are exposed to many factors such as coloring foods and beverages, saliva, dental plaque, and bacteria in the oral flora, which can cause a color change in restorations. In our study, when the color changes of CAD/CAM materials with and without biofilm were examined after they were kept in tea, the samples in the coloring beverage with biofilm were statistically significantly more colored. However, no significant difference was found between the tea groups with and without biofilms when the materials were examined in terms of initial and post-brushing ΔE3 values. Although there was no significant difference in the samples kept in water for the Cerasmart ΔE3 value, the samples kept in water without biofilm for IPS e.max CAD and dental samples were more colored than with biofilm. In a study, adsorption to the pellicle-like protein layer by black tea and red wine components formed in vitro by the adsorption of unstimulated saliva on hydroxyapatite discs was investigated. According to some studies, the pellicle quickly absorbs the components of black tea, and subsequent contact with saliva increases the adsorption of salivary components overall [ 41 ]. In our study, Cerasmart (6.153) and IPS e.max CAD (7.787) materials, which were first coated with a pellicle using artificial saliva and on which biofilm was formed, were Cerasmart (2.265) and IPS e.max CAD (2.118) materials without pellicle and kept in tea were statistically significantly more colored. Detectable and acceptable color change ΔE threshold values ​​were determined as 0.8 and 1.8, respectively, by Paravina et al. [ 42 ] who evaluated changes in color and roughness properties of resin composites exposed to aging in water or red wine and brushed. Color and roughness measurements were made of the samples on the 5th, 10th, and 15th day. No significant color change was observed in the groups that were kept in water and brushed according to the detectable ΔE > 0.8 threshold value. Less color change was observed in the brushed groups compared to the non-brushing groups. They concluded that short-term brushing reduced color change and did not increase roughness, although the samples deteriorated significantly with red wine and brushing [ 43 ]. In our study, the fact that the brushing process mostly returns the initial colors of the materials supports this result. Conclusion The adhesion of biofilm to restorative dental materials plays an important role in the coloring of dental treatment. The presence of biofilm on CAD/CAM materials immersed in water caused an unacceptable degree of coloration (ΔE > 1.8), and immersion in tea caused a greater color change (ΔE < 0.8). The brushing process allowed CAD/CAM materials to return to their original color. Although there is no consensus on the selection of materials showing the minimum coloring and biofilm adhesion among restorative materials, in our study, the IPS e.max CAD group had less biofilm formation than Cerasmart group. Therefore, IPS e.max can be preferred over Cerasmart. However, there is no difference in the coloring process between the two materials with the formation of biofilm. Further studies are needed to understand these materials. Declarations Conflict of interest The authors declare no competing interests. Ethical approval This study was approved by the Ethics Committee of Kocaeli University (KOU KAEK 2021/222). Funding This work was supported by the writers. Author Contribution: Dilan Kopuz has been written this article, Sümeyra Topçu, Eda Yazıcı Özcelik, Fetiye Kolaylı, Neslihan Tekçe, Mustafa Demirci and Safa Tuncer has been done this research and conducted this manuscript. Informed consent: All participants were freely invited, and those who accepted signed an informed consent approved and stamped by the local ethics committee. References Pflughoeft KJ, Versalovic J. Human microbiome in health and disease. Annu Rev Pathol. 2012; 7:99–122. Montanaro L, Campoccia D, Rizzi S, et al. 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Clin Oral Implants Res. 2015; 26:1297–1301. Ionescu A, Brambilla E, Schneider-feyrer S, et al: Influence of surface properties of resin-based composites on in vitro Streptococcus mutans biofilm development. Eur J Oral Sci. 2012; 120:458–465. Ertas E, Güler AU, Yücel AC, et al. Color stability of resin composites after immersion in different drinks. Dent Mater. 2006; 25:361–6. Schlueter N, Glatzki J, Klimek J, et al. Erosive-abrasive tissue loss in dentine under simulated bulimic conditions. Arch Oral Biol. 2012; 57:1176–1182. Shellis RP, Ganss C, Ren Y, et al. Methodology and models in erosion research: discussion and conclusions. Caries Res. 2011; 45:69–77. Carter K, Landini G, Walmsley AD. Plaque removal characteristics of electric toothbrushes using an in vitro plaque model. J Clin Periodontol 2001; 28:1045–9. Murat S, Alp G, Alatalı C, et al: In vitro evaluation of adhesion of Candida albicans on CAD/CAM PMMA-based polymers. J Prosthodont. 2019; 28:873–9. Mota EG, Smidt LN, Fracasso LM, et al. The effect of milling and postmilling procedures on the surface roughness of CAD/CAM materials. J Esthet Restor Dent. 2017; 29:450–8. Quirynen M, Marechal M, Busscher HJ, et al: The influence of surface free energy and surface roughness on early plaque formation. An in vivo study in man. J Clin Periodontol. 1990;17:138–144. Bollen CM, Lambrechts P, Quirynen M: Comparison of surface roughness of oral hard materials to the threshold surface roughness for bacterial plaque retention: a review of the literature. Dent Mater. 1997; 13(4):258–69. Teughels W, Van Assche N, Sliepen I, et al: Effect of material characteristics and/or surface topography on biofilm development. Clin Oral Implants Res. 2006; 17:68–81. Dezelic T, Schmidlin PR: Multi-species biofilm formation on dental materials and an adhesive patch. Oral Health Prev Dent. 2009; 7:47–53. Eick S, Glockmann E, Brandl B, et al: Adherence of Streptococcus mutans to various restorative materials in a continuous flow system. J Oral Rehabil. 2004; 31:278–285. Antonson SA, Yazici AR, Kilinc E, et al: Comparison of different finishing/polishing systems on surface roughness and gloss of resin composites. J Dent. 2011; 39:9–17. Mörmann WH, Stawarczyk B, Ender A, et al: Wear characteristics of current aesthetic dental restorative CAD/CAM materials: two-body wear, gloss retention, roughness and Martens hardness. J Mech Behav Biomed Mater. 2013; 20:113–125. Jones CS, Billington RW, Pearson GJ. The in vivo perception of roughness of restorations. Br Dent J. 2004; 196:42–5. Ereifej NS, Oweis YG, Eliades G. The effect of polishing technique on 3-D surface roughness and gloss of dental restorative resin composites. Oper Dent. 2012; 38:9–20. Ono M, Nikaido T, Ikeda M, et al. Surface Properties of Resin Composite Materials Relative to Biofilm Formation. Dent Mater J. 2007; 26:613–22. Wang Z, Shen Y, Haapasalo M. Dental materials with antibiofilm properties. Dent Mater. 2014; 30:1–16. Ikeda M, Matin K, Nikaido T, et al. Effect of surface characteristics on adherence of S. mutans biofilms to indirect resin composites. Dent Mater J. 2007; 26:915–923. Carlén A, Nikdel K, Wennerberg A, et al. Surface characteristics and in vitro biofilm formation on glass ionomer and composite resin. Biomaterials. 2001; 22:481–7. Totiam P, González-Cabezas C, Fontana MR, et al. A new in vitro model to study the relationship of gap size and secondary caries. Caries Res. 2007; 41:467–473. Yoo SY, Park SJ, Jeong DK, et al. Isolation and characterization of the mutans streptococci from the dental plaques in Koreans. J Microbiol. 2007; 45:246–255. Nyvad B, Kilian M. Comparison of the initial streptococcal microflora on dental enamel in caries-active and in caries-inactive individuals. Caries Res. 1990; 24:267–272. McBain AJ. In vitro biofilm models: an overview. Adv Appl Microbiol. 2009; 69:99–132. Meier R, Hauser-Gerspach I, Lüthy H, et al. Adhesion of oral streptococci to all-ceramics dental restorative materials in vitro. J Mater Sci Mater Med. 2008; 19:3249–3253. Steinberg D, Eyal S. Early formation of Streptococcus sobrinus biofilm on various dental restorative materials. J Dent. 2002; 30:47–51. Joiner A, Muller D, Elofsson UM, et al. Adsorption from black tea and red wine onto in vitro salivary pellicles studied by ellipsometry. Eur J Oral Sci. 2003; 111:417–422. Paravina RD, Ghinea R, Herrera LJ, et al. Color difference thresholds in dentistry. J Esthet Restor Dent 2015; 27:1–9. Mozzaquatro LR, Rodrigues CS, Kaizer MR, et al. The Effect of Brushing and Aging on the Staining and Smoothness of Resin Composites. J Esthet Restor Dent. 2017; 29:44–55. Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-3221496","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":223385617,"identity":"6de42958-7bbf-4401-ac3d-40afd965655e","order_by":0,"name":"Sümeyra Topçu","email":"","orcid":"","institution":"Kocaeli University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Sümeyra","middleName":"","lastName":"Topçu","suffix":""},{"id":223385618,"identity":"cbb232a4-f284-4dd9-b8ef-dad2bb582cbd","order_by":1,"name":"Eda Yazıcı Özcelik","email":"","orcid":"","institution":"Kocaeli University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Eda","middleName":"Yazıcı","lastName":"Özcelik","suffix":""},{"id":223385619,"identity":"f3fdbd57-8e06-459a-8214-50b589d2c428","order_by":2,"name":"Dilan Kopuz","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA+ElEQVRIiWNgGAWjYDCCw8wNQDIBxGR82AARMyCghRGuhdkQyJSAaEnAo+UAQgubJFFa+I4zNn4uYEiTM5dIf1Y5o+JOHQN78zYJxh/3cGqRPMzYLD2DIcfYckZC2s0NZ55JMPAcK5NgSCjGqcUA6BdpHoaKxA03Eo7dfNh2WIJBIscMqAW3y4Bamn9DtCS2FT78B9Qi/4agljagLTlALclsjBsbQLbw4NcC9EubNY9BmrFlzzNmyRnHDku28aQVWySk4dbCd/7w4ds8Fcly5uzpDz/21Bzm52c/vPHGBxvcWqDOQ4pxNhBBSANM1ygYBaNgFIwC7AAAJ01RCCCTA+UAAAAASUVORK5CYII=","orcid":"","institution":"Istanbul Kent University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Dilan","middleName":"","lastName":"Kopuz","suffix":""},{"id":223385620,"identity":"94a9af9b-804b-45f5-838d-d05443ebae44","order_by":3,"name":"Fetiye Kolaylı","email":"","orcid":"","institution":"Kocaeli University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Fetiye","middleName":"","lastName":"Kolaylı","suffix":""},{"id":223385621,"identity":"f711df5b-cd27-4098-b5ee-3638dffe8391","order_by":4,"name":"Neslihan Tekçe","email":"","orcid":"","institution":"Kocaeli University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Neslihan","middleName":"","lastName":"Tekçe","suffix":""},{"id":223385622,"identity":"3993140e-3631-43ff-a6df-29fad95854ff","order_by":5,"name":"Mustafa Demirci","email":"","orcid":"","institution":"Istanbul University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Mustafa","middleName":"","lastName":"Demirci","suffix":""},{"id":223385623,"identity":"bf35857a-fc67-4a87-8d54-4287e45b973f","order_by":6,"name":"Safa Tuncer","email":"","orcid":"","institution":"Istanbul University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Safa","middleName":"","lastName":"Tuncer","suffix":""}],"badges":[],"createdAt":"2023-07-31 16:29:25","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3221496/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3221496/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":41197530,"identity":"621c261a-a799-46f5-9ec6-067c47fb8917","added_by":"auto","created_at":"2023-08-07 16:21:14","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":198381,"visible":true,"origin":"","legend":"\u003cp\u003eDistribution of materials in the subgroups\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-3221496/v1/c01a8f0fa03add9bb5bb361b.png"},{"id":41197531,"identity":"40a255f6-5b4e-416b-833a-d1d41dd57c64","added_by":"auto","created_at":"2023-08-07 16:21:15","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":789684,"visible":true,"origin":"","legend":"\u003cp\u003eThree-dimensional images of the samples\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-3221496/v1/05238616edc2f846d21b2884.png"},{"id":41598366,"identity":"4479443d-e61e-42c9-8c5a-0c977b9e7a7a","added_by":"auto","created_at":"2023-08-15 20:52:20","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1303356,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3221496/v1/fc8dc5cb-daa5-4871-9718-a17a2b27b5bd.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Effect of Biofilm Formation on the Color Properties of CAD/CAM Materials","fulltext":[{"header":"Introduction","content":"\u003cp\u003eBiofilms that are in equilibrium with the host under healthy conditions are complex ecosystems. As a result of changes in this balance, bacterial colonization and biofilm formation on teeth may cause dental plaque, then this process causes enamel demineralization, dental caries, periodontitis, or many diseases [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Adhesion is related to the material's surface structure and its surface roughness, which is the most important component in determining the quantity of plaque accumulation [\u003cspan additionalcitationids=\"CR3\" citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIt is very important to finish restorations with a smooth surface to achieve an esthetically and functionally successful restoration [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Cracks, grooves, and worn surfaces often lead to greater surface roughness, creating areas suitable for bacterial adhesion [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. These surface irregularities protect bacteria from shear forces in the oral cavity and cause bacteria to bind more strongly to the substrate [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. For these reasons, the choice of material for biofilm formation is important in restorative material selection.\u003c/p\u003e \u003cp\u003eThe preservation of color stability of the material is also a very important factor in evaluating the success of a restoration. Teeth and restoration materials are exposed to many factors such as biofilm, acidic by-products formed in the biofilm, and coloring foods and beverages [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. The results of in vitro testing of materials exposed to these conditions can be used to predict the clinical performance of restorations regarding longevity and esthetic results. Our study aimed to investigate the color changes that occurred in CAD/CAM materials and human teeth by forming a biofilm layer on the CAD/CAM materials used in the restoration of teeth and to what extent this color change was reversed.\u003c/p\u003e \u003cp\u003eThe null hypotheses created in the study:\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eIncreasing the degree of roughness of CAD/CAM materials does not increase the amount of live biomass formed on the material.\u003c/p\u003e\u003cp\u003eIncreasing the degree of roughness of CAD/CAM materials does not cause more coloration of the material.\u003c/p\u003e\u003cp\u003eThe presence of biofilm on CAD/CAM materials does not cause further coloration of the materials by the effect of coloring beverages.\u003c/p\u003e\u003cp\u003eBrushing would not influence discoloration.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cp\u003e106 CAD/CAM block sections of 2 mm thickness were prepared, Cerasmart (A2 HT, GC Corporation, Tokyo, Japan) (n\u0026thinsp;=\u0026thinsp;53) and IPS e.max CAD (A2 HT, Ivoclar Vivadent, Schaan, Liechtenstein) (n\u0026thinsp;=\u0026thinsp;53), and 53 extracted caries-free third-molar human teeth were collected for testing. The materials used in our study are given in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMaterials used in the study\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMaterial\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eContents\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eManufacturer\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eLOT\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCerasmart CAD/CAM Bloc\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e71% silica and barium glass nanoparticles, 20 nm silica, 300 nm barium glass nanoparticles, 29% Bis-MEPP, UDMA and DMA polymers\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eGC Corporation, Tokyo, Japan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2012151\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eIPS e.max CAD CAD/CAM Bloc\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(57\u0026ndash;80%) SiO\u003csub\u003e2\u003c/sub\u003e, (11\u0026ndash;19%) Li\u003csub\u003e2\u003c/sub\u003eO, (0\u0026ndash;13%) K\u003csub\u003e2\u003c/sub\u003eO, (0\u0026ndash;11%) P\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e, (0\u0026ndash;8%) ZrO\u003csub\u003e2\u003c/sub\u003e, (0\u0026ndash;8%) ZnO, (% 0\u0026ndash;12) other oxides\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eIvoclar Vivadent, Schaan, Liechtenstein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eZ00FTN\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"4\" nameend=\"c4\" namest=\"c1\"\u003e \u003cp\u003e\u003cem\u003eAbbreviations\u003c/em\u003e: Bis-MEPP: 2,2-Bis (4-methacryloxy polyethoxy phenyl) propane, UDMA: Urethane Dimethacrylate, DMA: Dimethacrylate, SiO\u003csub\u003e2\u003c/sub\u003e: Silicon Dioxide, Li\u003csub\u003e2\u003c/sub\u003eO: Lithium oxide, K\u003csub\u003e2\u003c/sub\u003eO: Potassium oxide, P\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e5\u003c/sub\u003e: Diphosphorus penta oxide, ZrO\u003csub\u003e2\u003c/sub\u003e: Zirconium oxide, ZnO: Zinc oxide\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eFive samples from each group (IPS e.max CAD, Cerasmart, teeth) were separated for the measurement of live biomass in the biofilm. The remaining 48 samples from each group were divided into 4 experimental subgroups (n\u0026thinsp;=\u0026thinsp;12 in each): group 1, kept in water without forming a dental biofilm; group 2, kept in tea without forming a dental biofilm; group 3, kept in water after the dental biofilm was formed; group 4, kept in the tea after the dental biofilm was formed (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe surfaces of the samples were sanded for 60 seconds with 600, 800, and 1000-grit silicon carbide (SIC) sandpaper to provide standardization before the polishing process.