Abstract The aim of this study was to evaluate the effect of different die materials used in the indirect resin composite restorative technique on the fracture resistance and failure mode of restored molars and compare it with the direct resin composite restorative technique. Two flexible die silicone materials for dental models (Die Silicone – Voco and Scan die – Yller) and a type IV dental stone material (Fujirock EP – GC) were evaluated. Sixty third molars were selected and divided into four groups: indirect resin composite restoration – Die silicone (IRCR-DS); indirect resin composite restoration – Scan die (IRCR-SD); indirect resin composite restoration – Fujirock EP (IRCR-FR), and direct resin composite restoration (DRCR). Class II MOD cavities were prepared with 5 mm of buccolingual width and depth. The specimens were restored and subjected to an axial compression load until fracture, and the data were analyzed by one-way ANOVA and Tukey’s HSD test (α=.05). The fracture mode was classified into restorable and unrestorable fractures. Fracture resistance values were influenced by the die material used for the IRCR fabrication and by the restorative technique (p<.001). Fracture resistance mean values and standard deviation were: IRCR-DS: 1835.5 ± 324.0 A; IRCR-SD: 1732.5 ± 384.1 AB; IRCR-FR: 1419.3 ± 318.8 BC; and DRCR: 1100.6 ± 224.9 C. Restorable fracture was more prevalent. IRCR with flexible die casts promoted higher fracture resistance and lower prevalence of unrestorable fractures.

Abstract The genetic basis of oral epithelial (OED) is unknown, and there is no reliable method for evaluating the risk of malignant transformation. Somatic mutations are responsible for the transformation of dysplastic mucosa to invasive cancer. In addition, these genomic variations could represent objective markers of the potential for malignant transformation. We performed whole-exome sequencing of 10 OED samples from Brazilian and Chilean patients. Using public genetic repositories, we identified 41 deleterious variants that could produce high-impact changes in the amino acid structures of 38 genes. In addition, the variants were filtered according to normal skin and Native American genome profiles. Finally, 13 genes harboring 15 variants were found to be exclusively related to OED. High-grade epithelial dysplasia samples showed a tendency to accumulate highly deleterious variants. We observed that 62% of 13 OED genes identified in our study were also found in head and neck squamous cell carcinoma. Among the shared genes, eight were not identified in oral squamous cell carcinoma. To our knowledge, we have described for the first time 13 genes that are found in OED in a Latin American population, of which five genes have already been observed in oral squamous cell carcinoma. Through this study, we identified genes that may be related to basal biological functions in OED.

Abstract The aim of the present study was to assess whether mothers’ sense of coherence (SOC) was a predictor of decline in oral health-related quality of life (OHRQoL) of preschoolers. A 3-year cohort study was conducted in Diamantina, Brazil. At baseline, 162 preschoolers aged one to three years were randomly selected from among children registered in local Primary Healthcare Units. In the first stage, mothers completed a sociodemographic questionnaire, the Brazilian version of the Early Childhood Oral Health Impact Scale (B-ECOHIS), and the Brazilian short version of the Sense of Coherence Scale (SOC-13). The total score obtained from the SOC-13 was used to select exposed and unexposed children. Clinical examinations were performed to detect the presence of dental caries, traumatic dental injury, and malocclusion. At follow-up, mothers completed the sociodemographic questionnaire and the B-ECOHIS again. The incidence of severe dental caries and adherence to the proposed treatment at baseline were evaluated. A decline in OHRQoL was considered if there was an increase in the B-ECOHIS score of at least one unit. The chi-square test and Poisson regression were performed. A total of 151 preschoolers participated in the study, among whom 37.7% showed a decline in OHRQoL. Mothers’ SOC was not associated with a negative impact on OHRQoL (RR = 1.24; 95%CI = 0.81–1.88), while the incidence of severe dental caries had a greater impact on the decline in OHRQoL (RR = 2.02; 95%CI = 1.29–3.16). Mothers’ low SOC was not a predictor of decline in the OHRQoL of preschoolers after a 3-year follow-up period.

