Oral Health and Preventive Dentistry

Purpose: To determine if an interactive electric toothbrush and smartphone application (app) can reduce self-reported gingival bleeding and promote better brushing behaviour based on global, in-market usage data. Materials and Methods: Anonymised data were collected worldwide between July 2020 and January 2021 from users of interactive oscillating-rotating electric toothbrushes and app (Oral-B Genius, GeniusX and iO). Self-reported gingival bleeding and brushing behaviour data captured via the app were sent to Google Firebase and Google BigQuery to aid processing and analysis. Results: Data from 16.7 million brushing sessions were analysed. 439,481 new users responded at least once to the app question: ‘Do you have gum bleeding?’ Of users answering the question over their first two weeks of app use (153,201), the proportion reporting bleeding decreased statistically significantly from week 1 to 2 (28.8% to 17.1%, p < 0.0001). Of users answering the question over each of the first five weeks (43,060) a further statistically significant decrease in those reporting bleeding was seen in each consecutive week, with the week-5 rate being 12.7% (p < 0.0001 vs any previous week). Decreases in duration of excessive pressure (i.e. > 2.5 N – 3.0 N depending on the handle) decreased the proportion of self-reported gingival bleeding (p < 0.0001). Users brushed longer and with less overpressure (p < 0.0001) with vs without live feedback from the app, and showed 94.4% average coverage with live feedback. Conclusion: The interactive oscillating-rotating electric toothbrushes and app, particularly with live feedback, promote good brushing behaviour. Self-reported gingival bleeding occurred less frequently the longer the system was used. Keywords: compliance, gum bleeding, in-market evaluation, interactive electric toothbrush

Purpose: To evaluate the use of hydrogen peroxide as an adjunct to ultrasonication (US) in biofilm removal and whether it can limit the spread of viable microorganisms in the aerosol. Materials and Methods: Multi-species biofilms were formed on dentin disks and titanium disks fixed on a plastic surface. After placing the specimens in a periodontal pocket model, an ultrasonic scaler was applied for 30 s, in part combined with 0.25% or 0.5% H2O2. After treatment, the remaining biofilm was analysed for bacterial counts (colony forming units [CFU]), biofilm quantity and metabolic activity. Further, the cytotoxic effect of hydrogen peroxide on periodontal ligament fibroblasts was assessed and the spread of bacteria in aerosol was quantified. Results: Ultrasonication reduced bacterial counts in biofilm, biofilm mass and metabolic activity on both dentin and titanium disks. Adjunctive use of 0.25% and 0.5% H2O2 more effectively reduced the viable bacteria in biofilm than ultrasonication alone; this was also found on both dentin and titanium. The different concentrations of H2O2 did not lead to corresponding differences in bacterial mass and metabolic activity. The spread of bacteria through aerosols was statistically significantly reduced when adjunctive H2O2 was used. However, a certain cytotoxic effect on periodontal ligament fibroblasts by H2O2 could not be ruled out. Conclusions: Irrigating with H2O2 during periodontal instrumentation with an ultrasonic scaler increases the reduction of viable bacteria within biofilms. It might limit bacterial spreading via aerosols. Keywords: aerosol, biofilm, hydrogen peroxide, periodontal therapy

Purpose: To compare the analgesic efficacy of orally administered Curcuma longa (curcumin) and mefenamic acid (MA) after surgical periodontal therapy (SPT). Materials and Methods: Seventy-six periodontitis patients were randomly divided into two groups. In the test group, patients received curcumin capsules (200 mg), and in the control group, patients received MA (500 mg). All patients underwent post-operative antibiotic therapy using 500 mg amoxicillin and 400 mg metronidazole for 7 days. Post-operative pain and discomfort were evaluated using the numerical rating scale (NRS) and verbal rating scale (VRS), respectively. Evaluation were performed after 24 (T1), 48 (T2), and 72 h (T3). Group comparisons were done using Student’s t-test and the Mann-Whitney U-test. The level of statistical significance was established at p < 0.05. Results: All patients had stage 3/grade C periodontitis. The mean age of individuals in the test and control groups were 58.4 ± 7.3 and 57.2 ± 5.2 years, respectively. A family history of periodontal diseases was reported by 37.5% and 47.4% individuals in the test and control groups, respectively. In the test and control groups, the total mean duration of periodontal surgery was 168.2 ± 12.2 and 173.4 ± 10.7 min, respectively. There was no statistically significant difference in the mean NRS and VRS scores among patients in the test and control groups. In both groups, there was no statistically significant difference in the change in NRS scores at any time point. Conclusions: Compared with MA, curcumin is ineffective for pain and discomfort management after SPT. The possibility of the results being biased due to lack of operator blinding cannot be overlooked. Keywords: curcumin, mefenamic acid, pain, periodontal flap surgery

