Effect of photobiomodulation on inflammatory cytokines produced by HaCaT keratinocytes.

Objective: to evaluate the effects of the red and near-infrared wavelength lasers in isolated and simultaneous way on the modulation of inflammatory cytokines produced by human keratinocytes (HaCaT) challenged by cytokines of human monocytes stimulated by lipopolysaccharide from Escherichia coli. Design: HaCaT cells was previously exposed to the laser with wavelengths red (660 nm), near-infrared (808 nm). Then, HaCat cells were stimulated with the supernatant of lipopolysaccharide-challenged peripheral blood cells. The cytokines expressed by HaCat cells were measured using multiplex CBA assay. Results: HaCaT cells increased the production of inflammatory cytokines when stimulated with infrared laser compared to the control group (IFN-α2, IFN-γ, TNF-α, MCP-1, IL-6, IL-8, IL-10, IL -12p70, IL -17A, IL-23, IL-33), the red laser group (IFN-γ and IL-23) and the group of two lasers used simultaneously (IFN-α2, IFN-γ, IL-6 and IL-8, IL-17A, IL-18 and IL-23) (p < 0.05). The red laser also stimulated an increase in the expression of IFN-α2 by HaCaT cells in relation to the control group (p < 0.05). Conclusion: Infrared laser, with an energy density of 5 J/cm2, appear to be able to modulate inflammatory cytokines produced by HaCaT cells challenged by human monocyte cytokines.

Exploring the association between a standardized extract of pequi peels (Caryocar brasiliense Cambess) and blue light as a photodynamic therapy for treating superficial wounds.

Natural products derived from plants can be used as photosensitizers for antimicrobial photodynamic therapy (aPDT) combining key therapeutic strategies for tissue repair while controlling microorganisms' growth. We investigated a standardized extract of pequi peels (Caryocar brasiliense Cambess) as a brownish natural photosensitizer for aPDT using blue light. Three concentrations of the pequi extract (PE; 10, 30, or 90 µg/mL) were tested solely or associated with blue laser (445 nm, 100 mW, 138 J/cm2 , 6 J, 60 s). In vitro, we quantified reactive oxygen species (ROS), assessed skin keratinocytes (HaCat) viability and migration, and aPDT antimicrobial activity on Streptococcus or Staphylococcus strains. In vivo, we assessed wound closure for the most active concentration disclosed by the in vitro assay (30 µg/mL). Upon aPDT treatments, ROS were significantly increased in cell monolayers regardless of PE concentration. PE at low doses stimulates epithelial cells. Although PE stimulated cellular migration, aPDT was moderately cytotoxic to skin keratinocytes, particularly at the highest concentration. The antimicrobial activity was observed for PE at the lowest concentration (10 µg/mL) and mostly at PE 10 µg/mL and 30 µg/mL when used as aPDT photosensitizers. aPDT with PE 30 µg/mL presents antimicrobial activity without compromising the initial phases of skin repair.

Effects of bisphenol A on murine salivary glands and human tumor cell lines.

Bisphenol A (BPA) is an endocrine-disrupting chemical with a potential role in endocrine cancers. However, the effects of BPA on the salivary glands have been barely explored. We investigated the impact of in vivo sub-chronic exposure to BPA and its in vitro effects on human salivary gland mucoepidermoid carcinoma cell lines. Male and female mice were exposed to BPA (30 mg/kg/day). Sublingual and submandibular salivary glands from an estrogen-deficiency model were also analyzed. BPA concentration in salivary glands was evaluated by gas chromatography coupled to ion trap mass spectrometry. Immunohistochemical analysis using anti-p63 and anti-α-SMA antibodies was performed on mouse salivary gland tissues. Gene expression of estrogen receptors alpha and beta, P63 and α-SMA was quantified in mouse salivary gland and/or mucoepidermoid (UM-HMC-1 and UM-HMC-3A) cell lines. Cell viability, p63 and Ki-67 immunostaining were evaluated in vitro. BPA disrupted the tissue architecture of the submandibular and sublingual glands, particularly in female mice, and increased the expression of estrogen receptors and p63, effects that were accompanied by significant BPA accumulation in these tissues. Conversely, ovariectomy slightly impacted BPA-induced morphological changes. In vitro, BPA did not affect the proliferation of neoplastic cells, but augmented the expression of p63 and estrogen receptors. The present data highlight a potential harmful effect of BPA on salivary gland tissues, particularly in female mice, and salivary gland tumor cells. Our findings suggest that estrogen-dependent pathways may orchestrate the effects of BPA in salivary glands.

