RESUMO
The interest of the scientific community on sex and gender differences in health and disease has increased substantially over the past 25 to 30 y as a result of a long process of events and policies in the biomedical field. This is crucial as compelling evidence from human and animal model studies has demonstrated that sex and gender influence health, molecular and cellular processes, and response and predisposition to disease. The present scoping review aims to provide a synthesis of sex differences in oral diseases, ranging from periodontal disease to orofacial pain conditions, from risk of caries development to apical periodontitis. Overall, findings from this review further support a role for sexual dimorphism influencing disease predisposition and/or progression in oral diseases. Of note, this review also highlights the lack of consideration of additional factors such as gender and other psychosocial and external factors potentially influencing oral health and disease. New conceptual frameworks capable of capturing multiple fundamental domains and measurements should be developed in clinical and preclinical studies to inform sex-based individualized preventive and treatment strategies.
Assuntos
Cárie Dentária , Doenças Periodontais , Animais , Humanos , Masculino , Feminino , Doenças Periodontais/prevenção & controle , Saúde Bucal , Cárie Dentária/prevenção & controle , Suscetibilidade a Doenças , Caracteres SexuaisRESUMO
Mutations in WNT10A have frequently been reported as etiologic for tooth agenesis (TA). However, the effects of WNT10A variation on gene/protein function and contribution to TA phenotypes remain poorly understood. Here, we performed bioinformatic and functional characterization analysis of WNT10A variants. In silico prediction of variant function was performed with VIPUR for all WNT10A missense variants reported in the Exome Aggregation Consortium database. Functional characterization experiments were then performed for selected WNT10A variants previously associated with TA. Expression vectors for wild-type and mutant WNT10A were made and transfected into stem cells from human exfoliated deciduous teeth (SHED) for evaluation of gene/protein function, WNT signaling activity, and effects on expression of relevant genes. While 75% of WNT10A variants were predicted neutral, most of the TA-associated variants received deleterious scores by potentially destabilizing or preventing the disulfide bond formation required for proper protein function. WNT signaling was significantly decreased with 8 of 13 variants tested, whereas wild-type-like activity was retained with 4 of 13 variants. WNT10A-mutant cells (T357I, R360C, and R379C mutants) showed reduced or impaired binding affinity to FZD5, suggesting a potential mechanism for the decreased WNT signaling. Mutant cells also had decreased WNT10A protein expression in comparison to wild-type cells. mRNA expression of PAX9, MSX1, AXIN2, and RUNX2 (known tooth development genes) was perturbed in mutant cells and quite significantly for PAX9 and RUNX2. Transcriptome analysis of wild-type and T357I-mutant cells identified 36 differentially expressed genes (26 downregulated, 10 upregulated) involved in skeletal system development and morphogenesis and pattern specification. WNT10A variants deemed pathogenic for TA likely affect protein folding and/or stabilization, leading to decreased WNT signaling and concomitant dysregulated expression of relevant genes. These findings may allow for improved interpretation of TA phenotypes upon clinical diagnosis while providing important insights toward the development of future tooth replacement therapies.