Abstract The aim of this study was to develop and validate a questionnaire that allows a systematic evaluation of the impact of self-perceived oral health on expectations of getting a job on adult population seeking dental care, and to describe its associations with demographic characteristics, job related, and health coverage variables. We designed a descriptive cross-sectional study including men and women aged 18 to 65 years from a population seeking dental services in a walk-in clinic. In a first stage we design and validated an instrument on a sample of 100 subjects. The questionnaire was registered in a Likert scale, with higher scores represented higher impact of the oral status self-perception on employability. We calculated internal consistency, construct validity, and domains validation. The final instrument consisted in an 18-item questionnaire (Cronbach α = 0.814), grouped into two domains based on exploratory and confirmatory factor analysis. The total variance explained with values >1 was 66 percent, grouping questions into six components. One domain refers to oral health status and importance of dental aesthetics, while the other refers to specific job-seeking elements. In a second stage we applied the questionnaire on 800 participants from the same population of reference. Women, people who intended to change jobs, those younger than 40 years old, having health insurance, and higher educational level showed statistically significant higher scores than their counterparts (p<0.001). We developed a tool that enables evaluating the impact of self-perceived oral health on expectations of getting a job for adults seeking emergency care in a dental clinic.

Abstract The aim of this study is to report an original case series of synchronous jawbone diseases. Data of patients seen over 13 years were extracted from the files of three Oral Radiology and Pathology diagnostic centers in Brazil. The clinical, radiographic, and laboratory characteristics were tabulated and analyzed by the authors; the patients were described according to lesion type. Seventy-two synchronous jawbone diseases were included in this study. Florid osseous dysplasia, Gorlin-Goltz syndrome, and cherubism were the most frequent disorders reported in this case series. In addition, the posterior mandible area was the main site of manifestation. Florid osseous dysplasia and Gorlin-Goltz syndrome represented two-thirds of our samples. With the utilization of adequate demographic, clinical, and radiologic information, it is possible to diagnose most of the synchronous lesions of jawbones. Sometimes, however, we need complementary exams, such as histopathologic and biochemical analysis or dosing of calcium, phosphorus, and alkaline phosphatase.

Abstract This study evaluated the bioactive potential of a macro-porous chitosan scaffold incorporated with calcium hydroxide (CH-Ca) and functionalized with bioactive doses of simvastatin (SV) for bone tissue regeneration. Initially, the bioactive dose of SV in osteoblastic cells (SAOS-2) was determined. For the direct contact experiment, SAOS-2 cells were plated on scaffolds to assess cell viability and osteogenic differentiation. The second assay was performed at a distance using extracts from scaffolds incubated in culture medium to assess the effect of conditioned medium on viability and osteogenic differentiation. The initial screening showed that 1 μM SV presented the best biostimulating effects, and this dose was selected for incorporation into the CH-Ca and pure chitosan (CH) scaffolds. The cells remained viable throughout the direct contact experiment, with the greatest cell density in the CH-Ca and CH-Ca-SV scaffolds because of their higher porosity. The CH-Ca-SV scaffold showed the most intense bio-stimulating effect in assays in the presence and absence of osteogenic medium, leading to an increased deposition of mineralized matrix. There was an increase in the viability of cells exposed to the extracts for CH-Ca, CH-SV, and CH-Ca-SV during the one-day period. There was an increase in ALP activity in the CH-Ca and CH-Ca-SV; however, the CH-Ca-SV scaffold resulted in an intense increase in the deposition of mineralized nodules, approximately 56.4% at 7 days and 117% at 14 days, compared with CH (control). In conclusion, functionalization of the CH-Ca scaffold with SV promoted an increase in bioactivity, presenting a promising option for bone tissue regeneration.