Noncarious cervical lesions in the aging population analyzed through digital scanning: occlusal risk factors, dimensions, and morphology.

OBJECTIVES: To evaluate noncarious cervical lesions (NCCL) in older adults by analyzing each lesion's morphology, dimension, and occlusal risk factors using intraoral scanning and clinical evaluation. MATERIALS AND METHODS: Individuals over 60 treated at the School of Dentistry in Minas Gerais, Brazil, were invited to participate in this cross-sectional study. We clinically evaluated 1245 teeth for the presence of NCCL and dentin hypersensitivity (DH) in 53 individuals. Three-dimensional models were obtained using Omnicam, and occlusal risk factors were evaluated through the presence of wear facets, intense occlusal contacts, and interferences. The 3Shape 3DViewer software was used to classify NCCL morphology and to measure their height. NCCL depth, width, and area measurements were determined using ImageJ. Descriptive analyses were performed. The Kolmogorov-Smirnov test showed that quantitative data exhibited non-normal distribution. For bivariate analyses of quantitative data, the Mann-Whitney test was employed. The results were reported with mean, standard deviation, median, minimum, and maximum. For bivariate analyses of categorical data, the Pearson chi-square test was used. The results were reported with frequencies (counts) and percentages. A regression model evaluating the association between occlusal risk factors and the absence/presence of NCCL was built. RESULTS: Most participants were female (58.5%), with a mean age of 66. The frequency of NCCL was 28.27%, corresponding to 352 teeth. Of these, 68.18% exhibited saucer morphology and 31.82% wedge-shaped morphology. The frequency of DH was 13.92% in teeth with NCCL. Saucer NCCL exhibited significantly greater height (p = 0.02), while wedge-shaped lesions showed greater depth (p < 0.001). Also, teeth with NCCL had a higher proportion of wear facets (66.8%; p < 0.001), intense occlusal contacts (39.8%; p = 0.008), and occlusal interferences (21.9%; p = 0.05). The regression model showed that teeth with wear facets were 1.50 times more likely to exhibit NCCL than teeth without wear facets (p < 0.001). Teeth with intense occlusal contact were 1.22 times more likely to exhibit NCCL than teeth without intense occlusal contact (p = 0.031). CONCLUSIONS: NCCL is a common dental condition in the older adults evaluated in this study. Wedge-shaped NCCL exhibited greater depth, while saucer NCCL had more expressive height. Occlusal risk factors may represent a crucial mechanism for the occurrence of NCCL in this population. CLINICAL RELEVANCE: Understanding the mechanisms involved in developing NCCL is helpful in diagnostic and preventive practices. The study showed the importance of dental occlusion and characteristics of different NCCL morphologies that help the clinician in decision-making.

The participation of tumor residing pericytes in oral squamous cell carcinoma.

Pericytes are perivascular cells related to vessel structure and angiogenesis that can interact with neoplastic cells, interfering with cancer progression and outcomes. This study focused on the characterization of pericytes in oral squamous cell carcinoma (OSCC) using clinical samples and a transgenic mouse model of oral carcinogenesis. Nestin-/NG2+ (type-1) and nestin+/NG2+ (type-2) pericytes were analyzed by direct fluorescence after induction of oral carcinogenesis (4-nitroquinoline-1-oxide). Gene expression of neuron glial antigen-2 (NG2), platelet-derived growth factor receptor beta (PDGFR-ß), and cluster of differentiation 31 (CD31) was examined in human OSCC tissues. The protein expression of von Willebrand factor and NG2 was assessed in oral leukoplakia (i.e., oral potentially malignant disorders) and OSCC samples. Additionally, clinicopathological aspects and survival data were correlated and validated by bioinformatics using The Cancer Genome Atlas (TCGA). Induction of carcinogenesis in mice produced an increase in both NG2+ pericyte subsets. In human OSCC, advanced-stage tumors showed a significant reduction in CD31 mRNA and von Willebrand factor-positive vessels. Low PDGFR-ß expression was related to a shorter disease-free survival time, while NG2 mRNA overexpression was associated with a reduction in overall survival, consistent with the TCGA data. Herein, oral carcinogenesis resulted in an increase in NG2+ pericytes, which negatively affected survival outcomes.

