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Item Artificial Caries Lesion Characteristics after Secondary Demineralization with Theobromine-Containing Protocol(MDPI, 2021-01-08) Nassar, Hani M.; Lippert, Frank; Cariology, Operative Dentistry and Dental Public Health, School of DentistryDeveloping artificial caries lesions with varying characteristics is needed to adequately study caries process in vitro. The objective of this study was to investigate artificial caries lesion characteristics after secondary demineralization protocol containing theobromine and fluoride. Sixty bovine enamel slabs (4 × 3 mm) were demineralized using a Carbopol-containing protocol for 6 days. A baseline area (2 × 3 mm) was protected with acid-resistant nail varnish, after which specimens were exposed for 24 h to a secondary demineralization protocol containing acetic acid plus one of four fluoride/theobromine combinations (n = 15): theobromine (50 or 200 ppm) and fluoride (0 or 1 ppm). Specimens were sectioned and analyzed using transverse microradiography for changes in mineral content, lesion depth, and surface layer mineralization. Data was analyzed using paired t-test and analysis of variance followed by Bonferroni test at 0.05 significance level. After secondary demineralization, fluoride-containing groups had significantly deeper lesions (p = 0.002 and 0.014) compared to the group with 0 ppm fluoride and 50 ppm theobromine. Mineral content and lesion depth were significantly different compared to baseline for all groups. Theobromine did not show an added effect on mineral uptake. Theobromine-containing groups exhibited particularly deep lesions with a more uniform mineral profile in the presence of fluoride.Item Associations between Oral Health and Cannabis Use among Adolescents and Young Adults: Implications for Orthodontists(MDPI, 2022-11-18) Le, Austin; Khoo, Edmund; Palamar, Joseph J.; Orthodontics and Oral Facial Genetics, School of DentistryCannabis use is prevalent among adolescents and young adults in the US. Virtually all modes of cannabis consumption involve the oral cavity, and previous studies have linked cannabis use with poorer oral health. We sought to identify associations between cannabis use and various oral health outcomes and behaviors among individuals 12–25 years of age, and to discuss implications for orthodontists who largely interact with this age group over an extended period of treatment time. We examined data from patient electronic health records (N = 14,657) obtained between 2015 and 2021. Associations between lifetime and current self-reported cannabis use and several oral health outcomes or related behaviors that reflect periodontal health, caries status, oral lesions, and physical integrity of tooth structure and restorations were examined in a bivariable and multivariable manner, controlling for patient age, sex, and self-reported tobacco and alcohol use. Reporting lifetime cannabis use was associated with higher risk for having oral lesions (aPR = 1.41, 95% CI: 1.07–1.85), bruxism (aPR = 1.31, 95% CI: 1.09–1.58), and frequent consumption of sugary beverages and snacks (aPR = 1.27, 95% CI: 1.12–1.41). Reporting current cannabis use was associated with higher risk for oral lesions (aPR = 1.45, 95% CI: 1.03–2.06) and frequent consumption of sugary beverages and snacks (aPR = 1.26, 95% CI: 1.07–1.48). Cannabis users aged 12–25 are at increased risk for bruxism, oral lesions, and frequent consumption of sugary beverages and snacks. Orthodontists and other dental professionals should probe for drug use and be cognizant of increased risk for oral health problems in patients that report actively using cannabis.Item Bioactivity of Dental Restorative Materials: FDI Policy Statement(Elsevier, 2023) Schmalz, Gottfried; Hickel, Reinhard; Price, Richard Bengt; Platt, Jeffrey A.; Biomedical Sciences and Comprehensive Care, School of DentistryThe term bioactivity is being increasingly used in medicine and dentistry. Due to its positive connotation, it is frequently utilised for advertising dental restorative materials. However, there is confusion about what the term means, and concerns have been raised about its potential overuse. Therefore, FDI decided to publish a Policy Statement about the bioactivity of dental restorative materials to clarify the term and provide some caveats for its use in advertising. Background information for this Policy Statement was taken from the current literature, mainly from the PubMed database and the internet. Bioactive restorative materials should have beneficial/desired effects. These effects should be local, intended, and nontoxic and should not interfere with a material's principal purpose, namely dental tissue replacement. Three mechanisms for the bioactivity of such materials have been identified: purely biological, mixed biological/chemical, or strictly chemical. Therefore, when the term bioactivity is used in an advertisement or in a description of a dental restorative material, scientific evidence (in vitro or in situ, and preferably in clinical studies) should be provided describing the mechanism of action, the duration of the effect (especially for materials releasing antibacterial substances), and the lack of significant adverse biological side effects (including the development and spread of antimicrobial resistance). Finally, it should be documented that the prime purpose, for instance, to be used to rebuild the form and function of lost tooth substance or lost teeth, is not impaired, as demonstrated by data from in vitro and clinical studies. The use of the term bioactive dental restorative material in material advertisement/information should be restricted to materials that fulfil all the requirements as described in the FDI Policy Statement.Item Combined Effects of Soda Drinks and Nicotine on Streptococcus Mutans Metabolic Activity and Biofilm Activity(2019) Mokeem, Lamia Sami; Gregory, Richard; Cook, Norman Blaine; Windsor, Jack; Eckert, GeorgeItem Comparison of Two Types of Pit and Fissure Sealants in Reducing the Incidence of Dental Caries Using a Split-Mouth Design(2021) Fernandez-Barrera, Miguel Angel; Saucedo-Mlina, Teresita de Jesus; Scougall-Vilchis, Rogelio Jose; Marquez-Corona, Maria de Lourdes; Medina-Solis, Carlo Eduardo; Maupome, GerardoObjectives: To evaluate the effectiveness of two pit and fissure sealants (PFS) in reducing the incidence of dental caries in schoolchildren. Material and Methods: A randomized split-mouth experimental design was used in a sample of 140 subjects assigned to two groups. The sealants used were Clinpro©3M and BeautiSealant©Shofu placed in first permanent molars (FPMs). Each sealant was compared to molars in the controls to determine effectiveness over a period of 6 months. The study had a 12.9% loss to follow-up. No statistically significant differences (p>0.05) were observed for sex, age, baseline dmft, or type of sealants. Nonparametric tests were used for statistical analysis. Results: Average dmft index at baseline was 4.10±3.16. Lower incidence of caries was observed in FPMs with pit and fissure sealants (p<0.01), regardless of the type used. When sealants remained intact there was a lower caries incidence compared to sealants partially or completely missing – but differences were only significant for FPM 16. Caries incidence was higher for BeautiSealant sealed teeth than for Clinpro’s, but only statistically significant in FPMs 16, 36 and 46 (p< 0.05). Caries incidence was higher in those cases with a higher baseline dmft but it only reached statistical significance in FPMs 26 and 36. Relative risks for dental caries were lower in sealed teeth (p<0.01). Conclusions: Pit and fissures sealants are an effective preventive treatment to reduce caries during a 6-month follow-up in schoolchildren 6 to 8 years of age, regardless of the type of sealant used. The sealant brand that showed greater effectiveness in terms of prevention and retention was Clinpro©3M.Item The effect of cigarette smoking on the virulence of streptococcus mutans caries and cardiovascular diseases-epidemiological analysis and in vitro studies(2010) Zheng, Cunge; Gregory, Richard L.; Windsor, L. Jack; Kowolik, Michael J.; Steele, Gregory K.; Holt, Robert G.The impact of tobacco smoking on human health is well documented. The influence of smoking on tooth loss and cardiovascular diseases was investigated in the current study via both epidemiology and in vitro studies. From analyzing the 2006 Behavioral Risk Factor Surveillance System (2006 BRFSS) database, we confirmed that smoking was significantly associated with the number of teeth lost in a dose-dependent manner and smoking cessation reduced the risk when compared to those subjects continuing to smoke. In addition, the virulence factors related to caries were compared between Streptococcus mutans and Streptococcus gordonii in response to cigarette smoking condensate (CSC) treatment. We observed that S. gordonii was more susceptible to CSC treatment than S. mutans. CSC significantly enhanced S. mutans sucrose-dependent and independent adherence. Western blot assays revealed that several bacterial surface proteins including glucosyltransferase (GTF), glucan-binding proteins and antigen I/II, were significantly upregulated for the treated S. mutans. These findings suggested that the oral environment with CSC may favor a cariogenic dominant composition, which may increase the risk for smokers to develop caries. We also found that smoking and oral health status modified each other and synergistically increased the risk of CVD and this joint effect was more pronounced among the youngest age group using the 2006 BRFSS database. To further understand the joint effect, we conducted an in vitro study to investigate bacterial attachment to fibronectin and endothelial cells in response to smoking condensate treatment. Our study clearly demonstrated CSC significantly enhanced S. mutans attachment to both soluble and immobilized fibronectin as well as endothelial cells. Furthermore, our data suggested that bacteria possessed several adhesins that bound to host tissues and endothelial cells also had multiple receptors for bacterial attachment. Among these adhesins, antigen I/II seemed essential for bacterial attachment to endothelial cells without CSC. The knowledge of bacterial attachment to host tissues in the presence of CSC may help in developing different preventive or therapeutic strategies against attachment and colonization of the host by S. mutans.Item Effect of nicotine on streptococcus mutans(2014-11) Huang, Ruijie; Gregory, Richard L.; Tu, Wanzhu; Windsor, L. Jack; Wu, Christine; Song, FengyuStreptococcus mutans is a key contributor to dental caries. Smokers have increased caries, but the association between tobacco, nicotine, caries and S. mutans growth is little investigated. In the first section, seven S. mutans strains were used for screening. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and minimum biofilm inhibitory concentration (MBIC) were 16 mg/ml (0.1 M), 32 mg/ml (0.2 M), and 16 mg/ml (0.1 M), respectively, for most of the S. mutans strains. Growth of planktonic S. mutans cells was significantly repressed by 2.0-8.0 mg/ml nicotine concentrations. Biofilm formation and metabolic activity of S. mutans was increased in a nicotine-dependent manner up to 16.0 mg/ml. Scanning electron microscopy (SEM) revealed higher nicotine-treated S. mutans had thicker biofilm and more spherical bacterial cells than lower concentrations of nicotine. In the second section, confocal laser scanning microscopy (CLSM) results demonstrated that both biofilm bacterial cell numbers and extracellular polysaccharide (EPS) synthesis were increased by nicotine. Glucosyltransferase (Gtf) and glucan binding protein A (GbpA) protein expression of S. mutans planktonic cells were upregulated, while GbpB protein expression of biofilm cells were downregulated by nicotine. The mRNA expression of those genes were mostly consistent with their protein results. Nicotine was not directly involved in S. mutans LDH activity. However, since it increased the total number of bacterial cells in biofilm; total LDH activity of S. mutans biofilm was increased. In the third section, a PCR-based multiple species cell counting (PCR-MSCC) method was designed to investigate the effect of nicotine on S. mutans in a ten mixed species culture. The absolute S. mutans number in mixed biofilm culture was increased but the percentage of S. mutans in the total number of bacterial cells was not changed. In conclusion, nicotine enhanced biofilm formation and biofilm metabolism of S. mutans, through stimulating S. mutans planktonic cell Gtfs and Gbps expression. This leads to more planktonic cells attaching to dental biofilm. Increased S. mutans cell numbers, in biofilms of single species or ten mixed species, resulted in higher overall LDH activity. More lactic acid may be generated and contribute to caries development in smokers.Item The Effect of Silver Diamine Fluoride on Caries Lesion Remineralization as a Function of Lesion Baseline Mineral Distribution(2022) AlMady, Fahad; Lippert, Frank; Hara, Anderson; Al Dehailan, Laila; Capin, OrianaPurpose: The primary objective of this laboratory study was to investigate whether the ability of SDF to remineralize caries lesion is modulated by their baseline mineral distribution. The exploratory objective was to determine whether the staining caused by SDF is modulated by lesion mineral distribution. Materials and methods: This in vitro study utilized bovine permanent enamel specimens. SDF was compared to the clinical reference standard, 5% sodium fluoride varnish (FV), and deionized water (DIW) was the negative control. By utilizing three lesion creation protocols (methylcellulose [MeC], hydroxyethylcellulose [HEC], Carbopol [C907]), artificial caries lesions with different mineral distributions (n=20 per subgroup) were created in the enamel specimens Interventions were applied and the lesions remineralized using artificial saliva. Changes in mineral loss, lesion depth, mineral density of the surface zone, and lesion mineral distribution were determined using transverse microradiography. Throughout the study, color of the lesion and changes thereof were measured using a spectrophotometer. Data were analyzed by using two-way ANOVA. Pair-wise comparisons were performed using Fisher’s Protected Least Significant Differences to control the overall significance level at 5%. Results: For changes in mineral loss, DIW in MeC showed significantly greater change (more remineralization) than both SDF (p<0.01) and FV (p=0.01), which were not different from one another (p=0.13). There were no statistically significant differences between SDF and FV in the other lesions (C907 – p=0.18; HEC – p=0.56). For changes in lesion depth, there was no statistically significant interaction between study factors lesion protocol and treatments (p=0.23) as well as the individual factors lesion protocols (p=0.08) and treatments (p=0.82). For changes in surface zone mineral density, SDF showed significant change in mineral density compared to FV (p=0.02); however, SDF was not different from with DIW (p=0.25). For lesion mineral distribution, MeC exhibited the greater mineral loss in the lesion body and lowest mineral density at the surface zone. HEC lesions were the deepest but exhibited modest differences in mineral loss between the lesion body and the