Purpose: To reach a consensus on a consistent strategy to adopt when screening patients for the clinical management of dentin hypersensitivity. Materials and Methods: A panel consisting of members of the Advanced Technology in Oral Hygiene Sciences Academy (ATASIO) was formed to start a review process on dentin hypersensitivity (DH) and subsequently elaborate a decision tree to manage DH, from diagnosis to prognosis. The panel employed the RAND in their deliberations. After an initial systematic literature review, it became evident that a consensually validated protocol for the management of patients affected by dentin hypersensitivity has to be considered mandatory by all dental professionals. However, the outcome of the systematic review made it evident that the treatment options to be provided, as well as their prognosis and timing, had never been defined. The panel produced documents that addressed the topic and were subsequently used to generate a questionnaire. A workshop of expert dental professionals was organised to reach consensus on the main steps of the decision tree. Each member completed the questionnaire independently, and then a panel discussion was held to reach a consensus. Results: A high level of agreement was reached regarding all the items on the questionnaire, and each of the clinical questions formulated was answered. A clinical decision threshold was created. Conclusions: The dissemination of the information to a wide dental audience should commence upon publication of this consensus document. The authors hope that this consensus will become a model for the development of a dedicated protocol to manage DH. Keywords: consensus, dentin hypersensitivity, diagnosis, Italy, prognosis

Purpose: To evaluate the knowledge and practices of Swiss women regarding oral health during pregnancy. Materials and Methods: Self-reported questionnaires were attributed to 385 women from 4 public hospitals in the French speaking part of Switzerland from February 2015 to June 2016.The questionnaire consisted of 32 questions including demographic characteristics, oral health habits and awareness of oral changes during pregnancy. Results: The majority of women (64%) were in the 3rd trimester of pregnancy and had a university education (41%). Oral health was considered very important for half of the women (52%) and moderately important for 38% of them. 71% of the women did not notice any change concerning their oral health conditions. Of the remaining 29%, gingival bleeding was the main symptom reported, followed by gingival redness and oedema. Pain and sensitivity were also reported by a few participants. Most of the pregnant women attended dental appointments during their pregnancy, but not on a regular basis, mainly because of lack of time. Almost half of the population had an adequate oral hygiene routine and adapted their eating habits to a much healthier pattern during pregnancy. A large proportion of the participants (71%) was aware that pregnancy renders teeth and gums more vulnerable and that oral health is related to adverse pregnancy outcomes. However, this information was rarely imparted to them by health professionals. Conclusion: Although pregnant women in the French speaking part of Switzerland seem to be moderately informed about the importance of oral health during pregnancy, health-care professionals do not seem to participate actively. Health professionals need to more actively inform pregnant patients about the importance of preventive oral health measures and oral health care during pregnancy. Keywords: education, oral health, pregnancy, self-care

Purpose: To investigate inflammatory responses in peri-implant crevicular fluid (PICF) in comparison to periodontal tissue. Materials and Methods: Nineteen participants with healthy implants restored with titanium or gold-casting abutment were included. PICF and gingival crevicular fluid (GCF) were collected for inflammatory cytokine detection by ELISA. Cytokine levels in PICF or GCF of the same individual were compared using the paired t-test, and those from titanium or gold-casting (UCLA) abutment were compared using the independent t-test. Human gingival fibroblast responses to PICF and GCF were then evaluated with one-way ANOVA. Results: The results demonstrated that IL-6, IL-8, TNFα, and IFNγ expressed in PICF are similar to GCF in the same individual. However, IL-1β (p = 0.032) and IL-1α (p = 0.030) was statistically significantly higher in PICF than in GCF. IL-8 level was statistically significantly higher with gold-casting than with titanium abutments (p = 0.003). PICF statistically significantly stimulated higher expression of RANKL, IL-1β, IL-6, and IL-8 mRNA in human gingival fibroblasts (HGF), while focal adhesion kinase (FAK) suppressed mRNA. Conclusion: The inflammatory cytokines, including IL-1α and IL-1β, are higher in healthy peri-implant tissues. Abutment materials may also influence the level of inflammatory cytokines in PICF. Inflammatory mediators in crevicular fluid may affect HGF inflammatory responses and peri-implant tissue integration. Keywords: cytokines, fibroblast, gold alloy, inflammatory response, peri-implant fluid, titanium

Purpose: To determine acid-formation potential of saliva and evaluate whether this method corresponds with microbiome composition of individuals with and without caries. Materials and Methods: A clinical, controlled pilot study was performed with two groups: individuals without caries (n = 25; DMFT = 0) and individuals with at least one active carious lesion (n = 25; DMFT>0). A detailed intraoral examination was performed, and the gingival bleeding index (GBI) and plaque index (PI) were recorded. The acid-formation potential was measured (ΔpH) after 1 h. Streptococcus mutans (SM) and lactobacilli (LB) were also quantified. Intergroup comparisons were made using the Mann-Whitney U-test. The diagnostic value was evaluated using the receiver operating characteristics (ROC) method and area under the curve (AUC) values were calculated. The saliva microbiome was analysed by 16S rDNA next-generation sequencing. Results: A statistically significant difference was found in ΔpH, with the ‘caries’ group showing a higher mean value after 1 h (‘healthy’ = 1.1,’caries’ = 1.4; p = 0.035). The AUC values were moderate to good (ΔpH = 0.67; SM = 0.83; LB = 0.83;1 = ideal). Streptococcus mutans and Lactobacilli were more frequently detected in the ‘caries’ group (p < 0.001), as were statistically significantly higher GBI (p = 0.006) and PI (p = 0.001). The saliva microbiome had a higher α-diversity and greater richness in individuals with active caries. The incidence of the genera Alloprevotella, Prevotella, Campylobacter and Veillonella was statistically significantly higher in the ‘healthy’ group. The incidence of the genera Fretibacterium, Lactobacillus, and Leptotrichia, as well as the phyla Spirochaetes and Synergistetes, was statistically significantly higher in the ‘caries’ group. Conclusion: Further studies must be carried out to determine the extent to which both tests are suitable for predicting future caries development. Keywords: acid formation, bacterial tests, caries, oral microbiome, saliva

