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Abstract Objectives This review highlights the existence and association of Acinetobacter baumannii with the oro-dental diseases, transforming this systemic pathogen into an oral pathogen. The review also hypothesizes possible reasons for the categorization of this pathogen as code blue due to its stealthy entry into the oral cavity. Methodology Study data were retrieved from various search engines reporting specifically on the association of A. baumannii in dental diseases and tray set-ups. Articles were also examined regarding obtained outcomes on A. baumannii biofilm formation, iron acquisitions, magnitude of antimicrobial resistance, and its role in the oral cancers. Results A. baumannii is associated with the oro-dental diseases and various virulence factors attribute for the establishment and progression of oro-mucosal infections. Its presence in the oral cavity is frequent in oral microbiomes, conditions of impaired host immunity, age related illnesses, and hospitalized individuals. Many sources also contribute for its prevalence in the dental health care environment and the presence of drug resistant traits is also observed. Its association with oral cancers and oral squamous cell carcinoma is also evident. Conclusions The review calls for awareness on the emergence of A. baumannii in dental clinics and for the need for educational programs to monitor and control the sudden outbreaks of such virulent and resistant traits in the dental health care settings.

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Abstract Conventional views associate microbial biofilm with demineralization in root caries (RC) onset, while research on their collagenases role in the breakdown of collagen matrix has been sporadically developed, primarily in vitro. Recent discoveries, however, reveal proteolytic bacteria enrichment, specially Porphyromonas and other periodontitis-associated bacteria in subgingivally extended lesions, suggesting a potential role in RC by the catabolism of dentin organic matrix. Moreover, genes encoding proteases and bacterial collagenases, including the U32 family collagenases, were found to be overexpressed in both coronal and root dentinal caries. Despite these advancements, to prove microbial collagenolytic proteases’ definitive role in RC remains a significant challenge. A more thorough investigation is warranted to explore the potential of anti-collagenolytic agents in modulating biofilm metabolic processes or inhibiting/reducing the size of RC lesions. Prospective treatments targeting collagenases and promoting biomodification through collagen fibril cross-linking show promise for RC prevention and management. However, these studies are currently in the in vitro phase, necessitating additional research to translate findings into clinical applications. This is a comprehensive state-of-the-art review aimed to explore contributing factors to the formation of RC lesions, particularly focusing on collagen degradation in root tissues by microbial collagenases.

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Abstract In the era of ultra-connectivity, the proliferation of speculative notions driven by personal emotions eclipses the credibility of scientific evidence. This trend has led to an alarming surge in information pollution, particularly by the pervasive influence of social media platforms. Consequently, this overflow of falsehoods poses a significant threat to public health and overall societal well-being. In this sense, this critical review aims to present the harmful impacts of the health information pollution on society, health professionals, and health science, as well as strategies for their mitigation. The management of information pollution requires coordinated efforts to develop and implement multiple effective preventive and debunking strategies, such as the regulation of big tech companies’ actions and algorithm data transparency, the education of health professionals on responsible social media use, and the establishment of a novel academic culture, shifting from the valorization of productivism to socially relevant scientific production. By acknowledging the complexities of this contemporary issue and drawing insights from distinct perspectives, it is possible to safeguard the integrity of information dissemination and foster a more informed and resilient community.

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Abstract This narrative review critically examines some protocols of biomimetic restorative dentistry (BRD), which supposedly outperforms traditional adhesive techniques. This review explores the origins of BRD, introduces cognitive biases influencing the adoption of BRD protocols without evidence scrutiny, and discusses nine BRD protocols. For this, we searched randomized clinical trials and systematic reviews in the literature on the PubMed, Embase, and Cochrane Library CENTRAL databases, which lead to the following conclusions about the revised protocols: 1) The use of dyes excessively removes carious dentin; 2) Aluminum oxide air abrasion contributes to overtreatment and may pose long-term health risks to dental professionals; 3) Beveling enamel in posterior teeth is technically difficult and leads to unnecessary loss of adjacent sound enamel with no evidence of its use outperforming butt-joint preparations; 4) Deactivating matrix metalloproteinases with chlorhexidine shows no clinical evidence of improving restoration longevity. 5) “Elected“ gold-standard adhesive systems perform no better than other good performing available systems; 6) Immediate dentin sealing and resin coating result in similar post-operative sensitivity and longevity of indirect fillings as delayed dentin sealing; 7) Deep margin elevation is a viable alternative to manage subgingival margins in occlusoproximal cavities; 8) The process of “decoupling” with time lacks scientific evidence to support its use; 9) Placing fiber inserts on the pulpal floor and/or axial wall to minimize stress offers no benefits over current alternatives. In conclusion, more rigorous research is needed to validate BRD protocols, focusing on important clinical outcomes that impact in the longevity of the restoration, such as fracture, debonding, post-operative sensitivity, esthetic quality, presence of caries lesions adjacent to restorations and patients’ satisfaction need to be thoroughly investigated. Reliance on anecdotal evidence, clinical experience, and common sense propagates myths and undervalues the need for a critical approach in evaluating dental techniques.

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Abstract Aim: Polymorphisms in the COMT gene can alter enzymatic functions, raising levels of endogenous catecholamines, which stimulates beta-adrenergic receptors related to pain. This study aimed to evaluate whether a polymorphism in the COMT gene (rs4818) is associated with dental pain in children. Methodology: A cross-sectional study was conducted with a representative sample of 731 pairs of children and parents randomly selected from a population-based sample of eight-year-old children. Reports of dental pain was evaluated using a question directed at the parents and self-reported pain using the Faces Pain Scale – Revised. Dental caries experience was determined using the Decayed, Missing, and Filled Teeth (DMFT) index. For genetic analysis, DNA was obtained from oral mucosa epithelial cells of 352 children randomly selected from the initial sample. Results: Children with the CC genotype had higher odds of reporting moderate to intense pain than those with the GG genotype (OR=3.60; 95% CI=0.80–16.20; p=0.03). These same children had greater odds of parental reports of pain (OR=1.93; 95% CI=0.91-4.08; p=0.02). Moreover, lower schooling of parents/guardians and caries experience in the primary dentition were significantly associated with greater odds of a parental report of dental pain (OR=2.06; 95% CI=1.47–2.91; p<0.001; OR=6.26; 95% CI=4.46–8.78; p<0.001). Conclusions: The rs4818 polymorphism of the COMT gene is associated with dental pain. Children with the C allele are more likely to report higher levels of pain. Clinical Relevance: Even though the experience of pain is subjective and multifactorial, this study raises the hypothesis that there is a genetic predisposition to dental pain that should be considered in clinical practice.

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Abstract During oral surgery and temporomandibular joint repositioning, pain hypersensitivity often occurs due to irritation or inflammation of the nerve endings in the orofacial region. Objective: This study aimed to investigate the effects of ECa 233, a Centella asiatica–standardized extract, on the development of mechanical hyperalgesia and allodynia induced by chronic constriction injury of the infraorbital nerve in mice. Methodology: The right infraorbital nerves of the mice were ligated. Oral carbamazepine (20 mg/kg) or ECa 233 (30, 100, or 300 mg/kg) was administered daily for 21 days. Von Frey and air-puff tests were performed on both sides of the whisker pad on days 0, 7, 14, and 21. Thereafter, the expression of purinergic receptor subtype 3 (P2X3) and voltage-gated sodium channel 1.7 (NaV1.7), a transmembrane protein, in the trigeminal ganglion and c-fos immunoreactivity-positive neurons in the trigeminal nucleus caudalis was assessed. Results: After 21 days of infraorbital nerve ligation, the mice showed allodynia- and hyperalgesia-like behavior, P2X3 and NaV1.7 were upregulated in the trigeminal ganglion, and nociceptive activity increased in the trigeminal nucleus caudalis. However, the oral administration of carbamazepine (20 mg/kg), ECa 233 (100 mg/kg), or ECa 233 (300 mg/kg) mitigated these effects. Nevertheless, ECa 233 failed to affect NaV1.7 protein expression. Conclusion: Carbamazepine and ECa 233 can prevent pain hypersensitivity in mice. Considering the side effects of the long-term use of carbamazepine, ECa 233 monotherapy or combined ECa 233 and carbamazepine therapy can be used as an alternative for regulating the development of hypersensitivity in trigeminal pain. However, further detailed clinical studies should be conducted to provide comprehensive information on the use of ECa 233.

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Abstract Objective This retrospective study aimed to analyze the clinical efficacy of two regenerative surgical methods — Bio-Oss granules combined with barrier membranes and Bio-Oss Collagen alone — and to help clinicians achieve better periodontal regeneration outcomes in the specific periodontal condition. Methodology Patients who underwent periodontal regeneration surgery from January 2018 to April 2022 were retrospectively screened, and their clinical and radiographic outcomes at 6 months postoperatively were analyzed. The probing depth (PD), clinical attachment level (CAL), bleeding on probing (BOP), gingival recession (GR), distance from the cemento-enamel junction to the bottom of the bone defect (CEJ-BD), and depth of intrabony defects (INFRA) were recorded before the operation (T0) and 6 months after it (T1), and subsequently compared. Results In total, 143 patients were included — 77 were placed in the Bio-Oss group and 66 were placed in the Bio-Oss Collagen group. All indicators, including PD and CAL at T1, showed significant differences compared to baseline, for both groups (P<0.001). PD reduction was greater in the group receiving the Bio-Oss Collagen treatment (P=0.042). Furthermore, in cases when the baseline PD range was 7-11 mm and the age range was 35-50 years, PD reduction was more significant for patients receiving the Bio-Oss Collagen treatment (P=0.031, 0.023). A linear regression analysis indicated that postoperative PD and CAL were positively correlated with baseline values, and that the efficacy tended to decrease with increasing age. Conclusion Both the use of Bio-Oss Collagen alone and the use of Bio-Oss granules combined with barrier membranes resulted in significant effects in the treatment of periodontal intrabony defects. The Bio-Oss Collagen treatment generated more improvements in PD than the Bio-Oss granules combined with barrier membranes, particularly within the baseline PD range of 7-11 mm and the 35-50 years age group. Additionally, age was the main factor influencing the effectiveness of regenerative surgery for intrabony defects: older individuals exhibited fewer improvements.

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Abstract Objective: In this study, a comparative evaluation of the physicochemical properties of Cention N and other direct restorative materials was performed. Three restorative materials—a resin-modified glass ionomer (Fuji II LC), an alkasite-based resinous material (Cention N), and a resin composite (Tetric N Ceram)—were characterized in terms of degree of conversion, Knoop hardness number (KHN) ratio, flexural strength, elastic modulus, water sorption, water solubility, microshear bond strength to dentin, immediate microleakage, and radiopacity. Methodology: The microshear bond strength to dentin and microleakage of Cention N were evaluated with and without the application of an adhesive system (Tetric N Bond Universal). A one-way ANOVA test was used to analyze the data in terms of degree of conversion, KHN ratio, water sorption, water solubility, microshear bond strength to dentin, and radiopacity. A two-way ANOVA test (carried out considering the material type and ethanol aging as factors) was used to analyze the data in terms of flexural strength and elastic modulus. The Kruskal–Wallis test was used to statistically analyze the data on microleakage. A significance level of α=0.05 was used for all tests. Results: Fuji II LC was found to have the highest degree of conversion, water sorption, and microleakage, as well as the lowest flexural strength. Cention N had the highest solubility; when used with an adhesive system, it achieved bond strength and microleakage similar to those of the Tetric N Ceram composite. Tetric N Ceram had the highest degree of conversion, KHN ratio, and radiopacity. Conclusion: The properties of Cention N validate its efficacy as an alternative direct restorative material when used in conjunction with an adhesive system.

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Abstract Objectives This study aimed to analyze the cost-effectiveness of whitening toothpastes and at-home bleaching for the treatment of tooth discoloration. Methodology A cost-effectiveness economic analysis was conducted, and eight randomized clinical trials were selected based on the whitening agent product used: blue covarine dentifrices (BCD), hydrogen peroxide dentifrices (HPD), dentifrices without bleaching agents (CD, negative control), and 10% carbamide peroxide (CP10, positive control) for at-home bleaching. The consumer/patient perspective was adopted, macro-costing techniques were used and a decision tree model was performed considering the costs in the American and Brazilian markets. The color change evaluation (ΔE*ab) was used to calculate the effectiveness of tooth bleaching. A probabilistic analysis was performed using a Monte Carlo simulation and incremental cost-effectiveness ratios were obtained. Results CP10 resulted in the highest cost-effectiveness compared to the use of dentifrices in both markets. In Brazil, HPD was more cost-effective than BCD and CD. In the US, the increased costs of HPD and BCD did not generate any whitening benefit compared to CD. Conclusions CP10 was more cost-effective than BCD and HPD for tooth bleaching from the perspectives of the Brazilian and American markets. Decision-making should consider the use of CP10 for treating tooth discoloration.

