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The effects of different concentrations of titanium dioxide (TiOTiOWIThe 5% TiOHP with 5% TiO

Using genetic, clinical, biochemical, and radiographic assessment and bioinformatic approaches, we present an unusual case of adult HPP caused by a novel de novo heterozygous nonsense mutation in the alkaline phosphatase (ALPL). Hypophosphatasia (HPP) is caused by genetic alterations of the ALPL gene, encoding the tissue-nonspecific isozyme of alkaline phosphatase (TNSALP). Here, the purpose was to perform clinical and molecular investigation in a 36-year-old Caucasian woman suspected to present adult HPP. Medical and dental histories were obtained for the proposita and family members, including biochemical, radiographic, and dental assessments. ALPL mutational analysis was performed by the Sanger sequencing method, and the functional impact prediction of the identified mutations was assessed by bioinformatic methods. We identified a novel heterozygous nonsense mutation in the ALPL gene (NM_000478.6:c.768G>A; W[TGG]>*[TGA]) associated with spontaneous vertebral fracture, severe back pain, musculoskeletal pain, low bone density, and short-rooted permanent teeth loss. Functional prediction analysis revealed that the Trp256Ter mutation led to a complete loss of TNSALP crown domain and extensive loss of other functional domains (calcium-binding domain, active site vicinity, and zinc-binding site) and over 60% loss of homodimer interface residues, suggesting that the mutant TNSALP molecules are nonfunctional and form unstable homodimers. Genotyping of the ALPL in the proposita’s parents, sister, and niece revealed that in this case, HPP occurred due to a de novo mutation. The present study describes a novel genotype-phenotype and structure-function relationship for HPP, contributing to a better molecular comprehension of HPP etiology and pathophysiology.

The purpose of this study was: 1) to analyze the physical-chemical properties of hydrogen peroxide (HP) agents at 7.5% (HP7) and 35% (HP35), and the association with or without TiO80 bovine incisors were treated according to groups (n=10): HP35; HP35 + VL; HP35T (HP35 + TiOThe pH of HP35 thickener was higher when associated to TiOThe incorporation of TiO

Amelogenin isoforms, including full-length amelogenin (AMEL) and leucine-rich amelogenin peptide (LRAP), are major components of the enamel matrix, and are considered as signaling molecules in epithelial-mesenchymal interactions regulating tooth development and periodontal regeneration. Nevertheless, the molecular mechanisms involved are still poorly understood. The aim of the present study was to identify novel binding partners for amelogenin isoforms in the cementoblast (OCCM-30), using an affinity purification assay (GST pull-down) followed by mass spectrometry and immunoblotting. Protein-protein interaction analysis for AMEL and LRAP evidenced the plasminogen activation system (PAS) as a potential player regulating OCCM-30 response to amelogenin isoforms. For functional assays, PAS was either activated (plasmin) or inhibited (e-aminocaproic acid [aminocaproic]) in OCCM-30 cells and the cell morphology, mineral nodule formation, and gene expression were assessed. PAS inhibition (EACA 100 mM) dramatically decreased mineral nodule formation and expression of OCCM-30 differentiation markers, including osteocalcin (Bglap), bone sialoprotein (Ibsp), osteopontin (Spp1), tissue-nonspecific alkaline phosphatase (Alpl) and collagen type I (Col1a1), and had no effect on runt-related transcription factor 2 (Runx2) and Osterix (Osx) mRNA levels. PAS activation (plasmin 5 µg/µl) significantly increased Col1a1 and decreased Bglap mRNA levels (p < .05). Together, our findings shed new light on the potential role of plasminogen signaling pathway in the control of the amelogenin isoform-mediated response in cementoblasts and provide new insights into the development of targeted therapies.

Background Cellular cementum, a mineralized tissue covering apical tooth roots, grows by apposition to maintain the tooth in its occlusal position. We hypothesized that resident cementocytes would show morphological changes in response to cementum apposition, possibly implicating a role in cementum biology. Methods Mandibular first molars were induced to super-erupt (EIA) by extraction of maxillary molars, promoting rapid new cementum formation. Tissue and cell responses were analyzed at 6 and/or 21 days post-procedure (dpp). Results High-resolution micro-computed tomography (micro-CT) and confocal laser scanning microscopy showed increased cellular cementum by 21 dpp. Transmission electron microscopy (TEM) revealed that cementocytes under EIA were 50% larger than control cells, supported by larger pore sizes detected by micro-CT. Cementocytes under EIA displayed ultrastructural changes consistent with increased activity, including increased cytoplasm and nuclear size. We applied EIA to Hyp mutant mice, where cementocytes have perilacunar hypomineralization defects, to test cell and tissue responses in an altered mechanoresponsive milieu. Hyp and WT molars displayed similar super-eruption, with Hyp molars exhibiting 28% increased cellular cementum area versus 22% in WT mice at 21 dpp. Compared to control, Hyp cementocytes featured well-defined, disperse euchromatin and a thick layer of peripherally condensed heterochromatin in nuclei, indicating cellular activity. Immunohistochemistry (IHC) for cementum markers revealed intense dentin matrix protein-1 expression and abnormal osteopontin deposition in Hyp mice. Both WT and Hyp cementocytes expressed gap junction protein, connexin 43. Conclusion This study provides new insights into the EIA model and cementocyte activity in association with new cementum formation.

Cementum is a mineralized tissue that covers tooth roots and functions in the periodontal attachment complex. Cementocytes, resident cells of cellular cementum, share many characteristics with osteocytes, are mechanoresponsive cells that direct bone remodeling based on changes in loading. We hypothesized that cementocytes play a key role during orthodontic tooth movement (OTM). To test this hypothesis, we used 8-week-old male Wistar rats in a model of OTM for 2, 7, or 14 days (0.5 N), whereas unloaded contralateral teeth served as controls. Tissue and cell responses were analyzed by high-resolution micro-computed tomography, histology, tartrate-resistant acid phosphatase staining for odontoclasts/osteoclasts, and transmission electron microscopy. In addition, laser capture microdissection was used to collect cellular cementum, and extracted proteins were identified by liquid chromatography coupled to tandem mass spectrometry. The OTM model successfully moved first molars mesially more than 250 μm by 14 days introducing apoptosis in a small number of cementocytes and areas of root resorption on mesial and distal aspects. Cementocytes showed increased nuclear size and proportion of euchromatin suggesting cellular activity. Proteomic analysis identified 168 proteins in cellular cementum with 21 proteins found only in OTM sites and 54 proteins only present in control samples. OTM-down-regulated several extracellular matrix proteins, including decorin, biglycan, asporin, and periostin, localized to cementum and PDL by immunostaining. Furthermore, type IV collagen (COL14A1) was the protein most down-regulated (−45-fold) by OTM and immunolocalized to cells at the cementum-dentin junction. Eleven keratins were significantly increased by OTM, and a pan-keratin antibody indicated keratin localization primarily in epithelial remnants of Hertwig's epithelial root sheath. These experiments provide new insights into biological responses of cementocytes and cellular cementum to OTM.

