Publications by authors named "Paulo Nelson-Filho"

177 Publications

Periodontal ligament repair after active splinting of replanted dogs' teeth.

Dent Traumatol 2021 Jul 1. Epub 2021 Jul 1.

Department of Pediatric Dentistry, School of Dentistry of Ribeirão Preto, University of São Paulo, São Paulo, Brazil.

Background/aim: The high rate of root resorption resulting from tooth replantation represents a serious clinical problem. In order to prevent ankylosis and replacement resorption, the contemporary literature highlights the importance of using a flexible stabilization for traumatized teeth. For this purpose, orthodontic devices may be promising for obtaining a better prognosis and periodontal repair. The aim of this study was to evaluate the effect of an active splinting protocol with controlled force in dog's teeth following replantation.

Material And Methods: Sixty premolar roots from three dogs were used. They were submitted to endodontic treatment, hemisected, atraumatically extracted and subsequently replanted. They were divided into four groups: Passive Stabilization (n = 20)-after 20 min in a dry medium; Active Stabilization (n = 20)-after 20 min in a dry medium; Negative control (n = 10)-immediate replantation and passive Stabilization; and Positive control (n = 10)-90 min of extra-alveolar time and passive Stabilization. The samples were collected and submitted to histologic processing. They were then evaluated for the count of inflammatory cells, expression of neurotrophin 4, osteoclasts, apoptotic cells and collagen fibres. The results were submitted to ANOVA or Kruskal-Wallis statistical tests followed by Tukey or Dunn post-tests (α = 5%).

Results: Passive Stabilization with orthodontic brackets without traction used after replantation had the highest number of inflammatory cells (p = .0122), osteoclasts (p = .0013) and percentage of collagen fibres in the periodontal ligament (p < .0001) when compared to Active Stabilization with orthodontic brackets applying amild tensile force. Neurotrophin 4 had no statistically significant difference (p = .05), regardless of the treatment. The apoptotic cells count revealed statistical differences (p < .0001) between Active Stabilization (189.70 ± 47.99) and Positive Control (198.90 ± 88.92) when compared to Passive Stabilization (21.19 ± 32.94).

Conclusion: The active splinting protocol using orthodontic appliances generating a light and controlled force favoured periodontal ligament repair of replanted teeth.
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http://dx.doi.org/10.1111/edt.12698DOI Listing
July 2021

An Epigallocatechin-3-gallate Formulation Developed for Endodontic Use: A Physicochemical and Biological Evaluation.

J Endod 2021 Jun 23. Epub 2021 Jun 23.

Department of Pediatric Dentistry, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.

Introduction: Although epigallocatechin-3-gallate (EGCG) from green tea has been successfully used in the prevention and treatment of several infectious and immunoinflammatory diseases because of its proven anti-inflammatory, antioxidant, antimicrobial, and antiresorptive role, its use as an intracanal dressing has not been proposed. The aim of this study was to develop a formulation based on EGCG for endodontic use by assessing its physicochemical and biological properties.

Methods: Initially, physicochemical characterization of EGCG was performed by ultraviolet-visible and fluorescence spectroscopy to evaluate if the properties were maintained in acidic pH and time (1-6, 24, and 27 hours). After that, biological studies evaluated the developed formulation of EGCG at different concentrations (1.25, 5, 10, and 20 mg/mL). The tissue compatibility with subcutaneous tissue of mice was evaluated by plasma leakage after 24 hours and the examination of macroscopic and microscopic features at 7, 21, and 63 days after the insertion of polyethylene tubes containing the formulations. The repair of experimentally induced periapical lesions in dog's teeth by radiographic and histopathologic analysis was also evaluated. The scores were statistically analyzed by the chi-square and Fisher exact test. Analysis of variance followed by the Tukey posttest were used for the quantitative analysis. The significance level was 5%.

Results: The physicochemical characterization performed under ultraviolet-visible spectrophotometry showed that the EGCG properties remained unaltered in acid pH and function of time, keeping its wavelength to 274 nm. Macroscopic parameters evaluated at 7, 21, and 63 days showed that all concentrations presented no epithelial ulceration or presence of mild superficial tissue necrosis, edema, or vascularization with no significant difference in the control group. During all periods of microscopic examination, all groups presented the absence of abscess foci and edema and the presence of fibrous capsule and neovascularization. The presence of reparative tissue with a gentle presence of neutrophilic inflammatory cells was also observed for all groups, except for the calcium hydroxide paste group, which presented a more pronounced inflammation and tissue necrosis at days 7 and 21 (P < .001). At day 63, all groups presented an absence of inflammatory infiltrate and necrosis. The evaluation of dog teeth showed that treatment with the EGCG formulation provided a reduction of the periapical radiolucent area and allowed the repair of apical and periapical tissues (P > .05).

Conclusions: The developed formulation based on EGCG from green tea presented physicochemical stability and tissue compatibility and provided the repair of periapical lesions when used as an intracanal dressing.
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http://dx.doi.org/10.1016/j.joen.2021.06.010DOI Listing
June 2021

Gene and Genes Involved in the Maintenance of Vitamin D Levels Association with Mandibular Retrognathism.

J Pers Med 2021 May 2;11(5). Epub 2021 May 2.

Department of Orthodontics, University of Regensburg, Franz-Josef-Strauss-Allee 11, 93053 Regensburg, Germany.

In this study we evaluated whether single nucleotide polymorphisms (SNPs) in the genes encoding PTH, VDR, CYP24A1, and CYP27B1 were associated with mandibular retrognathism (MR). Samples from biologically-unrelated Brazilian patients receiving orthodontic treatment were included in this study. Pre-orthodontic lateral cephalograms were used to determine the phenotype. Patients with a retrognathic mandible were selected as cases and those with an orthognathic mandible were selected as controls. Genomic DNA was used for genotyping analysis of SNPs in (rs694, rs6256, and rs307247), (rs7975232), (rs464653), and (rs927650). Chi-squared or Fisher's tests were used to compare genotype and allele distribution among groups. Haplotype analysis was performed for the SNPs in . The established alpha was < 0.05. Multifactor dimensionality reduction (MDR) was used to identify SNP-SNP interactions. A total of 48 (22 males and 26 females) MR and 43 (17 males and 26 females) controls were included. The linear mandibular and the angular measurements were statistically different between MR and controls ( < 0.05). In the genotype and allele distribution analysis, the SNPs rs694, rs307247, and rs464653 were associated with MR ( < 0.05). MDR analyses predicted the best interaction model for MR was rs694-rs927650, followed by rs307247-rs464653-rs927650. Some haplotypes in the gene presented statistical significance. Our results suggest that SNPs in , , , and genes are associated with the presence of mandibular retrognathism.
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http://dx.doi.org/10.3390/jpm11050369DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8147469PMC
May 2021

Effects of estrogen deficiency during puberty on maxillary and mandibular growth and associated gene expression - an μCT study on rats.