Abstract The diagnosis of irreversible pulpitis (IP) depends on clinical data, especially the chief complaint of the patient, visual inspection, response to the application of stimuli, and radiographic examination. The characterization of nerve fibers (NF) in IP may contribute to better interpret painful symptoms, but has been barely explored. This study sought to characterize the density and integrity of NF in 16 samples of IP and in five healthy pulps (HP) using S-100 and PGP 9.5 markers. Immunohistochemistry was performed to determine the density/mm2 of S-100+ and PGP 9.5+ in NF. The amount of degenerated NF was obtained by subtracting the total NF density from the amount of intact NF. Associations between NF density and integrity and symptomatology were calculated. All samples were positive for S-100 and PGP 9.5. Compared to HP samples (38.20/mm2), IP samples had a lower density of intact NF (6.24/mm2). A significantly higher density of degenerated NF was found in IP samples with spontaneous pain (39.59/mm2) compared to those with provoked pain (23.96/mm2) (p = 0.02). No association was observed between intensity of the inflammatory infiltrate and NF density and integrity (p > 0.05). The findings of this study suggest that pulpitis may involve different stages of degeneration and may be more advanced in cases with spontaneous pain. The symptoms reported by affected individuals do not appear to depend on the intensity of the inflammatory infiltrate, but rather on the integrity of NF.

Abstract Fanconi anemia is a rare autosomal recessive disease. In this disease, cytokine pathways can induce the bone marrow failure that is observed in individuals with Fanconi anemia. Interleukin IL-17 exhibits a protective effect in organisms because it induces neutrophil recruitment and shows a pathological role in several models of autoimmune diseases, periodontal disease, cancer, allograft rejection, and graft versus host disease. Polymorphisms in the IL17A and IL17RA genes were evaluated from DNA in saliva, comparing individuals with or without Fanconi anemia, using models of genotypic transmission (additive, dominant, and recessive). Polymorphisms in the IL17A and IL17RA genes (rs2241044 [C allele], rs879577 [C allele], rs9606615 [T allele], and rs2241043 [C allele]) were risk factors for developing Fanconi anemia. We also performed an analysis of gene markers with clinical variables in the Fanconi group. Polymorphisms in the IL17A gene (rs3819025 [A allele] and rs2275913 [G allele], respectively) were associated with an age of less than 20 years (p = 0.026; RP 0.65) and the female sex (p = 0.043; RP 0.88). The IL17RA gene was also associated with age and the presence of leukoplakia (a potentially malignant oral disorder). An age of less than 20 years was associated with rs917864 (T allele; p = 0.036; RP 0.67). The presence of leukoplakia was associated with rs17606615 (T allele; p = 0.042; RP 0.47). To our knowledge, this is the first study that associates IL17A and IL17RA gene polymorphisms with Fanconi anemia and examines rs2241044 polymorphisms in scientific literature thus far.

Abstract Oral cancer is a public health problem worldwide with approximately 300,000 new cases diagnosed every year and more than 170,000 deaths annually. Squamous cell carcinoma (SCC) accounts for approximately 90% of all oral malignancies and it is frequently preceded by lesions known as oral potentially malignant disorders (OPMDs). Screening programs for early detection of oral lesions have been conducted. Therefore, the objective of this research was to carry out an active search in a screening program in the city of Piracicaba, Brazil. High-risk patients were identified at the city’s health center through their medical records and referred for dental consultation. Other patients who opportunistically sought dental care were also seen and if they did not present risk factors for SCC, they were considered low-risk. A total of 756 patients were examined, and 445 met the criteria for the high-risk group and 311 for the low-risk group. It was possible to diagnose 27 OPMDs and six SCCs – 21 OPMDs and six SCCs occurred in high-risk patients and six OPMDs in low-risk patients. A chi-square test was applied and a statistically significant value (p = 0.006) was obtained for the detection of OPMD and SCC in patients of the high-risk group. Screening of high-risk patients through active search proved to be an effective program for diagnosing OPMD and SCC. Therefore, we encourage its implementation on a large scale to reduce the current scenario of this disease.