Efficacy of antiseptics and chemomechanical methods for dentin caries lesions: A systematic review with GRADE approach.

Objectives: Selective caries removal aims to remove carious tissue in deep dentin lesions. However, a discussion stands on the value of antiseptics and chemomechanical adjuvant methods to reduce the bacterial load on residual caries lesions. This systematic review has addressed two main clinical questions to compare the antimicrobial efficacy of available methods using (1) antiseptic or (2) chemomechanical agents before restoring dentin carious lesions. Methods: We included randomized and non-randomized controlled trials (RCTs/ NRCTs). We searched eight databases from inception to October 2021. Paired reviewers independently screened studies, extracted data, and assessed the risk of bias. The primary outcome was the reduction in the number of total bacterial in dentin, whereas secondary outcomes were reduction in the number of Lactobacillus and Streptococcus. We used the ratio of ratio of post-treatment to baseline means between two interventions in the logarithmic scale as a proper effect measure. Certainty of evidence was assessed with the Grading of Recommendations, Assessment, Development and Evaluation approach. Results: We included 14 RCTs and 9 NRCTs, with nine interventions. Regardless the method, the number of bacteria at baseline was similar or exceeded that after the intervention, particularly in NRCTs. The evidence was inconclusive for most comparisons. Among antiseptic agents, chlorhexidine (CHX) resulted in an average of 1.14 times [95% confidence interval (CI): 1.08-1.21] more total bacterial than photodynamic therapy in RCTs. Among NRCTS, the natural agents resulted in five times more total bacterial than CHX (95% CI: 2-11). For chemomechanical methods, the control resulted in eight times (95% CI: 4-17) more total bacterial than Carisolv (SHAA). Conclusions: The certainty of the evidence was very low for all comparisons showing uncertainty whether one treatment could be more effective than another for dentin disinfection. So far, exclusively removing soft carious dentin would be enough to reduce the bacterial count.

Release of leachable products from resinous compounds in the saliva of children with anterior open bite treated with spur.

BACKGROUND: To evaluate the release of bisphenol-A glycidyl methacrylate (BisGMA), triethylene glycol dimethacrylate (TEGDMA), bisphenol A (BPA), and phthalates of the composite resin used in the bonding of spurs applied in the treatment of children with anterior open bite and its effects on human keratinocytes. METHODOLOGY: Saliva samples of 22 children were collected before spur attachment (baseline) and 30 minutes (min) and 24 hours (h) after spur bonding. Analysis was performed using high-performance liquid chromatography (HPLC) coupled to tandem mass spectrometry (HPLC-MS/MS) and gas chromatography coupled to mass spectrometry (GC-MS). Standardized resin increments were added to three different dilutions of the cell culture medium. Keratinocytes (HaCaT) were cultivated in the conditioned media and evaluated for cell viability (MTT) and cell scratch assay. RESULTS: The levels of BisGMA (1.74±0.27 µg/mL), TEGDMA (2.29±0.36 µg/mL), and BPA (3.264±0.88 µg/L) in the saliva after 30 min, in comparison to baseline (0±0 µg/mL, 0±0 µg/mL, and 1.15±0.21 µg/L, respectively), presented higher numbers. After 24 h, the levels of the monomers were similar to the baseline. Phthalates showed no significant difference among groups. HaCat cells showed increased viability and reduced cell migration over time after exposure to methacrylate-based resin composites. CONCLUSION: Resin composites, used to attach spurs in children with anterior open bite during orthodontic treatment, release monomers after polymerization and can influence the behavior of human keratinocytes, even at very low concentrations. Orthodontists should be aware of the risks of the resinous compounds release and preventive procedures should be held to reduce patient exposure.