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Abstract Associations between the WNT5A rs566926 variant and non-syndromic orofacial cleft (NSOC) have been reported in different populations. Objective This study aimed to investigate the role of the rs566926 single nucleotide polymorphism (SNP) in WNT5A and its interactions with SNPs in BMP4, FGFR1, GREM1, MMP2, and WNT3 in the occurrence of NSOC in a Brazilian population. Methodology A case-control genetic association study was carried out involving participants from four regions of Brazil, totaling 801 patients with non-syndromic cleft lip with or without cleft palate (NSCL±P), 273 patients with cleft palate only (NSCPO), and 881 health volunteers without any congenital condition (control). Applying TaqMan allelic discrimination assays, we evaluated WNT5A rs566926 in an ancestry-structured multiple logistic regression analysis, considering sex and genomic ancestry as covariates. Interactions between rs566926 and variants in genes involved in the WNT5A signaling pathway (BMP4, FGFR1, GREM1, MMP2, and WNT3) were also explored. Results WNT5A rs566926 was significantly associated with an increased risk of NSCL±P, particularly due to a strong association with non-syndromic cleft lip only (NSCLO), in which the C allele increased the risk by 32% (OR: 1.32, 95% CI: 1.04–1.67, p=0.01). According to the proportions of European and African genomic ancestry, the association of rs566926 reached significant levels only in patients with European ancestry. Multiple interactions were detected between WNT5A rs566926 and BMP4 rs2071047, GREM1 rs16969681 and rs16969862, and FGFR1 rs7829058. Conclusion The WNT5A rs566926 polymorphism was associated with NSCL±P, particularly in individuals with NSCLO and high European ancestry. Epistatic interactions involving WNT5A rs566926 and variants in BMP4, GREM1, and FGFR1 may contribute to the risk of NSCL±P in the Brazilian population.

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Abstract Objective: We aimed to investigate the regulatory effects of HMGB1/TLR4 signaling pathway on the expression of IL-10 and VEGF in human bone marrow mesenchymal stem cells. Methodology: Human JBMSCs were isolated and cultured. Then, HMGB1 was added into the JBMSCs culture medium, and the protein and mRNA expression levels of IL-10 and VEGF were assessed. Moreover, cells were pretreated with a specific TLR4 inhibitor (TAK-242), and the expression changes of IL-10 and VEGF were compared. Results: Compared with the control group, exposure to HMGB1 in human JBMSCs up-regulated TLR4, IL-10, and VEGF secretion at both protein and mRNA levels (P<0. 05). In addition, the increased expression of IL-10 and VEGF could be restrained in TAK-242 group compared with the HMGB1 group (P<0.05). Conclusions: The results indicated that HMGB1 activate TLR4 signaling pathway in Human JBMSCs, which plays a regulatory role in cytokines expression.

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Abstract Objective: This study aimed to investigate the effects of systemic administration of P. eurycarpa Yalt. plant extract on alveolar bone loss and oxidative stress biomarkers in gingival tissue in a rat model of experimental periodontitis. Methodology: 32 male Wistar albino rats, weighing 200–250 g, were divided into four groups (n=8): Healthy control (HC), Experimental periodontitis control (EPC), Experimental periodontitis 400 mg/kg (EP400), Experimental periodontitis 800 mg/kg (EP800). Experimental periodontitis was induced using the ligating method. Distilled water was administered to the HC and EPC groups and the plant extract was administered to the EP400 and EP800 groups by oral gavage at doses of 400 mg/kg and 800 mg/kg, respectively. The rats were sacrificed on the 15th day. The values of glutathione peroxidase GSH-Px, malondialdehyde (MDA), superoxide dismustase (SOD), interleukin-1β (IL-1β), interleukin-10 (IL-10), total antioxidant status (TAS), total oxidant status (TOS), oxidative stress index (OSI) in the gingival tissues were analyzed by ELISA tests. Alveolar bone loss was assessed using micro-CT images of the maxilla. Results: Although the IL-1β, TOS, OSI results of the healthy control group were lower than those of the other groups, the TAS values were higher (p<0.05). No significant difference was found in the biochemical parameters among the EPC, EP400, and EP800 groups (p>0.05). Alveolar bone loss was significantly reduced in the extract groups compared to the EPC group (p<0.001). Conclusion: Within the limitations of this study, it was observed that the systemic P. eurycarpa extract application reduced alveolar bone loss in a rat model of experimental periodontitis. Further studies are needed to elucidate the beneficial effects of P. eurycarpa.

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Abstract Objective: The aim of this population-based retrospective study was to compare the osteogenic effect of newly formed bone after maxillary sinus floor elevation (MSFE) and simultaneous implantation with or without bone grafts by quantitatively analyzing trabecular bone parameters. Methodology: A total of 100 patients with missing posterior maxillary teeth who required MSFE and implantation were included in this study. Patients were divided into two groups: the non-graft group (n=50) and the graft group (n=50). Radiographic parameters were measured using cone beam computed tomography (CBCT), and the quality of newly formed bone was analyzed by assessing trabecular bone parameters using CTAn (CTAnalyzer, SkyScan, Antwerp, Belgium) software. Results: In the selected regions of interest, the non-graft group showed greater bone volume/total volume (BV/TV), bone surface/total volume (BS/TV), trabecular number (Tb. N), and trabecular thickness (Tb. Th) than the graft group (p<0.001). The non-graft group showed lower trabecular separation (Tb. Sp) than the graft group (p<0.001). The incidence of perforation and bleeding was higher in the graft group than in the non-graft group (p<0.001), but infection did not significantly differ between groups (p>0.05). Compared to the graft group, the non-graft group showed lower postoperative bone height, gained bone height and apical bone height (p<0.001). Conclusion: MSFE with and without bone grafts can significantly improve bone formation. In MSFE, the use of bone grafts hinders the formation of good quality bone, whereas the absence of bone grafts can generate good bone quality and limited bone mass.

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Abstract Objective The present in vitro study incorporated niobium oxyhydroxide fillers into an experimental high-viscosity bulk-fill resin composite to improve its mechanical performance and provide it a bioactive potential. Methodology Scanning electron microscopy synthesized and characterized 0.5% niobium oxyhydroxide fillers, demonstrating a homogeneous morphology that represented a reinforcement for the feature. Fillers were weighed, gradually added to the experimental resin composite, and homogenized for one minute, forming three groups: BF (experimental high-viscosity bulk-fill resin composite; control), BF0.5 (experimental high-viscosity bulk-fill resin composite modified with 0.5% niobium oxyhydroxide fillers), and BFC (commercial bulk-fill resin composite Beautifil Bulk U, Shofu; positive control). In total, 10 specimens/groups (8 × 2 × 2 mm) underwent flexural strength (FS) tests in a universal testing machine (Instron) (500N). Resin composites were also assessed for Knoop hardness (KH), depth of cure (DoC), degree of conversion (DC), elastic modulus (E), and degree of color change (ΔE). The bioactive potential of the developed resin composite was evaluated after immersing the specimens into a simulated body fluid in vitro solution and assessing them using a Fourier-transformed infrared spectroscope with an attenuated total reflectance accessory. One-way ANOVA, followed by the Tukey’s test (p<0.05), determined FS, DC, KH, and ΔE. For DoC, ANOVA was performed, which demonstrated no significant difference between groups (p<0.05). Conclusions The high-viscosity bulk-fill resin composite with 0.5% niobium oxyhydroxide fillers showed promising outcomes as reinforcement agents and performed well for bioactive potential, although less predictable than the commercial resin composite with Giomer technology.

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Abstract Repositioning guides are commonly employed in clinical studies to ensure consistent tooth color measurements. Yet, their influence on measured color remains uncertain. Objective This study evaluated the impact of repositioning guides’ color and usage on tooth color measurement using a clinical spectrophotometer. Methodology In total, 18 volunteers participated in this study, in which the color of their upper left central incisor and upper left canine was measured with or without repositioning guides (control). The guides were made from pink, blue, or translucent silicone, as well as an acetate-based bleaching tray. Tooth color was measured in triplicates using a clinical spectrophotometer based on the CIELAB system. The standard deviations of these readings were used to estimate reproducibility, and color differences (ΔE00) between the measurements with guides and the control were calculated. Results Repositioning guides had a minimal effect on L* values and no effect on b* values. The use of pink silicone increased a* values, whereas blue or translucent silicone reduced them. Irrespective of the evaluated tooth, the lowest ΔE00 values were observed for the translucent silicone and bleaching tray. The usage of guides only affected data variability for the L* color coordinate. Conclusion Using repositioning guides can significantly impact the precision of tooth color measurement with a clinical spectrophotometer.

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Abstract Objective This study aimed to analyze the longitudinal bond strength of a universal adhesive and chemically characterize the dentin substrate under different acid etching protocols. Methodology Dentin samples were etched with polyacrylic acid 25% (PAA) for 10 seconds (n=3) and phosphoric acid 32% (PA) for 15 seconds (n=3) and analyzed by Fourier transform infrared spectroscopy – attenuated total reflectance (FTIR-ATR) before and after treatment. For collagen degradation, samples (n=12) were divided into 3 groups: PAA, PA, and Deionized water (control), and analyzed by the quantity of solubilized type I collagen C-terminal cross-linked telopeptides and solubilized C-terminal peptide in relation to total protein concentration (ICTPtp and CTXtp) and by their ultimate tensile strength (UTS). For the adhesive interface analysis, dentin samples (n=72) were divided into 3 groups: PAA, PA, and Self-etch (SE), and subdivided into 2 groups: 24 h (baseline) and 1 year. The following tests were performed: microtensile bond strength (μTBS) (n=48), scanning electron microscopy (SEM) (n=12), and nanoleakage (n=12). Results The FTIR of PAA showed lower reduction of the peaks in the phosphate group when compared to PA. For ICTPtp, PA showed a significantly higher value. For CTXtp, PA and PAA groups failed to statically differ from each other. UTS was significantly lower for PA. For μTBS, storage time significantly affected bond strength. The results were unaffected by the etching protocol. For SEM, after 1 year, PA had little evidence of degradation in the upper third of the adhesive interface in comparison to the other groups. Nanoleakage showed no considerable silver impregnation after 1 year in the SE group. Conclusion The use of PAA prior to a universal adhesive (when compared to PA) represents a less aggressive type of etching to dentin. However, self-etching still seems to be the best option for universal adhesive systems that have functional monomers in their composition.

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Abstract Denture biofilm acts as a potential reservoir for respiratory pathogens, considerably increasing the risk of lung infections, specifically aspiration pneumonia, mainly 48h after hospital admission. The establishment of a straightforward, affordable, and applicable hygiene protocol in a hospital environment for the effective control of denture biofilm can be particularly useful to prevent respiratory infections or reduce the course of established lung disease. Objectives To evaluate the anti-biofilm effectiveness of denture cleaning protocols in hospitalized patients. Methodology The maxillary complete dentures (MCDs) of 340 hospitalized participants were randomly cleaned once using one of the following 17 protocols (n=20): brushing with distilled water, toothpaste, or neutral liquid soap (controls); immersion in chemical solutions (1% sodium hypochlorite, alkaline peroxide, 0.12% or 2% chlorhexidine digluconate), or microwave irradiation (650 W for 3 min) combined or not with brushing. Before and after the application of the protocols, the biofilm of the intaglio surface of the MCDs was evaluated using two methods: denture biofilm coverage area (%) and microbiological quantitative cultures on blood agar and Sabouraud Dextrose Agar (CFU/mL). Data were subjected to the Wilcoxon and Kruskal-Wallis tests (α=0.05). Results All 17 protocols significantly reduced the percentage area of denture biofilm and microbial and fungal load (P<0.05). The highest percentage reductions in the area of denture biofilm were observed for 1% hypochlorite solution with or without brushing and for 2% chlorhexidine solution and microwave irradiation only in association with brushing (P<0.05). The greatest reductions in microbial and fungal load were found for the groups that used solutions of 2% chlorhexidine and 1% hypochlorite and microwave irradiation, regardless of the association with brushing (P<0.05). Conclusions A single immersion for 10 min in 1% sodium hypochlorite, even in the absence of brushing, proved to be a straightforward, rapid, low-cost, and effective protocol for cleaning the dentures of hospitalized patients.

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Abstract Objective the aim of this study was to analyze the influence of ozone therapy (OZN) on peri-implant bone repair in critical bones by installing osseointegrated implants in the tibia of ovariectomized rats. Methodology ovariectomy was performed on 30 Wistar rats, aged six months (Rattus novergicus), and, after 90 days, osseointegrated implants were installed in each tibial metaphysis. The study groups were divided into the animals that received intraperitoneal ozone at a concentration of 700 mcg/kg — OZ Group (n=15) — and a control group that received an intraperitoneal saline solution and, for this reason, was named the SAL group (n=15). The applications for both groups occurred during the immediate post-operative period on the 2nd, 4th, 6th, 8th, 10th, and 12th day post-surgery. At various stages (14, 42, and 60 days), the animals were euthanized, and tests were performed on their tibiae. These tests include histomorphometric and immunohistochemical analyses, computerized microtomography, sampling in light-cured resin for calcified sections, and confocal microscopy. The obtained data were then analyzed using One-way ANOVA and the Shapiro-Wilk, Kruskal-Wallis, and student t-tests (P<0.05). Results our findings indicate that the OZ group (3.26±0.20 mm) showed better cellular organization and bone neoformation at 14 days (SAL group, 0.90±1.42 mm) (P=0.001). Immunohistochemistry revealed that osteocalcin labeling was moderate in the OZ group and mild in the SAL group at 14 and 42 days post-surgery. The data from the analysis of calcified tissues (microtomography, histometric, and bone dynamism analysis) at 60 days showed no statistically significant differences between the groups (P=0.32). Conclusion it was concluded that ozone therapy anticipated the initial phases of the peri-implant bone repair process.