Purpose: A number of disinfectants and sanitisers are used in dentistry, and there are numerous commercial solutions available. Nonetheless, because each cleaning solution has its own set of indications and limits, there is no one-size-fits-all approach for processing all types of dental equipment. Functional water, such as electrolysed hypochlorite microbubbled water, efficiently eliminates and sterilises biofilms. The objective of the study was to evaluate whether ozonated water could be used to sterilise and disinfect dental-unit water pipelines (DUWP) that had been contaminated with micro-organisms, including Gram-positive and Gram-negative bacilli and cocci. Materials and Methods: Three different groups were formed: group A – ozonated water (Cantoosh); group B – 1% povidine iodine; and group C: conventional distilled water. Group A was the test group, group B the control group, and group C was the positive control group. The water sterilising system was replaced with the appropriate sterilising agent as per the allocated group classification, with 2 min of purging, so that the complete DUWP was filled with the water sterilising system. Samples were collected and analysed, along with a 2-min purge after 24 h, 7 days and 21 days, at the 3 outlet (OL) points: the 3-way syringe at the dental tray(OL1), the cup filler (OL2), and the 3-way syringe of the assistant zone (OL3). Repeated measures ANOVA was used to test for statistical significance between colony-forming units of control and experimental groups (p < 0.05). Results: The cup filler yielded higher counts than did the 3-way syringe at the dental tray (OL1) (6.40 and 8.05 on the log scale, respectively). A statistically significant difference in the CFUs was also observed between samples taken after 24 h vs 21 days between groups A, B and C. Conclusion: The findings showed that exposing DUWP tube systems to ozonated water for an extended length of time drastically lowered the number of microorganisms adhering to their surfaces. Keywords: biofilms, dental-unit water pipelines, disinfection, microbial contamination, ozonated water

Purpose: The intention of the study was to evaluate whether Sub-Saharan schoolteachers may play a key role in oral health promotion of primary schoolchildren, particularly in terms of prevention of HIV/AIDS-related oral health problems. Materials and Methods: The study examined the level of knowledge about oral disease and the prevention of HIV/AIDS oral lesions amongst schoolteachers of Tanzania and Burkina Faso, identified their sources of information, and evaluated their ability for HIV/AIDS intervention. A multi-centre cross-sectional study was implemented in the two Sub-Saharan African countries. Participants comprised primary school teachers selected by stratified cluster sampling: 261 teachers from Tanzania and 313 teachers from Burkina Faso. All participants completed a structured questionnaire prepared for self-administration. The questions were designed from a standardised questionnaire developed by the World Health Organization. Results: Most teachers knew about the principal causes of the major oral diseases and the means of disease prevention. Nearly all teachers (95.6%) were aware of HIV/AIDS and they knew (92.3%) a virus was the direct cause of AIDS. Teachers were well informed of the general symptoms of HIV/AIDS, although oral symptoms were mentioned less often. In all, 17.6% of schoolteachers reported that children suffering from HIV/AIDS were found in their classes and 10.3% of teachers were conscious of students receiving anti-retroviral therapy (ART). Knowledge about the disease seemed to reflect mass media as a source, while teacher colleagues and health personnel played a somewhat lesser role in communication. In total, 83.2% emphasised that they should teach children about HIV/AIDS and the mouth. Schoolteachers from Tanzania (70.5%) were more often engaged in classroom-based oral health education than were the Burkinabe teachers (53.9%). Conclusion: The study confirms that schoolteachers may be a relevant source in the fight against HIV/AIDS among children. However, they would benefit from interaction with health personnel. Keywords: HIV/AIDS oral lesions, oral disease prevention, schoolteachers, public health

Purpose: Fixed retainers have been advocated for the prevention of anterior mandibular crowding after orthodontic treatment. However, limited data is available to help clinicians choose a retention protocol that is acceptable in terms of stability, emergencies, and side effects in the long term. It was the aim of this study to assess survival and alignment stability of the 0.016” x 0.022” stainless steel wire compared to more common protocols. Materials and Methods: Three different mandibular fixed retention protocols were compared in 600 consecutive patients: 1. 0.0215” multistrand wire (MW) with separate curing of resin and composite; 2. 0.016” x 0.022” stainless-steel wire with simultaneous curing of resin and composite (SS1C); and 3. 0.016” x 0.022” stainless-steel wire with separate curing of resin and composite (SS2C). The hazard rate for detachment across wire groups was assessed with a Cox frailty model. Results: Incisor alignment was maintained with all retention wires. One incisor with unexpected torque change was observed in group MW. The average annual emergency rate was below 2% for all three protocols. Fewer emergency visits were found in patients with solid steel wires than with multistrand wires. Detachment of the wire is the most common cause of emergency visits with no difference between wire types. Multistrand wires were more often damaged than were solid steel wires. There was no evidence that direct application of the composite on the uncured primer influenced retainer adhesion to the enamel. Conclusions: The mandibular anterior teeth can be predictably stabilised with a 0.016” x 0.022” stainless steel wire. Keywords: adverse effects, bond failure, enamel adhesion, fixed retainers, relapse, retainer failure, retention, stability of orthodontic treatment