A multilevel analysis model for dental caries determinants in independently-living elderly

To evaluate the prevalence of untreated caries and its association with biological, individual, and environmental variables in independently-living elderly people. Methods: This cross-sectional study included 72 elderly (≥60 years) patients of a university dental clinic in Belo Horizonte, Brazil. Sociodemographic data, systemic diseases, medications, and free sugar intake were collected. Visible plaque, Decayed, Missing, and Filled Teeth (DMFT), and Decayed and Filled Root (DFR) indexes were assessed through clinical examination. Unstimulated saliva was collected to determine salivary flow, pH, and buffering capacity. Descriptive analysis and multilevel logistic regression analysis were performed following a dental caries theoretical model (p <0.05, 95% CI). Results: The mean DMFT and DFR were 24.44 (SD=4.59) and 3.21 (SD=2.93), respectively. The prevalence of untreated caries was 61.11%. In the adjusted multilevel regression model involving 1639 teeth, untreated dental caries was significantly associated with the presence of biofilm (OR = 1.84; 95% CI: 1.24­2.74), salivary buffering capacity (OR = 0.87; 95% CI: 0.77­0.99) and per capita income (OR = 0.06; 95% CI: 0.004­0.74). Conclusion: The experience of dental caries was widespread among independently-living elderly patients, and its variability was best explained by the presence of biofilm, reduced salivary buffering capacity, and low per capita income. A comprehensive assessment is needed of the biological, individual, and environmental factors related to the presence of dental caries in independently-living elderly people

Adjunctive therapies for in vitro carious lesions: Antimicrobial activity, activation of dentin metalloproteinases and effects on dental pulp cells.

BACKGROUND: Adjunctive therapies used before dental restorative procedures may encourage carious tissue removal. Beyond promising antimicrobial properties, treatments could positively modulate the dentin-pulp complex while not interfering with restoration survival. Herein, we evaluated a set of substances and their effects on carious lesions and the underlying dentin or pulp cells. METHODS: Artificial caries lesions were developed in bovine teeth cavities immersed in Streptococcus mutans and Lactobacillus casei co-cultures. The cavities were treated according to the following groups: Phosphate Buffer Saline (PBS), Chlorhexidine (CHX), Papacárie® (Papain gel), Ozone (O3), and antimicrobial Photodynamic Therapy (aPDT). After treatments, samples were cultivated to count isolated microbial colonies. The zymography assay evaluated the activity of dentin metalloproteinases (MMP-2 and MMP-9). Cell viability was indirectly assessed on human dental pulp cells after 24, 72, or 120 h, whereas the odontodifferentiation potential was evaluated after ten days of cell culture. RESULTS: CHX and aPDT led to around 1 log bacterial load reduction. PBS, CHX, and aPDT showed the eventual expression of MMP-2 and MMP-9. Cell viability was reduced (< 30%) after 120 h for all groups compared to the control. CHX, O3, and aPDT induced greater odontodifferentiation (≈ 20% higher) than PBS and papain gel. CONCLUSION: Adjunctive therapies presented little or no biological significance in reducing bacterial load in artificial carious lesions. Although the activation of endogenous metalloproteinases may represent a possible concern for adhesive restorations, some of these treatments may have a positive role in dental pulp tissue repair.

Enhanced mechanical properties, anti-biofilm activity, and cytocompatibility of a methacrylate-based polymer loaded with native multiwalled carbon nanotubes.