Head Face Med 2021 Apr 22;17(1):14. Epub 2021 Apr 22.

Department of Orthodontics, University of Regensburg, Franz-Josef-Strauss-Allee 11, 93053, Regensburg, Germany.

Background: Estrogen is a well-known and important hormone involved in skeletal homeostasis, which regulates genes involved in bone biology. Some studies support that estrogen is important for craniofacial growth and development. Therefore this in vivo animal study aimed to investigate, whether and in which way low estrogen levels in the prepubertal period affect craniofacial development in the postpubertal stage and to quantify the gene expression of RANK, RANKL and OPG in cranial growth sites in ovariectomized estrogen-deficient rats during puberty.

Methods: Control (sham-operated, n = 18) and ovariectomy (OVX, n = 18) surgeries were performed on 21-days-old female Wistar rats. Animals euthanized at an age of 45 days (pubertal stage) were used for gene expression analyses (n = 6 per group) and immunohistochemistry of RANK, RANKL and OPG. Animals euthanized at 63 days of age (post-pubertal stage) were used for craniofacial two-dimensional and three-dimensional craniofacial measurements using μCT imaging (n = 12 per group).

Results: In the μCT analysis of the mandible and maxilla many statistically significant differences between sham-operated and OVX groups were observed, such as increased maxillary and mandibular bone length in OVX animals (p < 0.05). Condylar volume was also significantly different between groups (p < 0.05). The sham-operated group showed a higher level of RANK expression in the midpalatal suture (p = 0.036) and the RANKL:OPG ratio levels were higher in the OVX group (p = 0.015).

Conclusions: Our results suggest that estrogen deficiency during the prepubertal period is associated with alterations in the maxillary and mandibular bone length and condylar growth.
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http://dx.doi.org/10.1186/s13005-021-00265-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8061017PMC
April 2021

Effect of non-steroidal anti-inflammatory drugs on pulpal and periapical inflammation induced by lipopolysaccharide.

Clin Oral Investig 2021 Apr 1. Epub 2021 Apr 1.

School of Dentistry of Ribeirão Preto, University of São Paulo, Avenida do Café, s/n. CEP 14040-904, Ribeirão Preto, São Paulo, Brazil.

Objectives: The objective of this study was to evaluate the effect of non-steroidal anti-inflammatory drugs (NSAIDs) in controlling pulpal and periapical inflammation in vivo as a potential coadjutant systemic therapy for pulpitis.

Materials And Methods: A suspension containing E. coli lipopolysaccharide (LPS; 1.0 μg/μL) was inoculated into the pulp chamber of the first molars of C57BL/6 mice (n = 72), and the animals were treated daily with indomethacin or celecoxib throughout the experimental periods. After 7, 14, 21, and 28 days, the tissues were removed for histopathological, histoenzymology, histometric, and immunohistochemical evaluation.

Results: Inoculation of LPS into the pulp chamber induced the synthesis of the enzyme cyclooxygenase-2 (COX-2) in dental pulp and periapical region. Indomethacin and celecoxib treatment changed the profile of inflammatory cells recruited to dental pulp and to the periapex, which was characterized by a higher mononuclear cell infiltrate, compared to LPS inoculation alone which recruited a higher amount of polymorphonuclear neutrophils. Administration of indomethacin for 28 days resulted in the development of apical periodontitis and increased osteoclast recruitment, unlike celecoxib.

Conclusions: NSAIDs indomethacin and celecoxib changed the recruitment of inflammatory cells to a mononuclear profile upon inoculation of LPS into the pup chamber, but indomethacin enhanced periapical bone loss whereas celecoxib did not.

Clinical Relevance: Celecoxib, a selective COX-2 inhibitor, can change the profile of inflammatory cells recruited to the dental pulp challenged with LPS and might a be potential systemic coadjutant for treatment of pulpitis.
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http://dx.doi.org/10.1007/s00784-021-03919-3DOI Listing
April 2021

Multifaceted effect of caffeic acid against Streptococcus mutans infection: microbicidal and immunomodulatory agent in macrophages.

Arch Microbiol 2021 Aug 26;203(6):2979-2987. Epub 2021 Mar 26.

Department of Pediatric Clinics, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.

The aim of the present study was to investigate the effects of caffeic acid in the interface between the antimicrobial and anti-inflammatory function in macrophage response against S. mutans. S. mutans (10 cfu/mL) were incubated with caffeic acid to determinate the half-maximal inhibitory concentration (IC) and macrophage cells were incubated with caffeic acid to determinate cell viability and toxicity. Anti-inflammatory effects were measured by nitrite accumulation, TNF-α and PGE production, and NF-kB phosphorylation, and S. mutans survival following internalization by macrophages was investigated. We found that caffeic acid presented antimicrobial activity against S. mutans (IC = 2.938 ± 0.1225 mM) without exerting cytotoxicity. Caffeic acid inhibited nitrite, TNF-α and PGE production by the NF-kB dependent pathway, indicating an immunomodulatory property. Caffeic acid also contributed to macrophage bacteria clearance activity. In summary, caffeic acid presented antimicrobial activity against S. mutans and anti-inflammatory effects in macrophages.
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http://dx.doi.org/10.1007/s00203-021-02290-xDOI Listing
August 2021

Esthetic elastomeric ligatures: Quantification of bacterial endotoxin in vitro and in vivo.

Am J Orthod Dentofacial Orthop 2021 May 10;159(5):660-665. Epub 2021 Mar 10.

Department of Pediatric Clinic, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.

Introduction: The objective of this study was to evaluate in vitro and in vivo bacterial endotoxin (LPS) adhesion in polyurethane and silicone esthetic elastomeric orthodontic ligatures. The null hypotheses tested were: (1) there is no LPS adhesion in esthetic elastomeric orthodontic ligatures; and (2) there is no difference in the LPS adhesion between different brands of these ligatures.

Methods: For the in vitro study, 4 types of esthetic elastomeric ligatures were used (Sani-Ties and Sili-Ties [Dentsply GAC, Islandia, NY;] and Mini Single Case Ligature Stick and Synergy low-friction ligatures [Rocky Mountain Orthodontics, Denver, Colo]), contaminated or not with endotoxin solution. Replicas of twisted wire and cast stainless steel ligatures were used as control. For the in vivo study, 10 male and 10 female patients, aged 15-30 years, received the same 4 types of ligatures, 1 of each inserted in the maxillary and mandibular canines, randomly. Twenty-one days later, the ligatures were removed, and endotoxin quantification was performed using the Limulus amebocyte lysate test. Data were analyzed (α = 0.05) using the Kruskal-Wallis test and Dunn's posttest or analysis of variance and Tukey's posttest.

Results: GAC silicone group had the lowest median contamination (1.15 endotoxin units/mL; P <0.0001) in vitro. In the in vivo study, the GAC silicone group had the lowest mean contamination (0.577 endotoxin units/mL; P <0.001). In both studies, the other groups did not present a significant difference when compared with each other (P >0.05).