Purpose: To identify the gaps in the Jordanian population’s knowledge about oral cancer, screening and attitudes toward screening, in addition to determining the barriers to oral cancer screening. Materials and Methods: A cross-sectional web-based study was conducted. The first section of the questionnaire employed collected the participants’ sociodemographic data. A question about whether patients had heard about oral cancer was then included, and those who answered ‘no’ were instructed to submit the questionnaire. The subsequent parts evaluated the participants’ knowledge of oral cancer and screening, attitudes toward screening, and barriers against screening. ANOVA and chi-squared tests were conducted to investigate the sample characteristics associated with the participants’ unfamiliarity with oral cancer. Binary regression was conducted to predict the variables associated with the participants’ knowledge and attitudes. Results: The questionnaire was filled by 1307 participants (1011 females). Most of the participants (70.01%) stated that they had heard of oral cancer, and the variables associated with awareness of oral were sex, monthly income, health insurance coverage, working status, and educational level. Sources of information and age were significantly associated with knowledge and attitude levels. The most ‘agree/strongly agree’ responses about barriers were lack of knowledge and lack of awareness. Conclusion: The study participants had moderate knowledge about oral cancer and satisfactory attitudes toward screening. Nearly all barriers to screening can be overcome by the joint efforts of healthcare providers and health authorities. Keywords: attitudes, barriers, Jordan, knowledge, oral cancer, screening

Purpose: This cross-sectional observational study evaluated the frequency of dental erosion in 12-year-old schoolchildren in Hungary and its connection to gender, geographical region, eating/drinking habits, and to socioeconomic factors, such as the educational level of their mothers. Materials and Methods: 579 randomly selected children aged 12 (287 boys and 292 girls) were examined in our cross-sectional study from 14 different regions in Hungary. Clinical examinations were carried out by the same examiner, using the ‘Basic Erosive Wear Examination’ (BEWE) index. A self-administered questionnaire was filled in by each child, surveying their oral hygiene, nutritional habits and socioeconomic status. Results: 21.2% of the children showed dentitions with signs of erosion. We found statistically significantly higher BEWE scores in urban than in rural areas (p = 0.0058). There was no difference between genders. Among children drinking carbonated soft drinks once or more daily, the prevalence of BEWE score < 3 was statistically significantly lower than among those who consumed these kinds of beverages less frequently (83.6% vs 90%, respectively, p = 0.034). Children of mothers with a highschool diploma had a BEWE score ≥ 3 statistically significantly less frequently than those whose mothers had not graduated from highschool (8.4% vs 22.5%, respectively, p = 0.000). Conclusions: The prevalence of dental erosion among 12-year-old children in Hungary is not as high as reported previously in Western European countries. A positive correlation was observed between the consumption of carbonated soft drinks, the educational level of the mothers and the level of erosion. These factors statistically significantly affected the prevalence and severity of erosive dental lesions. Keywords: dental erosion, national survey, prevalence, risk factors, 12-year-old-children

Purpose: To compare the levels of salivary IGF-1, IGFBP-3, and CTX with periodontal status among patients belonging to various skeletal maturity groups. Materials and Methods: This cross-sectional study was conducted on 80 participants 6 to 25 years of age. Based on skeletal maturity, the participants were categorised into 3 different stages: prepubertal, pubertal, and post-pubertal stages. The periodontal status of the participants was assessed using the simplified oral hygiene index (OHI-S), bleeding on probing (BOP), probing pocket depth (PPD), clinical attachment loss (CAL), and community periodontal index (CPI). The saliva samples were examined for IGF-1, IGFBP-3, and CTX using the respective ELISA kits. One-way ANOVA was used to determine statistically significant differences of means across the study groups for continuous variables. Results: The study demonstrated statistically significant differences for the parameters OHI-S, bleeding on probing, PPD, CPI, and CAL (p < 0.05) depending on skeletal maturity stage. ANOVA test showed a statistically significant difference by stage in IGF-1, IGFPB3, and CTX (p < 0.01). Conclusion: An association exists between periodontal status and levels of salivary IGF-1, IGFBP-3, and CTX in patients belonging to various skeletal maturity groups. Keywords:  C-terminal telopeptide region of type I collagen, IGF-binding protein-3, insulin-like growth factors, periodontitis, skeletal maturity