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Abstract Research would be important for obtaining a better understanding of voice complaints among patients with temporomandibular disorders (TMD). Objective The identification of predictors of voice disorders associated with TMD pain was made according to Diagnostic Criteria for TMD (DC/TMD) Axis I. Methodology Functional limitations were measured using the Jaw Functional Limitation Scales for mastication (JFLS-M), jaw mobility (JFLS-JM), and verbal and emotional expression (JFLS-VEE). Patients also completed the Hospital Anxiety and Depression Scale (HADS). The primary outcome was social-emotional and physical functioning as indicated by scores on the Voice-Related Quality of Life (V-RQOL) questionnaire. Multiple linear regression was used to model the relationship between the domains on the V-RQOL questionnaire and scores on the HADS and JFLS after adjusting for age, gender, DC/TMD diagnosis, pain intensity, and time since pain onset. Results The HADS-D (B=-1.15; 95% CI, -1.718 to -0.587; p<.001) and JFLS-VEE (B=-0.22; 95% CI, -0.40 to -0.06; p=.008) were significant predictors of scores on the V-RQOL questionnaire. Conclusion Higher scores on depression measures and limitations in verbal and emotional expression could exacerbate voice problems among TMD pain patients. Future research should promote multidisciplinary treatments for TMD pain-related voice disorders.

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Abstract Objective To determine whether intra-mucosal injection of injectable platelet-rich fibrin (i-PRF) can promote healing after Diode Laser Gingival Depigmentation (DLGD). Methodology A total of 20 arch sites of hyperpigmented gingiva of 10 patients underwent DLGD. For each patient, two arch sites were randomly assigned for either intra-mucosal injection of i-PRF (G1-i-PRF) (n=10 sites) or no treatment (G2-Control): (n=10 sites). Wound Healing Score (WHS), patient satisfaction, and Pigmentation Index (DOPI) were measured at 1 week and 1 and 3 months postoperatively. Histological assessment of tissue specimens was performed at baseline and 1 week. Results The percentage change in WHS at 1 week was significantly higher in G1 (58.34±15.43) compared to G2 (37.50±11.79). At day 1, 50% of patients in G1 were pain free compared with 75% in G2, who had mild pain. Mean DOPI decreased significantly at 3 months in both groups (P-value <0.001), without significant differences between groups. G1 specimens showed significantly higher epithelial thickness (P-value <0.001), as well as a higher number of blood vessels and less percentage of inflammatory cells. Conclusions i-PRF demonstrated better clinical and histological healing potential and less patient discomfort compared to sites without treatment after DLGD. Registered at https://clinicaltrials.gov/ as (NCT05283668).

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Abstract At low concentrations used for in-office bleaching gels, such as 6% HP, gingival barrier continues to be performed. If we take into account that, in the at-home bleaching technique, no barrier is indicated, it seems that the use of a gingival barrier fails to make much sense when bleaching gel in low concentration is used for in-office bleaching. Objective This double-blind, split-mouth, randomized clinical trial evaluated the gingival irritation (GI) of in-office bleaching using 6% hydrogen peroxide (HP) with and without a gingival barrier in adolescents, as well as color change and the impact of oral condition on quality of life. Methodology Overall, 60 participants were randomized into which side would or would not receive the gingival barrier. In-office bleaching was performed for 50 minutes with 6% HP in three sessions. The absolute risk and intensity of GI were assessed with a visual analogue scale. Color change was assessed using a digital spectrophotometer and color guides. The impact of oral condition on quality of life was assessed using the Brazilian version of the Oral Health Impact Profile (α=0.05). Results The proportion of patients who presented GI for the “with barrier” group was 31.6% and for the “without barrier” group, 30% (p=1.0). There is an equivalence for the evaluated groups regarding GI intensity (p<0.01). Color change was detected with no statistical differences (p>0.29). There was a significant impact of oral condition on quality of life after bleaching (p<0.001). Conclusions The use or not of the gingival barrier for in-office bleaching with 6% HP was equivalent for GI, as well as for bleaching efficacy, with improvement in the impact of oral condition on quality of life.

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Abstract Objective Platelet-rich fibrin (PRF) contains a variety of growth factors and bioactive molecules that play crucial roles in wound healing and angiogenesis. We aimed to evaluate the effects of PRF on tissue thickness and vascularization of the palatal donor site by ultrasound (USG) following subepithelial connective tissue harvesting. Methodology A subepithelial connective tissue graft was harvested from the palatal region with a single incision for root coverage in 20 systemically healthy patients. In the test group (n = 10), the PRF membrane was placed at the donor site, whereas no material was applied in the control group (n=10). Palatal tissue thickness (PTT) and pulsatility index (PI) were evaluated by USG at baseline and on the 3rd, 7th, 14th, 30th, and 90th days after surgery. The early healing index (EHI) was used to evaluate donor site healing for 30 days. Results PTT was significantly higher in the PRF group on the 3rd and 14th days after surgery when compared to the controls. In the PRF-treated group, PI levels were significantly higher than in the controls, especially on the 14th day. PTT increased significantly 90 days after surgery compared to the test group baseline, but controls showed a significant decrease. The PRF group showed statistically significant improvements in EHI scores compared to controls on days 3, 7, and 14. This study found a negative correlation between PI values and EHI scores on postoperative days three and seven in the test group. Conclusion USG is a non-invasive, objective method to radiographically evaluate the regenerative effects of PRF on palatal wound healing after soft tissue harvesting. To overcome graft inadequacy in reharvesting procedures, PRF application may enhance clinical success and reduce possible complications by increasing tissue thickness and revascularization in the donor area.

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Abstract Studies evaluating the roughness, wettability and microbial adhesion of 3D-printed resins for complete denture bases and teeth are scarce. Objective This study evaluated the surface roughness, wettability and adhesion of multispecies biofilms (Candida albicans, Staphylococcus aureus and Streptococcus mutans) on 3D-printed resins for complete denture bases and teeth compared to conventional resins (heat-polymerized acrylic resin; artificial pre-fabricated teeth). Methodology Circular specimens (n=39; 6.0 mm Ø × 2.0 mm) of each group were subjected to roughness (n=30), wettability (n=30) and biofilm adhesion (n=9) tests. Three roughness measurements were taken by laser confocal microscopy and a mean value was calculated. Wettability was evaluated by the contact angle of sessile drop method, considering the mean of the three evaluations per specimen. In parallel, microorganism adhesion to resin surfaces was evaluated using a multispecies biofilm model. Microbial load was evaluated by determining the number of Colony Forming Units (CFU/mL) and by scanning electron microscopy (SEM). Data were subjected to the Wald test in a generalized linear model with multiple comparisons and Bonferroni adjustment, as well as two-way ANOVA (α=5%). Results The roughness of the conventional base resin (0.01±0.04) was lower than that of the conventional tooth (0.14±0.04) (p=0.023) and 3D-printed base (0.18±0.08) (p<0.001). For wettability, conventional resin (84.20±5.57) showed a higher contact angle than the 3D-printed resin (60.58±6.18) (p<0.001). Higher microbial loads of S. mutans (p=0.023) and S. aureus (p=0.010) were observed on the surface of the conventional resin (S. mutans: 5.48±1.55; S. aureus: 7.01±0.57) compared to the 3D-printed resin (S. mutans: 4.11±1.96; S. aureus: 6.42±0.78). The adhesion of C. albicans was not affected by surface characteristics. The conventional base resin showed less roughness than the conventional dental resin and the printed base resin. Conclusion The 3D-printed resins for base and tooth showed less hydrophobicity and less adhesion of S. mutans and S. aureus than conventional resins.

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Abstract Oral soft tissue lesions require a precise diagnosis by oral biopsy with the ability to recognize these lesions within histopathological levels, so the instrument used for the incisions should be safe and cause little to no harm to the surrounding tissue. Objective This study compared a dual-wavelength diode laser and an Er, Cr:YSGG laser in oral soft tissue incisions to determine the most effective and safest laser system at the histopathological level. Methodology The (810 and 980 nm) dual-wavelength diode laser was used at 1.5 W and 2.5 W (CW) power settings, and the (2780 nm) Er, Cr:YSGG laser was used at 2.5 W and 3.5 W (PW) power settings. Both laser systems were used to incise the tissues of freshly dissected sheep tongue pieces to obtain the following histopathological criteria: epithelial tissue changes, connective tissue changes, and lateral thermal damage extent by optical microscopy. Results The epithelial and connective tissue damage scores were significantly higher in the dual-wavelength diode laser groups than in the Er, Cr:YSGG laser groups (P<0.001), and there was a significant difference between some groups. The extent of lateral thermal damage was also significantly higher in the diode laser groups than in the Er, Cr: YSGG laser groups (P<0.001), and there was a significant difference between groups. Group 2 (2.5 W) of the diode laser was the highest for all three criteria, while group 3 (2.5 W) of the Er, Cr:YSGG laser was the lowest. Conclusion The Er, Cr:YSGG laser with an output power of 2.5 W is, histologically, the most effective and safest laser for oral soft tissue incision. The dual-wavelength diode laser causes more damage than the Er, Cr:YSGG laser, but it can be used with a low output power and 1 mm safety distance in excisional biopsy.

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Abstract Objective To evaluate whether antimicrobial photodynamic therapy (aPDT) repairs bisphosphonate-related osteonecrosis of the jaw (BRONJ) modulated by the reduction of NF-kB protein in a murine model. Methodology Male Wistar rats (N=30) were divided into the following groups (n=6/group): negative control (NC); experimental osteonecrosis (ONE); ONE + photosensitizer (PS); ONE + photobiomodulation (PBM); and ONE + aPDT. Over 8 weeks, ONE was induced by zoledronic acid 250 µg/kg injections, except in the NC group, which received sterile 0.9% saline, followed by extraction of the lower left first molar. Red light laser irradiation (wavelength ~660 nm, power 50 mW, energy of 2 J, energy dose of 66.67 J/cm2 for 40 s) was performed once a week for 4 weeks. Methylene blue 0.3% was used as PS. The animals were euthanized and examined macroscopically for the presence of exposed bone and epithelial repair and microscopically by histochemical (hematoxylin-eosin and Masson’s trichrome staining) and immunohistochemical (anti-NF-kB) methods. Macroscopic and histomorphometric data were analyzed by one-way ANOVA and Tukey’s post-test (p<0.05). Results Mucosal repair, viable osteocytes, and NF-kB immunostaining were observed in the NC, ONE+PS, ONE+PBM, and ONE+aPDT groups. The ONE group showed no mucosal repair, showing empty lacunae and multifocal immunostaining for NF-kB. The ONE+PBM and ONE+aPDT groups had greater deposition of extracellular matrix and less necrotic bone tissue (p<0.05). Conclusion PBM and aPDT treatments for BRONJ were effective for bone and epithelial repair, in addition to reducing inflammation mediated by the decrease of NF-kB protein in the irradiated regions.

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Abstract Specific products containing natural resources can contribute to the innovation of complete denture hygiene. Objective: To conduct an in vitro evaluation of experimental dentifrices containing essential oils of Bowdichia virgilioides Kunth (BvK), Copaifera officinalis (Co), Eucalyptus citriodora (Ec), Melaleuca alternifolia (Ma) and Pinus strobus (Ps) at 1%. Methodology: The variables evaluated were organoleptic and physicochemical characteristics, abrasiveness (mechanical brushing machine) simulating 2.5 years, and microbial load (Colony Forming Units - CFU/mL), metabolic activity (XTT assay) and cell viability (Live/Dead® BacLight™ kit) of the multispecies biofilm (Streptococcus mutans: Sm, Staphylococcus aureus: Sa, Candida albicans: Ca and Candida glabrata: Cg). Specimens of heat-polymerized acrylic resins (n=256) (n=96 specimens for abrasiveness, n=72 for microbial load count, n=72 for biofilm metabolic activity, n=16 for cell viability and total biofilm quantification) with formed biofilm were divided into eight groups for manual brushing (20 seconds) with a dental brush and distilled water (NC: negative control), Trihydral (PC: positive control), placebo (Pl), BvK, Co, Ec, Ma or Ps. After brushing, the specimens were washed with PBS and immersed in Letheen Broth medium, and the suspension was sown in solid specific medium. The organoleptic characteristics were presented by descriptive analysis. The values of density, pH, consistency and viscosity were presented in a table. The data were analyzed with the Wald test in a generalized linear model, followed by the Kruskal-Wallis test, Dunn's test (mass change) and the Bonferroni test (UFC and XTT). The Wald test in Generalized Estimating Equations and the Bonferroni test were used to analyze cell viability. Results: All dentifrices showed stable organoleptic characteristics and adequate physicochemical properties. CN, Ec, Ps, Pl and PC showed low abrasiveness. There was a significant difference between the groups (p<0.001) for microbial load, metabolic activity and biofilm viability. Conclusions: It was concluded that the BvK, Ec and Ps dentifrices are useful for cleaning complete dentures, as they have antimicrobial activity against biofilm. The dentifrices containing Bowdichia virgilioides Kunth showed medium abrasiveness and should be used with caution.

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Abstract Studies have highlighted numerous benefits of ozone therapy in the field of medicine and dentistry, including its antimicrobial efficacy against various pathogenic microorganisms, its ability to modulate the immune system effectively, reduce inflammation, prevent hypoxia, and support tissue regeneration. However, its effects on dental extraction healing remain to be elucidated. Objective Therefore, this study aimed to evaluate the effects of systemically administered ozone (O3) at different doses in the healing of dental extraction sockets in rats. Methodology To this end, 72 Wistar rats were randomly divided into four groups after extraction of the right upper central incisor: Group C – control, no systemic treatment; Group OZ0.3 – animals received a single dose of 0.3 mg/kg O3; Group OZ0.7 – a single dose of 0.7 mg/kg O3; and Group OZ1.0 – a single dose of 1.0 mg/kg O3, intraperitoneally. In total, six animals from each group were euthanized at 7, 14, and 21 days after the commencement of treatment. Bone samples were harvested and further analyzed by descriptive histology, histomorphometry, and immunohistochemistry for osteocalcin (OCN) and tartrate-resistant acid phosphatase (TRAP) protein expression. Results All applied doses of O3 were shown to increase the percentage of bone tissue (PBT) after 21 days compared to group C. After 14 days, the OZ0.7 and OZ1.0 groups showed significantly higher PBT when compared to group C. The OZ1.0 group presented the most beneficial results regarding PBT among groups, which denotes a dose-dependent response. OCN immunostaining was higher in all groups at 21 days. However, after seven and 14 days, the OZ1.0 group showed a significant increase in OCN immunostaining compared to C group. No differences in TRAP+ osteoclasts were found between groups and time points. Conclusion Therefore, O3 therapy at higher doses might be beneficial for bone repair of the alveolar socket following tooth extraction.