Conclusions: LPS exhibited an affinity for all the tested polyurethane and silicone elastomeric ligatures. GAC silicone ligatures presented with lower amounts of LPS attached to their surfaces. Thus, both null hypotheses were rejected.
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http://dx.doi.org/10.1016/j.ajodo.2020.02.015DOI Listing
May 2021

Osteoclast formation, inflammation, and matrix metalloproteinase-9 are downregulated in bone repair following root canal treatment in dogs teeth.

Clin Oral Investig 2021 Jul 15;25(7):4699-4707. Epub 2021 Jan 15.

Department of Pediatric Clinics, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.

Objectives: The aim of this study was to investigate the inflammatory infiltrate, osteoclast formation, and expression of MMP-9 during the healing phase following root canal treatment in teeth with apical periodontitis.

Materials And Methods: Apical periodontitis was induced in dogs teeth, and root canal treatment was performed in a single visit or using calcium hydroxide as intracanal medication. One hundred and eighty days following treatment the presence of inflammation was examined, and the tissues were stained to detect osteoclasts by means of a tartrate resistant alkaline phosphatase (TRAP) assay. Synthesis of MMP-9 was detected using Western blotting and immunohistochemistry.

Results: Teeth with apical periodontitis that had root canal therapy performed in a single visit presented a higher synthesis of MMP-9 compared with root canal treatment using calcium hydroxide. Treatment with calcium hydroxide resulted in a reduced amount of inflammatory cells and MMP-9 positive cells. Osteoclast formation, the number of MMP-9 positive osteoclasts and cementocytes, was reduced following root canal treatment, regardless of the root canal treatment protocol used.

Conclusion: Root canal treatment reduced the amount of inflammatory cells and osteoclasts in periapical area. The use of calcium hydroxide as intracanal medication resulted in a lower synthesis of MMP-9, though the number of osteoclasts and MMP-9 positive osteoclasts were similar between the groups.

Clinical Relevance: Periapical bone repair following root canal treatment is impacted by therapy performed either in single visit or using calcium hydroxide dressing measured by inflammatory cell recruitment, osteoclast formation, and MMP-9 synthesis.
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http://dx.doi.org/10.1007/s00784-021-03784-0DOI Listing
July 2021

Alteration of the oral microbiota may be a responsible factor, along with estrogen deficiency, by the development of larger periapical lesions.

Clin Oral Investig 2021 Jun 14;25(6):3651-3662. Epub 2020 Nov 14.

Department of Pediatric Dentistry, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brasil.

Objectives: To answer the questions: (1) Does reducing estrogen levels influence the microbial composition of the oral cavity? (2) Does the presence of periapical lesion (PL) cause changes in the oral microbiota? (3) Since estrogen deficiency alters the oral microbiota, can this be one of the factors that contribute to the increase of the PL?

Materials And Methods: Thirty-six rats were divided into four groups: sham (control), ovariectomy (OVX), control with PL (Sham + PL), and OVX + PL. After 9 weeks of OVX, the lower first molars were submitted to PL induction. After 21 days, the microbiological collection of the oral cavity was performed, and the animals were euthanized. The contents were evaluated by the checkerboard DNA-DNA hybridization method, to verify the prevalence of 40 bacterial species (divided into 7 microbial complexes). The blocks containing the lower first molars were submitted to histotechnical processing and staining with hematoxylin and eosin (HE), for the measurement of the periapical lesion area. The results were submitted to ANOVA and Kruskal-Wallis tests and Tukey and Dunn post-tests, with a significance level of 5%.

Results: In conditions of estrogen deficiency, there was alteration of the oral microbiota. The OVX groups had a higher amount of bacteria compared to the SHAM group in most of the microbial complexes (p < 0.001). The animals in the control group (with or without lesion) did not present a statistically significant difference (p > 0.001) in any of the microbial complexes. The PLs in OVX animals were significantly higher compared to SHAM animals (p < 0.001).

Conclusions: Hypoestrogenicity conditions interfere in the oral microbiota by increasing the amount of bacteria in the saliva and influencing the progression of periapical lesions.

Clinical Relevance: This inedited study shows that deficiency of estrogen leads to alteration of the oral microbiota.
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http://dx.doi.org/10.1007/s00784-020-03688-5DOI Listing
June 2021

Photosensitizers attenuate LPS-induced inflammation: implications in dentistry and general health.

Lasers Med Sci 2021 Jun 4;36(4):913-926. Epub 2020 Nov 4.

Departamento de Cirurgia e Traumatologia Buco-Maxilo-Facial e Periodontia, Faculdade de Odontologia de Ribeirão Preto, Universidade de São Paulo, Av. do Café, s/n, Monte Alegre, 14040-904, Ribeirão Preto, São Paulo, Brazil.

Antimicrobial photodynamic therapy (aPDT) is a complementary therapeutic modality for periodontal and endodontic diseases, in which Gram-negative bacteria are directly involved. Currently, there are few evidences regarding the effects of aPDT on bacterial components such as lipopolysaccharide (LPS) and it would represent a major step forward in the clinical use of this therapy. In this context, this study aimed to evaluate the efficacy of different photosensitizers (PSs) used in aPDT in LPS inhibition. Four PSs were used in this study: methylene blue (MB), toluidine blue (TBO), new methylene blue (NMB), and curcumin (CUR). Different approaches to evaluate LPS interaction with PSs were used, such as spectrophotometry, Limulus amebocyte lysate (LAL) test, functional assays using mouse macrophages, and an in vivo model of LPS injection. Spectrophotometry showed that LPS decreased the absorbance of all PSs used, indicating interactions between the two species. LAL assay revealed significant differences in LPS concentrations upon pre-incubation with the different PSs. Interestingly, the inflammatory potential of LPS decreased after previous treatment with the four PSs, resulting in decreased secretion of inflammatory cytokines by macrophages. In vivo, pre-incubating curcumin with LPS prevented animals from undergoing septic shock within the established time. Using relevant models to study the inflammatory activity of LPS, we found that all PSs used in this work decreased LPS-induced inflammation, with a more striking effect observed for NMB and curcumin. These data advance the understanding of the mechanisms of LPS inhibition by PSs.
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http://dx.doi.org/10.1007/s10103-020-03180-2DOI Listing
June 2021

Calcium and Phosphorus Levels in Saliva are Influenced by Genetic Polymorphisms in Estrogen Receptor Alpha and Microrna17.

Braz Dent J 2020 Sep-Oct;31(5):466-470

School of Health Sciences, UP - Universidade Positivo, Curitiba, Brazil.