\u003c/p\u003e \u003cp\u003eAccording to the manufacturer's instructions, the IPS e.max CAD blocks were fired to crystallize and were glazed in the Ivoclar Programat P300 oven. Cerasmart blocks were finished and polished using a two-stage Sof-Lex Diamond Polishing system with a slow-speed micromotor running at 15,000\u0026ndash;20,000 rpm. A beige-colored disc was used for pre-polishing, and a pink-colored disc was used for polishing. These procedures were performed underwater for 15 seconds. The samples were cleaned using deionized water in an ultrasonic cleaner for 10 minutes following the completion of the surface treatments.\u003c/p\u003e \u003cp\u003eAfter surface finishing applications were completed, roughness measurements of all samples were performed using a Nanovea 3D Non-Contact Profilometer PS50 (Nanovea, 6 Morgan Ste 156, Irvine, CA, USA). The purpose of this measurement was to compare the effect of surface roughness on surface discoloration.\u003c/p\u003e \u003cp\u003eColor measurements of the samples were performed using a VITA Easyshade Compact device (VITA Zahnfabrik, Bad S\u0026auml;ckingen, Germany). Each measurement began with the calibration of the device. The measurements were taken on a standard white background and under constant light for the elimination of the background effect [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. During the measurements, the fiber optic end of the device was held perpendicular to the samples and parallel to the ground. Each measurement was repeated three times and the average of L, a, b, C, and H values ​​were recorded. A CIEDE2000 system (ΔE\u003csub\u003e00\u003c/sub\u003e) was used to calculate the color change. Then all samples were sterilized using an ethylene oxide gas sterilization system (Steris Amsco Eagle, Mentor, AND).\u003c/p\u003e \u003cp\u003eTo ensure bacterial attachment and to create a pellicle layer on the samples, artificial saliva was created. For 2 liters of artificial saliva, 8.4 mg NaF, 2560 mg NaCl, 332.97 mg CaCl\u003csub\u003e2\u003c/sub\u003e, 250.00 mg MgCl\u003csub\u003e2\u003c/sub\u003e (6H\u003csub\u003e2\u003c/sub\u003eO), 189.48 mg KCl, 0.1 mL H\u003csub\u003e3\u003c/sub\u003ePO\u003csub\u003e4\u003c/sub\u003e (85%), and 0.1 mmol NaOH were used. All ingredients were mixed and the pH of the resulting mixture was measured using a pH meter. To adjust the pH of the mixture between 6.5-7, 30 mL of 0.1 M NaOH was added [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. The mixture was sterilized by passing a 0.22 \u0026micro;m syringe tip through a PES filter (BIOSORFA, China) and then 140 mg of Type II mucin (Sigma-Aldrich Chemie GmbH, Deisenhofen, Germany) was added under aseptic conditions at a rate of 140 mg per 100 mL.\u003c/p\u003e \u003cp\u003eA biofilm culture medium was made for the bacteria Streptococcus mutans (S. mutans) HF76, Streptococcus sanguinis (S. sanguinis) ATCC 10556, and Candida albicans (C. albicans) ATCC 90028 strains. S. mutans and S. sanguinis strains were removed from the freezer and inoculated on 5% sheep blood agar (SBA) medium and incubated at 37\u0026deg;C in 5% CO\u003csub\u003e2\u003c/sub\u003e. C. albicans were seeded on Sabouraud dextrose agar (SDA) medium and incubated at 37\u0026deg;C for 24 hours. After incubation, S. mutans, S. sangiunis, and C. albicans were subcultured. The subcultured tubes were centrifuged at 3000 \u003cem\u003eg\u003c/em\u003e at 19\u0026deg;C for 5 minutes and the supernatant was discarded. Then, sterile phosphate-buffered saline (PBS) (pH 7.0) was added and washed, and this process was repeated twice. To form a biofilm, a bacterial density of 0.5 McFarland (1.5x108 cfu/mL) S. mutans and S. sangiunis was prepared by suspending brain heart infusion (BHI) broth with 3% sucrose, 0.5 McFarland C. albicans SDB [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn the biofilm formation phase of the study, e.max CAD (n\u0026thinsp;=\u0026thinsp;29) and Cerasmart (n\u0026thinsp;=\u0026thinsp;29) CAD/CAM blocks, and caries-free third-molar human teeth (n\u0026thinsp;=\u0026thinsp;29) which previously sterilized using ethylene oxide were used. Sterile CAD/CAM samples were placed in 24-well plates and dental samples were placed in tubes and incubated at 37\u0026deg;C for 1 hour by adding artificial saliva until the material was completely covered to ensure pellicle formation. After incubation, artificial saliva was removed from the environment. For 48 hours, the CAD/CAM samples and dental samples were incubated at 37\u0026deg;C in an environment containing 5% CO\u003csub\u003e2\u003c/sub\u003e by inoculating equal amounts of the prepared bacterial suspension and yeast suspension on plates [\u003cspan additionalcitationids=\"CR13\" citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eMeasurement of biofilm mass through Live Biomass Assessment (MTT method) was performed with reference to previous studies [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eCAD/CAM block samples on which biofilm was formed were placed in new 24-well plates and the teeth were placed in new tubes and washed 3 times with PBS (VWR USA) to remove microorganisms that were not in the biofilm structure. Two different solutions, distilled water and tea, were prepared for coloring the samples. Two tea bags (Yellow Label Tea, Lipton, Rize, Turkey) (2 x 2 g) were dissolved in 300 mL of boiling distilled water for 10 minutes to create the tea solution.\u003c/p\u003e \u003cp\u003eTea was added to half (n\u0026thinsp;=\u0026thinsp;12) of the biofilm-formed CAD/CAM samples and teeth until they were completely covered, and water was added to the other half. The control group CAD/CAM samples, on which no biofilm was formed, were placed in 24-well plates, and teeth were placed in tubes; tea was added to half of the samples (n\u0026thinsp;=\u0026thinsp;12) until they were completely covered, and distilled water was added to the other half, and all samples were kept at 37℃ for 24 hours [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. After the samples were kept in solutions for 24 hours, second color measurements were performed.\u003c/p\u003e \u003cp\u003eTo remove the plaque on the samples, a rechargeable Braun Oral B Ultra Plaque Remover toothbrush (Braun D9, Braun AG, Kronberg, Germany) was used and fixed to a stand to simulate brushing under 2 N load, according to ISO 14569-1 [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. The experiment samples were brushed under a 2 N load for 10 seconds, using toothpaste (ProNamel, Sensodyne) containing 1450 ppm sodium fluoride with a relative dentin abrasivity (RDA) value of 34, and a slurry prepared by mixing homogeneously with distilled water in a 1:1 ratio [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. After brushing, the samples were washed in distilled water and cleaned, then color measurements were made again.\u003c/p\u003e \u003cp\u003eThe IBM SPSS 20.0 (IBM Corp., Armonk, NY, USA) package program was used for statistical evaluation. Differences between groups were examined using the independent sample t-test and one-way analysis of variance (ANOVA) for normally distributed variables, and the Mann-Whitney U and Kruskal-Wallis tests for non-normally distributed data. Tukey and Dunn tests were used for multiple comparisons (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eAfter applying polishing procedures, the initial surface roughness of the tooth, Cerasmart, IPS e.max CAD materials were measured, and Ra, Rz, and Sa values were compared. The values are shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. Three-dimensional images of the samples are shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eSurface roughness values (Ra, Rz, Sa \u0026micro;m) (median values, 25\u0026ndash;75% percentile, mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"13\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMaterial\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003eRa\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c9\" namest=\"c6\"\u003e \u003cp\u003eRz\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c13\" namest=\"c10\"\u003e \u003cp\u003eSa\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eCerasmart\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eMedian\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0,132\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eMedian\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0,729\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e \u003cp\u003eMedian\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0,353\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eF\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePercentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e%25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0,120\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ePercentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e%25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0,635\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003ePercentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e%25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0,319\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e%75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0,145\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e%75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0,790\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e%75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0,497\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0,181\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0,700\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0,402\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eStandard deviation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026plusmn;\u0026thinsp;0,186\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eStandard deviation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026plusmn;\u0026thinsp;0,099\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e \u003cp\u003eStandard deviation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u0026plusmn;\u0026thinsp;0,108\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eIPS e.max CAD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eMedian\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0,410\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eMedian\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2,102\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e \u003cp\u003eMedian\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e4,122\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePercentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e%25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0,302\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ePercentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e%25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1,637\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003ePercentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e%25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e2,960\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e%75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0,592\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e%75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2,918\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e%75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e5,286\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0,427\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2,209\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e4,827\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eStandard deviation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026plusmn;\u0026thinsp;0,141\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eStandard deviation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026plusmn;\u0026thinsp;0,732\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e \u003cp\u003eStandard deviation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u0026plusmn;\u0026thinsp;2,623\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eTooth\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eMedian\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0,233\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eAB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eMedian\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0,767\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e \u003cp\u003eMedian\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e4,441\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eG\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePercentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e%25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0,166\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003ePercentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e%25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0,595\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003ePercentile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e%25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e3,792\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e%75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0,298\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e%75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1,116\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e%75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e6,181\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0,238\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0,838\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e4,811\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eStandard deviation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026plusmn;\u0026thinsp;0,080\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eStandard deviation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026plusmn;\u0026thinsp;0,261\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c11\" namest=\"c10\"\u003e \u003cp\u003eStandard deviation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u0026plusmn;\u0026thinsp;1,553\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"13\" nameend=\"c13\" namest=\"c1\"\u003e \u003cp\u003eWhile the different capital letters indicated indicate a statistically significant difference within the same column, the same capital letters indicate that there is no statistically significant difference within the same column.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eWhen the Ra values ​​of the samples were examined, the median values ​​of Cerasmart, IPS e.max CAD, and tooth groups were 0.132, 0.410, and 0.233, respectively. Accordingly, the lowest roughness was observed in Cerasmart samples, and the highest roughness values ​​were observed in IPS e.max CAD samples. In the pairwise comparison of the groups, there was a significant difference between Cerasmart-IPS e.max CAD (p\u0026thinsp;=\u0026thinsp;0.001). No statistically significant difference was found between the Cerasmart blocks and teeth (p\u0026thinsp;=\u0026thinsp;0.067) and the teeth and IPS e.max CAD blocks (p\u0026thinsp;=\u0026thinsp;0.078).\u003c/p\u003e \u003cp\u003eWhen the Rz values ​​of the groups were examined, the median values ​​of Cerasmart, IPS e.max CAD, and the teeth were 0.729, 2.102, and 0.767, respectively. Accordingly, the lowest roughness was observed in Cerasmart samples, and the highest roughness values ​​were observed in IPS e.max CAD samples. In the pairwise comparison of the groups, there was a significant difference between Cerasmart-IPS e.max CAD (p\u0026thinsp;=\u0026thinsp;0.001) and dental-IPS e.max CAD (p\u0026thinsp;=\u0026thinsp;0.001). No statistically significant difference was found between the Cerasmart samples and teeth (p\u0026thinsp;=\u0026thinsp;0.737).\u003c/p\u003e \u003cp\u003eWhen the Sa values ​​of the groups were examined, the median values ​​of Cerasmart, IPS e.max CAD, and tooth groups were 0.353, 4.122, and 4.441, respectively. Accordingly, the lowest roughness was observed in Cerasmart samples, and the highest roughness values ​​were observed in the teeth. In the pairwise comparison of the groups, there was a significant difference between Cerasmart and IPS e.max CAD (p\u0026thinsp;=\u0026thinsp;0.001), and Cerasmart and teeth (p\u0026thinsp;=\u0026thinsp;0.001). No statistically significant difference was found between the teeth and IPS e.max CAD (p\u0026thinsp;=\u0026thinsp;0.999).