Background and objectives Dental cementum (DC) is a mineralized tissue covering tooth roots that plays a critical role in dental attachment. Differences in deciduous vs. permanent tooth DC have not been explored. We hypothesized that proteomic analysis of DC matrix would identify compositional differences in deciduous (DecDC) vs. permanent (PermDC) cementum that might reflect physiological or pathological differences, such as root resorption that is physiological in deciduous teeth but can be pathological in the permanent dentition. Methods Protein extracts from deciduous (n = 25) and permanent (n = 12) teeth were pooled (five pools of DecDC, five teeth each; four pools of PermDC, three teeth each). Samples were denatured, and proteins were extracted, reduced, alkylated, digested, and analyzed by liquid chromatography-mass spectrometry (LC-MS/MS). The beta-binomial statistical test was applied to normalized spectrum counts with 5% significance level to determine differentially expressed proteins. Immunohistochemistry was used to validate selected proteins. Results A total of 510 proteins were identified: 123 (24.1%) exclusive to DecDC; 128 (25.1%) exclusive to PermDC; 259 (50.8%) commonly expressed in both DecDC and PermDC. Out of 60 differentially expressed proteins, 17 (28.3%) were detected in DecDC, including myeloperoxidase (MPO), whereas 43 (71.7%) were detected in PermDC, including decorin (DCN) and osteocalcin (BGLAP). Overall, Gene Ontology (GO) analysis indicated that all expressed proteins were related to GO biological processes that included localization and response to stress, and the GO molecular function of differentially expressed proteins was enriched in cell adhesion, molecular binding, cytoskeletal protein binding, structural molecular activity, and macromolecular complex binding. Immunohistochemistry confirmed the trends for selected differentially expressed proteins in human teeth. Conclusions Clear differences were found between the proteomes of DecDC and PermDC. These findings may lead to new insights into developmental differences between DecDC and PermDC, as well as to a better understanding of physiological/pathological events such as root resorption.

Made available in DSpace on 2020-12-12T01:55:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-03-01 Objectives: The aim of this study was to determine the physico-mechanical properties of a high viscosity glass ionomer cement (GIC) reinforced with TiO2 nanotubes (TiO2-nt). Methods: TiO2-nt was incorporated into the GIC powder components (Ketac Molar EasyMix™) in concentrations of 0% (control group), 3%, 5%, 7% by weight. Compressive strength (n = 10/group), three point bending for flexural strength (n = 18/group), microshear bond strength to dentin and failure mode (n = 20/group), and surface roughness and weight loss before and after brushing simulation (30,000 cycles) (n = 8/group) were evaluated. Data were submitted to Shapiro-Wilk, ANOVA, Tukey and Chi-square tests (α ≤ 0.05). Results: Addition of 5% of TiO2-nt into GIC presented the highest values for compressive strength and differed from the control, 3% and 7% groups (p = 0.023). There were no significant differences in flexural strength (p = 0.107) and surface roughness before and after the dental brushing (p = 0.287) among the groups. GIC added with 5% TiO2-nt showed the lowest weight loss values (p = 0.01), whereas the control, 3% or 5% TiO2-nt groups presented similar microshear bond strength values (p ≥ 0.05). The 5% TiO2-nt group featured higher microshear bond strength than the 7% TiO2-nt group (p = 0.034). Cohesive in material was the most representative failure mode for all groups. Significance: The incorporation of TiO2-nt did not affect GIC's adhesiveness to dentin, but improved its compressive strength at 5%. Furthermore, TiO2-nt decreased the percentage of weight loss after GIC's surface wear. Faculdade São Leopoldo Mandic - SLMANDIC Campinas Department of Pediatric Dentistry Piracicaba Dental School University of Campinas, Piracicaba - UNICAMP Department of Physics School of Science São Paulo State University - UNESP, Bauru Department of Biostatistics Dental School São Paulo University - UNIP Department of Dentistry Endodontic and Dental Materials Bauru Dental School University of São Paulo - USP, Bauru Department of Prosthodontics and Periodontics Division of Periodontics Piracicaba Dental School University of Campinas - UNICAMP, Piracicaba Department of Physics School of Science São Paulo State University - UNESP, Bauru

Background The aim of the present study was to compare indirect methods to assess the clinical performance of pit and fissure sealants and validate the use of 3D scanners. Material and Methods Sample consisted of 58 plaster models of upper and lower first permanent molars, sealed with resin sealants, as well as photographs obtained during the 18-month follow-up. Pre-established criteria were applied to categorize the sealant presence/absence and marginal integrity. Two calibrated examiners performed the evaluations, independently, using Scanning Electron Microscopy (SEM; gold-standard), Photography, 3D (CEREC In Lab) and Stereomicroscope analysis. Results The intra-examiner Spearman correlation was 94% e 97%, respectively, and the inter-examiner was 96%. Data was submitted to Kappa test, Spearman correlation and Receiver Operating Characteristic Curve (ROC). 3D and SEM presented good concordance; Stereomicroscope showed regular concordance with SEM and 3D (p0.05). SEM had a significant positive correlation with 3D and Stereomicroscope (r=0.76 and 0.71, respectively; p