Homeostasis between salivary calcium and phosphorus is important for maintaining oral health. The aim of this study was to evaluate if polymorphisms in ESR1 (Estrogen Receptor Alpha), ESR2 (Estrogen Receptor Beta) and miRNA17 (microRNA17) are associated with calcium and phosphorus levels in saliva. Saliva from 276 12-year-old children were collected by masticatory stimulation and calcium and phosphorus levels were determined by Mass Spectrometry. Genomic DNA was extracted from remaining saliva and genetic polymorphisms in ESR1 (rs12154178, rs1884051, rs9340799 and rs2234693), in ESR2 (rs4986938 and rs1256049) and in miRNA17 (rs4284505) were genotyped using TaqMan chemistry and a real-time PCR equipment. Statistical differences in genotype and allele distributions between 'low' and 'high' calcium and phosphorus levels were determined using chi-square or Fisher´s exact tests. The analysis was also adjusted by sex (alpha of 5%). ESR1 rs9340799 had the less common genotype associated with higher calcium levels (p=0.03). The less common allele of ESR1 rs1884051 was associated with lower phosphorus levels (p=0.005) and there was an excess of heterozygotes for miRNA17 rs4284505 among individuals with lower calcium levels (p=0.002), both adjusted by sex. This study provides evidence that genetic polymorphisms in ESR1 and miRNA17 are involved in determining salivary calcium and phosphorus levels.
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http://dx.doi.org/10.1590/0103-6440202002934DOI Listing
November 2020

The role of postnatal estrogen deficiency on cranium dimensions.

Clin Oral Investig 2021 May 25;25(5):3249-3255. Epub 2020 Oct 25.

Department of Orthodontics, University of Regensburg, Franz-Josef-Strauss-Allee 11, 93053, Regensburg, Germany.

Objectives: The aim of this study was investigate the cranium dimensions of adult female rats, who suffered estrogen deficiency during the prepubertal stage, to assess the impact of estrogen deficiency on craniofacial morphology.

Material And Methods: Twenty-two female Wistar rats were divided into ovariectomy (OVX) (n = 11) and sham-operated control (n = 11) groups. Bilateral ovariectomy were performed in both groups at 21 days old (prepubertal stage), and rats were euthanized at an age of 63 days (post-pubertal stage). Micro-CT scans were performed with rat skulls, and the cranium morphometric landmark measurements were taken in the dorsal, lateral, and ventral view positions. Differences in measurements between the OVX and sham control groups were assessed using t test with an established alpha error of 5%.

Results: The measures of the rats' skull showed that the inter-zygomatic arch width and anterior cranial base length were significantly larger in OVX group (p = 0.020 and p = 0.050, respectively), whereas the length of parietal bone was significantly higher in the sham group (p = 0.026). For the remaining measurements no significant differences between groups were detected (p > 0.05).

Conclusion: This study provides evidence that ovariectomized rats had alterations in cranial bone dimensions, demonstrating that estrogens during puberty are important for skull morphology.

Clinical Relevance: To understand the role of estrogen on the postnatal cranium development will impact the clinical diagnose and therapy during childhood and adolescence.
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http://dx.doi.org/10.1007/s00784-020-03655-0DOI Listing
May 2021

Potential interactions among single nucleotide polymorphisms in bone- and cartilage-related genes in skeletal malocclusions.

Orthod Craniofac Res 2021 May 2;24(2):277-287. Epub 2020 Nov 2.

Department of Orthodontics, University Medical Centre of Regensburg, Regensburg, Germany.

Objective: To investigate SNPs in bone- and cartilage-related genes and their interaction in the aetiology of sagittal and vertical skeletal malocclusions.

Settings And Sample Population: This study included 143 patients and classified as follows: skeletal class I (n = 77), class II (n = 47) and class III (n = 19); maxillary retrusion (n = 39), protrusion (n = 52) and well-positioned maxilla (n = 52); mandibular retrognathism (n = 50), prognathism (n = 50) and well-positioned mandible (n = 43); normofacial (n = 72), dolichofacial (n = 55) and brachyfacial (n = 16).

Materials And Methods: Steiner's ANB, SNA, SNB angles and Ricketts' NBa-PtGn angle were measured to determine the skeletal malocclusion and the vertical pattern. Nine SNPs in BMP2, BMP4, SMAD6, RUNX2, WNT3A and WNT11 were genotyped. Chi-squared test was used to compare genotypes among the groups. Multifactor dimensionality reduction (MDR) and binary logistic regression analysis, both using gender and age as co-variables, were also used. We performed Bonferroni correction for multiple testing.

Results: Significant associations at P < .05 were observed for SNPs rs1005464 (P = .042) and rs235768 (P = .021) in BMP2 with mandibular retrognathism and for rs59983488 (RUNX2) with maxillary protrusion (P = .04) as well as for rs708111 (WNT3A) with skeletal class III (P = .02; dominant model), rs1533767 (WNT11) with a brachyfacial skeletal pattern (P = .01, OR = 0.10; dominant model) and for rs3934908 (SMAD6) with prognathism (P = .02; recessive model). After the Bonferroni correction, none of the SNPs remained associated. The MDR predicted some interaction for skeletal class II, dolichofacial and brachyfacial phenotypes.

Conclusion: Our results suggest that SNPs in BMP2, BMP4, SMAD6, RUNX2, WNT3A and WNT11 could be involved in the aetiology of sagittal and vertical malocclusions.
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http://dx.doi.org/10.1111/ocr.12433DOI Listing
May 2021

GHR and IGF2R genes may contribute to normal variations in craniofacial dimensions: Insights from an admixed population.

Am J Orthod Dentofacial Orthop 2020 Nov 29;158(5):722-730.e16. Epub 2020 Sep 29.

Department of Pediatric Dentistry, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil, and Department of Dentistry, Positivo University, Curitiba, Paraná, Brazil. Electronic address:

Introduction: This study aimed to determine whether single nucleotide polymorphisms in the growth hormone receptor (GHR) and insulin-like growth factor 2 receptor (IGF2R) genes are associated with different craniofacial phenotypes.

Methods: A total of 596 orthodontic and 98 orthognathic patients from 4 cities in Brazil were included for analyses. Angular and linear cephalometric measurements were obtained, and phenotype characterizations were performed. Genomic DNA was collected from buccal cells and single nucleotide polymorphisms in GHR (rs2910875, rs2973015, rs1509460) and IGF2R (rs2277071, rs6909681, rs6920141) were genotyped by polymerase chain reactions using TaqMan assay. Genotype-phenotype associations were assessed in the total sample (statistical significance was set at P <8.333 × 10) and by a meta-analytic approach implemented to calculate the single effect size measurement for the different cohorts.

Results: Rare homozygotes for the GHR rs2973015 showed increased measurements for the lower anterior facial height (ANS-Me) and mandibular sagittal lengths (Co-Gn and Go-Pg). In contrast, common homozygotes for the IGF2R rs6920141 presented reduced measurements for these dimensions (ANS-Me and Go-Pg). Furthermore, the less common homozygotes for IGF2R rs2277071 had reduced maxillary sagittal length (Ptm'-A'). The meta-analytical approach replicated the associations of rs2973015 with ANS-Me, rs2277071 with Ptm'-A', and rs6920141 with Go-Pg.