\u003c/p\u003e \u003cp\u003eThe mean (\u0026plusmn;\u0026thinsp;standard deviation) and median (mean value, 25th percentile-75th percentile) color change values of the CAD/CAM materials and tooth samples after dental biofilm was formed on them are shown in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eΔE values mean (mean, \u0026plusmn; standard deviation) and median (mean value, 25th percentile-75th percentile)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"11\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" morerows=\"1\" nameend=\"c2\" namest=\"c1\" rowspan=\"2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c5\" namest=\"c3\"\u003e \u003cp\u003eΔE1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c8\" namest=\"c6\"\u003e \u003cp\u003eΔE2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c11\" namest=\"c9\"\u003e \u003cp\u003eΔE3\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003cp\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;Std dev\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMedian\u003c/p\u003e \u003cp\u003ePer(%25\u0026ndash;75)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003cp\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;Std dev\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMedian\u003c/p\u003e \u003cp\u003ePer(%25\u0026ndash;75)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003cp\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;Std dev\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003cp\u003e\u0026thinsp;\u0026plusmn;\u0026thinsp;Std dev\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003e\u003cb\u003eCERASMART\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eG1\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0,893\u0026thinsp;\u0026plusmn;\u0026thinsp;0,634\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0,641\u003c/p\u003e \u003cp\u003e(0,306-1,668)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0,832\u0026thinsp;\u0026plusmn;\u0026thinsp;0,332\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0,749\u003c/p\u003e \u003cp\u003e(0,559-1,111)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0,696\u0026thinsp;\u0026plusmn;\u0026thinsp;0,509\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0,566\u003c/p\u003e \u003cp\u003e(0,375-0,822)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eAa\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eG2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2,265\u0026thinsp;\u0026plusmn;\u0026thinsp;0,780\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2,132\u003c/p\u003e \u003cp\u003e(1,681-2,791)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eBa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2,244\u0026thinsp;\u0026plusmn;\u0026thinsp;0,843\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1,990\u003c/p\u003e \u003cp\u003e(1,659-2,834)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eBa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0,496\u0026thinsp;\u0026plusmn;\u0026thinsp;0,231\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0,538\u003c/p\u003e \u003cp\u003e(0,335-0,592)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eAa\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eG3\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2,917\u0026thinsp;\u0026plusmn;\u0026thinsp;0,438\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2,928\u003c/p\u003e \u003cp\u003e(2,600-3,222)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eCa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2,897\u0026thinsp;\u0026plusmn;\u0026thinsp;0,412\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2,953\u003c/p\u003e \u003cp\u003e(2,544-3,154)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eCa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0,553\u0026thinsp;\u0026plusmn;\u0026thinsp;0,218\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0,559\u003c/p\u003e \u003cp\u003e(0,342-0,721)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eAa\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eG4\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6,153\u0026thinsp;\u0026plusmn;\u0026thinsp;1,365\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6,185\u003c/p\u003e \u003cp\u003e(4,972-7,217)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eDa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6,214\u0026thinsp;\u0026plusmn;\u0026thinsp;1,325\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6,447\u003c/p\u003e \u003cp\u003e(4,922-7,368)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eDab\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0,479\u0026thinsp;\u0026plusmn;\u0026thinsp;0,322\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0,334\u003c/p\u003e \u003cp\u003e(0,227-0,729)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eAa\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003e\u003cb\u003eIPS e-max CAD\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eG1\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0,444\u0026thinsp;\u0026plusmn;\u0026thinsp;0,103\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0,427\u003c/p\u003e \u003cp\u003e(0,379-0,480)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0,652\u0026thinsp;\u0026plusmn;\u0026thinsp;0,167\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0,648\u003c/p\u003e \u003cp\u003e(0,569-0,694)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAb\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0,636\u0026thinsp;\u0026plusmn;\u0026thinsp;0,165\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0,606\u003c/p\u003e \u003cp\u003e(0,546-0,720)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eAa\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eG2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2,118\u0026thinsp;\u0026plusmn;\u0026thinsp;1,019\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2,050\u003c/p\u003e \u003cp\u003e(1,303-2,406)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eBa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1,629\u0026thinsp;\u0026plusmn;\u0026thinsp;0,599\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1,619\u003c/p\u003e \u003cp\u003e(1,049\u0026thinsp;\u0026minus;\u0026thinsp;2,256)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eBa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0,948\u0026thinsp;\u0026plusmn;\u0026thinsp;1,282\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0,335\u003c/p\u003e \u003cp\u003e(0,256-0,947)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eAa\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eG 3\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2,722\u0026thinsp;\u0026plusmn;\u0026thinsp;0,883\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2,583\u003c/p\u003e \u003cp\u003e(2,279-2,893)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eCa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3,009\u0026thinsp;\u0026plusmn;\u0026thinsp;0,940\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2,886\u003c/p\u003e \u003cp\u003e(2,410-3,321)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eCa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0,432\u0026thinsp;\u0026plusmn;\u0026thinsp;0,120\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0,416\u003c/p\u003e \u003cp\u003e(0,326-0,498)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eBa\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eG 4\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7,787\u0026thinsp;\u0026plusmn;\u0026thinsp;4,563\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e7,716 (3,596\u0026thinsp;\u0026minus;\u0026thinsp;10,510)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eDa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7,863\u0026thinsp;\u0026plusmn;\u0026thinsp;4,508\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e7,759\u003c/p\u003e \u003cp\u003e(3,767\u0026thinsp;\u0026minus;\u0026thinsp;10,667)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eDb\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0,347\u0026thinsp;\u0026plusmn;\u0026thinsp;0,217\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0,284\u003c/p\u003e \u003cp\u003e(0,217-0,422)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eAa\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003e\u003cb\u003eTOOTH\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eG1\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3,575\u0026thinsp;\u0026plusmn;\u0026thinsp;1,228\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3,510 (2,407-4,926)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAb\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1,567\u0026thinsp;\u0026plusmn;\u0026thinsp;1,033\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1,205\u003c/p\u003e \u003cp\u003e(0,703-2,246)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3,725\u0026thinsp;\u0026plusmn;\u0026thinsp;1,003\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e3,959\u003c/p\u003e \u003cp\u003e(2,631-4,618)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eAb\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eG 2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6,373\u0026thinsp;\u0026plusmn;\u0026thinsp;5,492\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4,015\u003c/p\u003e \u003cp\u003e(2,634-9,306)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eABb\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3,731\u0026thinsp;\u0026plusmn;\u0026thinsp;5,076\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1,543\u003c/p\u003e \u003cp\u003e(1,032\u0026thinsp;\u0026minus;\u0026thinsp;3,776)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eABa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3,996\u0026thinsp;\u0026plusmn;\u0026thinsp;2,594\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e3,058\u003c/p\u003e \u003cp\u003e(2,568-5,215)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eACb\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eG 3\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2,683\u0026thinsp;\u0026plusmn;\u0026thinsp;1,204\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2,272\u003c/p\u003e \u003cp\u003e(1,710-3,682)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1,496\u0026thinsp;\u0026plusmn;\u0026thinsp;0,957\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1,351\u003c/p\u003e \u003cp\u003e(0,636-2,591)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAb\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e2,680\u0026thinsp;\u0026plusmn;\u0026thinsp;1,290\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e3,029\u003c/p\u003e \u003cp\u003e(1,627-3,547)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eBb\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eG 4\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6,581\u0026thinsp;\u0026plusmn;\u0026thinsp;4,634\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5,580\u003c/p\u003e \u003cp\u003e(2,297-9,829)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eBa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3,892\u0026thinsp;\u0026plusmn;\u0026thinsp;2,026\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3,213\u003c/p\u003e \u003cp\u003e(2,340-4,894)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eBa\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e5,436\u0026thinsp;\u0026plusmn;\u0026thinsp;2,080\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e5,212\u003c/p\u003e \u003cp\u003e(3,724-7,446)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eCb\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"11\" nameend=\"c11\" namest=\"c1\"\u003e \u003cp\u003eWhile the different capital letters indicated indicate a statistically significant difference between the groups of the same material, the same capital letters indicate that there is no statistically significant difference between the groups. While the different lowercase letters indicated show a statistically significant difference between the same groups of materials, the same lowercase letters indicate that there is no statistically significant difference.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe color values of the materials were recorded at the beginning, after the biofilm test, and after the brushing process. The color change value obtained after the initial biofilm test was ΔE1, the color change value obtained after the biofilm-brushing process was ΔE2, and the color change value obtained after the initial brushing process is expressed as ΔE3.\u003c/p\u003e \u003cp\u003eIn the biofilm formed on the teeth and Cerasmart and IPS e.max CAD/CAM materials, live biomass evaluation was made using the MTT method and the obtained data are shown in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eLive Biomass Evaluation\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMaterial\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"5\" nameend=\"c6\" namest=\"c2\"\u003e \u003cp\u003eLive Biomass Evaluation\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eCerasmart\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0,883\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eStandard deviation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026plusmn;\u0026thinsp;0,235\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eA\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eMedian\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0,779\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePercentile %25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0,748\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePercentile %75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1,069\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eIPS e.max CAD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0,352\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eStandard deviation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026plusmn;\u0026thinsp;0,065\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eB\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eMedian\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0,316\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePercentile %25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0,306\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePercentile %75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0,418\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eTooth\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0,496\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eStandard deviation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026plusmn;\u0026thinsp;0,054\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eAB\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eMedian\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0,491\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePercentile %25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0,447\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePercentile %75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0,548\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"6\" nameend=\"c6\" namest=\"c1\"\u003e \u003cp\u003eWhile the different capital letters indicated indicate a statistically significant difference within the same column, the same capital letters indicate that there is no statistically significant difference within the same column.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eCerasmart had the lowest roughness measurement. Similar results were obtained in the teeth and IPS e.max CAD groups. The highest amount of viable biomass was observed in Cerasmart, and the lowest was in IPS e.max CAD. Group 4 had the highest ΔE1 value for all materials and group 1 had the lowest ΔE1 value. Brushing enabled the materials to return to their initial color or to reduce the color change in most groups. In the group 1 ΔE1 comparison between materials, the greatest color change was observed in the teeth, and the least in the IPS e.max CAD material. In the group 2 ΔE1 comparison between materials, the greatest color change was observed in the teeth, and the least was in the IPS e.max CAD material. Group 3 ΔE1 and group 4 ΔE1 comparisons between materials showed similar results.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eTo obtain restorative materials with reduced secondary caries formation, materials should be evaluated from a microbiologic perspective [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. (i) Our hypothesis that increasing the degree of roughness of CAD/CAM materials would not increase the amount of live biomass formed on the material was accepted because although Cerasmart material had the lowest roughness values, it showed the highest live biomass value; (ii) increasing the roughness degree of CAD/CAM materials would not cause more coloration of the material was accepted; (iii) the presence of biofilm on CAD/CAM materials would not cause further coloration of the materials by the effect of coloring beverages was rejected. In this study, CAD/CAM materials on which biofilm was formed were more colored. (iv) Brushing would not influence discoloration was rejected.\u003c/p\u003e \u003cp\u003eTo achieve an esthetically and functionally successful restoration, it is important to finish restorations with a smooth surface. Rough restoration surfaces can cause discoloration, gingival infections, plaque accumulation, and secondary caries development over time [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Quirynen \u003cem\u003eet al.\u003c/em\u003e concluded that roughness took a more active role in the development of bacterial adhesion than other surface properties. It has been reported that an increase in surface roughness above a Ra value of 0.2 \u0026micro;m leads to a rise in in-vivo biofilm formation [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Bollen \u003cem\u003eet al.