OBJECTIVES: We aimed to optimize the mechanical and biological properties of a conventional methacrylate-based dental polymer by loading it with double- and triple-walled carbon nanotubes as growth (DTWCNTG). METHODS: A formulation of bisphenol A-glycidyl methacrylate and triethylene glycol dimethacrylate (mass ratio = 2:1) was mixed with DTWCNTG at concentrations of 0.0% (control), 0.001%, 0.005%, and 0.010%. The concentrations were physicochemical and morphologically evaluated, and antibacterial activity was assessed by seeding a Streptococcus mutans strain (ATCC 25175) on the experimental polymeric surfaces. Cellular survival and osteodifferentiation were evaluated in epithelial (HaCat) and preosteoblast cells (MC3T3-E1). RESULTS: The 0.001% DTWCNTG concentration yielded higher compressive strength, elastic modulus, flexural strength, flexural modulus, water sorption, and solubility than the control. The degree of conversion and color did not significantly change with a low amount of DTWCNTG incorporated into the polymer. Antibacterial activity significantly improved when tested on the 0.001% DTWCNTG discs. No groups showed cytotoxicity in a short-term analysis and adding DTWCNTG favored MC3T3-E1 mineralization over the control, particularly in the 0.001% formulation. SIGNIFICANCE: The micro-addition of 0.001% DTWCNTG confers mechanical resistance, antimicrobial properties, and bioactivity to methacrylate-based polymers without significantly compromising color. Incorporating DTWCNTG improved dental composite properties and could be a biomodified material for minimally invasive procedures.

Equisetum hyemale-derived unprecedented bioactive composite for hard and soft tissues engineering.

Although Bioactive Glasses (BGs) have been progressively optimized, their preparation often still involves the use of toxic reagents and high calcination temperatures to remove organic solvents. In the present work, these synthesis related drawbacks were overcome by treating the ashes from the Equisetum hyemale plant in an ethanol/water solution to develop a bioactive composite [glass/carbon (BG-Carb)]. The BG-Carb was characterized by scanning electron microscopy, and transmission electron microscopy; and its chemical composition was assessed by inductively coupled plasma-optical emission spectroscopy. Brunauer-Emmett-Teller gas adsorption analysis showed a specific surface area of 121 m2 g-1. The formation of hydroxyapatite (HA) surface layer in vitro was confirmed by Fourier-transform infrared spectroscopy analysis before and after immersion in simulated body fluid (SBF) solution. The Rietveld refinement of the XRD patterns and selected area electron diffraction analyses confirmed HA in the sample even before immersing it in SBF solution. However, stronger evidences of the presence of HA were observed after immersion in SBF solution due to the surface mineralization. The BG-Carb samples showed no cytotoxicity on MC3T3-E1 cells and osteo-differentiation capacity similar to the positive control. Altogether, the BG-Carb material data reveals a promising plant waste-based candidate for hard and soft tissue engineering.

Grafting biomaterials associated to topical glucocorticoid: effects on pre-osteoblastic cells (MC3T3-E1).

The topical glucocorticoid budesonide has been prescribed before and after sinus lift surgery as adjuvant drug treatment for maxillary sinus membrane inflammation. However, there is no study on the effects of budesonide on the regenerative process of bone grafting biomaterials. We investigated the effect of the association of budesonide with some biomaterials on the growth and differentiation capacity of pre-osteoblastic cells (MC3T3-E1 subclone 4). Xenogeneic (Bio-Oss and Bio-Gen) and synthetic hydroxyapatites (Osteogen, Bonesynth, and HAP-91) were tested in conditioned medium (1% w/v). The conditioned medium was then supplemented with budesonide (0.5% v/v). Cell viability was assessed using the MTT assay (48, 96, and 144 h), and mineralized nodules were quantified after 14 days of culture using the Alizarin Red Staining. Alkaline phosphatase activity was assessed through the release of thymolphthalein at day seven. All biomaterials showed little or no cytotoxicity. The Bio-Gen allowed significantly less growth than the control group regardless of the experimental time. Regarding differentiation potential of MC3T3-E1, the HAP-91-conditioned medium showed remarkable osteoinductive properties. In osteodifferentiation, the addition of budesonide favored the formation of mineral nodules when cells were cultured in medium conditioned with synthetic materials, whereas it weakened the mineralization potential of cells cultured in xenogeneic medium. Regardless of whether budesonide was added or not, Osteogen and Bio-Oss showed higher alkaline phosphatase activity than the other groups. Budesonide may improve bone formation when associated with synthetic biomaterials. Conversely, the presence of this glucocorticoid weakens the mineralization potential of pre-osteoblastic cells cultured with xenogeneic hydroxyapatites.