A adição de flúor nas águas de abastecimento público e seu monitoramento são meios importantes na prevenção à cárie dentária e controle de fluorose dentária. Essa pesquisa buscou analisar e monitorar os teores de fluoreto nas águas de abastecimento no município de Penedo- AL. Abordagem quantitativa foi realizada por meio de: 1. Levantamento epidemiológico em 2.335 escolares (7-12 anos), grau de fluorose classificada de acordo com Dean (0=normal a 5=severo), realizados por (dentistas, agentes comunitários de saúde e auxiliares de saúde bucal) treinada e calibrada (Kappa = 0,95); 2. Levantamento da concentração de íons fluoretos no período de 2014 a 2018 no serviço de abastamento de água e esgoto (SAAE); e 3. Monitoramento mensal da água de abastecimento de 11 escolas durante 6 meses consecutivos (2019 e 2020). Análise de fluoretos foi determinada pelo método do eletrodo específico (ppmF). Análise de íons fluoretos entre as escolas foi realizada pelos testes de Kruskal-Wallis. Verificou-se que 56,1% dos escolares apresentou algum grau de fluorose, sendo predominante o grau I e II (p=0,103). As concentrações de íons fluoretos apresentados pelo SAEE (0,8428 – 0,9025 mg F/mL) e pelo pesquisador (0,9372 – 1,0394 mg F/mL) oscilaram significativamente (p=0,0103). A prevalência de fluorose dentária em Penedo-AL é elevada nos escolares de 7-12 anos. Sendo os graus leve e muito leve os de maior ocorrência. A concentração de íons fluoretos na água de abastecimento público está acima do preconizado para o município.

Tissue engineering is a contemporary field of science, which aims to create conditions based on principles of cell and molecular biology, bioengineering and biomaterials to regenerate tissues. Mesenchymal stem cells present high proliferation rates and are able to differentiate into multilineages under certain conditions, suggesting that they have great potential to act in regeneration field. Tooth derived stem cells are a suitable alternative source of mesenchymal cells once they are easily accessible and have poor morbidity to the donor. Studies showed that they have been isolated and characterized from diverse tissues such as dental pulp, exfoliated deciduous teeth, periodontal ligament, gingiva, dental follicle and apical papilla. However studies show that there is heterogeneity among these populations and there is no standard method to select the most appropriate tooth derived stem cells for regenerative procedures. The aim of this review is to present the current perspective of the multiple types of tooth-derived stem cells and to discuss the basis for their use in periodontal tissue engineering. RESUMO A engenharia de tecidos é um campo contemporâneo da ciência, que visa criar condições baseadas em princípios de biologia celular e molecular, bioengenharia e biomateriais para regenerar tecidos. As células tronco mesenquimais apresentam altas taxas de proliferação e são capazes de se diferenciar, sob certas condições, em multi-linhagens, sugerindo que elas têm grande potencial para atuar no campo da regeneração. As células tronco derivadas de tecidos dentais são uma fonte alternativa adequada de células mesenquimais uma vez que são de fácil acesso e têm baixa morbidade para o doador. Estudos demonstraram que elas já foram isoladas e caracterizadas a partir de diversos tecidos tais como polpa dentária, dentes decíduos esfoliados, ligamento periodontal, gengiva, folículo dental e papila apical. Entretanto, os estudos demonstram que há heterogeneidade entre essas populações e não existe um método padrão para selecionar as células-tronco dentais mais apropriadas para procedimentos regenerativos. O objetivo desta revisão é apresentar o conhecimento atual dos vários tipos de células-tronco derivadas de dentes e discutir as novas perspectivas para seu uso na engenharia de tecidos periodontais.

Somatic activating mutations in the GNAQ have been recently associated with several congenital genetic disorders and tumors; however, the molecular mechanism/etiology that leads to GNAQ somatic mosaic mutation are unknown. Here, we reported a case of Sturge-Weber Syndrome (SWS) manifesting cutaneous vascular malformations (hemifacial Port-wine stain), cerebral and ocular vascular abnormalities (including epilepsy and glaucoma) and harboring a c.548G > A (p.R183Q) somatic mosaic mutation in GNAQ. Computational modeling studies were performed to assistant with the comprehension of the functional impact of p.R183Q and p.Q209L mutations in GNAQ, which encodes a G protein subunit alpha q (Gαq). The p.R183Q mutation was predicted to abolish hydrogen bonds between R183 residue and GDP molecule, destabilizing the inactive GDP-bound conformation of the Gαq mutants. Furthermore, replacement of R183 by Q183 residue was predicted to promote conformation changes in protein surface features affecting the switch I region, a key region that undergoes conformational changes triggered by receptor binding during signal transduction. In addition, replacement of Q209 by L209 residue was predicted to affect the molecular interaction between Gαq and Gβ subunit, impairing formation of the inactive heterotrimeric complex. These findings, in association with PPI network analysis, indicate that p.R183Q and p.Q209L mutations result in the over-activation of different downstream effectors, which in turn will determine the distinct cell responses and phenotype. These findings bring new insights on molecular etiology of vascular malformations associated to SWS and on different mechanisms underlying hyperactivation of downstream pathways to Gαq.

O objetivo do estudo foi avaliar o efeito da adicao de diferentes concentracoes de clorexidina (CHX) a um selante resinoso quanto as propriedades fisicas. Realizando teste de resistencia a flexao foi conduzido em maquina de ensaio universal Instron, com carga de 50N e velocidade de 0,5mm/min ate a fratura. Posteriormente tambem realizando o teste de amolecimento/softening test para a determinacao da dureza do material.