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Abstract Objective This study aims to develop a compound biomaterial to achieve effective soft tissue regeneration. Methodology Compound hyaluronic acid (CHA) and liquid horizontal-platelet-rich fibrin (H-PRF) were mixed at a ratio of 1:1 to form a CHA-PRF gel. Human gingival fibroblasts (HGFs) were used in this study. The effect of CHA, H-PRF, and the CHA-PRF gel on cell viability was evaluated by CCK-8 assays. Then, the effect of CHA, H-PRF, and the CHA-PRF gel on collagen formation and deposition was evaluated by qRT‒PCR and immunofluorescence analysis. Finally, qRT‒PCR, immunofluorescence analysis, Transwell assays, and scratch wound-healing assays were performed to determine how CHA, H-PRF, and the CHA-PRF gel affect the migration of HGFs. Results The combination of CHA and H-PRF shortened the coagulation time of liquid H-PRF. Compared to the pure CHA and H-PRF group, the CHA-PRF group exhibited the highest cell proliferation at all time points, as shown by the CCK-8 assay. Col1a and FAK were expressed at the highest levels in the CHA-PRF group, as shown by qRT‒PCR. CHA and PRF could stimulate collagen formation and HGF migration, as observed by fluorescence microscopy analysis of COL1 and F-actin and Transwell and scratch healing assays. Conclusion The CHA-PRF group exhibited greater potential to promote soft tissue regeneration by inducing cell proliferation, collagen synthesis, and migration in HGFs than the pure CHA or H-PRF group. CHA-PRF can serve as a great candidate for use alone or in combination with autografts in periodontal or peri-implant soft tissue regeneration.

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Abstract Objective To compare the effect of submucosal cryotherapy using cold saline to dexamethasone sodium phosphate and diclofenac sodium injections on substance P and interleukin 6 release in experimentally induced pulpal inflammation in rabbits’ molar teeth. Methodology Fifteen rabbits were randomly classified into 3 groups according to the submucosal injection given: cold saline, dexamethasone sodium phosphate, and diclofenac sodium. A split-mouth design was adopted, the right mandibular molars were experimental, and the left molars served as the control without injections. Intentional pulp exposures were created and left for 6 hours to induce pulpitis. Pulpal tissue was extracted and examined for SP and IL-6 levels using ELISA. Within each group, the level of cytokines released was measured for both control and experimental groups for intragroup comparison to determine the effect of injection. The percentage reduction of each mediator was calculated compared with the control side for intergroup comparison then the correlation between SP and IL-6 levels was analyzed using Spearman’s rank order correlation coefficient. Statistical analysis was performed, and the significance level was set at p<0.05. Results Submucosal cryotherapy, dexamethasone sodium phosphate, and diclofenac sodium significantly reduced SP and IL-6 pulpal release. Submucosal cryotherapy significantly reduced SP more than and IL-6 more than dexamethasone sodium phosphate and diclofenac sodium. Pulpal reduction of SP and IL-6 showed a strong positive significant correlation. Conclusions Submucosal cryotherapy reduces the pulpal release of SP and IL-6 and could be tested as an alternative to premedication to potentiate the effect of anesthesia and control postoperative endodontic pain.

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Abstract This study aimed to compare the quality of root canal obturation (ratio of area occupied by gutta-percha (G), sealer (S), and presence of voids (V)) in different anatomical irregularities (intercanal communications, lateral irregularities, and accessory canals) located at different thirds of the root canal system of mandibular molar replicas. Sixty-seven 3D printed replicas of an accessed mandibular molar were prepared using ProGlider and ProTaper Gold rotatory systems. Three specimens were randomly selected to be used as controls and did not receive further treatment. The rest were randomly distributed in 4 experimental groups to be obturated using either cold lateral compaction (LC), continuous wave of condensation (CW), and core-carrier obturation (ThermafilPlus (TH) or GuttaCore (GC)) (n=16 per group). AHPlus® sealer was used in all groups. The three controls and a specimen from each experimental group were scanned using micro-computed tomography. The rest of the replicas were sectioned at the sites of anatomical irregularities and examined at 30× magnification. The G, S, and V ratios were calculated dividing the area occupied with each element by the total root canal area and then compared among groups using the Kruskal-Wallis test. Voids were present in all obturation techniques with ratios from 0.01 to 0.15. CW obtained a significantly higher G ratio in the irregularity located in the coronal third (0.882) than LC (0.681), TH (0.773), and GC (0.801) (p<0.05). TH and GC achieved significantly higher G ratios in those located in the apical third (p<0.05). The worst quality of obturation was observed in the loop accessory canal with all obturation techniques. Whitin the limitations of this study, it can be concluded that CW and core-carrier obturation are respectively the most effective techniques for obturating anatomical irregularities located in the coronal and the apical third.

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Abstract Objective To explore the feasibility of injectable platelet-rich fibrin (i-PRF) in regenerative endodontics by comparing the effect of i-PRF and platelet-rich fibrin (PRF) on the biological behavior and angiogenesis of human stem cells from the apical papilla (SCAPs). Methodology i-PRF and PRF were obtained from venous blood by two different centrifugation methods, followed by hematoxylin-eosin (HE) staining and scanning electron microscopy (SEM). Enzyme-linked immunosorbent assay (ELISA) was conducted to quantify the growth factors. SCAPs were cultured with different concentrations of i-PRF extract (i-PRFe) and PRF extract (PRFe), and the optimal concentrations were selected using the Cell Counting Kit-8 (CCK-8) assay. The cell proliferation and migration potentials of SCAPs were then observed using the CCK-8 and Transwell assays. Mineralization ability was detected by alizarin red staining (ARS), and angiogenesis ability was detected by tube formation assay. Real-time quantitative polymerase chain reaction (RT-qPCR) was performed to evaluate the expression of genes related to mineralization and angiogenesis. The data were subjected to statistical analysis. Results i-PRF and PRF showed a similar three-dimensional fibrin structure, while i-PRF released a higher concentration of growth factors than PRF ( P <.05). 1/4× i-PRFe and 1/4× PRFe were selected as the optimal concentrations. The cell proliferation rate of the i-PRFe group was higher than that of the PRFe group ( P <.05), while no statistical difference was observed between them in terms of cell mitigation ( P >.05). More importantly, our results showed that i-PRFe had a stronger effect on SCAPs than PRFe in facilitating mineralization and angiogenesis, with the consistent result of RT-qPCR ( P <.05). Conclusion This study revealed that i-PRF released a higher concentration of growth factors and was superior to PRF in promoting proliferation, mineralization and angiogenesis of SCAPs, which indicates that i-PRF could be a promising biological scaffold for application in pulp regeneration.

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Abstract Objective This study aimed to validate the integrated correlation between the buccal bone and gingival thickness of the anterior maxilla, and to gain insight into the reference plane selection when measuring these two tissues before treatment with implants. Methodology Cone beam computed tomography (CBCT) and model scans of 350 human subjects were registered in the coDiagnostiX software to obtain sagittal maxillary incisor sections. The buccal bone thickness was measured at the coronal (2, 4, and 6 mm apical to the cementoenamel junction [CEJ]) and apical (0, 2, and 4 mm coronal to the apex plane) regions. The buccal gingival thickness was measured at the supra-CEJ (0, 1mm coronal to the CEJ) and sub-CEJ regions (1, 2, 4, and 6 mm apical to the CEJ). Canonical correlation analysis was performed for intergroup correlation analysis and investigation of key parameters. Results The mean thicknesses of the buccal bone and gingiva at different levels were 0.64~1.88 mm and 0.66~1.37 mm, respectively. There was a strong intergroup canonical correlation between the thickness of the buccal bone and that of the gingiva (r=0.837). The thickness of the buccal bone and gingiva at 2 mm apical to the CEJ are the most important indices with the highest canonical correlation coefficient and loadings. The most and least prevalent subgroups were the thin bone and thick gingiva group (accounting for 47.6%) and the thick bone and thick gingiva group (accounting for 8.6%). Conclusion Within the limitations of this retrospective study, the thickness of the buccal bone is significantly correlated with that of the buccal gingiva, and the 2 mm region apical to the CEJ is a vital plane for quantifying the thickness of these two tissues

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Abstract Objective To evaluate the effect of the labiolingual diameter and construction of an endodontically treated (ET) anterior tooth with crown restoration on stress distribution and biomechanical safety under occlusal loading. Methodology Three-dimensional finite element models were generated for maxillary central incisors with all-ceramic crown restorations. The labiolingual diameters of the tooth, defined as the horizontal distance between the protrusion of the labial and lingual surfaces, were changed as follows: (D1) 6.85 mm, (D2) 6.35 mm, and (D3) 5.85 mm. The model was constructed as follows: (S0) vital pulp tooth; (S1) ET tooth; (S2) ET tooth with a 2 mm ferrule, restored with a fiber post and composite resin core; (S3) ET tooth without a ferrule, restored with a fiber post and composite resin core. A total of 12 models were developed. In total, two force loads (100 N) were applied to the crown’s incisal edge and palatal surface at a 45° oblique angle to the longitudinal axis of the teeth. The Von Mises stress distribution and maximum stress of the models were analyzed. Results Regardless of the loading location, stress concentration and maximum stress (34.07~66.78MPa) in all models occurred in the labial cervical 1/3 of each root. Both labiolingual diameter and construction influenced the maximum stress of the residual tooth tissue, with the impact of the labiolingual diameter being greater. A reduction in labiolingual diameter led to increased maximum stress throughout the tooth. The ferrule reduced the maximum stress of the core of S2 models (7.15~10.69 MPa), which is lower compared with that of S3 models (19.45~43.67 MPa). Conclusion The labiolingual diameter exerts a greater impact on the biomechanical characteristics of ET anterior teeth with crown restoration, surpassing the influence of the construction. The ferrule can reduce the maximum stress of the core and maintain the uniformity of stress distribution.

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Abstract Aim To investigate iron-deficiency anemia as a risk factor for dental pulp disease in children from the central Peruvian jungle. Methodology A case-control study was carried out with 270 children, of which 90 referred to cases and 180, to controls. Patients with pulp disease were diagnosed according to the criteria of the Association of Endodontists and the American Board of Endodontics. A specific questionnaire was used to assess ferrous sulfate consumption, maternal education level, maternal age, occupation, and household income. Data were analyzed using Pearson’s correlation coefficient and a binary logistic regression. Results Iron deficiency anemia offers a risk factor for pulp disease in children (OR 7.44, IC 95% 4.0-13.8). According to multivariate analysis using binary logistic regression, ferrous sulfate consumption (OR 13.8, IC 95% 5.6.33.9), maternal education level (OR 2.4, IC 95% 1.1-5.3), maternal age (OR 7.5, IC 95% 2.9-19.4), household income (OR 4.0, IC 95% 1.6-9.6), and caries (OR 10.7, IC 95% 4.5-25.7) configured independent factors that were statistically associated with pulp disease. Conclusion Iron deficiency anemia, ferrous sulfate consumption, maternal education level, maternal age, household income, and dental caries were positively associated with pulp disease in children

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Abstract The prevalence of gingivitis is substantial within the general population, necessitating rigorous oral hygiene maintenance. Objective This study assessed a Garcinia indica (GI) fruit extract-based mouthrinse, comparing it to a 0.1% turmeric mouthrinse and a 0.2% Chlorhexidine (CHX) mouthrinse. The evaluation encompassed substantivity, staining potential, antimicrobial efficacy and cytocompatibility. Methodology The study employed 182 tooth sections. For antimicrobial analysis, 64 extracted human teeth coated with a polymicrobial biofilm were divided into four groups, each receiving an experimental mouthrinse or serving as a control group with distilled water. Microbial reduction was assessed through colony forming units (CFU). Substantivity was evaluated on 54 human tooth sections using a UV spectrophotometer, while staining potential was examined on 64 tooth sections. Cytocompatibility was tested using colorimetric assay to determine non-toxic levels of 0.2% GI fruit extract, 0.1% Turmeric, and 0.2% CHX. Results Data were analysed with one-way ANOVA (α=0.05). Cell viability was highly significant (p<0.001) in the 0.2% GI group (64.1±0.29) compared to 0.1% Turmeric (40.2±0.34) and 0.2% CHX (10.95±1.40). For antimicrobial activity, both 0.2% GI (20.18±4.81) and 0.2% CHX (28.22±5.41) exhibited no significant difference (P>0.05) at end of 12 hours. However, 0.1% Turmeric showed minimal CFU reduction (P<0.001). Substantivity results at 360 minutes indicated statistically significant higher mean release rate in 0.1%Turmeric (12.47±5.84 ) when compared to 0.2% GI (5.02±3.04) and 0.2% CHX (4.13±2.25) (p<0.001). The overall discoloration changes (∆E) were more prominent in the 0.2% CHX group (18.65±8.3) compared to 0.2% GI (7.61±2.4) and 0.1% Turmeric (7.32±4.9) (P<0.001). Conclusion This study supports 0.2% GI and 0.1% Turmeric mouth rinses as potential natural alternatives to chemical mouth rinses. These findings highlight viability of these natural supplements in oral healthcare.