\u003c/em\u003e showed that 0.2 \u0026micro;m Ra was a threshold value at which a decrease in bacterial adhesion was not expected [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. Some studies also did not associate this factor with biofilm formation in dental materials [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e], explaining that long-term incubation times reduce the importance of the roughness of the surface and bacterial adhesion on the existing biofilm instead of directly on the material surface [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Eick \u003cem\u003eet al.\u003c/em\u003e showed that to investigate the adhesion of S. mutans in the biofilm formed after 48 hours on restorative materials, no correlation was found between bacterial adhesion and surface roughness [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. In other studies, it was reported that the surface roughness threshold for plaque formation was Ra\u0026thinsp;=\u0026thinsp;0.2 \u0026micro;m [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e], the roughness value that the patient could feel with their tongue was Ra\u0026thinsp;=\u0026thinsp;0.28 \u0026micro;m [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e], and Ra\u0026thinsp;\u0026lt;\u0026thinsp;1 \u0026micro;m was perceived as smooth, clinically [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eBesides the polishing and finishing procedures, differences in the chemical composition of resin-based materials affect bacterial adhesion and biofilm formation [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. These materials generally consist of a hydrophobic resin matrix containing hydrophilic fillers and an intermediate binding agent (silane) between the two phases. It should be considered that materials with such composition cannot form homogeneity, and topographic and chemical differences can be seen on the surface [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. In our study, in line with the study of Wang \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e], it was observed that the composite-based material Cerasmart hybrid CAD/CAM material (although less roughness) caused more viable biomass formation than the IPS e.max CAD material in lithium disilicate glass-ceramic structure.\u003c/p\u003e \u003cp\u003eCovering teeth and restorative materials with a salivary pellicle layer is the first step for bacterial colonization. Oral bacterias bind to host-derived receptors in the pellicle [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. In our study, after forming a pellicle with artificial saliva on the samples, biofilm was formed on the pellicle layer, and biofilm biomass values ​​were obtained independent of the roughness, thus supporting the hypothesis that the surface roughness could not be transferred from the pellicle. On the other hand, it was shown that an increase in surface roughness, even in the presence of salivary pellicle, causes an increase in bacterial adhesion [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. In light of these indicators, it can be thought that the physicochemical properties of the surfaces are partially balanced by the presence of saliva.\u003c/p\u003e \u003cp\u003eThe choice of culture medium depends on the type of inoculum, and the needs and purpose of the study. It has been noted that monocultures are more accurate for assessing specific physiologic aspects of biofilm. For monoculture biofilms, S. mutans has been the preferred microorganism [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. S. mutans has been identified as one of the more cariogenic strains present in oral biofilms [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. In the study, S. mutans and S. sanguinis were included because they are the main colonizers of hard surfaces in the oral cavity, functioning as a bridge between other bacteria that attach to and grow on the substrate surface.\u003c/p\u003e \u003cp\u003eLike several other Streptococcal species, they participate in the initial colonization of tooth enamel [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. In multi-culture-based models of maintaining clinical oral conditions, bacterial growth occurs in the presence of nutrient-limited media. In this case, bacterial growth is faster in the oral cavity and exhibits an entirely unnatural behavior [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. This may explain why multicultural studies take a shorter time. Like some other studies using multi-culture media, a 48-hour biofilm evaluation was performed in our study [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eTo evaluate the pathogenicity of dental plaque, the amount of bacterial viability in the biofilm is important. In a study, it was determined that there was a correlation between the glass content of the materials and bacterial viability. It has been reported that the contents of ceramics affect bacterial viability during adhesion. It was determined that the number of S.oralis and S.sanguinis was low on tetragonal stabilized zircon, zirconia-reinforced glass ceramics, and glass alumina ceramics [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. Again, in a study investigating bacterial viability, it was reported that although there was high bacterial accumulation in amalgam, the number of viable bacteria was low [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. In our study, although the roughness rate was higher, a lower amount of live biomass was detected in the IPS e.max CAD compared with the Cerasmart material.\u003c/p\u003e \u003cp\u003eThe correct color selection is important for a successful restoration, restorative materials in the oral cavity are exposed to many factors such as coloring foods and beverages, saliva, dental plaque, and bacteria in the oral flora, which can cause a color change in restorations. In our study, when the color changes of CAD/CAM materials with and without biofilm were examined after they were kept in tea, the samples in the coloring beverage with biofilm were statistically significantly more colored. However, no significant difference was found between the tea groups with and without biofilms when the materials were examined in terms of initial and post-brushing ΔE3 values. Although there was no significant difference in the samples kept in water for the Cerasmart ΔE3 value, the samples kept in water without biofilm for IPS e.max CAD and dental samples were more colored than with biofilm.\u003c/p\u003e \u003cp\u003eIn a study, adsorption to the pellicle-like protein layer by black tea and red wine components formed in vitro by the adsorption of unstimulated saliva on hydroxyapatite discs was investigated. According to some studies, the pellicle quickly absorbs the components of black tea, and subsequent contact with saliva increases the adsorption of salivary components overall [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]. In our study, Cerasmart (6.153) and IPS e.max CAD (7.787) materials, which were first coated with a pellicle using artificial saliva and on which biofilm was formed, were Cerasmart (2.265) and IPS e.max CAD (2.118) materials without pellicle and kept in tea were statistically significantly more colored.\u003c/p\u003e \u003cp\u003eDetectable and acceptable color change ΔE threshold values ​​were determined as 0.8 and 1.8, respectively, by Paravina \u003cem\u003eet al.\u003c/em\u003e [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e] who evaluated changes in color and roughness properties of resin composites exposed to aging in water or red wine and brushed. Color and roughness measurements were made of the samples on the 5th, 10th, and 15th day. No significant color change was observed in the groups that were kept in water and brushed according to the detectable ΔE\u0026thinsp;\u0026gt;\u0026thinsp;0.8 threshold value. Less color change was observed in the brushed groups compared to the non-brushing groups. They concluded that short-term brushing reduced color change and did not increase roughness, although the samples deteriorated significantly with red wine and brushing [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. In our study, the fact that the brushing process mostly returns the initial colors of the materials supports this result.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe adhesion of biofilm to restorative dental materials plays an important role in the coloring of dental treatment. The presence of biofilm on CAD/CAM materials immersed in water caused an unacceptable degree of coloration (ΔE\u0026thinsp;\u0026gt;\u0026thinsp;1.8), and immersion in tea caused a greater color change (ΔE\u0026thinsp;\u0026lt;\u0026thinsp;0.8). The brushing process allowed CAD/CAM materials to return to their original color. Although there is no consensus on the selection of materials showing the minimum coloring and biofilm adhesion among restorative materials, in our study, the IPS e.max CAD group had less biofilm formation than Cerasmart group. Therefore, IPS e.max can be preferred over Cerasmart. However, there is no difference in the coloring process between the two materials with the formation of biofilm. Further studies are needed to understand these materials.