O estudo?in vitro?objetivou?investigar a influencia da incorporacao de diferentes concentracoes de nanotubos de?dioxido de titânio?(n-?TiO2)?nas propriedades fisicas do cimento de ionomero de vidro (CIV).?Ao?Ketac?Molar?EasyMix?foi incorporado concentracoes de?n-TiO2?(0%,?3%; 5%; 7% em peso), sintetizados pelo metodo alcalino (20?nm). O?tempo de reacao de presa inicial e final, apos a aglutinacao do?CIV?foi avaliado apos?90?s?da aglutinacao usando?as agulhas?maior e menor calibre de?Gillmore,?respectivamente?(n=12). Enquanto o?tempo da?perda de?brilho foi?calculado?utilizando um cronometro digital TIMEX.?As avaliacoes foram realizadas seguindo as especificacoes da?ISO 9917-1?e o experimento foi repetido?por duas vezes.?Os dados foram submetidos a ANOVA?e?teste de?Tukey?(?=0,05).? O CIV contendo 7% de TiO2 exibiu o valor mais elevado de tempo de presa inicial (344,16 ± 6,84 s), diferindo estatisticamente do grupo controle (334,25 ± 7,47 s) e do grupo contendo 3% de TiO2 (331,75 ± 7,23 s) (p

The current study aimed at the determination of the impact of obesity on the salivary microbiome in adolescents. Sixty subjects ranging 14–17 years old were enrolled (obese: n = 30–50% females, and normal weight: n = 30–50% females). Stimulated saliva was collected for denaturing gradient gel electrophoresis (DGGE) band patterns and massive 16S rRNA gene sequencing using the Ion Torrent platform. Overall, data analysis revealed that male subjects harbored a higher diverse salivary microbiome, defined by a significant higher richness (32.48 versus 26.74) and diversity (3.36 versus 3.20), higher Simpson values (0.96 versus 0.95) and distinct bacterial community structure considering either sex or condition (p

The aim of this study was to determine the physico-mechanical properties of a high viscosity glass ionomer cement (GIC) reinforced with TiOTiOAddition of 5% of TiOThe incorporation of TiO

O objetivo do estudo foi avaliar o efeito da adição de diacetato de clorexidina (CHX) a 0,1% e 0,2% a um selante resinoso comercial, na propriedade de sorção e solubilidade. Espécimes de material foram distribuídos nos grupos: S - selante resinoso (Fluorshield®- controle), S1 - selante resinoso + 0,1% de CHX e S2 - selante resinoso + 0,2% de CHX.

Hypophosphatasia (HPP) is an inherited metabolic disorder that causes defective skeletal and dental mineralization. HPP exhibits a markedly heterogeneous range of clinical manifestations caused by dysfunction of the tissue-nonspecific isozyme of alkaline phosphatase (TNSALP), resulting from loss-of-function mutations in the ALPL gene. HPP has been associated with predominantly missense mutations in ALPL, and a number of compound heterozygous genotypes have been identified. Here, we describe a case of a subject with adult-onset HPP caused by a novel combination of missense mutations p.Gly473Ser and p.Ala487Val, resulting in chronic musculoskeletal pain, myopathy, persistent fatigue, vomiting, and an uncommon dental phenotype of short-rooted permanent teeth. Pedigree and biochemical analysis indicated that severity of symptoms was correlated with levels of residual ALP activity, and co-segregated with the p.Gly473Ser missense mutation. Bioinformatic analysis to predict the structural and functional impact of each of the point mutations in the TNSALP molecule, and its potential contribution to the clinical symptoms, revealed that the affected Gly473 residue is localized in the homodimer interface and predicted to have a dominant negative effect. The affected Ala487 residue was predicted to bind to Tyr479, which is closely located the N-terminal α-helix of TNSALP monomer 2, suggesting that both changes may impair dimer stability and catalytic functions. In conclusion, these findings assist in defining genotype-phenotype associations for HPP, and further define specific sites within the TNSALP molecule potentially related to neuromuscular manifestations in adult HPP, allowing for a better understanding of HPP pathophysiology.

Periodontal ligament (PDL) has been reported to be a source of mesenchymal stem cells (MSCs).New vascular networks from undifferentiated cells are essential for repair/regeneration of specialized tissues, including PDL. The current study aims to determine potential of CD105(+)-enriched cell subsets of periodontal ligament cells (PDLSCs) to differentiate into endothelial cell (EC)-like cells and to give insights into the mechanism involved.CD105(+)-enriched PDLSCs were induced to EC differentiation by endothelial growth medium 2 (EGM-2) for 3, 7, 14, and 21 days, with mRNA/protein levels and functional activity assessed by: 1) real-time polymerase chain reaction; 2) Western blotting; 3) fluorescence-activated cell sorting; 4) immunohistochemistry; 5) immunofluorescence; 6) matrigel; and 7) small interfering RNA assays.Data analyses demonstrated that EGM-2 treated PDLSCs presented increased expression of EC markers, including: 1) CD105; 2) kinase domain-containing receptor; and 3) Ulex europaeus agglutinin 1, and were able to form cord/tube-like structures. Gene and protein expression analysis showed that neuropilin 2 (NRP2), a key factor for vascular development, was significantly downregulated during EC differentiation. NRP2 was constitutively expressed in mouse PDL tissues by immunohistochemistry analysis, and NRP2 knockdown in CD105(+)-enriched PDLSCs resulted in increased cord/tube-like structures in a matrigel assay.These findings demonstrated the potential of CD105(+)-enriched PDLSCs to support angiogenesis, and NRP2 as a pivotal factor regulating this process.

Basic, pre-clinical, and clinical studies have documented the potential of amelogenin, and its variants, to affect cell response and tissue regeneration. However, the mechanisms are unclear. Thus, the aim of the present study was to identify, in cementoblasts, novel binding partners for an alternatively spliced amelogenin form (Leucine-Rich Amelogenin Peptide-LRAP), which is supposed to act as a signaling molecule in epithelial-mesenchymal interactions. LRAP-binding protein complexes from immortalized murine cementoblasts (OCCM-30) were achieved by capture affinity assay (GST pull down) and proteins present in these complexes were identified by mass spectrometry and immunoblotting. Flotillin-1, which functions as a platform for signal transduction, vesicle trafficking, endocytosis, and exocytosis, was identified and confirmed by co-precipitation and co-localization assays as a protein-binding partner for LRAP in OCCM-30 cells. In addition, we found that exogenously added GST-LRAP recombinant protein was internalized by OCCM-30 cells, predominantly localized in the perinuclear region and, that inhibition of flotillin1-dependent functions by small interference RNA (siRNA) methodology significantly affected LRAP uptake and its biological properties on OCCM-30 cells, including LRAP effect on the expression of genes encoding osteocalcin (Ocn), bone sialoprotein (Bsp), and runt-related transcription factor 2 (RunX2). In conclusion, LRAP uptake by cementoblast involves flotillin-assisted endocytosis, which suggests an involvement of LRAP in lipid-raft-dependent signaling pathways which are mediated by flotillin-1. J. Cell. Physiol. 232: 556-565, 2017. © 2016 Wiley Periodicals, Inc.