Purpose: Early childhood caries (ECC) and childhood obesity are among the most prevalent health conditions affecting children. ECC is associated with obesity through the common risk factor of sugar consumption. The present study aimed to assess the association between ECC and obesity in preschool children. Materials and Methods: A cross-sectional study was conducted among 1250 preschool children (698 girls, 552 boys; mean age: 4.3 [1.1] years). The children’s body mass index was determined (BMI: weight/height in kg/m2). The World Health Organization criteria were used for the diagnosis of caries. Multivariable logistic regression was used to analyse the relationship between ECC prevalence and childhood obesity. Results: ECC was detected in 929 (74.3%) children. The mean dmft and dmfs was 5.91 (1.13) and 8.92 (2.07), respectively. The multiple regression model showed a statistically significant association between ECC prevalence and obesity with an adjusted odds ratio (OR) of 2.59 (95% CI: 1.88 – 3.57; P = 0.001). The logistic regression model showed that in children with a monthly family income > $2666, sugar consumption, preterm low birth-weight/full-term low birth-weight (PTLBW/FTLBW), and toothbrushing frequency ≤ 1 time/day were statistically significantly associated with ECC prevalence. Conclusion: ECC was positively associated with obesity. Keywords: ECC, obesity, preschool, prevalence

Purpose: To evaluate the performance of sealing materials used in the screw-access holes of screw-retained implant final superstructures in vivo and in vitro. Materials and Methods: Twenty-one screw-access holes in the final superstructures were randomly divided into three groups (each group, n = 7). Following disinfection and isolation, all access holes were initially filled with sterilised cotton pellets of the same weight. Depending on the group, the access holes were finally sealed with either provisional composite restorations (group A), self-curing resin for provisional sealing (group B), or acrylic resin (group C). After one month of the functional period, the inner cotton pellets were collected as bacterial reservoirs. Results: Total aerobic bacteria and total gram-negative anaerobic bacteria were measured after bacterial culture for 48 h and 72 h, respectively. In vitro evaluation of porosity using scanning electron microscopy (SEM) was also performed. Samples from superstructures sealed with provisional composite restorations showed fewer bacteria and less porosity than samples from superstructures sealed with self-curing resin for provisional sealing and acrylic resin. In this study, provisional composite restorations showed the best sealing properties. Provisional composite restorations may prevent bacterial invasion of the access holes of the final superstructures. Conclusion: In this study, provisional composite restorations showed the best sealing properties. Provisional composite restorations may prevent bacterial invasion of the access holes of the final superstructures. Keywords: access hole, dental prosthesis, implant restoration, microbiological evaluation, prevention of peri-implantitis, sealing material, sealing properties

Purpose: To the assess whole salivary cotinine and interleukin 1β (IL-1β) levels among individuals involuntarily exposed to vapor from electronic nicotine delivery systems (ENDS) (test group) and unexposed individuals (control group). Materials and Methods: Demographic data and information related to ENDS vapor exposure were collected using a questionnaire. Unstimulated whole saliva samples were collected, unstimulated whole-saliva flow rate (UWSFR) was calculated, and cotinine and IL-1β levels were determined using enzyme-linked immunosorbent assay. Sample-size estimation and statistical analysis were performed. Regression analysis was performed to determine the correlation between whole salivary cotinine and IL-1β levels. Statistical significance was set at p < 0.05. Results: Forty-eight individuals (24 and 24 in test and control groups, respectively) were included. Mean ages of individuals in the test and control groups were comparable. In the test group, the mean duration for which the individuals inhaled vapor from ENDS in each session was 22.3 ± 9.5 min and they were exposed to ENDS vapor 12.2 ± 2.4 times daily. There was no difference in the UWSFR between patients in the test (0.21 ± 0.02 ml/min) and control (0.22 ± 0.04 ml/min) groups. Whole salivary cotinine (p < 0.001) and IL-1β (p < 0.001) levels were significantly higher in the test than control group. Conclusion: Young adults involuntarily exposed to vapor from ENDS express elevated whole salivary cotinine and IL-1β levels. Long-term exposure to ENDS vapor may potentially predispose vulnerable populations to oral and systemic inflammatory diseases. Keywords: cotinine, electronic nicotine delivery systems, interleukin 1 beta, unstimulated whole saliva, vaping

Purpose: Regenerative periodontal therapy using platelet-rich plasma (PRP) and bone substitutes with guided tissue regeneration (GTR) have been proposed as a therapeutic method to enhance the outcome of regenerative surgery. This includes light microscopic evaluation of retrieved ePTFE membranes to assess formation of new connective tissue attachment, and following the regeneration process. The objectives of this study were to evaluate the histological findings of retrieved ePTFE membranes using PRP and bone substitutes, the effect of PRP on graft materials, and the correlation of the findings with the clinical outcomes. Materials and Methods: Seventy-two (72) patients with chronic periodontitis, each of whom had one deep intrabony defect, were randomly included in two studies and treated using the same type of membrane and different bone substitutes. In the first study, 17 cases were treated with a natural bone mineral and a non-resorbable membrane (NBM + GTR), and 17 cases were treated with PRP + NBM + GTR. In the second study, 19 patients were treated with β-tricalcium phosphate and a non-resorbable membrane (β-TCP + GTR), and 19 patients were treated with PRP + β-TCP + GTR. In both studies, tissue integration of the retrieved ePTFE membranes and attached remnants were evaluated histologically. Results: Histological scores showed that membranes combined with NBM are better integrated than membranes combined with β-TCP; the difference between the two decreased with the addition of PRP. The application of PRP had no significant effect on the quality of membrane integration combined with NBM, whilst significantly improving the integration quality when combined with β-TCP. No correlations were detected between the histological scores and the clinical attachment level (CAL) gain in any of the groups. Conclusions: The present results indicate that: a) application of β-TCP and PRP may enhance membrane integration and periodontal healing, and b) histological examination of retrieved membranes may provide valuable additional information with regard to the clinical findings. Keywords: β-tricalcium phosphate, guided tissue regeneration, histological evaluation, natural bone mineral, platelet-rich plasma