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eConflict\u0026nbsp;of interest\u003c/strong\u003e The authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval\u003c/strong\u003e This study was approved by the Ethics Committee of\u0026nbsp;Kocaeli\u0026nbsp;University (KOU KAEK 2021/222).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003eThis work was supported by the writers.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contribution:\u0026nbsp;\u003c/strong\u003eDilan Kopuz has been written this article, S\u0026uuml;meyra Top\u0026ccedil;u, Eda Yazıcı \u0026Ouml;zcelik, Fetiye Kolaylı, Neslihan Tek\u0026ccedil;e, Mustafa Demirci and Safa Tuncer\u0026nbsp;has been done this research and conducted this manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformed consent:\u003c/strong\u003e All participants were freely invited, and those who accepted signed an informed consent approved and stamped by the local ethics committee.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003ePflughoeft KJ, Versalovic J. 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J Prosthodont. 2019; 28:873\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMota EG, Smidt LN, Fracasso LM, et al. The effect of milling and postmilling procedures on the surface roughness of CAD/CAM materials. J Esthet Restor Dent. 2017; 29:450\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eQuirynen M, Marechal M, Busscher HJ, et al: The influence of surface free energy and surface roughness on early plaque formation. An in vivo study in man. J Clin Periodontol. 1990;17:138\u0026ndash;144.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBollen CM, Lambrechts P, Quirynen M: Comparison of surface roughness of oral hard materials to the threshold surface roughness for bacterial plaque retention: a review of the literature. Dent Mater. 1997; 13(4):258\u0026ndash;69.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTeughels W, Van Assche N, Sliepen I, et al: Effect of material characteristics and/or surface topography on biofilm development. Clin Oral Implants Res. 2006; 17:68\u0026ndash;81.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDezelic T, Schmidlin PR: Multi-species biofilm formation on dental materials and an adhesive patch. Oral Health Prev Dent. 2009; 7:47\u0026ndash;53.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEick S, Glockmann E, Brandl B, et al: Adherence of Streptococcus mutans to various restorative materials in a continuous flow system. J Oral Rehabil. 2004; 31:278\u0026ndash;285.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAntonson SA, Yazici AR, Kilinc E, et al: Comparison of different finishing/polishing systems on surface roughness and gloss of resin composites. J Dent. 2011; 39:9\u0026ndash;17.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eM\u0026ouml;rmann WH, Stawarczyk B, Ender A, et al: Wear characteristics of current aesthetic dental restorative CAD/CAM materials: two-body wear, gloss retention, roughness and Martens hardness. J Mech Behav Biomed Mater. 2013; 20:113\u0026ndash;125.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJones CS, Billington RW, Pearson GJ. The in vivo perception of roughness of restorations. Br Dent J. 2004; 196:42\u0026ndash;5.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEreifej NS, Oweis YG, Eliades G. The effect of polishing technique on 3-D surface roughness and gloss of dental restorative resin composites. Oper Dent. 2012; 38:9\u0026ndash;20.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOno M, Nikaido T, Ikeda M, et al. Surface Properties of Resin Composite Materials Relative to Biofilm Formation. Dent Mater J. 2007; 26:613\u0026ndash;22.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang Z, Shen Y, Haapasalo M. Dental materials with antibiofilm properties. Dent Mater. 2014; 30:1\u0026ndash;16.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIkeda M, Matin K, Nikaido T, et al. Effect of surface characteristics on adherence of S. mutans biofilms to indirect resin composites. Dent Mater J. 2007; 26:915\u0026ndash;923.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCarl\u0026eacute;n A, Nikdel K, Wennerberg A, et al. Surface characteristics and in vitro biofilm formation on glass ionomer and composite resin. Biomaterials. 2001; 22:481\u0026ndash;7.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTotiam P, Gonz\u0026aacute;lez-Cabezas C, Fontana MR, et al. A new in vitro model to study the relationship of gap size and secondary caries. 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J Mater Sci Mater Med. 2008; 19:3249\u0026ndash;3253.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSteinberg D, Eyal S. Early formation of Streptococcus sobrinus biofilm on various dental restorative materials. J Dent. 2002; 30:47\u0026ndash;51.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJoiner A, Muller D, Elofsson UM, et al. Adsorption from black tea and red wine onto in vitro salivary pellicles studied by ellipsometry. Eur J Oral Sci. 2003; 111:417\u0026ndash;422.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eParavina RD, Ghinea R, Herrera LJ, et al. Color difference thresholds in dentistry. J Esthet Restor Dent 2015; 27:1\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMozzaquatro LR, Rodrigues CS, Kaizer MR, et al. The Effect of Brushing and Aging on the Staining and Smoothness of Resin Composites. J Esthet Restor Dent. 2017; 29:44\u0026ndash;55.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"CAD/CAM, Color, Biofilm Formation, Biofilm, Surface Roughness","lastPublishedDoi":"10.21203/rs.3.rs-3221496/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3221496/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eObjectives\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo investigate the effect of biofilm formation on the coloration properties of CAD/CAM materials.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMaterials and Methods\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e106 samples of 2-mm thickness were prepared from two different CAD/CAM materials (IPS e.max CAD; Cerasmart) and a total of 53 samples were prepared from extracted caries-free human-teeth. Five samples from each sample group were used to measure the amount of alive biomass in the biofilm. The remaining 48 samples in each group were divided into four subgroups: group-1, kept in water without the formation of dental biofilm; group-2, kept in tea without the formation of dental biofilm; group-3, kept in water after the formation of dental biofilm; group-4: kept in tea after the formation of dental biofilm (n=12). After finishing and polishing, color and surface roughness measurements were made. After baseline measurements, a biofilm layer was formed in groups-3 and 4, and the measurements were repeated. Afterward, all samples were brushed, and a third measurement was performed. Data were statistically analyzed (p\u0026lt;0.05).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe lowest roughness value was observed in Cerasmart. Tooth-IPS e.max CAD gave similar results. The Cerasmart material had the most viable biomass, whereas the IPS e.max CAD material had the least. Group-4 had the highest ΔE1 value for all materials and group-1 had the lowest. The presence of biofilm on CAD/CAM materials immersed in water caused an unacceptable degree of coloration (ΔE\u0026gt;1.8), and immersion in tea caused much more color change.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe brushing process allowed CAD/CAM materials to return to their original color.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical Relevance\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe adhesion of biofilm to restorative dental materials plays an important role in the coloring of dental treatment.\u003c/p\u003e","manuscriptTitle":"Effect of Biofilm Formation on the Color Properties of CAD/CAM Materials","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-08-07 16:21:09","doi":"10.21203/rs.3.rs-3221496/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"4b20c8d7-43a7-4148-9f39-702d991d4921","owner":[],"postedDate":"August 7th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2023-08-15T20:44:12+00:00","versionOfRecord":[],"versionCreatedAt":"2023-08-07 16:21:09","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3221496","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3221496","identity":"rs-3221496","version":["v1"]},"buildId":"ApUGefWb6u5IBVtyqm6d5","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}