This case discusses a multi-disciplinary approach to oral functional and esthetic rehabilitation on a high-caries-risk child with prematurely lost primary teeth due to endodontic complications associated with wide-spread dental caries. The patient was diagnosed with an anterior open bite and atypical swallowing. He exhibited anxiety and low self-esteem due to esthetic impairment. A combination of targeted clinical procedures, a focus on oral home care and active parental involvement was integral to the treatment process. The patient was examined monthly by a pediatric dentist regarding dietary habits, fluoride exposure, biofilm presence, caries prevalence, and malocclusion. Improvement was evident in patient’s masticatory function, facial esthetics, and psychological behavior at a 12-month follow-up evaluation.KeywordsDeciduous; Dental Caries; Mouth Rehabilitation; Tooth.

Objective The present study assessed the effect of smoking on clinical, microbiological and immunological parameters in an experimental gingivitis model. Material and Methods Twenty-four healthy dental students were divided into two groups: smokers (n = 10); and nonsmokers (n = 14). Stents were used to prevent biofilm removal during brushing. Visible plaque index (VPI) and gingival bleeding index (GBI) were determined 5- on day -7 (running phase), baseline, 21 d (experimental gingivitis) and 28 d (resolution phase). Supragingival biofilm and gingival crevicular fluid were collected and assayed by checkerboard DNA–DNA hybridization and a multiplex analysis, respectively. Intragroup comparison was performed by Friedman and Dunn's multiple comparison tests, whereas the Mann–Whitney U-test was applied for intergroup analyses. Results Cessation of oral hygiene resulted in a significant increase in VPI, GBI and gingival crevicular fluid volume in both groups, which returned to baseline levels 7 d after oral hygiene was resumed. Smokers presented lower GBI than did nonsmokers (p < 0.05) at day 21. Smokers had higher total bacterial counts and higher proportions of red- and orange complex bacteria, as well as lower proportions of Actinomyces spp., and of purple- and yellow-complex bacteria (p < 0.05). Furthermore, the levels of key immune-regulatory cytokines, including interleukin (IL)-8, IL-17 and interferon-γ, were higher in smokers than in nonsmokers (p < 0.05). Conclusion Smokers and nonsmokers developed gingival inflammation after supragingival biofilm accumulation, but smokers had less bleeding, higher proportions of periodontal pathogens and distinct host-response patterns during the course of experimental gingivitis.

With the evolution of bacterial resistance over conventional antimicrobials and the development of new antimicrobials, the search for new compounds of natural origin has intensified, since plants with therapeutic potential constitute a source of new biologically active compounds, encouraging the development of new therapeutic options. The objective of this study was to evaluate the antibacterial and anti-adherent activity of the ethanolic extract of Praxelis clematidea (Griseb.) R.M. King & Robinson (Asteraceae) on strains of Staphylococcus aureus, Streptococcus pneumoniae, Klebsiella pneumoniae, Pseudomonas aeruginosa and Enterococcus faecalis. Tests were carried out to determine the MIC and MBC by means of broth microdilution in 96-well plates, and to determine the MICA, a technique with inclined tubes in the presence of 5% sucrose was used. It was observed that the ethanolic extract of P. clematidea has strong bactericidal activity on K. pneumoniae, and moderate bactericidal activity on S. pneumoniae and E. faecalis. Being able to inhibit adherence to K. pneumoniae strains. Thus, the ethanolic extract of P. clematidea proves to be effective as an antimicrobial in the control and prevention of infections by S. pneumoniae, K. pneumoniae and E. faecalis. It also has an effective anti-adherent effect on K. pneumoniae strains. Con la evolución de la resistencia bacteriana sobre los antimicrobianos convencionales y el desarrollo de nuevos antimicrobianos, se ha intensificado la búsqueda de nuevos compuestos de origen natural, ya que las plantas con potencial terapéutico son fuente de nuevos compuestos biológicamente activos, favoreciendo el desarrollo de nuevas opciones terapéuticas. El objetivo de este estudio fue evaluar la actividad antibacteriana y antiadherente del extracto etanólico de Praxelis clematidea (Griseb.) R.M. King & Robinson (Asteraceae) sobre cepas de Staphylococcus aureus, Streptococcus pneumoniae, Klebsiella pneumoniae, Pseudomonas aeruginosa y Enterococcus faecalis. Se realizaron pruebas para determinar la CMI y CBM mediante microdilución en caldo en placas de 96 pocillos, y para determinar la CIMA se utilizó una técnica con tubos inclinados en presencia de sacarosa al 5%. Se observó que el extracto etanólico de P. clematidea tiene una fuerte actividad bactericida sobre K. pneumoniae y una moderada actividad bactericida sobre S. pneumoniae y E. faecalis. Ser capaz de inhibir la adherencia a cepas de K. pneumoniae. Así, el extracto etanólico de P. clematidea demuestra ser eficaz como antimicrobiano en el control y prevención de infecciones por S. pneumoniae, K. pneumoniae y E. faecalis. También tiene un efecto antiadherente eficaz sobre las cepas de K. pneumoniae. Com a evolução da resistência bacteriana sobre os antimicrobianos convencionais e o desenvolvimento de novos antimicrobianos, a procura por novos compostos de origem natural tem se intensificado, uma vez que plantas que apresentam potencial terapêutico constituem uma fonte de novos compostos biologicamente ativos, incentivando o desenvolvimento de novas opções terapêuticas. O objetivo deste estudo foi avaliar a atividade antibacteriana e antiaderente do extrato etanólico de Praxelis clematidea (Griseb.) R.M. King & Robinson (Asteraceae) sobre cepas de Staphylococcus aureus, Streptococcus pneumoniae, Klebsiella pneumoniae, Pseudomonas aeruginosa e Enterococcus faecalis. Foram realizados os ensaios para determinação da CIM e CBM por meio de técnica de microdiluição em caldo em placas de 96 poços, e para determinação da CIMA foi utilizada técnica com tubos inclinados na presença de 5% de sacarose. Observou-se que o extrato etanólico de P. clematidea apresenta forte atividade bactericida sobre K. pneumoniae, e moderada atividade bactericida sobre S. pneumoniae e E. faecalis. Sendo capaz de inibir a aderência das cepas de K. pneumoniae. Com isso, o extrato etanólico de P. clematidea demonstra ser eficaz como antimicrobiano no controle e prevenção de infecções por S. pneumoniae, K. pneumoniae e E. faecalis. Também apresentando eficaz efeito antiaderente sobre as cepas de K. pneumoniae.