Conclusions: Our results provide further evidence that GHR contributes to the determination of mandibular morphology. In addition, we report that IGF2R is a possible gene associated with variations in craniofacial dimensions. Applying meta-analytical approaches to genetic variation data originating from likely underpowered samples may provide additional insight regarding genotype and/or phenotype associations.
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http://dx.doi.org/10.1016/j.ajodo.2019.10.020DOI Listing
November 2020

Radiotherapy Activates Matrix Metalloproteinases in the Dentinoenamel Junction of Primary Teeth.

J Dent Child (Chic) 2020 May;87(2):83-89

Dr. Paula-Silva is an associate professor, Department of Pediatric Clinics, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.

Radiation-related caries is characterized by enamel delamination near the dentinoenamel junction (DEJ). We investigated the activity and expression of the matrix metalloproteinases (MMPs) -2 and -9 in order to understand disease pathogenesis in teeth submitted or not to radiotherapy (RT).
In situ zymography and immunofluorescence assays were performed to evaluate the activity and expression of MMPs -2 and -9, respectively. Twelve primary second molars were randomly assigned into two experimental subgroups: irradiated and nonirradiated. Dental fragments were exposed to radiation at a dose fraction of two Gy for five consecutive days until reaching the total dose of 60 Gy. The percentage of fluorescence in the DEJ was evaluated in three distinct regions of the tooth (cervical, cusp, and pit). The regions were photographed under fluorescence microscopy at 1.25× and 5× magnification.
The intensity of fluorescence per mm 2 in the DEJ was higher in the cervical region of irradiated primary teeth ( <0.05) versus nonirradiated ones. In these areas, immunofluorescence revealed expression of MMPs -2 and -9.
Radiotherapy can increase the activity of MMPs -2 and -9 in the cervical region of the DEJ of primary teeth.
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May 2020

Radiotherapy Impairs Adhesive Bonding in Primary Teeth: An In Vitro Study.

J Dent Child (Chic) 2020 May;87(2):69-76

Dr. Queiroz, Professor; Department of Pediatric Dentistry, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil;, Email:

To evaluate the morphological alterations in enamel and dentin of primary teeth following radiotherapy (RT) and to determine the best adhesive technique and time to carry out restorative procedures.
Enamel and dentin fragments of primary teeth were randomly assigned into four groups (n=30): G1 (control)-non-irradiated, only restorative procedure; G2- restorative procedure immediately before RT; G3-restorative procedure 24 hours after RT; and G4-restorative procedure six months after RT. Each group was divided into one of two subgroups according to the adhesive system used for restoration: (1) AdperSingle Bond 2 (SB); and (2) ClearfillSE Bond (CL). The specimens were submitted to fractionated RT until they reached the final dose of 60 Gy. They were then subjected to confocal microscopy and the shear bond strength test. Data were analyzed using two-way analysis of variance followed by Tukey's tests ( α = five percent).
Morphological changes were first observed in enamel and dentin after 40 Gy of irradiation. G4 bond strength values were similar to G1 in the CL and SB groups for enamel and in the CL group for dentin ( >0.05). G2 showed the lowest values for enamel and dentin ( <0.05). In G3, CL presented the highest strength values in enamel; for G4, the highest values were found in dentin ( <0.05).
Radiotherapy affected the morphological surface of enamel and dentin. The restorations placed immediately after RT had the weakest shear bond strength, and the restorations placed six months after RT had similar means of bond strength compared to the nonirradiated teeth in enamel, regardless of the adhesive system used. In dentin, CL showed better performance than SB.
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May 2020

Effect of root surface treatment with denusomab after delayed tooth replantation.

Clin Oral Investig 2021 Mar 29;25(3):1255-1264. Epub 2020 Jul 29.

Department of Pediatric Dentistry, School of Dentistry of Ribeirão Preto, University of São Paulo, Av. do Café s/n, Ribeirão Preto, SP, 14040-904, Brazil.

Objectives: To evaluate denosumab, a human monoclonal antibody that mimics the effects of osteoprotegerin in bone metabolism, as a topical treatment of root surface to be used prior to delayed tooth replantation.

Materials And Methods: Thirty-six rats' right incisors were used. Teeth were extracted and divided into: delayed replantation without root surface treatment (control); delayed replantation with root surface treatment with denosumab 60 mg/mL and 30 mg/mL, respectively, for 10 min both experimentals groups. After that, the root canals were filled with calcium hydroxide and replanted. After 15 and 60 days, the animals were euthanized, and the samples were collected and processed for microscopic analysis. Histological sections were performed, and stained with HE to describe the dental characteristics, measure ankylosis, replacement resorption, and dental resorption by conventional microscopy. Also, was performed Brown & Brenn staining and immunohistochemistry for RANKL, OPG, and periostin.

Results: Denosumab 60 mg/mL reducted ankylosis (p < 0.0001), replacement resorption (p < 0.0001), and tooth resorption, 60 days after replantation, compared to untreated replanted teeth (p < 0.005). Lower bacterial contamination in root surface in the denosumab treatment groups was found, regardless of the concentration used (p < 0.001). Also, denosumab treatment inhibited the expression of RANKL without modulating OPG. Periostin was observed in periodontal ligament of replanted tooth, although this labelling was absent in the ankylosis areas, in both experimental periods.

Conclusion: Treatment of the root surface with denosumab at 60 mg/mL of rat teeth before delayed replantation reduced dental root resorption compared with the untreated teeth after 60 days.

Clinical Relevance: Survival of a replanted tooth has been a challenge in clinical practice. The use of a medication, such as denosumab, to limit dental root resorption represents an important therapeutical approach.
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http://dx.doi.org/10.1007/s00784-020-03430-1DOI Listing
March 2021

Bruxism Throughout the Lifespan and Variants in , and .

J Pers Med 2020 May 27;10(2). Epub 2020 May 27.

Department of Oral Biology, University of Pittsburgh School of Dental Medicine, 412 Salk Pavilion, 335 Sutherland Drive, Pittsburgh, PA 15261, USA.

Bruxism is a masticatory muscle activity characterized by grinding of the teeth and clenching of the jaw that causes tooth wear and breakage, temporomandibular joint disorders, muscle pain, and headache. Bruxism occurs in both adults and children. Clinical characteristics and habits were evaluated in an adult sample. Moreover, we used DNA samples from 349 adults and 151 children to determine the presence of association with specific genes. Genomic DNA was obtained from saliva. The markers and (metalloproteinase 2 ()), and (metalloproteinase 9 ()), and (cathecol-o-methyltransferase ()) were genotyped. Data were submitted to statistical analysis with a significance level of 0.05. In adults, in univariate logistic regression, presence of caries, attrition, and use of alcohol were increased in bruxism individuals ( < 0.05). In addition, in adults, there was an association between bruxism and (, = 0.0001) and bruxism and (, = 0.003). In children, a borderline association was observed for (, = 0.08). When we performed multivariate logistic regression analyses in adults, the same clinical characteristics remained associated with bruxism, and orthodontic treatment was also associated, besides , in the AG genotype ( = 0.015, OR: 3.40 (1.27-9.07)). For the first time, we provide statistical evidence that these genes are associate with bruxism.
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http://dx.doi.org/10.3390/jpm10020044DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7354525PMC
May 2020

Association between craniofacial morphological patterns and tooth agenesis-related genes.