surface zone. C907 lesions were somewhat between MeC and HEC. SDF in MeC had the highest mineral gain in the surface zone, while DIW resulted in the highest mineral gain in the lesion body. SDF in HEC showed the highest mineral gain in the surface area compared to FV and DIW, with all treatments resulting in the largest mineral gain to a similar extent in the lesion body. In C907, SDF showed the most mineral gain in the lesion body compared to DIW and FV, while differences in the surface zone between treatments were less pronounced. For color changes post intervention, SDF showed more darkening in C907 and HEC lesions compared to MeC (p<0.01) and compared to FV and DIW. For post remineralization, SDF treated C907 lesions became significantly whiter (p<0.01) compared to SDF in MeC and HEC which continued to get darker. Conclusion: SDF did exhibit different remineralization abilities and behaviors and the modulation was based on lesion baseline mineral distribution. Staining resulting from SDF treatment varied significantly based on lesion mineral distribution.Item Effects of silver diamine fluoride on demineralization protection after a secondary acid challenge(University of Sao Paulo, 2023-11-03) Tudares, Mauro A.; Eckert, George J.; Lippert, Frank; Biomedical and Applied Sciences, School of DentistryObjective: This investigation describes the effects of 5% sodium fluoride varnish and 38% silver diamine fluoride on demineralization protection of human enamel lesions of three different severities after a secondary acid challenge. Study design: Specimens underwent color and enamel surface microhardness change measurements after demineralization and treatment events. Transverse microradiography was conducted following the secondary demineralization. Results: After treatments, enamel surface microhardness change showed that 24-hour lesions treated with fluoride varnish had less rehardening than 24-hour lesions treated with silver diamine fluoride (p<0.05), whereas 144-hour lesions from both treatment groups showed a beneficial decrease in surface microhardness change that was markedly better in samples treated with silver diamine fluoride (p<0.05). After the secondary demineralization, 24- and 144-hour lesions treated with silver diamine fluoride showed a sustained beneficial decrease in enamel surface microhardness change when compared to fluoride varnish-treated samples of the corresponding lesion severity (p<0.05). Transverse microradiography showed no difference between fluoride varnish- and silver diamine fluoride-treated samples of any corresponding lesion severity, indicating that remineralization in both fluoride varnish- and silver diamine fluoride-treated samples was proportional to each other after a secondary acid challenge. Conclusions: Using silver diamine fluoride may have comparable benefits to fluoride varnish in mineral loss prevention.Item In vitro Cariostatic effects of cinnamon water extract on nicotine-induced Streptococcus mutans biofilm(BMC, 2020-02-11) Alshahrani, Abdulaziz M.; Gregory, Richard L.; Cariology, Operative Dentistry and Dental Public Health, School of DentistryBackground: Dental caries is one of the most prevalent chronic oral diseases worldwide. Dental caries is mainly associated with Streptococcus mutans and the Lactobacillus species. A specific relationship was found between nicotine and S. mutans growth as the presence of nicotine increased S. mutans biofilm formation. Nicotine is able to increase the number of S. mutans and extracellular polysaccharide (EPS) synthesis. Among the widely used herbs and spices is cinnamon which demonstrated a strong antibacterial activity against a wide variety of bacteria including S. mutans and showed the ability to inhibit S. mutans biofilm formation. Cinnamon essential oil, obtained from the leaves of C. zeylanicum, has been demonstrated to be effective against S. mutans and Lactobacillus acidophilus, which are partially responsible for dental plaque formation and caries development. The aim of this study was to identify the effects of nicotine exposure on the inhibitory effects of cinnamon water extract on S. mutans biofilm formation. Materials and methods: A 24-h culture of S. mutans UA159 in microtiter plates was treated with varying nicotine concentrations (0-32 mg/ml) in Tryptic Soy broth supplemented with 1% sucrose (TSBS) with or without a standardized concentration (2.5 mg/ml) of cinnamon water extract. A spectrophotometer was used to determine total growth absorbance and planktonic growth. The microtiter plate wells were washed, fixed and stained with crystal violet dye and the absorbance measured to determine biofilm formation. Results: The presence of 2.5 mg/ml cinnamon water extract inhibits nicotine-induced S. mutans biofilm formation from 34 to 98% at different concentrations of nicotine (0-32 mg/ml). Conclusion: The results demonstrated nicotine-induced S. mutans biofilm formation is decreased from 34 to 98% in the presence of 2.5 mg/ml cinnamon water extract. This provides further evidence about the biofilm inhibitory properties of cinnamon water extract and reconfirms the harmful effects of nicotine.