BACKGROUND Physiological roles for the periodontal ligament (PDL) include tooth eruption and anchorage, force absorption, and provision of proprioceptive information. Despite the advances in understanding the biology of PDL cells, there is a lack of information regarding the molecular signature of deciduous (DecPDL) and permanent (PermPDL) PDL tissues. Thus, the present study was designed to characterize the membrane proteome of DecPDL and PermPDL cells. METHODS Primary PDL cells were obtained (n = 6) and a label-free quantitative proteome of cell membrane-enriched components was performed. Proteome findings were validated by quantitative polymerase chain reaction and Western blot assays in fresh human tissues (n = 8) and primary cell cultures (n = 6). In addition, confocal microscopy was used to verify the expression of target factors in the PDL cell cultures. RESULTS Comparative gene ontology enrichment analysis evidenced that most stickling differences involved "endomembrane system" (PICALM, STX4, and LRP10), "hydrolase activity" (NCSTN and XRCC6), "protein binding" (PICALM, STX4, GPNMB, VASP, extended-synaptotagmin 2 [ESYT2], and leucine-rich repeat containing 15 [LRRC15]), and "isomerase activity" (FKBP8). Data are available via ProteomeXchange with identifier PXD010226. At the transcript level, high PICALM in DecPDL and ESYT2 and LRRC15 in PermPDL were confirmed in fresh PDL tissues. Furthermore, Western blot analysis confirmed increased levels of PICALM, LRRC15, and ESYT2 in cells and/or fresh tissues, and confocal microscopy confirmed the trends for PICALM and LRRC15 expression in PDL cells. CONCLUSION We report the first comprehensive characterization of the membrane protein machinery of DecPDL and PermPDL cells, and together, we identified a distinct molecular signature for these cell populations, including unique proteins for DecPDL and PermPDL.

Bone sialoprotein (BSP) is a multifunctional extracellular matrix protein found in mineralized tissues, including bone, cartilage, tooth root cementum (both acellular and cellular types), and dentin. In order to define the role BSP plays in the process of biomineralization of these tissues, we analyzed cementogenesis, dentinogenesis, and osteogenesis (intramembranous and endochondral) in craniofacial bone in Bsp null mice and wild-type (WT) controls over a developmental period (1-60 days post natal; dpn) by histology, immunohistochemistry, undecalcified histochemistry, microcomputed tomography (microCT), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and quantitative PCR (qPCR). Regions of intramembranous ossification in the alveolus, mandible, and calvaria presented delayed mineralization and osteoid accumulation, assessed by von Kossa and Goldner's trichrome stains at 1 and 14 dpn. Moreover, Bsp(-/-) mice featured increased cranial suture size at the early time point, 1 dpn. Immunostaining and PCR demonstrated that osteoblast markers, osterix, alkaline phosphatase, and osteopontin were unchanged in Bsp null mandibles compared to WT. Bsp(-/-) mouse molars featured a lack of functional acellular cementum formation by histology, SEM, and TEM, and subsequent loss of Sharpey's collagen fiber insertion into the tooth root structure. Bsp(-/-) mouse alveolar and mandibular bone featured equivalent or fewer osteoclasts at early ages (1 and 14 dpn), however, increased RANKL immunostaining and mRNA, and significantly increased number of osteoclast-like cells (2-5 fold) were found at later ages (26 and 60 dpn), corresponding to periodontal breakdown and severe alveolar bone resorption observed following molar teeth entering occlusion. Dentin formation was unperturbed in Bsp(-/-) mouse molars, with no delay in mineralization, no alteration in dentin dimensions, and no differences in odontoblast markers analyzed. No defects were identified in endochondral ossification in the cranial base, and craniofacial morphology was unaffected in Bsp(-/-) mice. These analyses confirm a critical role for BSP in processes of cementogenesis and intramembranous ossification of craniofacial bone, whereas endochondral ossification in the cranial base was minimally affected and dentinogenesis was normal in Bsp(-/-) molar teeth. Dissimilar effects of loss of BSP on mineralization of dental and craniofacial tissues suggest local differences in the role of BSP and/or yet to be defined interactions with site-specific factors.

The periodontal complex is essential for tooth attachment and function and includes the mineralized tissues, cementum and alveolar bone, separated by the unmineralized periodontal ligament (PDL). To gain insights into factors regulating cementum-PDL and bone-PDL borders and protecting against ectopic calcification within the PDL, we employed a proteomic approach to analyze PDL tissue from progressive ankylosis knock-out ( Ank −/− ) mice, featuring reduced PP i , rapid cementogenesis, and excessive acellular cementum. Using this approach, we identified the matrix protein osteopontin ( Spp1 /OPN) as an elevated factor of interest in Ank −/− mouse molar PDL. We studied the role of OPN in dental and periodontal development and function. During tooth development in wild-type (WT) mice, Spp1 mRNA was transiently expressed by cementoblasts and strongly by alveolar bone osteoblasts. Developmental analysis from 14 to 240 days postnatal (dpn) indicated normal histological structures in Spp1 −/− comparable to WT control mice. Microcomputed tomography (micro-CT) analysis at 30 and 90 dpn revealed significantly increased volumes and tissue mineral densities of Spp1 −/− mouse dentin and alveolar bone, while pulp and PDL volumes were decreased and tissue densities were increased. However, acellular cementum growth was unaltered in Spp1 −/− mice. Quantitative PCR of periodontal-derived mRNA failed to identify potential local compensators influencing cementum in Spp1 −/− vs. WT mice at 26 dpn. We genetically deleted Spp1 on the Ank −/− mouse background to determine whether increased Spp1 /OPN was regulating periodontal tissues when the PDL space is challenged by hypercementosis in Ank −/− mice. Ank −/− ; Spp1 −/− double deficient mice did not exhibit greater hypercementosis than that in Ank −/− mice. Based on these data, we conclude that OPN has a non-redundant role regulating formation and mineralization of dentin and bone, influences tissue properties of PDL and pulp, but does not control acellular cementum apposition. These findings may inform therapies targeted at controlling soft tissue calcification.