Prog Orthod 2020 Apr 6;21(1). Epub 2020 Apr 6.

School of Dentistry, Fluminense Federal University, Rua São Paulo 28, Campus do Valonguinho, Niterói, RJ, 24020-150, Brazil.

Background: The aim of the present study was to assess if genetic polymorphisms in tooth agenesis (TA)-related genes are associated with craniofacial morphological patterns.

Methods: This cross-sectional, multi-center, genetic study evaluated 594 orthodontic Brazilians patients. The presence or absence of TA was determined by analysis of panoramic radiography. The patients were classified according to their skeletal malocclusion and facial growth pattern by means of digital cephalometric analysis. Genomic DNA was extracted from squamous epithelial cells of buccal mucosa and genetic polymorphisms in MSX1 (rs1042484), PAX9 (rs8004560), TGF-α (rs2902345), FGF3 (rs1893047), FGF10 (rs900379), and FGF13 (rs12838463, rs5931572, and rs5974804) were genotyped by polymerase chain reaction using TaqMan chemistry and end-point analysis.

Results: Genotypes (p = 0.038) and allele (p = 0.037) distributions for the FGF3 rs1893047 were significantly different according to the skeletal malocclusion. Carrying at least one G allele increased in more than two times the chance of presenting skeletal class III malocclusion (OR = 2.21, CI 95% = 1.14-4.32; p = 0.017). There was no association between another skeletal craniofacial pattern and some polymorphism assessed in the present study.

Conclusion: Our results suggest that the genetic polymorphism rs1893047 in FGF3 might contribute to variations in the craniofacial sagittal pattern.
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http://dx.doi.org/10.1186/s40510-020-00309-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7131971PMC
April 2020

Effect of ovariectomy on maxilla and mandible dimensions of female rats.

Orthod Craniofac Res 2020 Aug 12;23(3):342-350. Epub 2020 Apr 12.

Department of Pediatric Dentistry, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, Brazil.

Objective: The role of oestrogen in craniofacial growth still remains unclear. Therefore, the present study aimed to assess the effect of oestrogen deficiency on maxilla and mandible dimensions.

Setting And Sample Population: The study was conducted at the Department of Pediatric Dentistry at the School of Dentistry of Ribeirão Preto, University of São Paulo, and used forty female Wistar rats.

Material And Methods: Ovariectomy (OVX) and placebo surgery (Sham) were performed when animals were twenty-one days old (prepubertal stage). Dimensions of the maxilla and mandible were assessed by craniometric analysis using radiographs, during and after puberty of the animals (45 and 63 days old, respectively). Quantitative real-time PCR and immunohistochemical analyses were performed to determine the expression and localization, respectively, of oestrogen receptor alpha (ERα) and oestrogen receptor beta (ERβ) in different growth sites of the evaluated structures at puberty. The differences between the groups for each outcome were evaluated using the t test with an established alpha error of 5%.

Results: There were significant differences between the OVX and Sham groups for horizontal and vertical linear measurements in the maxilla and the mandible at both pubertal and post-pubertal stages (P < .05). The ovariectomized rats showed significantly greater measures for all dimensions assessed. No differences in gene expression of ERα and ERβ were identified at the different growth sites between the OVX and Sham groups (P > .05). Immunohistochemical analyses revealed the presence of both oestrogen receptors in osteoblasts and chondrocytes in the midpalatal suture and mandibular condyle, respectively, in the OVX and Sham groups.

Conclusion: Our results suggest that oestrogen deficiency from the prepubertal stage might increase the growth of the maxilla and mandible in female rats.
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http://dx.doi.org/10.1111/ocr.12376DOI Listing
August 2020

Factors involved in the treatment sought immediately after traumatic dental injuries in Brazilian children.

Indian J Dent Res 2020 Jan-Feb;31(1):109-112

Department of Pediatric Dentistry, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brasil.

Aim: The aim of this study was to evaluate demographic and clinical factors involved in the immediate seeking of care after traumatic dental injury (TDI) in Brazilian children.

Materials And Methods: Records from 74 patients, age ranged 1-11 years, who sought treatment at the School of Dentistry of Ribeirão Preto at University of São Paulo, Brazil, were collected. Data was analyzed using the Epi Info 7.0 software by t-test, odds ratio calculation, Chi-square, or Fisher's exact tests.

Results: Twenty-three (31.1%) sought dental treatment immediately and 51 (68.9%) did not seek dental treatment immediately. The most common type of trauma was lateral luxation (44.6%). In primary teeth, 31 cases (60.78%) involved the soft tissue and 16 (39.2%) involved hard tissue injuries. While in permanent teeth, 20 cases (40%) involved soft tissue and 24 (60%) involved hard tissue injuries had more traumas in the hard tissue (P = 0.04). The type of injury and dentition was not associated with the time that the guardians sought dental treatment (P > 0.05). None of the factors were involved in immediately seeking care after TDI.

Conclusion: Moreover, the majority of parents/caregivers did not immediately seek dental treatment after TDI, regardless of the type of injury.
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http://dx.doi.org/10.4103/ijdr.IJDR_19_17DOI Listing
April 2020

Root canal conicity estimation of primary maxillary central and lateral incisors-A study by Nano-CT.

Int J Paediatr Dent 2020 Nov 20;30(6):764-774. Epub 2020 Apr 20.

Department of Pediatric Dentistry, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, Brazil.

Aim: The objective of the study was to estimate the conicity of the root canals of maxillary central and lateral incisors by computed nanotomography (Nano-CT).

Design: This in vitro study included nine extracted primary maxillary central incisors and 12 maxillary lateral incisors, which were subjected to Nano-CT analysis. The resulting images of each tooth were reconstructed using the OnDemand3D software, and root canal area, volume, and taper analysis were performed using the free FreeCAD 0.18 software for the 3D computer-aided design (CAD) model. Data were statistically analysed using the Stata v14.0 software, adopting a significance level of 5%.

Results: The results presented the mean value of the diameter and area of the root canal of primary central and lateral incisors. In addition, the taper values for both dental groups between points D0-D5, D5-D7, and D7-D10 were determined. Considering the diameters obtained over the entire length of the root, with a length of 12 mm, a conical model was constructed.

Conclusion: Detailed knowledge of root morphology of maxillary central and lateral incisors of primary dentition by means of Nano-CT is important to achieve faster, more accurate, and efficient endodontic treatments.
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http://dx.doi.org/10.1111/ipd.12642DOI Listing
November 2020

Negative Pressure Irrigation Presents Mineralizing Potential in Dogs' Immature Teeth with Periapical Lesion.