Evaluation of periodontal parameters and gingival crevicular fluid cytokines in children with anterior open bite receiving passive orthodontic treatment with a spur.

Objective: To evaluate clinical parameters and gingival crevicular fluid (GCF) cytokines in children with anterior open bite receiving passive orthodontic treatment with spurs. Methods: Twenty children with indications for interceptive orthodontic treatment, an anterior open bite, and good oral hygiene and periodontal health were included in this study. GCF samples were collected from the mandibular and maxillary central incisors before (baseline) and 24 hours and 7 days after spur bonding. Clinical and periodontal examinations and cytokine analyses were performed. Results: At 7 days after spur attachment, gingival bleeding in the mandibular group was increased relative to that in the maxillary group. Visible plaque was correlated with gingival bleeding at 7 days and the GCF volume at 24 hours after spur attachment. Compared with those at baseline, interleukin (IL)-8 levels in the maxillary group and IL-1ß levels in both tooth groups increased at both 24 hours and 7 days and at 7 days, respectively. At 24 hours, IL-8, IL-1ß, and IL-6 levels were higher in the maxillary group than in the mandibular group. Cytokine production was positively correlated with increased GCF volume, but not with gingival bleeding, visible plaque, or probing depth. Conclusions: Although orthodontic treatment with spurs in children resulted in increased gingival bleeding around the mandibular incisors, IL levels were higher around the maxillary incisors and not correlated with periodontal parameters. Increased cytokine levels in GCF may be associated with the initial tooth movement during open bite correction with a passive orthodontic appliance in children.

Nestin and Neuron-glial antigen 2 transgenes unveil progenitor units in murine salivary glands.

OBJECTIVE: Undifferentiated cells play pivotal roles in sustaining tissue homeostasis during physiological turnovers and after tissue impairment. Nestin and Neuron-glial antigen 2 (NG2) are markers frequently deployed to distinguish progenitor populations. In the salivary gland scenario, these markers remain largely unknown. Particularly for a double-labeled group of progenitor cells (NG2+Nestin+), their phenotype and distribution have never been explored in freshly isolated tissues. Herein, we analyzed a subset of plastic cells that express Nestin and NG2 near the ducts and in the periacinar region of the major salivary glands of murine samples. DESIGN: The major salivary glands tissues of Nestin-GFP/NG2-DsRed mice were analyzed under a fluorescence microscope. The cells marked by GFP and DsRed were counted in the merged image component of random representative images obtained for each gland sample at × 20 magnification. RESULTS: In the parotid, submandibular, and sublingual glands, the population of cells exclusively expressing Nestin was more abundant. There was a predominance of Nestin, NG2, and double-labeled cells in the submandibular gland compared to the parotid gland, mainly near the ductal system. Of note, the sublingual and parotid glands had similar populations of Nestin+ and NG2+, especially in acini, and some positive cells were observed surrounding ducts. CONCLUSIONS: Collectively, our study revealed differential expression patterns of Nestin and NG2, alone or in combination, in the salivary gland subset during homeostasis.

Release of leachable products from resinous compounds in the saliva of children with anterior open bite treated with spur

Abstract To evaluate the release of bisphenol-A glycidyl methacrylate (BisGMA), triethylene glycol dimethacrylate (TEGDMA), bisphenol A (BPA), and phthalates of the composite resin used in the bonding of spurs applied in the treatment of children with anterior open bite and its effects on human keratinocytes. Methodology Saliva samples of 22 children were collected before spur attachment (baseline) and 30 minutes (min) and 24 hours (h) after spur bonding. Analysis was performed using high-performance liquid chromatography (HPLC) coupled to tandem mass spectrometry (HPLC-MS/MS) and gas chromatography coupled to mass spectrometry (GC-MS). Standardized resin increments were added to three different dilutions of the cell culture medium. Keratinocytes (HaCaT) were cultivated in the conditioned media and evaluated for cell viability (MTT) and cell scratch assay. Results The levels of BisGMA (1.74±0.27 μg/mL), TEGDMA (2.29±0.36 μg/mL), and BPA (3.264±0.88 μg/L) in the saliva after 30 min, in comparison to baseline (0±0 μg/mL, 0±0 μg/mL, and 1.15±0.21 μg/L, respectively), presented higher numbers. After 24 h, the levels of the monomers were similar to the baseline. Phthalates showed no significant difference among groups. HaCat cells showed increased viability and reduced cell migration over time after exposure to methacrylate-based resin composites. Conclusion Resin composites, used to attach spurs in children with anterior open bite during orthodontic treatment, release monomers after polymerization and can influence the behavior of human keratinocytes, even at very low concentrations. Orthodontists should be aware of the risks of the resinous compounds release and preventive procedures should be held to reduce patient exposure.