Chédiak-Higashi syndrome (CHS), a rare autosomal recessive disorder caused by mutations in the lysosomal trafficking regulator gene (LYST), is associated with aggressive periodontitis. It is suggested that LYST mutations affect the toll-like receptor (TLR)–mediated immunoinflammatory response, leading to frequent infections. This study sought to determine the periodontal status of patients with classic (severe) and atypical (milder) forms of CHS and the immunoregulatory functions of gingival fibroblasts in CHS patients. In contrast to aged-matched healthy controls, atypical (n = 4) and classic (n = 3) CHS patients presented with mild chronic periodontitis with no evidence of gingival ulceration, severe tooth mobility, or premature exfoliation of teeth. As a standard of care, all classic CHS patients had undergone bone marrow transplantation (BMT). Primary gingival fibroblasts obtained from atypical and BMT classic CHS patients displayed higher protein expression of TLR-2 (1.81-fold and 1.56-fold, respectively) and decreased expression of TLR-4 (−2.5-fold and −3.85-fold, respectively) at baseline when compared with healthy control gingival fibroblasts. When challenged with whole bacterial extract of Fusobacterium nucleatum, both atypical and classic CHS gingival fibroblasts failed to up-regulate TLR-2 and TLR-4 expression when compared with their respective untreated groups and control cells. Cytokine multiplex analysis following F. nucleatum challenge showed that atypical CHS gingival fibroblasts featured significantly increased cytokine expression (interleukin [IL]–2, IL-4, IL-5, IL-6, IL-10, IL-12, interferon-γ, tumor necrosis factor–α), whereas classic CHS cells featured similar/decreased cytokine expression when compared with treated control cells. Collectively, these results suggest that LYST mutations in CHS patients affect TLR-2 and TLR-4 expression/function, leading to dysregulated immunoinflammatory response, which in turn may influence the periodontal phenotype noted in CHS patients. Furthermore, our results suggest that atypical CHS patients and classic CHS patients who undergo BMT early in life are less susceptible to aggressive periodontitis and that hematopoietic cells play a critical role in mitigating the risk of aggressive periodontitis in CHS. Knowledge Transfer Statement: Results from this study can be used to create awareness among clinicians and researchers that not all CHS patients exhibit historically reported aggressive periodontitis, especially if they have atypical CHS disease or have received bone marrow transplantation. LYST mutations in CHS patients may affect TLR-2 and TLR-4 expression/function leading to dysregulated immunoinflammatory response, which in turn may influence the periodontal phenotype noted in CHS patients.

To evaluate the penetration ratio of filled and unfilled resin-based sealants on different enamel substrates and pit and fissure morphologies.Forty-eight occlusal enamel blocks obtained from impacted human third molars were randomly divided (n equals eight) according to enamel substrates (sound; caries-like lesion; caries-like lesion plus topical fluoride application) and sealant material (FluroShield; Helioseal Clear Chroma). Sealants were applied on the enamel surface. The specimens were stored in 100 percent humidity for 24 hours at 37 degrees Celsius, sectioned in a buccal-lingual direction (at approximately 50 μm), and examined to determine the sealant penetration ratio (b x 100/a; a equals total fissure length and b equals sealant penetration length) and pit and fissure morphology (V-, U-, or Y-shaped). Statistical analysis was performed using Friedman and Kruskal-Wallis tests (P0.05).Enamel substrate and sealant material did not affect the sealant penetration ratio, with no interactions between these factors. Moreover, the morphology significantly affected the sealant penetration, with the "Y"-shaped fissures presenting the lowest sealant penetration ratio compared to "U-"shaped (P=0.0001) and "V-" shaped fissures (P=0.0018).Pit and fissure morphology was a critical factor on sealant's penetration capacity; however, enamel substrate and sealant type did not affect sealant's penetration ratio.

Made available in DSpace on 2018-11-26T17:32:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-01-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The aim of this study was to assess the performance of glass ionomer cement (GIC) added with TiO2 nanotubes. TiO2 nanotubes [3%, 5%, and 7%(w/w)] were incorporated into GIC's (Ketac Molar EasyMix (TM)) powder component, whereas unblended powder was used as control. Physical-chemical-biological analysis included energy dispersive spectroscopy (EDS), surface roughness (SR), Knoop hardness (SH), fluoride-releasing analysis, cytotoxicity, cell morphology, and extracellular matrix (ECM) composition. Parametric or nonparametric ANOVA were used for statistical comparisons (alpha

This study evaluated the effects of chemical agents on the physical properties and structure of primary pulp chamber dentin using surface roughness, microhardness tests, and scanning electron microscopy (SEM). Twenty-five primary teeth were sectioned exposing the pulp chamber and were divided into five groups (n = 5): NT, no treatment; SH1, 1% sodium hypochlorite (NaOCl); SH1U, 1% NaOCl + Endo-PTC®; SH1E, 1% NaOCl + 17% EDTA; and E, 17% EDTA. After dentin treatment, the specimens were submitted to roughness, microhardness testing, and SEM analysis. Roughness and microhardness data were submitted to one-way ANOVA and Tukey's test (P 0.05). Microhardness values could not be obtained in the EDTA groups (SH1E and E). The presence of intertubular dentin with opened dentin tubules was observed in the NT, SH1, and SH1U groups. SH1E showed eroded and disorganized dentin with few opened tubules and the intertubular/peritubular dentin was partially removed. Considering the physical and structural approaches and the chemical agents studied, it can be concluded that NaOCl and NaOCl associated with Endo-PTC® were the agents that promoted the smallest changes in surface roughness, microhardness, and structure of the pulp chamber dentin of primary teeth. Microsc. Res. Tech. 77:52–56, 2014. © 2013 Wiley Periodicals, Inc.