Purpose: To analyse the epidemiological profile of dental caries in children aged 5 and 12 years in the city of Benguela, Angola. Materials and Methods: This was an observational, analytical, cross-sectional study conducted in 2019 with 190 12-year-old schoolchildren and 240 5-year-old schoolchildren from the public education system in Benguela, Angola. The relationship between dental caries and dental characteristics, sociodemographic factors, access to dental services, oral hygiene practices, and eating habits was analysed. Dental condition was evaluated using the dmft and decayed, missing, and filled teeth (DMFT) indices. Results: It was found that 62.63% (n = 119) of 12-year-old students and 42.08% (n = 101) of 5-year-old students were free from dental caries. The average DMFT was 0.76 + 1.35 and dmft was 2.19 + 2.95. The majority of children (56.51%) had never been to the dentist, had no dental elements restored, and none of the students used dental floss. The proportion of students who consumed sweets every day was higher at 5 years of age (46.25%) than at 12 years of age (22.63%). There was a statistically significant association (P = 0.01) between the higher incidence of dental caries and peri-urban location among 5-year-old schoolchildren. Conclusion: This study showed that the prevalence of dental caries in the permanent dentition of schoolchildren in Benguela is very low; however, the situation is critical in the primary dentition, especially in the peri-urban area. The limited access to dental surgeons and lack of treatment for affected teeth highlight the need to implement and develop public policies to promote oral health. Keywords: dental caries, child, Angola, health services accessibility

Summary: The Minamata Convention resulted in restrictions in the use of amalgam in daily dental practice. This opens up new discussions about the biocompatibility of amalgam, but also of composites as alternative materials. In the following review article, these issues will be discussed in more detail to provide dentists with a knowledge base for themselves and for communication with their patients. In addition to mercury in amalgam or monomers in composites, bisphenol A and nanoparticles generated during the grinding, polishing or removal of restorations must also be included in the biocompatibility evaluation. In laboratory tests, these substances cause toxic reactions, and bisphenol A also exhibits estrogen-like effects. However, it must be taken into account that the concentrations used in laboratory tests are much higher than in clinical practice. Thus, both amalgam and composite can be used in the general population. Nevertheless, for scientifically, politically and legally defined risk groups (e.g. dental personnel, allergic persons, pregnant or lactating women, children under 15 years of age, people with certain systemic diseases), indication restrictions and precautionary measures must be observed. The well-known amalgam discussion has taught us the importance of thorough and open risk communication with the patient. Keywords: bisphenol-A, child, lactation, mercury, pregnancy

Purpose: This study analysed the relationship between caregiver-related factors (dental knowledge, attitude, behaviour, and health status) and early childhood caries. It aimed to explore better intervention methods for reducing caries prevalence in preschool children. Materials and Methods: This cross-sectional investigation was carried out in Zhejiang, China. A total of 1344 guardians (parents and grandparents) paired with their children aged 3–5 years old were enrolled. The guardians completed structured questionnaires, which included their attitude, knowledge level and oral health status. The children received dental examinations. All of the data were analysed with R software. Chi-squared and Fisher’s exact tests were used to analyse different variables. Multinomial logistic regression with stepwise procedures and curve fitting was used to explore the relationship between guardians’ risk factors and the level of early childhood caries. Results: Guardians have a great influence on the incidence of early childhood dental caries. When guardians pay attention to oral health and have a good command of relevant knowledge, then the risk of early dental caries in children is low (p = 0.027). The guardians’ dental problems, e.g. dental caries (p = 0.0002), gingival bleeding problems (p = 0.049) and chewing discomfort experience (p = 0.049), demonstrated statistically significant correlations with early childhood caries levels. Conclusion: Guardians’ attitudes, knowledge, and oral health status had a statistically significant relationship with the level of early childhood caries in their children/grandchildren. Instead of instructing schoolchildren about oral health, multiple-level dental knowledge instruction of guardians is needed to prevent early childhood caries. Keywords: child health, dental caries, guardians, risk factors

Purpose: To investigate the anti-caries effects of the school-based fluoride varnish (FV) application at 3-month intervals and weekly fluoride mouthrinsing (FMR) on primary teeth and to evaluate the anti-microbial effects of FV or FMR on cariogenic bacteria among Myanmar children. Materials and Methods: A 6-month interventional study was conducted on 234 schoolchildren who were divided into three groups: group A received FV application at 3-month intervals, group B received weekly FMR, and group C received no fluoride application. A clinical oral examination and caries risk tests were performed at baseline and the 6-month follow-up. Results: Caries prevalence and the debris score did not change statistically significantly from baseline to the 6-month follow-up in all groups, whereas the dmfs score statistically significantly increased in group C (p = 0.001). The plaque and saliva scores of Dentocult SM statistically significantly decreased in group A (p = 0.049 and p = 0.006), but those scores statistically significantly increased in group C (p = 0.001 and p = 0.014) after six months. On the other hand, no statistically significant changes were observed in group B. Although the Cariostat scores decreased from baseline to the 6-month follow-up in group A and group B, but increased in group C, no statistically significant differences were observed in any of the groups. Conclusion: Better anti-microbial effects were obtained for children who received FV application than for those who received FMR, but no statistically significant difference existed between the anti-caries effects of these two approaches. Keywords: caries experience, caries risk tests, fluoride mouthrinse, fluoride varnish.