Abstract Scientific evidence about genetic and molecular changes in oral squamous cell carcinoma (OSCC) among smokers and non-smokers is inconclusive. This systematic review and meta-analysis assessed the effects of tobacco on the DNA of individuals with OSCC based on protein mutations. Electronic searches were conducted on PubMed, Ovid, Web of Science, and Scopus to identify observational studies published up to January/2022. The Joanna Briggs Institute tool was used for the critical appraisal of studies. The certainty of the evidence was evaluated. Twenty-three studies assessing 4,060 individuals (2,967 smokers vs. 1,093 non-smokers) were included in this review. Fifteen groups of proteins/genes were investigated. Analysis of the quality of articles revealed low risk of bias in most studies. The certainty of the evidence was very low. The meta-analysis confirmed no significant difference between smokers and non-smokers with respect to damage to GSTM1 (OR: 0.60; 95%CI: 0.30–1.18), GSTT1 (OR: 1.18; 95%CI:0.49–2.83), hydrolase proteins (Ku70 and Ku80) (OR: 0.74; 95%CI: 0.18–3.05), and transferase proteins (GSTM1, GSTT1, GSTM3) (OR: 0.74; 95%CI: 0.47–1.18). Most of the studies included showed that smokers are more likely to exhibit genetic instability. However, the meta-analysis revealed that smokers do not necessarily have more genetic alterations in the DNA than non-smokers.

Abstract This study aimed to translate and to perform the cultural adaptation of the instrument Illness Perception Questionnaire-Revised for Dental (IPQ-RD) into Brazilian Portuguese. The IPQ-RD consists of 34 items that assess the cognitive and emotional representation/perception of parents/caregivers of children with dental caries, with response options on a 5-point Likert scale, ranging from “Strongly agree” (score 1) to “Strongly disagree” (score 5). The higher the score, the lower the perception of the disease. The protocol consists of translation into Brazilian Portuguese, back-translation into English, revision by an Expert Review Committee, and pre-test (application in parents/caregivers of children in dental care). For some questions, the translated versions were identical (T1 = T2); for others, one version was preferred (T1 or T2); for still others, it was decided to modify terms to obtain greater clarity on the item (T3). In the first pre-test, three questions were misunderstood by more than 15% of the sample, after which the items were reviewed by the Expert Review Committee. In the second pre-test, the adapted version was applied to a new sample of parents/caregivers (n = 15) and the questions were understood by more than 85% of the participants. The Brazilian Portuguese version of the IPQ-RD was well understood by the evaluated population.

Abstract Protease-activated receptor-2 (PAR2) is associated with the pathogenesis of many chronic diseases with inflammatory characteristics, including periodontitis. This study aimed to evaluate how the activation of PAR2 can affect the osteogenic activity of human periodontal ligament stem cells (PDLSCs) in vitro. PDLSCs collected from three subjects were treated in osteogenic medium for 2, 7, 14, and 21 days with trypsin (0.1 U/mL), PAR2 specific agonist peptide (SLIGRL-NH2) (100 nM), and PAR2 antagonist peptide (FSLLRY-NH2) (100 nM). Gene (RT-qPCR) expression and protein expression (ELISA) of osteogenic factors, bone metabolism, and inflammatory cytokines, cell proliferation, alkaline phosphatase (ALP) activity, alizarin red S staining, and supernatant concentration were assessed. Statistical analysis of the results with a significance level of 5% was performed. Activation of PAR2 led to decreases in cell proliferation and calcium deposition (p < 0.05), calcium concentration (p < 0.05), and ALP activity (p < 0.05). Additionally, PAR2 activation increased gene and protein expression of receptor activator of nuclear factor kappa-Β ligand (RANKL) (p < 0.05) and significantly decreased the gene and protein expression of osteoprotegerin (p <0. 05). Considering the findings, the present study demonstrated PAR2 activation was able to decrease cell proliferation, decreased osteogenic activity of PDLSCs, and upregulated conditions for bone resorption. PAR2 may be considered a promising target in periodontal regenerative procedures.