Grafting biomaterials associated to topical glucocorticoid: effects on pre-osteoblastic cells (MC3T3-E1)

Abstract The topical glucocorticoid budesonide has been prescribed before and after sinus lift surgery as adjuvant drug treatment for maxillary sinus membrane inflammation. However, there is no study on the effects of budesonide on the regenerative process of bone grafting biomaterials. We investigated the effect of the association of budesonide with some biomaterials on the growth and differentiation capacity of pre-osteoblastic cells (MC3T3-E1 subclone 4). Xenogeneic (Bio-Oss and Bio-Gen) and synthetic hydroxyapatites (Osteogen, Bonesynth, and HAP-91) were tested in conditioned medium (1% w/v). The conditioned medium was then supplemented with budesonide (0.5% v/v). Cell viability was assessed using the MTT assay (48, 96, and 144 h), and mineralized nodules were quantified after 14 days of culture using the Alizarin Red Staining. Alkaline phosphatase activity was assessed through the release of thymolphthalein at day seven. All biomaterials showed little or no cytotoxicity. The Bio-Gen allowed significantly less growth than the control group regardless of the experimental time. Regarding differentiation potential of MC3T3-E1, the HAP-91-conditioned medium showed remarkable osteoinductive properties. In osteodifferentiation, the addition of budesonide favored the formation of mineral nodules when cells were cultured in medium conditioned with synthetic materials, whereas it weakened the mineralization potential of cells cultured in xenogeneic medium. Regardless of whether budesonide was added or not, Osteogen and Bio-Oss showed higher alkaline phosphatase activity than the other groups. Budesonide may improve bone formation when associated with synthetic biomaterials. Conversely, the presence of this glucocorticoid weakens the mineralization potential of pre-osteoblastic cells cultured with xenogeneic hydroxyapatites.

Toxicity of Carnoy's solution toward human keratinocytes: an in vitro study.

The present study aimed to characterize the chemical elements and cytotoxicity of Carnoy's solution (CS) by comparing two different trademarked products (one Brazilian [NCS] and another imported [ICS]) using inductively coupled plasma mass spectrometry (ICP-MS) and human keratinocyte (HaCaT) cultures. For performing ICP-MS, the solutions were diluted according to calibration curves, and the chemical elements were analyzed with a spectrometer. HaCaT cells were exposed to CS concentrations ranging from 0.10% to 20% for 3 or 5 min. Cell viability was evaluated immediately (T0), 24 h (T1), and 7 days (T2) after exposure to CS using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) reduction assay. Data were analyzed using a t-test for ICP-MS and analysis of variance followed by Tukey's post-hoc test for MTT assay, both considering statistical significance at p<0.05. ICP-MS results revealed that ICS presented significantly lower concentrations of 12 chemical elements than NCS. The results of MTT assay revealed that at T0, ICS was more cytotoxic than NCS regardless of the time of exposure (p < 0.05). At T1, the only difference between the groups was at a concentration of 0.10% after 5 min of exposure. At T2, at a concentration of 0.5%, ICS resulted in a significant reduction in cell viability compared to NCS (p < 0.05). Thus, the results showed that ICS was more cytotoxic than NCS. Collectively, our findings suggest that the individual compositions of different CS formulations should be investigated.

Carious lesions recurrence in children with developmental disabilities: a longitudinal study.