Purpose: Recently, increasing attention has been paid to the function of long non-coding RNAs (lncRNAs) in osteogenic differentiation (OD) of dental pulp stem cells (DPSCs). LINC01133 was reported to have a close relationship with tumorigenesis for multiple cancers, but no study has yet explored the role of LINC01133 in modulating OD of DPSCs. Materials and Methods: Alizarin red S (ARS) staining and alkaline phosphatase (ALP) staining were perfomed to assess the OD potential of DPSCs. Osteogenic markers including runt-related transcription factor 2 (RUNX2), osterix (OSX) and ALP expression levels in DPSCs were monitored by qRT-PCR and Western blot before and after cell transfection. Luciferase reporter gene assay detected the relationship between LINC01133 and miR-199b-5p. Results: The expression of LINC01133 was low, while miR-199b-5p was increasingly expressed during OD of DPSCs. Overexpression of LINC01133 in DPSCs resulted in decreased expression of RUNX2, OSX, ALP, DSPP and DMP1, whose expression was reversed in DPSCs after transfections of miR-199b-5p overexpression. Co-transfection of pcDNA3.1-LINC01133 and miR-199b-5p mimic led to elevated expression of RUNX2, OSX, ALP, DSPP and DMP1 compared with pcDNA3.1-LINC01133 transfection alone. LINC01133 served as a sponge of miR-199b-5p. AKT3 was verified as a downstream effector of miR-199b-5p in DPSCs. Conclusion: LINC01133 inhibits the OD of DPSCs by upregulating AKT3 via sponging miR-199b-5p, which may act as a potential diagnostic biomarker for dentin regeneration in the dental pulp. Keywords: AKT3, DPSCs, competitive endogenous RNA, LINC01133, miR-199b-5p, osteogenic differentiation

Purpose: The oral cavity is an important entry point for SARS-CoV-2 infection. This study tested whether four commercially available mouthrinses and dentifrices have in vitro virucidal activity against SARS-CoV-2 (≥4 log10 reduction in viral titer). Materials and Methods: One part of stock SARS-CoV-2 virus plus one part 0.3 g/l bovine serum albumin were mixed with eight parts of test product solution. After 30 s for the rinses, or 60 s for the dentifrices, the mixture was quenched in an appropriate neutralizer, serially diluted, and inoculated onto Vero E6 cells to determine viral titer. Triplicate runs were performed for each test condition with appropriate controls for test product cytotoxicity, viral interference, and neutralizer effectiveness. Test products included: 1.5% hydrogen peroxide (H2O2) rinse; 0.07% cetylpyridinium chloride (CPC) rinse; 0.454% stannous fluoride (SnF2) dentifrice A; and 0.454% SnF2 dentifrice B. Results: The 1.5% H2O2 rinse, 0.07% CPC rinse, SnF2 dentifrice A, and SnF2 dentifrice B all produced > 4 log10 reduction in SARS-CoV-2 titer. Conclusion: All four test products displayed potent virucidal activity in vitro. Clinical studies are warranted to determine what role, if any, these oral care products might play in preventing transmission of SARS-CoV-2 or in the management of patients currently diagnosed with COVID-19 illness. Keywords: COVID-19, dentifrice, mouthwash, SARS-CoV-2, virucide

Purpose: The effectiveness of using different laser therapy strategies in adjunct to scaling and root planing (SRP) for treatment of periodontitis remains unclear. This study compared the treatment outcome of SRP and its combination with Er:YAG laser and/or photobiomodulation on patients with severe periodontitis. Materials and Methods: A total of 50 patients were included, whose teeth were divided into 4 quadrants: 1. SRP+Er:YAG laser irradiation+photobiomodulation treatment; 2. SRP+Er:YAG laser irradiation treatment; 3. SRP+photobiomodulation treatment; 4. SRP treatment only. An Er:YAG laser at powers of 50 or 40 mJ and an Nd:YAG laser at 50 mJ were used. Patients were followed-up for 3 months. Periodontal clinical parameters (probing depth [PD], clinical attachment level [CAL], plaque index [PLI] and bleeding index [BI]), inflammation factors (melatonin and MMP-8 levels) and pain intensity were compared among the 4 quadrants. Results: The quadrant treated by SRP combined with Er:YAG laser irradiation and photobiomodulation had statistically significantly lower PD (p = 0.0081 at 1 month; p < 0.0001 at 3 months), CAL (p = 0.003), PLI (p = 0.0011) and BI levels (p = 0.005 at 1 month; p = 0.0236 at 3 months) compared with the other 3 quadrants. In addition, melatonin (p = 0.0006 at 1 month; p = 0.0002 at 3 months) and MMP-8 levels (p = 0.0012; only at 1 month) were also statistically significantly higher. Conclusion: Of the treatments tested here, SRP combined with Er:YAG laser irradiation and photobiomodulation had the best short-term outcome for severe periodontitis. Keywords: YAG laser, inflammation, laser therapy, periodontitis, photobiomodulation