Assuntos
Anodontia , Dente , Anodontia/genética , Humanos , Mutação/genética , Fenótipo , Sequenciamento do Exoma , Proteínas Wnt/genética , Via de Sinalização Wnt/genéticaRESUMO
Periodontal and periapical lesions are infectious inflammatory osteolitytic conditions in which a complex inflammatory immune response mediates bone destruction. However, the uncertainty of a lesion's progressive or stable phenotype complicates understanding of the cellular and molecular mechanisms triggering lesion activity. Evidence from clinical and preclinical studies of both periodontal and periapical lesions points to a high receptor activator of NF-κB ligand/osteoprotegerin (RANKL/OPG) ratio as the primary determinant of osteolytic activity, while a low RANKL/OPG ratio is often observed in inactive lesions. Proinflammatory cytokines directly modulate RANKL/OPG expression and consequently drive lesion progression, along with pro-osteoclastogenic support provided by Th1, Th17, and B cells. Conversely, the cooperative action between Th2 and Tregs subsets creates an anti-inflammatory and proreparative milieu associated with lesion stability. Interestingly, the trigger for lesion status switch from active to inactive can originate from an unanticipated RANKL immunoregulatory feedback, involving the induction of Tregs and a host response outcome with immunological tolerance features. In this context, dendritic cells (DCs) appear as potential determinants of host response switch, since RANKL imprint a tolerogenic phenotype in DCs, described to be involved in both Tregs and immunological tolerance generation. The tolerance state systemically and locally suppresses the development of exacerbated and pathogenic responses and contributes to lesions stability. However, immunological tolerance break by comorbidities or dysbiosis could explain lesions relapse toward activity. Therefore, this article will provide a critical review of the current knowledge concerning periodontal and periapical lesions activity and the underlying molecular mechanisms associated with the host response. Further studies are required to unravel the role of immunological responsiveness or tolerance in the determination of lesion status, as well as the potential cooperative and/or inhibitory interplay among effector cells and their impact on RANKL/OPG balance and lesion outcome.
Assuntos
Osteoprotegerina , Ligante RANK , Doença Crônica , Citocinas , Humanos , Células Th17RESUMO
INTRODUCTION: Coronavirus disease 2019 (COVID-19) is a highly contagious disease that quickly reached pandemic levels. Over 5 million COVID-19 cases and approximately 330,000 deaths have been recorded worldwide. Transmission is primarily spread through direct, indirect (through contaminated objects or surfaces), or close contact with infected people via respiratory droplets, the mouth, and/or nose secretions. Health care professionals (HCPs), including dental HCPs, are recognized to be at considerably high risk for infection due to the close proximity to patients and aerosol-generating procedures. During pregnancy, HCPs may be at even higher risk since pregnancy substantially increases the susceptibility to infectious diseases. OBJECTIVES: Here, we present the posed risks and potential effects of COVID-19 on maternal and fetal health. Current prevention and management strategies for COVID-19 on pregnant dental and HCPs are also discussed. RESULTS: Significant progress is being made in understanding the pathogenesis and clinical consequences of COVID-19. Pregnant women are affected more adversely with viral illnesses, although evidence of vertical transmission of COVID-19 is controversial. Based on the presence of atypical symptoms, the significant numbers of asymptomatic individuals who are COVID-19 positive, and the high susceptibility to viral diseases observed in pregnant women, recommendations have been put forth to limit the exposure of COVID-19-positive or even suspected cases to pregnant HCPs, and these are likely to evolve as new information becomes available. CONCLUSION: Pregnant HCPs require extra caution: not only are they considered a high-risk population, but their work at the frontline in a pandemic may expose them to additional risks. Complete awareness of the effects of COVID-19 on maternal and fetal/infant health, as well as prevention and management guidelines for pregnant HCPs, will allow for a safer work environment. Health care institutional policies aimed at protecting pregnant HCPs should consider avoiding their assignment as first responders, especially if equally trained staff are available. KNOWLEDGE TRANSFER STATEMENT: Dental and health care professionals can use the information in this review to improve their awareness of COVID-19 risks, signs, and symptoms and the associated effects on the health of pregnant health care professionals and their unborn/newborn children.
Assuntos
Infecções por Coronavirus , Coronavirus , Pandemias , Pneumonia Viral , Complicações Infecciosas na Gravidez , Betacoronavirus , COVID-19 , Feminino , Pessoal de Saúde , Humanos , Recém-Nascido , Gravidez , SARS-CoV-2RESUMO
Extraordinary women scientists-past, current, and elected presidents of the International Association for Dental Research (IADR)-showcase pathways for success and leadership. In this series of autobiographical essays, these women of various cultural backgrounds with diverse areas of research describe their journeys in the passionate pursuit of excellence, despite the frequent obstacles and challenges. Through interviews and in their own words, we recap highlights of their dental research journeys and inspirations, their career trajectories toward the IADR presidency, and the benefits and challenges that they faced in their careers and personal lives. The purpose of this special issue is to honor these women, their life journeys, and how they have contributed to oral health research.