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Abstract Objective To assess the efficacy of Phyllanthus emblica extract in alleviating halitosis and reducing the inflammatory response to halitosis-related bacteria. Methodology This investigation, using Phyllanthus emblica fruit extract (PE), involved four aspects. First, we evaluated the effect on growth and aggregation of halitosis-related bacteria, including Fusobacterium nucleatum, Porphyromonas gingivalis, and Solobacterium moorei, using a microdilution assay and scanning electron microscopy. Second, volatile sulfur compound (VSC) levels were measured on individuals with halitosis in randomized short-term (26 participants) and double-blind randomized long-term trials (18 participants in each group) after rinsing with PE for 3, 6, and 12 h, and 28 days. Third, we analyzed pro-inflammatory cytokine expression in TR146 cells using quantitative real-time PCR and enzyme-linked immunosorbent assays. Lastly, we assessed pro-inflammatory cytokine secretion and Toll-like receptor (TLR) 2 mRNA expression via the same experimental methods in a three-dimensional oral mucosal epithelial model (3D OMEM). Results PE extract dose-dependently inhibited the growth of F. nucleatum (50% inhibition concentration [IC50]=0.079%), P. gingivalis (IC50=0.65%), and S. moorei (IC50=0.07%) and effectively prevented bacterial aggregation. Furthermore, VSC contents decreased significantly at 3, 6, and 12 h after rinsing with 5% PE compared with those in the control. Long-term use of mouthwash containing 5% PE for 28 days led to a significant decrease in VSC contents. PE attenuated the F. nucleatum- or P. gingivalis-stimulated mRNA expression and protein release of interleukin (IL)-6 and IL-8 in TR146 cells. It also suppressed IL-8 and prostaglandin E2 secretion and TLR2 mRNA expression in F. nucleatum-induced OMEMs. Conclusion Our findings support the use of PE in oral care products to alleviate halitosis and it may reduce inflammation.

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Abstract Creating artificial caries-like lesions that mimic the complex changes observed in natural caries is essential for properly evaluating new strategies, dental materials, and devices designed to arrest their progression and avoid more costly and invasive treatments. Objective This study compared three protocols for developing artificial white spot lesions (WSL) using biofilm models. Methodology In total, 45 human enamel specimens were sterilized and allocated into three groups based on the biofilm model: Streptococcus sobrinus and Lactobacillus casei (Ss+Lc), Streptococcus sobrinus (Ss), or Streptococcus mutans (Sm). Specimens were incubated in filter-sterilized human saliva to form the acquired pellicle and then subjected to the biofilm challenge consisting of three days of incubation with bacteria (for demineralization) and one day of remineralization, which was performed once for Ss+Lc (four days total), four times for Ss (16 days total), and three times for Sm (12 days total). After WSL creation, the lesion fluorescence, depth, and chemical composition were assessed using Quantitative Light-induced Fluorescence (QLF), Polarized Light Microscopy (PLM), and Raman Spectroscopy, respectively. Statistical analysis consisted of two-way ANOVA followed by Tukey’s post hoc test (α=0.05). WSL created using the Ss+Lc protocol presented statistically significant higher fluorescence loss (ΔF) and integrated fluorescence (ΔQ) in comparison to the other two protocols (p<0.001). Results In addition, Ss+Lc resulted in significantly deeper WSL (137.5 µm), followed by Ss (84.1 µm) and Sm (54.9 µm) (p<0.001). While high mineral content was observed in sound enamel surrounding the WSL, lesions created with the Ss+Lc protocol showed the highest demineralization level and changes in the mineral content among the three protocols. Conclusion The biofilm model using S. sobrinus and L. casei for four days was the most appropriate and simplified protocol for developing artificial active WSL with lower fluorescence, higher demineralization, and greater depth.

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Abstract Objective To evaluate the protective effect of an experimental solution containing TiF4/NaF on the development of radiation-induced dentin caries lesions. Methodology bovine root samples were irradiated (70Gy) and distributed as following (n=12/group): Commercial Saliva (BioXtra), NaF (500 ppm F-), TiF4 (500 ppm F), TiF4/NaF (TiF4: 300 ppm F-, NaF: 190 ppm F-), and Phosphate buffer solution (PBS, negative control). Biofilm was produced using biofilm from irradiated patients and McBain saliva (0.2% of sucrose, at 37oC and 5% CO2) for five days. The treatments were applied 1x/day. Colony-forming units (CFU) were counted and demineralization was quantified by transversal microradiography. The ANOVA/Tukey test was applied for all parameters. Results All treatments reduced CFU for total microorganisms. TiF4 reduced Lactobacillus sp. (7.04±0.26 log10 CFU/mL) and mutans streptococci (7.18±0.28) CFU the most, when compared to PBS (7.58±0.21 and 7.75±0.17) and followed by NaF (7.12±0.31 and 7.34±0.22) and TiF4/NaF (7.16±0.35 and 7.29± 0.29). TiF4 and Commercial saliva showed the lowest integrated mineral loss (ΔZ-vol%.mm) (1977±150 and 2062±243, respectively) when compared to PBS (4540±335), followed by NaF (2403±235) and TiF4/NaF (2340±200). Commercial saliva was the only to significantly reduce mineral loss (LD-µm) (111±25) compared to PBS (153±24).Mean mineral loss (R-vol%) decreased by 35.2% for TiF4 (18.2±3.3) when compared to PBS (28.1±2.9) Conclusion: TiF4/NaF has a comparable anti-cariogenic effect to TiF4 and Commercial saliva under the model in this study.

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Abstract Hypertensive individuals present alterations in their calcium metabolism that consequently decrease the concentration of this mineral in the bone tissue. Objectives to evaluate the morphological and functional characteristics of the peri-implant bone tissue that was formed during the healing process by the placement implants using two different surface treatments: hydrophilic Acqua™ (ACQ) and rough NeoPoros™ (NEO), in spontaneously hypertensive (SHR) and normotensive rats (Wistar) whether or not treated with losartan. Methodology In total, 96 male rats (48 Wistar and 48 SHR) were divided into eight subgroups: absolute control rough (COA NEO), absolute control hydrophilic (COA ACQ), losartan control rough (COL NEO), losartan control hydrophilic (COL ACQ), SHR absolute rough (SHR NEO), SHR absolute hydrophilic (SHR ACQ), SHR losartan rough (SHRL NEO), and SHR losartan hydrophilic (SHRL ACQ). The rats medicated with losartan received daily doses of the medication. NeoPoros™ and Acqua™ implants were installed in the tibiae of the rats. After 14 and 42 days of the surgery, the fluorochromes calcein and alizarin were injected in the rats. The animals were euthanized 67 days after treatment. The collected samples were analyzed by immunohistochemistry, biomechanics, microcomputerized tomography, and laser confocal scanning microscopy analysis. Results The osteocalcin (OC) and vascular endothelium growth factor (VEGF) proteins had moderate expression in the SHRL ACQ subgroup. The same subgroup also had the highest implant removal torque. Regarding microarchitectural characteristics, a greater number of trabeculae was noted in the control animals that were treated with losartan. In the bone mineralization activity, it was observed that the Acqua™ surface triggered higher values of MAR (mineral apposition rate) in the COA, COL, and SHRL groups (p<0.05). Conclusion the two implant surface types showed similar responses regarding the characteristics of the peri-implant bone tissue, even though the ACQ surface seems to improve the early stages of osseointegration.

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Abstract Objective This study aims to explore the regulatory mechanism of long noncoding RNA X inactive specific transcript (lncRNA XIST) in the odontogenic differentiation of human dental pulp stem cells (hDPSCs). hDPSCs were obtained from freshly extracted third molars and identified by flow cytometry. Methodology Odontogenic differentiation was induced in mineralized culture medium, and hDPSCs were infected with shRNA lentivirus targeting XIST or fused in sarcoma (FUS), followed by detection of alkaline phoshpatase (ALP) activity, alizarin red staining of mineralized nodules, Real-Time Quantitative Reverse Transcription PCR (qRT-PCR) quantification of XIST expression, and Western blot analysis of FUS, ZBTB16, and odontogenic differentiation markers (DSPP and DMP1). IF-FISH was performed to detect the cellular localization of XIST and FUS. RIP assay validated the XIST and FUS binding. ZBTB16 mRNA stability was tested after actinomycin D treatment. hDPSCs were infected with oe-ZBTB16 lentivirus and further treated with sh-XIST for a combined experiment. Results LncRNA XIST was highly expressed in hDPSCs with odontogenic differentiation. Downregulation of XIST or FUS weakened the ALP activity of hDPSCs, reduced mineralized nodules, diminished DSPP and DMP1 expressions. XIST binds to FUS to stabilize ZBTB16 mRNA and promote ZBTB16 expression. ZBTB16 overexpression partially reversed the inhibitory effect of XIST silencing on odontogenic differentiation of hDPSCs. Conclusion In conclusion, XIST stabilizes ZBTB16 mRNA and promotes ZBTB16 expression by binding to FUS, thereby facilitating the odontogenic differentiation of hDPSCs.

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Abstract Aim To identify the phenotypic characteristics of individuals with temporomandibular disorders (TMD) who may benefit from adding duloxetine to self-management (SM) strategies. Methodology This was a post hoc exploratory analysis of a randomized, placebo-controlled clinical trial with SM-duloxetine (duloxetine 60 mg/day plus SM strategies for 12 weeks) in adult participants with painful TMD. The primary outcome was the proportion of responders to treatment (individuals with ≥ 30% reduction in pain intensity) in SM-duloxetine and SM-placebo group at week 12. For responder analysis, five phenotyping domains recommended by Initiative on Methods, Measurement, and Pain Assessment in Clinical Trials were assessed: pain, psychological, sleep, quantitative sensory testing, and conditioned pain modulation. Relative risk (RR), 95% confidence intervals (CI), and absolute risk reduction were calculated. Results Among participants treated with SM-duloxetine, severe pain intensity (RR 1.33, 95% CI: 0.56, 3.17), pain disability (RR 1.30, 95% CI: 0.63, 2.67), ≥ 1 painful comorbidity (RR 1.48, 95% CI: 0.57, 3.79), and anxiety symptoms (RR 1.80, 95% CI: 0.75, 4.34) were associated with greater likelihood of response to treatment. Among individuals treated with SM-placebo, only temporal summation of pain was associated with greater likelihood of response to treatment. Conclusion Personalized medicine may be implemented in painful TMD management, and phenotype characteristics related to pain and psychological domains may predict which individuals with painful TMD are more likely to respond to the addition of serotonin and norepinephrine reuptake inhibitors to SM strategies to clinically and significantly reduce pain intensity.

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Abstract Objective This study evaluated the color stability of enamel submitted to 10% hydrogen peroxide (HP) followed by antioxidants agents, and the pH and antioxidant activity (AA%) of these agents. Methodology Bovine enamel-dentin blocks were randomly distributed into groups (n=10/group): GNC (negative control: no treatment); GPC (positive control: bleaching only); TOC_10% (HP+10% α-tocopherol); GT_10% (HP+10% green tea extract); GS_5% (HP+5% grape seed extract); SA_10% (HP+10% sodium ascorbate); QUI_10% (HP+10% quinoa extract); and QC_1% (HP+1% quercetin). Color (ΔE00) and whiteness index (ΔWID) changes were analyzed using a digital spectrophotometer. The pH and AA% were determined using a pH meter and the DPPH method, respectively. Data were analyzed by ANOVA/Tukey’s and Dunnett’s tests (α=0.05). Results At 14 days post-bleaching, GNC promoted the lowest ΔWID and ΔE00 (p<0.05), and no differences were found between GPC and the remaining groups submitted to the antioxidant agents (p>0.05). QC_1% and QUI_10% exhibited acidic pH levels (3.64 and 4.75, respectively), whereas TOC_10% and GS_5% exhibited alkaline pH (7.07 and 7.64, respectively). No differences in AA% were found between the agents (p>0.05), ranging from 92.6 to 97.6%. Conclusion The antioxidant agents did not interfere in bleached enamel color stability, showing satisfactory antioxidant activity. However, QUI and QC gels displayed acidic pH. Clinical significance: The antioxidants evaluated showed high AA% and no impact on post-bleaching color stability, suggesting that their capacity to recover bond strength demonstrated elsewhere would not compromise the esthetic efficacy of tooth bleaching. However, those with acidic pH should be used with caution due to potential enamel damage.

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Abstract Objectives This study aimed to assess the effect of proanthocyanidin, palm oil and vitamin E against erosive and erosive+abrasive challenges in vitro after enamel pellicle formation in situ. Methodology Bovine enamel blocks (n=84) were obtained and divided into the following treatment groups: negative control (NC) - deionized water; positive control (PC) - SnCl2/NaF/AmF-containing solution; palm oil (PO); 2% proanthocyanidin (P2); vitamin E (VitE); 2% proanthocyanidin+palm oil (P2PO); and 2% proanthocyanidin+vitamin E (P2VitE). For 5 days, one half of the sample from each group was subjected to erosion and the other half was subjected to erosion+abrasion. The acquired enamel pellicle (AEP) was pre-formed in situ for 30 minutes. The specimens were then treated in vitro with solutions (500 µl, 30s for each group). Subsequently, the blocks were left in the oral cavity for another hour to obtain the modified AEP. The blocks were immersed in 0.5% citric acid (pH=2.5) for 90s, 4×/day. AEP formation and treatment were carried out before the first and third erosive challenges, and after these challenges, abrasive cycles (15s) were performed on half of the samples. Enamel wear was quantified by profilometry and data were analyzed by two-way ANOVA and Tukey’s test (p<0.05). Results All groups showed higher wear when exposed to erosion+abrasion than when exposed to erosion alone (p=0.0001). PO, P2VitE, P2, and P2PO showed enamel wear similar to the PC group, but only PC, PO and P2VitE differed from the NC group. The other groups behaved similarly to NC. Conclusion It was concluded that the combination of proanthocyanidin and vitamin E was effective in reducing wear in the face of in vitro erosive and erosive+abrasive challenges.