Braz Dent J 2020 Jan-Feb;31(1):37-43

Department of Pediatric Dentistry, University of Washington and Private Practice Limited to Endodontics in Everett WA, USA.

The objective of this in vivo study was to assess the effect of the root canal irrigation by negative and positive apical pressure on the expression of molecules that are an indicative of cell differentiation with mineralizing phenotype in teeth of dogs with incomplete rhizogenesis and induced periapical lesion. A total of 30 teeth (60 roots) were distributed into 3 groups (n=20): EndoVac®, Conventional and Control. After 90 days, the routine histotechnical procedures were performed and the sections were submitted to immunohistochemical technique for the staining of osteopontin (OPN), alkaline phosphatase (ALP) and the RUNX2 transcription factor in the apical and periapical regions of the roots. A semi-quantitative analysis of the positive immunostaining was performed and the intensity of the expression was classified in absent (0), mild (1), moderate (2), or intense (3). Scores data were statistically analyzed by the Kruskal-Wallis non-parametric test and Dunn post-test, and the significance level was set at 5%. RUNX2 immunostaining revealed that in the negative pressure group there was a significantly stronger (p<0.05) immunostaining in comparison to the control group. Regarding the OPN expression, it was not possible to detect a statistically significant difference between the groups (p>0.05). After analyzing ALP immunostaining, a statistically significant difference was observed between the groups (p<0.05), and the negative pressure group showed a markedly stronger mark immunostaining than the control group. The results of the present in vivo study allowed concluding that negative apical pressure irrigation presents mineralizing potential in immature teeth with apical periodontitis.
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http://dx.doi.org/10.1590/0103-6440201802764DOI Listing
March 2020

Effect of manual and electrical brushing on the enamel of sound primary teeth and teeth with induced white spot lesions.

Am J Dent 2020 Feb;33(1):25-28

Department of Pediatric Dentistry, School of Dentistry of Ribeirão Preto, University of São Paulo, São Paulo, SP, Brazil.

Purpose: To evaluate the effect of different electrical brushing systems on the surface roughness and wear profile of the enamel of sound primary teeth and teeth with induced white spot lesions.

Methods: 45 specimens were obtained from sound primary incisors, and the buccal surface was divided into four parts: sound enamel; enamel with white spot lesions; sound enamel with brushing; and enamel with white spot lesions and brushing. Specimens were randomly divided into three groups (n =15), according to the different brushing systems: Group 1 - Electric rotating toothbrush (Kid's Power Toothbrush - Oral B); Group 2 - Sonic electric toothbrush (Baby Sonic Toothbrush); and Group 3 - Manual toothbrush (Curaprox infantil) (control). The specimens were analyzed for surface roughness and wear profile. Data were analyzed by appropriate statistical tests, with a significance level of 5%.

Results: Regarding the surface roughness, no significant difference was observed between the groups. However, with respect to the wear profile, Group 1 caused significantly higher wear in the sound tooth enamel and in the presence of white spot lesions, in comparison to the other brushing systems (2 and 3) (P< 0.05), which did not cause wear. Manual and electric brushing (rotational and sonic) did not increase surface roughness in primary tooth enamel. However, the electric rotational brushing caused significant wear of the sound and demineralized enamel surface of primary teeth.

Clinical Significance: None of the toothbrushing systems tested caused significant alterations on sound dental enamel. However, rotational toothbrushing on enamel of primary teeth with white spot lesion increased wear.
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February 2020

Possible association between craniofacial dimensions and genetic markers in and .

J Orthod 2020 03 31;47(1):65-71. Epub 2020 Jan 31.

School of Dentistry of Ribeirão Preto, Department of Pediatric Dentistry, University of São Paulo, Ribeirão Preto, SP, Brazil.

Objective: To investigate the association of genetic markers in and with craniofacial measurements.

Design: Cross-sectional study.

Setting: School of Dentistry of Ribeirão Preto, University of São Paulo.

Participants: A total of 146 biologically unrelated, self-reported Caucasian Brazilians with no syndromic conditions were included.

Methods: Sagittal and vertical measurements (ANB, S-N, Ptm'-A', Co-Gn, Go-Pg, N-Me, ANS-Me, S-Go and Co-Go) from lateral cephalograms were examined for craniofacial evaluation. DNA was extracted from saliva and genetic markers in (rs2234693 and rs9340799) and in (rs1256049 and rs4986938) were analysed by real-time polymerase chain reaction. Hardy-Weinberg equilibrium was evaluated using the Chi-square test within each marker. The associations between craniofacial dimensions and genotypes were analysed by linear regression and adjusted by sex and age. The established alpha was 5%.

Results: Individuals carrying CC in rs2234693 had a decrease of -3.146 mm in ANS-Me ( = 0.044). In addition, rs4986938 in was associated with S-N measurement ( = 0.009/ ß = -3.465). This marker was also associated with Go-Pg measurement, in which the CC genotype had a decrease of -3.925 mm in the length of the mandibular body ( = 0.043).

Conclusion: The present study suggests that in and are markers for variations in the craniofacial dimensions. However, further research should confirm the results.
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http://dx.doi.org/10.1177/1465312520901725DOI Listing
March 2020

Assessing the Association between Dental Caries and Nutritional Status in Children from the Brazilian State of Amazonas.

Int J Clin Pediatr Dent 2019 Jul-Aug;12(4):293-296

Department of Pediatric Dentistry, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.

Aim: To evaluate the association between dental caries and nutritional status in a group of Brazilian schoolchildren, from Manaus. The studied population consisted of 197 students (10-12-year-olds) from public schools at Manaus, Amazonas, Brazil.

Materials And Methods: A clinical examination was carried out and the decay-missing-filled-teeth index for primary and permanent teeth (dmft and DMFT) was used to evaluate dental caries. Body mass index -score was calculated using variables such as individual height, weight, age, and gender. The nutritional status was classified as underweight, eutrophic, overweight, and obese. One-way ANOVA and Tukey's posttests were used for means' comparison between groups. The established alpha was 5%.

Results: Eighty-one (41.1%) children were caries-free. Five (2.5%) children were underweight; 127 (64.5%) were eutrophic; 49 (24.9%) were overweight; and 16 (8.1%) were obese. The mean dmft/DMFT index was 1.67 (2.05). Obese children had more caries experience than eutrophic and overweight children ( < 0.05).

Conclusion: Our study demonstrated that dental caries is associated with obesity in school children from Manaus.

How To Cite This Article: Vasconcelos K, Evangelista S, Assessing the Association between Dental Caries and Nutritional Status in Children from the Brazilian State of Amazonas. Int J Clin Pediatr Dent 2019;12(4):293-296.
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http://dx.doi.org/10.5005/jp-journals-10005-1638DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6898865PMC
December 2019

Tissue Response after Subcutaneous Implantation of Different Glass Ionomer-Based Cements.