Assuntos
Liderança , Sociedades Odontológicas , Escolha da Profissão , Feminino , Humanos , Sociedades Odontológicas/estatística & dados numéricosRESUMO
Gender inequality in science, medicine, and dentistry remains a central concern for the biomedical research workforce today. Although progress in areas of inclusivity and gender diversity was reported, growth has been slow. Women still face multiple challenges in reaching higher ranks and leadership positions while maintaining holistic success in these fields. Within dental research and academia, we might observe trends toward a more balanced pipeline. However, women continue to face barriers in seeking leadership roles and achieving economic equity and scholarship recognition. In an effort to evaluate the status of women in dental research and academia, the authors examined the role of the International Association for Dental Research (IADR), a global research organization, which has improved awareness on gender inequality. The goal of this article is to review five crucial issues of gender inequality in oral health research and academics-workforce pipeline, economic inequality, workplace harassment, gender bias in scholarly productivity, and work-life balance-and to discuss proactive steps that the IADR has taken to promote gender equality. Providing networking and training opportunities through effective mentoring and coaching for women researchers, the IADR has developed a robust pipeline of women leaders while promoting gender equality for women in dental academia through a culture shift. As knowledge gaps remained on the levels of conscious and unconscious bias and sexist culture affecting women advancement in academics, as well as the intersectionality of gender with race, gender identity, ability status, sexual orientation, and cultural backgrounds, the IADR has recognized that further research is warranted.
Assuntos
Pesquisa em Odontologia , Sociedades Odontológicas , Pesquisa em Odontologia/organização & administração , Pesquisa em Odontologia/estatística & dados numéricos , Pesquisa em Odontologia/tendências , Humanos , Liderança , Sociedades Odontológicas/tendênciasRESUMO
AIM: To investigate the expression of TP63 in apical periodontitis (AP) tissues and the association of single nucleotide polymorphisms (SNPs) in the TP63 gene with AP using a case-control dataset. METHODOLOGY: Expression of TP63 in human AP lesions (apical abscess, radicular cyst, periapical granuloma) was evaluated using immunohistochemistry. A case-control association study was performed to assess the association of TP63 polymorphisms in individuals having AP with or without associated pain. Cases were defined as subjects with deep caries and AP (n = 151) and subjects with symptomatic apical periodontitis or acute apical abscess (n = 124). Subjects without AP (n = 169) and asymptomatic (n = 196) were used as controls, respectively. Saliva samples were collected as source of genomic DNA. Twelve SNPs in the TP63 gene were selected for genotyping using Taqman chemistry in real-time PCR. Data analysis was performed using PLINK software. The Bonferroni method was applied to correct for multiple testing; α ≤ 0.004 indicates significant differences between groups. RESULTS: TP63 expression was evident in apical abscesses and radicular cysts, while weaker expression was observed in periapical granulomas. Positive expression was observed in mononuclear cells in the granulation tissues of all AP lesions. Regarding the presence of AP, a trend for allelic association was observed for rs16864812 and rs9810322 (P = 0.04) and rs9810322 genotypes were also nominally associated with AP under a dominant model (P = 0.04). When considering the presence of periapical pain, a trend for allelic and genotypic association was observed for rs10155037 (P = 0.03). Haplotypes were also associated with AP and periapical pain (P ≤ 0.05). CONCLUSIONS: Apical periodontitis is a complex multifactorial condition and it is likely that multiple genes and environmental effects may influence its susceptibility, progression or both. TP63 variants may play a role in AP pathogenesis and susceptibility, individually or interactively with other genes. Additional studies in other populations and functional studies are needed to improve understanding of the role of TP63 in AP.