This study investigated the factors associated with new carious lesions in one-to five-year-old children with developmental disabilities. This was a retrospective cohort that evaluated 64 dental charts of individuals with caries or fillings in their first dental appointment. The dependent variable was the occurrence of a new carious lesion or restoration. Gender, age, mother's education, sugar consumption, oral hygiene, mouth breathing, reports of xerostomia, gingival status, use of psychotropic drugs, use of asthma drugs, history of asthma, bronchitis, reflux or seizures and having at least one sibling were covariates. Cox proportional hazards regression model was used to estimate the raw and adjusted hazard ratios with their respective 95% confidence interval. The average time that individuals remained free of dental caries/restoration was 79.49 months (95%CI: 64.37 to 92.61). Increase in sucrose consumption increased the rate of caries recurrence (HR = 1.16; 95%CI: 1.04 to 1.30). Individuals who had poor oral hygiene had higher rate of new dental caries (HR = 3.88; 95%CI: 1.22 to 12.37) compared to those with good oral hygiene. The presence of mouth breathing decreased the rate of recurrence of the disease when compared to the nasal breathing (HR = 0.32; 95%CI: 0.15 to 0.70). Oral health-related behaviors and nasal respiration influenced the rate of dental caries recurrence in individuals with developmental disabilities.

Toxicity of Carnoy's solution toward human keratinocytes: an in vitro study

Abstract: The present study aimed to characterize the chemical elements and cytotoxicity of Carnoy's solution (CS) by comparing two different trademarked products (one Brazilian [NCS] and another imported [ICS]) using inductively coupled plasma mass spectrometry (ICP-MS) and human keratinocyte (HaCaT) cultures. For performing ICP-MS, the solutions were diluted according to calibration curves, and the chemical elements were analyzed with a spectrometer. HaCaT cells were exposed to CS concentrations ranging from 0.10% to 20% for 3 or 5 min. Cell viability was evaluated immediately (T0), 24 h (T1), and 7 days (T2) after exposure to CS using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) reduction assay. Data were analyzed using a t-test for ICP-MS and analysis of variance followed by Tukey's post-hoc test for MTT assay, both considering statistical significance at p<0.05. ICP-MS results revealed that ICS presented significantly lower concentrations of 12 chemical elements than NCS. The results of MTT assay revealed that at T0, ICS was more cytotoxic than NCS regardless of the time of exposure (p < 0.05). At T1, the only difference between the groups was at a concentration of 0.10% after 5 min of exposure. At T2, at a concentration of 0.5%, ICS resulted in a significant reduction in cell viability compared to NCS (p < 0.05). Thus, the results showed that ICS was more cytotoxic than NCS. Collectively, our findings suggest that the individual compositions of different CS formulations should be investigated.

Carious lesions recurrence in children with developmental disabilities: a longitudinal study

Abstract This study investigated the factors associated with new carious lesions in one-to five-year-old children with developmental disabilities. This was a retrospective cohort that evaluated 64 dental charts of individuals with caries or fillings in their first dental appointment. The dependent variable was the occurrence of a new carious lesion or restoration. Gender, age, mother's education, sugar consumption, oral hygiene, mouth breathing, reports of xerostomia, gingival status, use of psychotropic drugs, use of asthma drugs, history of asthma, bronchitis, reflux or seizures and having at least one sibling were covariates. Cox proportional hazards regression model was used to estimate the raw and adjusted hazard ratios with their respective 95% confidence interval. The average time that individuals remained free of dental caries/restoration was 79.49 months (95%CI: 64.37 to 92.61). Increase in sucrose consumption increased the rate of caries recurrence (HR = 1.16; 95%CI: 1.04 to 1.30). Individuals who had poor oral hygiene had higher rate of new dental caries (HR = 3.88; 95%CI: 1.22 to 12.37) compared to those with good oral hygiene. The presence of mouth breathing decreased the rate of recurrence of the disease when compared to the nasal breathing (HR = 0.32; 95%CI: 0.15 to 0.70). Oral health-related behaviors and nasal respiration influenced the rate of dental caries recurrence in individuals with developmental disabilities.