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Abstract A combination of peripheral blood mesenchymal stem cells (PBMSCs) and platelet rich fibrin matrix (PRFM) could be a probable periodontal regenerative material with the synergy of the added benefits of each material. Objective This randomized controlled clinical trial aimed to evaluate the regenerative capacity of supercell (PRFM and PBMSCs) compared with that of PRFM alone in human periodontal mandibular intraosseous defects (IOD). Methodology This study included 17 patients of both sexes (12 men, 5 women) aged 30-55 years (mean age = 37.7±4.4 years) who fulfilled the inclusion criteria (radiographic and clinical evaluation for bilateral IOD with probing pocket depth (PPD ≥ 6 mm). A split-mouth design was used in each patient. A total of 34 sites in the mandibular arch randomly received PRFM alone + open flap debridement (OFD) [Control sites] or supercell (PRFM+PBMSCs) + OFD [Test sites]. The clinical parameters plaque index (PI), gingival index (GI), PPD, clinical attachment level (CAL), and in the radiographic parameters; defect depth (DD) and defect fill percentage (DFP) were recorded at baseline, 3 and 6 months postoperatively. Early wound healing index (EHI) was used at 1 week to assess wound healing ability. Results At 6 months, radiographic parameters revealed significant reduction in DD (P<0.001) and significant DFP values in the test group compared with the control group. The supercell showed significant improvement in PPD and CAL at the end of 6 months (P<0.001). EHI scores at 1 week showed no statistically significant difference between the test and control groups. Conclusion Supercell can be considered a regenerative material in the treatment of periodontal IODs.

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Abstract Several materials have been developed to preserve pulp vitality. They should have ideal cytocompatibility characteristics to promote the activity of stem cells of human exfoliated deciduous teeth (SHED) and thus heal pulp tissue. Objective To evaluate the cytotoxicity of different dilutions of bioceramic material extracts in SHED. Methodology SHED were immersed in αMEM + the material extract according to the following experimental groups: Group 1 (G1) –BBio membrane, Group 2 (G2) - Bio-C Repair, Group 3 (G3) - MTA Repair HP, Group 4 (G4) – TheraCal LC, and Group 5 (G5) - Biodentine. Positive and negative control groups were maintained respectively in αMEM + 10% FBS and Milli-Q Water. The methods to analyze cell viability and proliferation involved MTT and Alamar Blue assays at 24, 48, and 72H after the contact of the SHED with bioceramic extracts at 1:1 and 1:2 dilutions. Data were analyzed by the three-way ANOVA, followed by Tukey’s test (p<0.05). Results At 1:1 dilution, SHED in contact with the MTA HP Repair extract showed statistically higher cell viability than the other experimental groups and the negative control (p<0.05), except for TheraCal LC (p> 0.05). At 1:2 dilution, BBio Membrane and Bio-C showed statistically higher values in intra- and intergroup comparisons (p<0.05). BBio Membrane, Bio-C Repair, and Biodentine extracts at 1:1 dilution showed greater cytotoxicity than 1:2 dilution in all periods (p<0.05). Conclusion MTA HP Repair showed the lowest cytotoxicity even at a 1:1 dilution. At a 1:2 dilution, the SHED in contact with the BBio membrane extract showed high cell viability. Thus, the BBio membrane would be a new non-cytotoxic biomaterial for SHED. Results offer possibilities of biomaterials that can be indicated for use in clinical regenerative procedures of the dentin-pulp complex.

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Abstract Bisphosphonates are prescribed to treat excessive bone resorption in patients with osteoporosis. However, its use is associated with potential adverse effects such as medication-related osteonecrosis of the jaw, prompting the introduction of the drug holiday concept in patients prior to dentoalveolar surgery. Furthermore, bisphosphonate discontinuation has been studied in vivo, in humans, and in animal models. However, it is not known whether this approach could affect bone cells in vitro. Therefore, the objective of this study was to investigate the potential effects of bisphosphonate discontinuation on pre-osteoblast and osteoblast activities in vitro. Methodology Pre-osteoblasts (MC3T3) and osteoblasts were treated with bisphosphonate (alendronate) at concentrations of 1, 5, and 10 µM. Alendronate was then withdrawn at different time points. The negative control consisted of untreated cells (0 µM), while the positive control consisted of cells incubated with alendronate throughout the experiment. Cell viability, cell adhesion, cell cytoskeleton, mineralization, and gene expressions were investigated. Results Pre-osteoblasts and osteoblasts showed a decrease in cell viability after treatment with 5-10 μM alendronate for 4 days or longer. Two days of alendronate discontinuation significantly increased cell viability compared with the positive control. However, these levels did not reach those of the negative control. Bone nodule formation was reduced by alendronate. Discontinuation of alendronate regained bone nodule formation. Longer periods of discontinuation were more effective in restoring nodule formation than shorter periods. Addition of alendronate resulted in an increase in the percentage of dead cells, which, in turn, decreased when alendronate was discontinued. Alendronate affected the cell cytoskeleton by disassembling actin stress fibers. Cell adhesion and cell morphological parameters were also affected by alendronate. Discontinuation of alendronate restored cell adhesion and these parameters. Overall, the highest improvement after alendronate discontinuation was seen at 10 µM. However, alendronate treatment and discontinuation did not affect osteoblast gene expression. Conclusion Discontinuation of alendronate helps to reverse the negative effects of the drug on cell viability, cell adhesion, and mineralization by restoring the cell cytoskeleton. Our data suggest the benefits of drug holiday and/or intermittent strategies for alendronate administration at the cellular level.

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Abstract Adhesion to dentin is a first step for a successful microbial root canal colonization. Cell hydrophobicity seems to have some influence in the Candida species adhesion to surfaces. Objective to measure cell surface hydrophobicity and to investigate the adherence ability to human dentin among Candida albicans strains isolated from root canal and lingual dorsum via an in vitro study. Methodology adhesion was quantified in function of dentin area covered by blastospores and/or hyphae presence detected by epifluorescence microscope. Cell surface hydrophobicity was estimated by assessing the percentage migration of cells from an aqueous phase to a hydrocarbon phase. Contact angles were measured by the sessile drop technique on the dentin surface using a contact angle measurements apparatus. We also examined the correlation between adhesion ability and hydrophobicity. Results although there was some intra-species variation in cell surface hydrophobicity, most isolates were characterized by moderate hydrophobicity. There was no significant difference in this parameter when the isolation niche was considered. Both root canal and lingual dorsum yeasts were able to adhere to dentin. No association was found between the strains’ site of isolation and adhesion. Moreover, cell surface hydrophobicity and adhesion ability were not correlated. Conclusion although hydrophobicity can influence Candida albicans virulence in many ways, this study suggests that this parameter by itself was not a good predictor of adhesion to dentin.

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Abstract Objective To analyze the effect of mast cells (MCs) in neurogenic inflammation and the neuroimmune response of trigeminal ganglia (TG) due to pulpitis and detect the regulatory effect of curcumin (Cur) on neuroimmune responses induced by pulpitis. Methodology Immunohistochemistry, toluidine blue staining (TB), and other methods were used to detect the dynamic changes of MCs, as well as tryptase expression changes and protease activated receptor 2 (PAR2) and calcitonin gene-related peptide (CGRP) levels in the neuroimmune response induced by pulpitis. After administering Cur by intraperitoneal injection, the expression levels of Toll-like receptor 4 (TLR4), CGRP, glial fibrillary acidic protein (GFAP), fractalkine (CX3CL1), Tumor necrosis factor (TNF-α), and other factors were examined in the TG of pulpitis-induced rats. Results After pulpitis induction, the expression of CGRP-positive neurons and GFAP-positive soluble guanylate cyclase (SGC) in the TG significantly increased. A large number of MCs underwent degranulation. MCs were scattered between the CGRP-positive nerve fibers. MCs showing a typical degranulated state within the TG significantly increased and tryptase-positive MCs surrounded the TG nerve fibers and neurons. After treatment with Cur, the inflammatory response in the periodontal bone induced by pulpitis decreased and promoted early tissue repair. The expression of TNF-α significantly decreased as did degranulation of MCs. In contrast, the expression of CGRP, TLR4-positive neurons, activated SGCs, and PAR2-positive TG neurons significantly decreased. MCs could participate in the neuroimmune response induced by pulpitis by the tryptase signaling pathway. Conclusion Importantly, Cur inhibited the degranulation of MCs, downregulated the expression of tryptase and PAR2 in the TG, and attenuated the activation response of osteoclasts in the apical periodontium.

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Abstract Objective To evaluate the effect of the addition of dynamic cervical electrical stimulation (electro-massage, ES) to manual therapy (MT), compared to MT by itself, in individuals with myofascial temporomandibular pain. Methodology A total of 46 participants with bilateral myofascial temporomandibular pain for at least three months were distributed into two groups. Group 1 (n=21) received local MT consisting of soft tissue mobilization and release techniques over the neck and temporomandibular regions. Group 2 (n=25) received an ES procedure in the cervical region combined with the same intervention as group 1. All participants underwent a 2-week protocol. The primary outcomes were pain intensity (Visual Analogue Scale), pressure pain threshold (PPT) at the masseter and upper trapezius muscles (algometer), and pain-free vertical mouth opening (manual gauge). The secondary outcome was active cervical range-of-movement. Measurements were taken at baseline, immediately after intervention, and at a 4-week follow-up. Results The ANOVA revealed significant changes over group*time, with better results for group 2 (large effect sizes) regarding pain intensity (p< 0.001; η2>0.14), pressure pain sensitivity and mouth opening (p<0.001; η2>0.14). Similar findings were observed for active cervical range-of-movement in all directions (p<0.001; η2>0.14), except rotation (p≥0.05). Conclusion Electrical stimulation therapy over the cervical region combined with a MT protocol over the neck and temporomandibular joint shows better clinical benefits than MT by itself in subjects with myofascial temporomandibular pain. Registration code: NCT04098952

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Abstract Objectives This study aimed to investigate the osseointegration of titanium (Ti) implants with micro-nano textured surfaces functionalized with strontium additions (Sr) in a pre-clinical rat tibia model. Methodology Ti commercially pure (cp-Ti) implants were installed bilaterally in the tibia of 64 Holtzman rats, divided into four experimental groups (n=16/group): (1) Machined surface - control (C); (2) Micro-nano textured surface treatment (MN); (3) Micro-nano textured surface with Sr2+ addition (MNSr); and (4) Micro-nano textured surface with a higher complementary addition of Sr2+ (MNSr+). In total, two experimental euthanasia periods were assessed at 15 and 45 days (n=8/period). The tibia was subjected to micro-computed tomography (μ-CT), histomorphometry with the EXAKT system, removal torque (TR) testing, and gene expression analysis by PCR-Array of 84 osteogenic markers. Gene expression and protein production of bone markers were performed in an in vitro model with MC3T3-E1 cells. The surface characteristics of the implants were evaluated by scanning electron microscopy (SEM), energy-dispersive spectroscopy (EDS), and laser scanning confocal microscopy. Results SEM, confocal, and EDS analyses demonstrated the formation of uniform micro-nano textured surfaces in the MN group and Sr addition in the MNSr and MNSr+ groups. TR test indicated greater osseointegration in the 45-day period for treated surfaces. Histological analysis highlighted the benefits of the treatments, especially in cortical bone, in which an increase in bone-implant contact was found in groups MN (15 days) and MNSr (45 days) compared to the control group. Gene expression analysis of osteogenic activity markers showed modulation of various osteogenesis-related genes. According to the in vitro model, RT-qPCR and ELISA demonstrated that the treatments favored gene expression and production of osteoblastic differentiation markers. Conclusions Micro-nano textured surface and Sr addition can effectively improve and accelerate implant osseointegration and is, therefore, an attractive approach to modifying titanium implant surfaces with significant potential in clinical practice.