ObjectivesThis study evaluated the effect of fluoride and non-fluoride sealants on hardness decrease (HD) and marginal adaptation (MA) on enamel substrates after cariogenic challenge.MethodsOcclusal enamel blocks, from human third molars, were randomly divided into six groups (n=12), according to occlusal fissures condition (S – sound; C – caries-like lesion; CF – caries-like lesion+topical fluoride) and sealants (F – FluroShield; H – Helioseal Clear Chroma). Lesion depths were 79.3±33.9 and 61.3±23.9 for C and CF groups, respectively. Sealants were placed on occlusal surface and stored at 100% humidity (37°C; 24h/d). HD was measured by cross-sectional microhardness analysis at the sealant margin distances: −1 (under sealant), 0 (sealant margin), 1, 2 (outer sealant). Sealant MA was observed by polarized light microscopy and scored according to: 0 – failure (no sealant MA or total sealant loss); 1 – success (sealant MA present). MA and HD were analysed by ANOVA-R and mixed model analysis, respectively.ResultsFor HD (ΔS), F values (6900.5±3686.6) were significantly lower than H values (8534.6±5375.3) regardless of enamel substrates and sealant margin distances. Significant differences were observed among sealant margin distances: −1 (5934.0±3282.6)

The aim of this study was to assess the performance of glass ionomer cement (GIC) added with TiO2 nanotubes. TiO2 nanotubes [3%, 5%, and 7% (w/w)] were incorporated into GIC's (Ketac Molar EasyMix™) powder component, whereas unblended powder was used as control. Physical-chemical-biological analysis included energy dispersive spectroscopy (EDS), surface roughness (SR), Knoop hardness (SH), fluoride-releasing analysis, cytotoxicity, cell morphology, and extracellular matrix (ECM) composition. Parametric or nonparametric ANOVA were used for statistical comparisons (α ≤ 0.05). Data analysis revealed that EDS only detected Ti at the 5% and 7% groups and that GIC's physical-chemical properties were significantly improved by the addition of 5% TiO2 as compared to 3% and GIC alone. Furthermore, regardless of TiO2 concentration, no significant effect was found on SR, whereas GIC-containing 7% TiO2 presented decreased SH values. Fluoride release lasted longer for the 5% and 7% TiO2 groups, and cell morphology/spreading and ECM composition were found to be positively affected by TiO2 at 5%. In conclusion, in the current study, nanotechnology incorporated in GIC affected ECM composition and was important for the superior microhardness and fluoride release, suggesting its potential for higher stress-bearing site restorations.

The effects of the non-steroidal anti-inflammatory drugs (NSAIDs) on bone quantity and quality were investigated for years. However, there is lack of information on the impact of NSAIDs on the quality of tooth-supporting alveolar bone in absence of periodontal inflammation. Thus, the aim of this study was to evaluate histometrically the influence of a selective COX-2 NSAID (Meloxicam) on the inter-radicular bone mineral density in rats. Forty-nine adult male Wistar rats were randomly divided into four experimental groups: Subcutaneous injection of 0.9% sterile saline for 15 days (G1; n=12) and 45 days (G2; n=11); and subcutaneous injection of Meloxicam for 15 days (G3; n=13) and 45 days (G4; n=13). Mineral density was histometrically determined in the inter-radicular area of the 1st mandibular molars and data analysis performed by two-way ANOVA (a=5%). Results showed no interaction between time and treatment (p>0.05) and that meloxicam did not affect the alveolar bone density. In contrast, it was found that inter-radicular alveolar bone density increased with time (91.88±3.08% and 92.86±2.38% for groups 15 and 45 days, respectively) (p0,05) e que o meloxicam não afetou a densidade óssea alveolar. Em contraste, foi encontrado que a densidade óssea alveolar inter-radicular aumentou ao longo do tempo (91,88±3,08% e 92,86±2,38% para os grupos 15 e 45 dias, respectivamente) (p

Basic, pre-clinical, and clinical studies have documented the potential of amelogenin, and its variants, to affect cell response and tissue regeneration. However, the mechanisms are unclear. Thus, the aim of the present study was to identify, in cementoblasts, novel binding partners for an alternatively spliced amelogenin form (Leucine-Rich Amelogenin Peptide-LRAP), which is supposed to act as a signaling molecule in epithelial-mesenchymal interactions. LRAP-binding protein complexes from immortalized murine cementoblasts (OCCM-30) were achieved by capture affinity assay (GST pull down) and proteins present in these complexes were identified by mass spectrometry and immunoblotting. Flotillin-1, which functions as a platform for signal transduction, vesicle trafficking, endocytosis, and exocytosis, was identified and confirmed by co-precipitation and co-localization assays as a protein-binding partner for LRAP in OCCM-30 cells. In addition, we found that exogenously added GST-LRAP recombinant protein was internalized by OCCM-30 cells, predominantly localized in the perinuclear region and, that inhibition of flotillin1-dependent functions by small interference RNA (siRNA) methodology significantly affected LRAP uptake and its biological properties on OCCM-30 cells, including LRAP effect on the expression of genes encoding osteocalcin (Ocn), bone sialoprotein (Bsp), and runt-related transcription factor 2 (RunX2). In conclusion, LRAP uptake by cementoblast involves flotillin-assisted endocytosis, which suggests an involvement of LRAP in lipid-raft-dependent signaling pathways which are mediated by flotillin-1. J. Cell. Physiol. 232: 556-565, 2017. © 2016 Wiley Periodicals, Inc.

It has been suggested that there are histological and functional distinctions between the periodontal ligament (PDL) of deciduous (DecPDL) and permanent (PermPDL) teeth. Thus, we hypothesized that DecPDL and PermPDL display differences in the constitutive expression of genes/proteins involved with PDL homeostasis. Primary PDL cell cultures were obtained for DecPDL (n = 3) and PermPDL (n = 3) to allow us to perform label-free quantitative secretome analysis. Although a highly similar profile was found between DecPDL and PermPDL cells, comparative secretome analysis evidenced that one of the most stickling differences involved cell adhesion molecules, including laminin subunit gamma 1 (LAMC1) and beta 2 (LAMB2). Next, total RNA and protein extracts were obtained from fresh PDL tissues of deciduous (n = 6) and permanent (n = 6) teeth, and Western blotting and qPCR analysis were used to validate our in vitro findings. Western blot analysis confirmed that LAMC1 was increased in DecPDL fresh tissues (p