Purpose: The role of developmental endothelial locus-1 (DEL-1) in Porphyromonas gingivalis (P. gingivalis)-induced periodontitis and the related molecular mechanisms are unclear. This study aimed to investigate the effect of DEL-1 on SH3 Domain Binding Protein 2 (SH3BP2) expression, and to explore the regulatory role of DEL-1 in periodontal inflammation. Materials and Methods: We constructed a P. gingivalis-induced rat experimental periodontitis model, and cultured P. gingivalis-stimulated THP-1 cells in vitro. THP-1 cell viability and cell cycle were examined by CCK-8 and flow cytometry. Rat gingival tissues were collected for hematoxylin-eosin staining. The expression of SH3BP2 and nicotinamide phosphoribosyltransferase (NAMPT) was examined using Western blot. Results: We found that the proliferation of P. gingivalis-infected THP-1 cells was increased by DEL-1. DEL-1 inhibited the expression of NAMPT and SH3BP2 in gingiva tissues of rats with periodontitis as well as in P. gingivalis-infected THP-1 cells. Conclusions: Overexpression of DEL-1 downregulated SH3BP2 expression and reduced gingival inflammation induced by P. gingivalis. DEL-1 presents some regulatory effects on gingival inflammation in a P. gingivalis-induced rat experimental periodontitis model, suggesting the therapeutic potential of DEL-1 in regulating periodontal inflammation. Keywords: developmental endothelial locus-1, periodontitis, Porphyromonas gingivalis, nicotinamide phosphoribosyl- transferase, SH3 domain binding protein 2

Purposes: The aim of this sytematic review was to evaluate the potential association of COVID-19 infection with oral health. Materials and Methods: Screening in different databases (PubMed/MEDLINE, Google Scholar, and Embase databases) was performed to identify relevant articles, focusing on the oral health of patients with COVID-19, and published up to November 2021. 5194 articles were identified, and 29 fulfilled the inclusion criteria. Results: Patients presenting more severe periodontal or dental diseases were at an increased risk of developing COVID-19 complications and being admitted to intensive care units. According to the included articles, U-shaped lingual papillitis and aphthous-like ulcers on the tongue are the most frequent lesions assessed in the oral cavity of COVID-19 patients, while xerostomia seems to be an early COVID-19 diagnostic symptom. Apart from the presence of the virus, the global lockdown had a detrimental impact on oral health. The occurrence of dental emergencies was augmented during this time due to the postponement of numerous non-emergency dental procedures. Conclusions: The presence of SARS-CoV-2 in periodontal tissues and salivary fractions may explain the presence of oral lesions during the infection. However, the virus’s direct or indirect effect on oral mucosa is unclear. It is important to consider that these manifestations might be attributed to underlying comorbidities, or co-existing or subsequent lesions produced by local irritants. Keywords: COVID-19, oral health, oral manifestation, periodontal diseases, SARS-CoV-2

Purpose: Cortisol levels (CL) in peri-implant sulcular fluid (PISF) samples in relation to type-2 diabetes mellitus (T2DM) and peri-implantitis remain unaddressed. It is hypothesised that PISF CL are higher in patients with type-2 diabetes and peri-implantitis than in healthy patients without and with peri-implantitis. The aim was to assess the PISF CL of peri-implantitis patients without and with T2DM. Materials and Methods: Peri-implantitis patients with T2DM (group 1), T2DM patients without peri-implantitis (group 2), non-diabetic patients with peri-implantitis (group 3) and non-diabetic patients without peri-implantitis (group 4) were included. Demographics were recorded; and patients’ medical and dental records were assessed. Peri-implant modified plaque-index (mPI), modified gingival index (mGI), and probing depth (PD) and crestal bone loss (CBL) were recorded. The PISF was collected and CL were determined. p < 0.01 was considered statistically significant. Results: Each of the four groups included 16 subjects (n = 64) with no difference in mean age. In groups 1 and 2, the mean duration of T2DM was 10.5 ± 0.8 and 10.6 ± 0.4 years, respectively. Mean HbA1c levels (p < 0.01) were higher and clinicoradiographic parameters (p < 0.001) were worse in group 1 than in the other groups. The median PISF volume and mean CL were higher in groups 1 (p < 0.01) and 3 (p < 0.01) than groups 2 and 4. There was a statistically significant correlation between PD and CL in group 3 (p < 0.001). Conclusion: Cortisol levels in the PISF are higher in T2DM and non-diabetic patients with peri-implantitis than in healthy individuals with and without peri-implantitis. Hyperglycemia did not influence peri-implant clinicoradiographic parameters and CL in the present patient population. Keywords: cortisol, dental implant, inflammation, peri-implantitis, unstimulated whole saliva