Braz Dent J 2019 Nov-Dec;30(6):599-606

Department of Pediatric Dentistry, School of Dentistry of Ribeirão Preto, USP - Universidade de São Paulo, Ribeirão Preto, SP, Brazil.

The aim of this study was to evaluate the subcutaneous connective tissue response of isogenic mice after implantation of different glass ionomer-based cements (EQUIA® Forte Fil, EQUIA® Fil and Ketac™ Universal Aplicap™). Eighty-seven isogenic BALB/c mice were allocated in 12 groups, 9 were considered as experimental groups (Ketac, E. Fil and E. Forte at 7, 21 and 63 days) and 3 controls (empty polyethylene tubes at 7, 21 and 63 days). After the experimental periods, the subcutaneous connective tissue surrounding the implanted material was removed and subjected to histotechnical processing and staining with hematoxylin and eosin. A histopathological description of the tissue reaction surrounding each material and a semi-quantitative analysis of collagen fiber formation and inflammatory infiltrate were performed. Additionally, the thickness of the granulomatous tissue in contact with each material was measured. Data were analyzed statistically (α=0.05) by the Kruskal-Wallis test, followed by Dunn post-test. Initially, the collagen fiber formation was not different among all the tested materials (p>0.05) but was different at 21 days with the control group presenting the most advanced stage of collagen fiber formation. At 63 days, EQUIA® Forte Fil group showed the most advanced stage of collagen fiber formation, compared to EQUIA® Fil group (p<0.05). The inflammatory infiltrate was not different among the tested materials in any experimental period (p>0.05). The thickness of the granulomatous tissue was greater in the E. Forte group, compared to control in all periods. All glass ionomer-based cements showed tissue compatibility, according to the evaluated parameters.
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http://dx.doi.org/10.1590/0103-6440201902619DOI Listing
December 2019

Morphological identification of Streptococcus mutans and Streptococcus sobrinus in SB-20M culture medium has efficiency comparable to proteomic identification by the MALDI-TOF mass spectrometry technique.

Arch Oral Biol 2020 Feb 31;110:104595. Epub 2019 Oct 31.

Department of Pediatric Dentistry, School of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.

Objectives: The objective of this study was to evaluate the efficiency of SB-20 M culture medium to perform differential morphological identification of S. mutans and S. sobrinus compared to biochemical identification and to proteomic identification by the MALDI-TOF mass spectrometry technique.

Material And Methods: Unstimulated saliva samples from 266 dental students were seeded on SB-20 M culture medium by the wooden spatula technique. After incubation, S. mutans and S. sobrinus colonies were identified by stereomicroscopy based on their differential morphological characteristics. Following these procedures, 135 colonies with characteristic morphology of S. mutans (89 colonies) and S. sobrinus (46 colonies) were randomly selected, submitted to biochemical identification (biotyping) and proteomic identification by the MALDI-TOF mass spectrometry technique. The results were compared using the Kappa test, with a 5% significance level.

Results: All (100%) S. mutans colonies were correctly identified after culture in SB-20 M medium compared to biotyping and proteomic identification. For S. sobrinus, morphological identification in SB-20 M medium was correct for 43 colonies (93.5%) compared to biotyping and proteomic identification. However, there was no statistically significant difference when comparing the capacity to identify S. mutans and S. sobrinus of the three techniques (p < 0.001; K = 0.951).

Conclusions: It was concluded that the SB-20 M culture medium for morphological identification of S. mutans and S. sobrinus was highly reliable, being comparable to the MALDI-TOF mass spectrometry technique.

Clinical Relevance: The efficiency evaluation of identification methods of S. mutans and S. sobrinus is clinically relevant in order to determine caries risk and activity of patients.
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http://dx.doi.org/10.1016/j.archoralbio.2019.104595DOI Listing
February 2020

Association between oestrogen receptors and female temporomandibular disorders.

Acta Odontol Scand 2020 Apr 24;78(3):181-188. Epub 2019 Oct 24.

Department of Dentistry, School of Health Sciences, Universidade Positivo, Curitiba, Brazil.

To evaluate if temporomandibular disorders (TMDs) are associated with genetic polymorphisms in and , which are genes encoding oestrogen receptor alpha () and beta (). Also, we included an animal model to check if and are expressed in the temporomandibular joint (TMJ) during adolescence. A total of 139 teenagers and 93 adults were diagnosed according to the (RDC/TMDs). The DNA was collected and the markers and were genotyped. Additionally, immunohistochemistry was performed in TMJ tissues from female rats during puberty. All data were submitted to statistical analysis with confidence interval of 95%. Teenagers presented more disc displacement and arthralgia than adults ( < .05). The genetic polymorphism in was associated with disc displacement ( = .040; OR = 10.50/95%CI 1.17-98.74) and arthralgia ( = .036; OR = 7.20/95%CI 1.10-46.88) in adults. The and are expressed in rat TMJ tissues. We provide evidence that is associated with TMD and could be a genetic marker for this condition in adult women. Furthermore, oestrogens receptors are presented in TMJ of adolescent female rats.
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http://dx.doi.org/10.1080/00016357.2019.1675904DOI Listing
April 2020

Quantification of pro-inflammatory cytokines and osteoclastogenesis markers in successful and failed orthodontic mini-implants.

J Appl Oral Sci 2019 7;27:e20180476. Epub 2019 Oct 7.

Universidade de São Paulo, Faculdade de Odontologia de Ribeirão Preto, Departamento de Clínica Infantil, Ribeirão Preto, São Paulo, Brasil.

Objectives: Miniscrew has been frequently used, considering that anchorage control is a critical point in orthodontic treatment, and its failure, the main adverse problem. Using two groups of stable (successful) and unstable (failed) mini-implants, this in vivo study aimed to quantify proinflammatory cytokines IL-1 α, IL-6, IL-17, and TNF-α and osteoclastogenesis marker RANK, RANKL, and OPG in gingival tissue, using the real-time polymerase chain reaction technique.

Methodology: Thirteen patients of both sexes (11-49 years old) under orthodontic treatment were selected, obtaining 11 successful and 7 failed mini-implants. The mini-implants were placed and removed by the same surgeon, in both jaws. The mean time of permanence in the mouth was 29.4 months for successful and 7.6 months for failed mini-implants. At removal time, peri-mini-implant gingival tissue samples were collected and processed for quantification of the proinflammatory cytokines and osteoclastogenesis markers. Nonparametric Wilcoxon rank-sum test considering the clusters and Kruskal-Wallis test were used for statistical analysis (α=0.05).

Results: No significant difference (p>0.05) was observed between the groups for either quantification of cytokines or osteoclastogenesis markers, except for IL-6 (p<0.05).

Conclusions: It may be concluded that the expression of IL-1α, IL-17, TNF-α, RANK, RANKL, and OPG in peri-implant gingival tissue were not determinant for mini-implant stability loss, but the higher IL-6 expression could be associated with mini-implant failure.
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http://dx.doi.org/10.1590/1678-7757-2018-0476DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6768117PMC
October 2019
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