Assuntos
Abscesso Periapical , Granuloma Periapical , Periodontite Periapical , Cisto Radicular , Humanos , Polimorfismo de Nucleotídeo Único , Fatores de Transcrição , Proteínas Supressoras de TumorRESUMO
AIM: To investigate the DNA methylation profiles of immune response-related genes in apical periodontitis (AP) lesions. METHODOLOGY: The methylation profiles on the cytosine-phosphate-guanine (CpG) regions of 22 gene promoters involved in inflammation and autoimmunity were assessed in 60 human AP lesions and 24 healthy periodontal ligaments (controls) using a pathway-specific real-time polymerase chain reaction array (EpiTect® Methyl Signature PCR Array Human Inflammatory Response). Differentially methylated genes were subsequently assessed for their mRNA expression. Data analyses (One-way anova, Tukey's multiple comparisons tests and Mann-Whitney tests) were performed using GraphPad Prism 6 software. P values ≤ 0.05 were considered statistically significant. RESULTS: Significant DNA hypermethylation was observed for CXCL3 and FADD gene promoters in AP lesions when compared to control tissues (P < 0.001) and among other genes (P < 0.05). In contrast, IL12B and IL4R were associated with significant hypomethylation in comparison to other genes (P < 0.05). IL12B, IL4R, CXCL3 and FADD had differential mRNA expression in AP lesions and controls (P < 0.001). CONCLUSIONS: Differential methylation profiles of immune response-related genes, such as FADD, CXCL3, IL12B and IL4R, may have an influence on individual AP susceptibility and patient treatment outcomes, through their potential contributions to altered expression of disease-relevant genes. Methylation and/or genetic variations in additional genes may also contribute to the dynamics of AP development and should be considered in future studies.
Assuntos
Metilação de DNA , Periodontite Periapical/genética , Periodontite Periapical/imunologia , Periodontite Periapical/metabolismo , Transcriptoma , Adolescente , Adulto , Idoso , Autoimunidade/genética , Brasil , Quimiocinas/genética , Quimiocinas CXC/genética , Citocinas/genética , Proteína de Domínio de Morte Associada a Fas/genética , Regulação da Expressão Gênica , Humanos , Inflamação , Subunidade p40 da Interleucina-12/genética , Subunidade alfa de Receptor de Interleucina-4/genética , Pessoa de Meia-Idade , Ligamento Periodontal , Regiões Promotoras Genéticas , RNA Mensageiro/biossíntese , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Citocinas/genética , Adulto JovemRESUMO
AIM: To investigate the effects of combinations of several irrigants on the roughness and wettability of dentine, adhesion of Enterococcus faecalis and Candida albicans and adsorption of chlorhexidine (CHX) to the dentine. METHODOLOGY: Bovine dentine samples were prepared and their surface roughness standardized. The samples were distributed in groups (n = 10) and subjected to one of the following irrigation protocols: G1 - saline solution; G2 - sodium hypochlorite (NaOCl); G3 - NaOCl + ethylenediaminetetraacetic acid (EDTA); G4 - NaOCl + peracetic acid (PAA); G5 - NaOCl + 1-hydroxyethylidene-1,1-diphosphonic acid (HEDP); G6 - NaOCl + EDTA + CHX; G7 - NaOCl + PAA + CHX; G8 - NaOCl + HEDP + CHX; and G9 - mixture of NaOCl + HEDP. After treatments, roughness and wettability were measured. In order to evaluate the adhesion of microorganisms to dentine, new dentine samples were prepared and after 2 h of contact with the microorganisms, were analysed using a confocal laser scanning microscope and the number of microorganisms adhering to the surfaces were determined. Absorption spectra were collected by attenuated total reflectance of Fourier-transform infrared spectroscopy before and after immersion of other dentine samples in each solution of G6, G7 and G8 and in a solution of 2% CHX at various time intervals. The areas of the band associated with CHX with the peak at 1492 cm-1 were calculated between 1479 and 1500 cm-1 of the spectral range. The data obtained in all experiments