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Abstract In children with poor oral hygiene, dental caries are the most common factor that causes periapical pathologies as a result of irreversible pulpal injuries and leads to the cessation of root development in immature permanent molar teeth. Objective The primary goal is to evaluate the effects of two different intracanal medicaments, calcium hydroxide [Ca(OH)2] and double antibiotic paste (DAP), on radiographic outcomes during regenerative endodontic procedures (REP) of immature permanent mandibular first molars with symptomatic irreversible pulpitis and symptomatic apical periodontitis (SIP/SAP). Additionally, the secondary goal was to evaluate MMP-8 levels during REP using two different intracanal medicaments. Methodology The study included 20 patients with immature mandibular first molars exhibiting SIP/SAP. Participants were randomly assigned into two groups based on the applied intracanal medicament. Ca(OH)2 (n=10) was prepared by mixing it with sterile distilled water, while the same amount of powdered metronidazole and ciprofloxacin were mixed and combined with sterile distilled water for DAP (n=10). MMP-8 in periapical samples were measured at baseline and on the 14th day using immunofluorometric assay. Image-J software with TurboReg plug-in was utilized to determine changes in root length, root width, radiographic root area (RRA) during the 12-month follow-up period. Data were analyzed by SPSS 25.0 (p<.05). Results Significant increase in MMP-8 on the 14th day compared to baseline in both groups (p<0.001). There was no significant difference between the two groups in terms of the increase in MMP-8 (p>0.05). Root length significantly increased in both groups (p=0.001), with Ca(OH)2 showing a greater increase (p=0.046). Root width and RRA increased similarly in both groups at 12th month. Conclusion Both Ca(OH)2 and DAP applications resulted in a significant increase in periapical MMP-8 levels. Increase in radiographic root width and root area was similar between two groups, but Ca(OH)2 led to a significantly greater increase in root length. Further studies with larger sample sizes are necessary to validate our findings during REP of vital immature permanent mandibular molars. Clinical Trials database: NCT05581706

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Abstract Objectives This study sought to determine effects of Thai propolis extract mixed in mineral trioxide aggregate (MTA) on matrix metalloproteinase-2 (MMP-2) expression and its activity in inflamed human dental pulp cells (HDPCs). Materials and Methods Interleukin-1β-primed HDPCs were treated with either the eluate of MTA mixed with distilled water, of MTA mixed with 0.75 mg/ml of the propolis extract, or of Dycal®, 0.75 mg/ml of the propolis extract, or 0.2% (v/v) of chlorhexidine for 24 or 72 h. The viability of HDPCs was determined by the PrestoBlue® cytotoxic assay. HDPCs’ lysates were analyzed for MMP-2 mRNA expression by RT-qPCR, while their supernatants were measured for MMP-2 activity by gelatin zymography. Results At 24 and 72 h, a non-toxic dose of the propolis extract at 0.75 mg/ml by itself or mixed in MTA tended to reduce MMP-2 expression upregulated by MTA, while it further decreased the MMP-2 activity as compared to that of MTA mixed with distilled water. The MMP-2 activity of interleukin-1β-primed HDPCs treated with the eluate of the propolis extract mixed in MTA was significantly lower than that of interleukin-1β-primed HDPCs at 24 h (p=0.012). As a control, treatment with chlorhexidine significantly inhibited MMP-2 expression induced by MTA and MMP-2 activity enhanced by interleukin-1β (p<0.05). Treatment with Dycal® caused a significant increase in HDPC’s death, resulting in a significant decrease in MMP-2 expression and activity (p<0.05). Conclusions MTA mixed with Thai propolis extract can reduce MMP-2 mRNA expression and activity when compared to MTA mixed with distilled water in inflamed HDPCs.

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Abstract There are many suitable strategies for addressing caries, which is an ongoing worldwide problem. Although white spot lesions (WSLs) can be either remineralized naturally or treated with non- or micro-invasive strategies, their whitish and opaque appearance may persist. Objective To evaluate the effects of tooth bleaching as a complement to fluoride-enhanced remineralization or resin infiltration in masking WSLs, as well as in enamel surface roughness relative to that of the adjacent enamel. Methodology Flattened rectangular bovine enamel fragments (6×3×~2.9 mm length, width and thickness) were divided into six groups (L/N, F/N, F.BL/BL, I/N, I.BL/BL, N/N; n=15). Treatments applied to the 3×3 mm left half included: L (Lesion) - WSL simulation with 50 mM acetate buffer, 96 hours, 37ºC; F (Fluoride) - WSL treatment with 2% NaF neutral gel, 1x/week, 8 weeks; I (Infiltration) - WSL treatment with H3PO4 37%/10 s; Icon®-Dry/30 s; Icon®-Infiltrant/3 min+1 min; N (Nothing) - sound enamel/control. Treatments applied to both halves after F and I included: BL (Bleaching) - Opalescence Boost 40%, 3×/20 min each; N (Nothing) - control. The differences in color (ΔE00, ΔL, Δa, Δb) and surface roughness (ΔRa) between the left and right halves were measured. Kruskal-Wallis/post-hoc tests were applied to ΔE00, ΔL, Δa and ΔRa, and 1-way ANOVA/Tukey tests to Δb (α=0.05). Results The factor under study significantly influenced ΔE00 (p=0.0001), ΔL (p=0.0024), Δb (p=0.0015), and ΔRa (p<0.001), but not Δa (p=0.1592). Both fluoride-enhanced remineralization and resin infiltration were able to mask WSL, regardless of subsequent bleaching. However, when bleaching was performed, ΔE00 median values did not exceed the acceptability threshold for color difference. Only resin infiltration reduced ΔRa between WSL and the adjacent enamel. Conclusions Both remineralization and infiltration, particularly if complemented by bleaching, fostered satisfactory esthetic results. Only infiltration without bleaching led to really good results in surface roughness.

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Abstract Objective this study sought to evaluate the effect of isobornyl methacrylate (IBOMA) as a diluent monomer on the physicochemical properties of experimental flowable resin composites. Methodology the organic resin matrix of a modal flowable resin composite was formulated with 50 wt.% of bisphenol-A-glycidyl methacrylate (Bis-GMA) and 50 wt.% of a diluent monomer, in which IBOMA was used as a combining or substituent diluent monomer to triethylene glycol dimethacrylate (TEGDMA). The resin matrices were filled with 55 wt.% particles, of which 10 wt.% was 0.05-μm fumed silica, and 45 wt.% was 0.7-μm BaBSiO2 glass. Polymerization shrinkage stress (PSS; n=10), degree of conversion (DC; n=3), maximum rate of polymerization (Rpmax; n=3), film thickness (FT; n=10), sorption (Wsp; n=10), solubility (Wsl; n=10), flexural strength (FS; n=10), flexural modulus (FM; n=10), Knoop microhardness (KH; n=10), and microhardness reduction after chemical softening (HR; n=10) were evaluated. Data were analyzed using one-way ANOVA, followed by Tukey’s test (α=0.05; β=0.2). Results the results showed that the substitution or addition of IBOMA reduced FT (p=0.001), PSS (p=0.013), Rpmax (p=0.001), DC (p=0.001), FM (p=0.006) Wsp (p=0.032), and Wsl (p=0.021). However, when used as a complete substituent, IBOMA demonstrated significantly lower FS (p=0.017) and KH (p=0.008), while TEGDMA demonstrated significantly lower HR (p=0.022). Conclusion the flowable composite containing IBOMA combined with TEGDMA showed no effect in KH and FS and effectively reduced the PSS, RP, FT, Wsp, and Wsl. However, it showed a reduction in DC, FS, and an increase in HR.

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Abstract Objective Uncooperative behavior in pediatric dentistry is one of the most common manifestations of dental anxiety. Managing anxious patients can be attained by moderate sedation. This study aimed to compare the effect of sedation by dexmedetomidine-ketamine combination (DEX-KET) versus dexmedetomidine (DEX) on behavior of uncooperative pediatric dental patients. Methodology In total, 56 uncooperative healthy children (3–5 years old) requiring dental treatment were divided randomly into two groups: Group I (study group), which received buccal dexmedetomidine (2 μg/kg) and ketamine (2 mg/kg), and Group II (control group), which received only buccal dexmedetomidine (4 μg/kg). Drugs effects were assessed in terms of hemodynamic parameters, patient’s drug acceptance, child behavior, postoperative effect of sedation, amnesic effect, incidence of adverse events, as well as procedural induced stress measured by salivary secretory immunoglobulin A (s-IgA). Results Hemodynamic results did not reveal a statistically significant difference between the two study groups (P>0.05). There was a significant difference in patient’s acceptance to sedative drug between both groups, favoring DEX (p=0.005). Children who received DEX-KET showed significantly better behavior than those who received DEX for local anesthesia (p=0.017) and during operative procedure (p=0.037). Adverse events, post-operative and amnesic effects of drugs were comparable in both groups (p>0.05). Moreover, the mean difference in the salivary s-IgA levels between initial and final value was not statistically significant between both groups (p=0.556). Conclusion Both DEX-KET combination and DEX alone are effective in providing hemodynamic stability. DEX-KET combination significantly improved the behavior of sedated children compared to DEX alone but the drug acceptance was decreased in the DEX-KET group. Both regimens did not have a negative effect on postoperative behavior of children and had comparable amnesic effect with no significant adverse events. Salivary s-IgA is not considered a potential stress biomarker in sedated children.

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Abstract Objective: This study sought to investigate the relationship between clinical response to nonsurgical periodontal therapy (NSPT) and serum changes in leukocyte count, fasting blood glucose, hemoglobin, hematocrit, creatinine, and uric acid in kidney transplant recipients (KTR). Methodology: A prospective study was performed on 20 KTRs. Periodontal and serum data were collected before and 90 days after NSPT, and delta values (Δ = after NSPT - before) were calculated. Periodontal assessment included periodontal probing depth (PPD), clinical attachment level (CAL), and bleeding on probing (BOP). Patients were classified based on the presence of periodontitis and then categorized into stages. Results: Patients showed a reduction in the percentage of sites with PPD≥3mm, PPD≥4 mm and BOP, after NSPT. There was a direct correlation between the deltas of leukocyte count and CAL ≥3 mm (r=0.645, P=0.002) and BOP (r=0.663, P=0.001), and the deltas of uric acid and CAL ≥3 mm (r=0.562, P=0.010). Conclusion: A good clinical response to NSPT may affect the reduction of serum levels of leukocyte count and uric acid, suggesting a beneficial effect on systemic health in KTR.

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Abstract Objective For treatment of medication-related osteonecrosis of the jaw, one proposed approach is the use of a topical agent to block entry of these medications in oral soft tissues. We tested the ability of phosphonoformic acid (PFA), an inhibitor of bisphosphonate entry through certain sodium-dependent phosphate contransporters (SLC20A1, 20A2, 34A1-3) as well as Dynasore, a macropinocytosis inhibitor, for their abilities to prevent zoledronate-induced (ZOL) death in human gingival fibroblasts (HGFs). Methodology MTT assay dose-response curves were performed to determine non-cytotoxic levels of both PFA and Dynasore. In the presence of 50 μM ZOL, optimized PFA and Dynasore doses were tested for their ability to restore HGF viability. To determine SLC expression in HGFs, total HGF RNA was subjected to quantitative real-time RT-PCR. Confocal fluorescence microscopy was employed to see if Dynasore inhibited macropinocytotic HGF entry of AF647-ZOL. Endosomal acidification in the presence of Dynasore was measured by live cell imaging utilizing LysoSensor Green DND-189. As a further test of Dynasore’s ability to interfere with ZOL-containing endosomal maturation, perinuclear localization of mature endosomes containing AF647-ZOL or TRITC-dextran as a control were assessed via confocal fluorescence microscopy with CellProfiler™ software analysis of the resulting photomicrographs. Results 0.5 mM PFA did not rescue HGFs from ZOL-induced viability loss at 72 hours while 10 and 30 μM geranylgeraniol did partially rescue. HGFs did not express the SLC transporters as compared to the expression in positive control tissues. 10 μM Dynasore completely prevented ZOL-induced viability loss. In the presence of Dynasore, AF647-ZOL and FITC-dextran co-localized in endosomes. Endosomal acidification was inhibited by Dynasore and perinuclear localization of both TRITC-dextran- and AF647-ZOL-containing endosomes was inhibited by 30 μM Dynasore. Conclusion Dynasore prevents ZOL-induced viability loss in HGFs by partially interfering with macropinocytosis and by inhibiting the endosomal maturation pathway thought to be needed for ZOL delivery to the cytoplasm.

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Abstract Antimicrobial photodynamic therapy (aPDT) is an adjuvant treatment to scaling and root planing (SRP) which improves periodontal health. It may be beneficial to patients with systemic diseases, such as type 1 diabetes mellitus. Objective This randomized clinical trial evaluated the adjunctive effect of aPDT on the periodontal treatment of patients with type 1 diabetes (T1D). Methodology 38 patients were included in the study and divided into four groups: DSRP – T1D patients treated with SRP; CSRP – normoglycemic patients treated with SRP; DPDT – T1D patients treated with SRP + aPDT (methylene blue and red laser); CPDT – normoglycemic patients treated with SRP + aPDT. , Periodontal clinical parameters and inflammatory cytokines in crevicular fluid were recorded at baseline and then after 1, 3 and 6 months. The clinical endpoint for treatment was evaluated after 6 months. Results Adjuvant aPDT treatment resulted in reduction of probing depth after 3 months (0.38 mm - p<0.05) on T1D patients and in control group after 6 months (0.66 mm - p<0.05). Reduction of clinical attachment levels was similar for both treatments in control patients (p>0.05). There was a significant reduction of TNF-α in crevicular fluid in both groups treated with aPDT (p<0.05). The T1D (65%) and normoglycemic (72%) groups achieved the clinical endpoint after both treatments (p>0.05). Conclusions Adjuvant aPDT provided additional benefits in improving periodontal clinical parameters and reducing inflammatory cytokines in both T1D and normoglycemic patients. However, normoglycemic patients showed greater clinical improvements compared to T1D patients following adjuvant aPDT treatment.