Abstract Data on clinical management options for sleep bruxism in the primary dentition are inconclusive. This umbrella review aimed to synthesize the available evidence from systematic reviews (SRs) on the associated factors and treatment approaches for clinical management of sleep bruxism in children. A search was conducted in the MEDLINE/PubMed, Web of Science, Embase, and OpenGrey databases up to March 2022. SRs published on sleep bruxism in children containing data on associated factors or treatment outcomes were included. The AMSTAR-2 tool was used to assess the methodological quality of SRs. The search identified 444 articles, of which six were included. Sleep conditions, respiratory changes, personality traits, and psychosocial factors were the associated factors commonly identified. Treatments included psychological and pharmacological therapies, occlusal devices, physical therapy, and surgical therapy. All SRs included presented a high risk of bias. Overlapping of the included studies was considered very high. The best evidence available to date for the management of sleep bruxism in children is based on associated factors, with sleep duration and conditions, respiratory changes, as well as personality traits and psychosocial factors being the most important factors commonly reported by studies. However, there is currently insufficient evidence to make recommendations for specific treatment options.

Abstract The purpose of this study was to compare the outcomes of a modified gingival graft technique, in which the released flap is positioned and sutured over the graft, with the conventional free gingival graft (FGG) procedure, when both are used for gingival augmentation. A 12-month, multicenter parallel randomized controlled trial was conducted. Subjects with buccal RT2 gingival recessions and keratinized tissue width (KTW) < 2 mm in at least one mandibular incisor were randomized to control group (n = 20; conventional FGG) or test group (n = 20; modified FGG; flap sutured over FGG using sling sutures). The primary outcome (KTW) was measured at baseline and after 3, 6 and 12 months, as was keratinized tissue thickness (KTT). Postoperative pain (POP) and analgesic intake were also recorded. Both techniques promoted a significant increase in KTW and KTT when compared to baseline (p < 0.05) with no significant differences between groups (KTW change of 6.1±1.5 mm and 5.4±1.6 mm, for control and test, respectively; p=0.16). However, test group patients reported less POP after 7 days and used less analgesic medication than control group patients (p < 0.05). We concluded that the modified FGG was comparable to conventional FGG in augmenting keratinized tissue width and thickness at mandibular incisors, but resulted in less patient morbidity.

Abstract The aim of this retrospective cross-sectional investigation was to perform a 3D analysis of craniofacial morphology of patients with unilateral cleft lip and palate (UCLP) at two stages of skeletal maturation. Cone-beam computed tomography (CBCT) scans of 52 UCLP patients (34 prepubertal; 18 pubertal) were collected from an outpatient referral center for the treatment of craniofacial deformities. In total 15 multiplanar craniofacial landmarks were identified, 3D virtual surface models were created, and 13 variables were measured to assess the 3D Euclidean distances between landmarks and spatial position of the landmarks in the projected X, Y and Z components. Maxillary and mandibular pitch (clockwise, counterclockwise) rotation relative to the cranial base was also evaluated. The significance level was set at 5%. Maxillary retrusion value relative to the cranial base was higher and statistically significant greater (p = 0.028) in pubertal (SNA, 77.4° ± 6.2; N-ANS Y, 3.3 mm ± 3.1) than in prepubertal patients (SNA 81.0° ± 5.2; N-ANS Y, 5.8 mm ± 2.7). The posterior cranial base length (S-Ba Y) was significantly longer (p = 0.013) in pubertal (20.7 mm ± 3.4) than in prepubertal patients (18.4 mm ± 2.7). The upper facial height (N-ANS Z) was significantly greater (p = 0.01) in pubertal (46.9 mm ± 4.5) than in prepubertal patients (43.4 mm ± 3.0). Prepubertal and pubertal UCLP patients presented distinct patterns of craniofacial morphology, mainly in the sagittal component of the maxilla and in the posterior cranial base length. Pubertal patients had greater maxillary retrusion and posterior cranial base length.