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Abstract Objective This study aims to explore the effects of miR-223-3p and miR-155-5p on epithelial-mesenchymal transition (EMT) and migration in oral squamous cell carcinoma (OSCC). Methodology EMT markers (E-cadherin, N-cadherin, P120 catenin (P120ctn), and vimentin) expression was determined by qRT-PCR and western blot analysis in SCC-9 cells which overexpress miR-155-5p and/or not express miR-223-3p. Scratch assays and Transwell migration assays were conducted to evaluate cell migration ability. Results When miR-223-3p was inhibited in OSCC cells, P120ctn and E-cadherin mRNA levels were dramatically downregulated (P<0.05), while N-cadherin levels were significantly upregulated, and the migration ability of OSCC cells increased. The overexpression of miR-155-5p in OSCC cells upregulated miR-223-3p significantly (34-fold) compared to the control group. It also led to significant downregulation of the mRNA of P120ctn and E-cadherin and significant upregulation of the mRNA of N-cadherin and Vimentin (P<0.05). Meanwhile, the migratory ability of OSCC cells significantly increased. When miR-155-5p was overexpressed while miR-223-3p was inhibited, the highest expression of E-cadherin and P120ctn mRNA and the lowest expression of N-cadherin(P<0.05) was observed. Simultaneously, tumor cell migration was significantly facilitated. Conclusion miR-223-3p inhibits the migration of OSCC cells, while miR-155-5p can elevate the miR-223-3p mRNA expression. The simultaneous miR-155-5p overexpression and miR-223-3p inhibition can activate pEMT, increasing OSCC migration in vitro. This provides a novel approach and potential target for the effective treatment of OSCC.

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Abstract Objectives: to evaluate inflammation and vascularization in a model of apical periodontitis in diabetic Wistar rats through histopathological examination of blood vessels and immunohistochemical examination of interleukin 1b (IL-1b) and tumor necrosis factor-a (TNF-a). Methodology: Diabetes was induced in 20 Wistar rats using multiple low-dose injections of streptozotocin (STZ) until blood glucose levels stabilized above 300 mg/dL, confirmed by glucometer. Under anesthesia, apical periodontitis was induced in the right mandibular first molars. After preparing the access cavity and extirpating pulp and canal, the teeth were left open. Apical periodontitis was projected seven days afterwards and the Wistar rats were assigned in random into four groups, each group consisting of five rats. The first group (C14) was euthanized 14 days post-induction, while the second group (C28) was euthanized 28 days later, serving as controls. The third group (T14) received mesenchymal stem cells from human umbilical cords and was euthanized after 14 days, while the fourth group (T28) received mesenchymal stem cells from human umbilical cord and was euthanized after 28 days. The number of blood vessels and the expressions of IL-1b and TNF-a were analyzed. Data were evaluated with ANOVA and by Tukey's HSD test, with significance at p<0.05. Results: Both control and treatment groups showed a significant increase in vascularization in the apical periodontal area of apical periodontitis in the control and treatment groups at 14 and 28 days (p<0.05). A significant reduction of IL-1b and TNF-a levels was found in the mesenchymal stem cells treatment groups when compared to control groups (p<0.05). Conclusion: Our findings support the use of mesenchymal stem cells from human umbilical cords to decrease inflammation and increase vascularization in an induced apical periodontitis model in diabetes mellitus Wistar rats.

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Abstract Background and objective Periodontitis is an inflammatory disease typically characterized by the destruction of periodontal tissues and complicated etiology. DNA methyltransferase 3A (DNMT3A) has been implicated in possessing pro-inflammatory properties. This study sought to explore the role of DNMT3A in periodontitis and its relevant mechanism. Methodology Lipopolysaccharide (LPS) was used to induce inflammation in human periodontal ligament stem cells (hPDLSCs). DNMT3A and KLF5 expressions were detected using RT-qPCR and western blot. The levels of inflammatory cytokines and inflammation-related proteins were detected using ELISA and western blot. NF-κB p65 expression was detected using immunofluorescence (IF) assay, while osteogenic differentiation was assessed using ALP assay and ARS staining. Western blot was used to measure the protein contents associated with osteogenic differentiation. DNMT3A activity was detected using luciferase report assay and chromatin immunoprecipitation (ChIP) was used to verify the interaction between KLF5 and DNMT3A. Results DNMT3A expression increased in LPS-induced hPDLSCs. Silencing DNMT3A suppressed the LPS-induced inflammation in hPDLSCs, while promoting osteogenic differentiation. It was also found that transcriptional factor KLF5 could bind to DNMT3A promoters and regulate DNMT3A expression. Rescue experiments showed that KLF5 interference partially counteracted the inhibitory impacts of DNMT3A deficiency on inflammation and the promotive effects on osteogenic differentiation in LPS-induced hPDLSCs. Conclusion DNMT3A, when transcriptionally downregulated by KLF5, could alleviate LPS-challenged inflammatory responses and facilitate osteogenic differentiation in hPDLSCs.

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Abstract Background: the escalating influx of patients with temporomandibular disorders and the challenges associated with accurate diagnosis by non-specialized dental practitioners underscore the integration of artificial intelligence into the diagnostic process of temporomandibular disorders (TMD) as a potential solution to mitigate diagnostic disparities associated with this condition. Objectives: In this study, we evaluated a machine-learning model for classifying TMDs based on the International Classification of Orofacial Pain, using structured data. Methodology: Model construction was performed by the exploration of a dataset comprising patient records from the repository of the Multidisciplinary Orofacial Pain Center (CEMDOR) affiliated with the Federal University of Santa Catarina. Diagnoses of TMD were categorized following the principles established by the International Classification of Orofacial Pain (ICOP-1). Two independent experiments were conducted using the decision tree technique to classify muscular or articular conditions. Both experiments uniformly adopted identical metrics to assess the developed model's performance and efficacy. Results: The classification model for joint pain showed a relevant potential for general practitioners, presenting 84% accuracy and f1-score of 0.85. Thus, myofascial pain was classified with 78% accuracy and an f1-score of 0.76. Both models used from 2 to 5 clinical variables to classify orofacial pain. Conclusion: The use of decision tree-based machine learning holds significant support potential for TMD classification.

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Abstract Diabetes mellitus (DM) delays wound healing, including those following tooth extractions. Curcumin (CCM) can promote soft tissue and bone healing. Objective: The present study investigates the healing effects of CCM on tooth extraction sockets in diabetic rats. Methodology: Ninety-six male Wistar rats were divided into the following four groups: Control+Corn Oil (CO), Control+CCM, DM+CO, and DM+CCM. Each group was subdivided into 7-, 14-, and 28-day time point subgroups comprising eight rats. All animals had their maxillary first molars extracted. CCM-treated rats received 100 mg/kg of CCM orally for 7, 14, and 28 days. The lesion area was evaluated using macroscopic analyses, whereas socket healing was assessed by hematoxylin and eosin staining. Keratinocyte growth factor (KGF), Runt-related transcription factor 2 (Runx2), and collagen type I (COL1) expression levels were obtained using quantitative polymerase chain reaction (qPCR). Bone healing was analyzed by means of microcomputed tomography (μCT). Results: After 7 days, the groups showed no significant differences in lesion area and by day 14, no lesions were present. CCM treatment increased KGF mRNA expression in diabetic rats; however, diabetic rats showed delayed bone healing unrelated to CCM. CCM treatment resulted in increased Runx2 mRNA expression only in control rats, whereas COL1 mRNA expression remained unaffected by CCM. Conclusion: CCM shows potential as a soft tissue healing enhancer in diabetic rats and could serve as an additional treatment to promote soft tissue repair in diabetic individuals. Although CCM did not impact alveolar bone healing, it may enhance bone healing in other skeleton regions.

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Periodontal dental ligament mesenchymal stem cells (PDLMSCs) play a major role in periodontal tissue regeneration by the neoformation of root cementum and alveolar bone. These cells are highly heterogeneous, and many present low potential to renovate the hard tissue damaged by periodontal disease. A previous study found that the low osteoblast/cementoblast (O/C) differentiation potential of PDLMSCs is related to high asporin (ASPN) expression, which was identified as a negative regulator of PDL cells differentiation and mineralization, suppressing BMP-2-induced O/C differentiation. Objective This study aimed to investigate whether 1,25(OH)2D3 treatment could stimulate the O/C differentiation of periodontal ligament mesenchymal progenitor cells characterized as low osteoblast potential (LOP), by asporin and bone morphogenetic protein-2 alteration. Methodology Three LOP cell populations were cultured in standard medium (CONTROL), osteogenic medium (OM), and osteogenic medium associated with 1 nM of 1,25(OH)2D3 (OM + VD). The following assays were performed: 1) MTT to evaluate metabolic activity; 2) gene expression for asporin (ASPN), bone morphogenetic protein-2 (BMP-2), runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin (OCN), and vitamin D receptor (VDR) using qRT-PCR; 3) BMP-2 extracellular expression; and 4) quantification of mineralized nodule deposition by Alizarin Red Staining. Data were subjected to two-way ANOVA and Tukey’s test (P<0.05). Results The results showed that the 1,25(OH)2D3 treatment did not affect the cell viability, as demonstrated by metabolic activity increase over the 10 days in culture. After 14 days of 1,25(OH)2D3 treatment, the mRNA levels for ASPN and VDR decreased (P<0.05), while BMP-2 transcripts and extracellular expression increased (P<0.05). In parallel, RUNX2, ALP, and OCN gene expression was upregulated by 1,25(OH)2D3 treatment, resulting in an increase of mineral nodule deposition in vitro (P<0.05). Conclusions These data show that 1,25(OH)2D3 improves osteoblast/cementoblast differentiation of low osteoblast potential accompanied by alterations in ASPN and BMP-2 expression.

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Abstract Cleft lip and palate are the most common congenital malformations in the craniofacial region, occurring at a rate of 1:700 births in Brazil. These conditions lead to functional impacts on patients, such as changes in breathing, teeth, speech, chewing, swallowing and sucking. Treatment begins with primary surgeries, including lip and palate repair, which aim to reconstruct the soft tissues. Secondary alveolar bone grafting (SABG) reconstructs the bone defect in the cleft region, with the main goal of supplying bone tissue to the cleft region and restore the continuity of the alveolar process. Objective: To measure the changes in cross-sectional areas (CSAs) and nasal volume in patients and their impact on the nasal cavity (NC) in the two-month postoperative period (PO2M). Methodology: This study included 15 patients with complete unilateral cleft lip and palate (U/CLP) indicated for alveolar bone grafting (ABG). Cone beam computed tomography scans obtained prior to SABG and at PO2M were compared. Nasal volumes and CSAs were measured by marking the masks delimiting the nasal cavity on CT scans using Mimics™ software. Results: NC volumes (total, right and left sides) were statistically lower at PO2M in patients with left-sided UCLP. In right-sided UCLP, these volumes were only significant for the total NC and left NC. The CSAs of the internal nasal valve in both groups showed significantly lower values compared to the preoperative period (p≤0.05). Conclusion: In the short term, alveolar bone graft surgery reduces the volume of nasal cavities and the cross-sectional areas of the right and left internal nasal valve as a whole, not only the cleft area where the graft material was placed.

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Abstract Objective: Although autogenous grafting is accepted as the gold standard in intraoral grafting, xenogenous grafts are frequently used in sinus lift surgeries due to their osteoinductive and osteoconductive properties. This study aimed to investigate the efficacy of fish spine-derived xenogenic grafts in sinus augmentation surgery. Material and Methods: In this study, a fish spine-derived xenogenic graft was produced for comparison with autogenous graft and bovine derived xenogenic grafts. Twenty-one New Zealand rabbits were used. Autogenous grafts (AG- Group 1), as well as bovine-derived (bHAP - Group 2) and fish spine-derived (fHAP - Group 3) xenogenic grafts were placed in the right and left sinuses of rabbits. The animals were sacrificed at the 4th and 8th weeks. New bone formation (NBF) was evaluated through histological examination, while bone volume (BV), new bone surface/bone volume (BS-BV), new bone surface/tissue volume (BS-TV), and trabecular separation (Tb-Sp) were assessed via Micro-CT. Statistical significance was considered at p<0.05. Results: Histological examination revealed a significant difference in NBF between AG-bHAP (p<0.001), as well as between fHAP-bHAP (p<0.001) in the fourth-week group. No significant difference was found in the eighth-week group (p=0.130). In the eighth-week group, a statistically significant difference was found between fHAP and bHAP in terms of BV. (p=0.007). Conclusion: Although both graft materials used in this study showed positive effects on bone regeneration, fHAP and AG presented similar effects on bone regeneration and were superior to bHAP.

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Abstract Objectives Considering the fact that resin infiltrants lack antibacterial activity, this study assessed the influence of the quaternary ammonium monomer dimethylaminohexadecyl methacrylate (DMAHDM) and amorphous calcium phosphate nanoparticles (NACP) on the physical and antibacterial properties of an experimental resin infiltrant (ERI). Methodology The following groups were established: ERI (75/25 wt.% TEGDMA/BISEMA), ERI + 2.5% DMAHDM (2.5DM), ERI + 5% DMAHDM (5DM), ERI + 2% NACP (NACP), ERI + 2.5% DMAHDM + 2% NACP (2.5DM_NACP), ERI + 5% DMAHDM + 2% NACP (5DM_NACP), and Icon® (IC), a commercial resin infiltrant. Degree of conversion (DC; n=4), sorption and solubility (SO/SOL; n=8), and contact angle (CA; n=10) tests were conducted. Biofilm biomass (BB; n=6) and bacterial metabolism (BM; n=8) were evaluated after Streptococcus mutans (UA159) cultivation for 48 h on material samples. Data were evaluated by one-way ANOVA and Tukey or Games-Howell post hoc tests (α=0.05). Results IC exhibited the highest DC, with no difference from 2.5DM and 5DM. IC showed the lowest CA. IC had the lowest SO, followed by ERI, which had the lowest SOL, with no difference from IC. 5DM_NACP showed the lowest biofilm biomass, similar to 2.5DM and 5DM. Resin infiltrants containing DMAHDM showed reduced bacterial metabolism. Conclusions DMAHDM, with or without NACP, demonstrated significant antibacterial activity, while NACP impaired DC. Both DMAHDM and NACP increased the contact angle, sorption, and solubility of the resin infiltrant, which may affect the material’s clinical performance.