were subjected to one-way ANOVA (α < 0.05). The values of the CHX band were also subjected to one-way repeated measures ANOVA (α < 0.05). RESULTS: Saline solution, NaOCl, HEDP and CHX did not alter the roughness of the dentine (P > 0.05), whilst EDTA and PAA did (P < 0.05). Dentine surface wettability increased after the use of all irrigants compared to saline solution (P < 0.05), with HEDP causing the greatest increases (P < 0.05). The adhesion of E. faecalis was favoured on surfaces treated with only saline solution and NaOCl, and on samples that had decalcifying agents as the final irrigant (P < 0.05). The adhesion of C. albicans was highest on surfaces treated with only saline solution and on surfaces that had NaOCl used as the last irrigant (P < 0.05). The use of CHX as a final irrigant reduced the adhesion of both microorganisms. The roughness and wettability did not influence the adhesion of the microorganisms tested. The adsorption of CHX to the dentine was significant after 1 min of immersion of the mineralized samples in the irrigant (P < 0.05), and the use of chelating agents prior to CHX potentiated this adsorption. CONCLUSIONS: The irrigation solutions had a variable effect on the properties of dentine, on the adhesion of E. faecalis and C. albicans and the adsorption of CHX to the dentine surface.
Assuntos
Aderência Bacteriana/efeitos dos fármacos , Clorexidina/farmacologia , Dentina/efeitos dos fármacos , Dentina/microbiologia , Dentina/patologia , Irrigantes do Canal Radicular/farmacologia , Adsorção/efeitos dos fármacos , Animais , Candida albicans/efeitos dos fármacos , Bovinos , Quelantes/farmacologia , Ácido Edético/farmacologia , Enterococcus faecalis/efeitos dos fármacos , Ácido Etidrônico/farmacologia , Teste de Materiais , Ácido Peracético/farmacologia , Hipoclorito de Sódio/farmacologia , Propriedades de Superfície/efeitos dos fármacosRESUMO
The chronic inflammatory immune response triggered by the infection of the tooth root canal system results in the local upregulation of RANKL, resulting in periapical bone loss. While RANKL has a well-characterized role in the control of bone homeostasis/pathology, it can play important roles in the regulation of the immune system, although its possible immunoregulatory role in infectious inflammatory osteolytic conditions remains largely unknown. Here, we used a mouse model of infectious inflammatory periapical lesions subjected to continuous or transitory anti-RANKL inhibition, followed by the analysis of lesion outcome and multiple host response parameters. Anti-RANKL administration resulted in arrest of bone loss but interfered in the natural immunoregulation of the lesions observed in the untreated group. RANKL inhibition resulted in an unremitting proinflammatory response, persistent high proinflammatory and effector CD4 response, decreased regulatory T-cell (Treg) migration, and lower levels of Treg-related cytokines IL-10 and TGFb. Anti-RANKL blockade impaired the immunoregulatory process only in early disease stages, while the late administration of anti-RANKL did not interfere with the stablished immunoregulation. The impaired immunoregulation due to RANKL inhibition is characterized by increased delayed-type hypersensitivity in vivo and T-cell proliferation in vitro to the infecting bacteria, which mimic the effects of Treg inhibition, reinforcing a possible influence of RANKL on Treg-mediated suppressive response. The adoptive transfer of CD4+FOXp3+ Tregs to mice receiving anti-RANKL therapy restored the immunoregulatory capacity, attenuating the inflammatory response in the lesions, reestablishing normal T-cell response in vivo and in vitro, and preventing lesion relapse upon anti-RANKL therapy cessation. Therefore, while RANKL inhibition efficiently limited the periapical bone loss, it promoted an unremitting host inflammatory response by interfering with Treg activity, suggesting that this classic osteoclastogenic mediator plays a role in immunoregulation.
Assuntos
Osteólise/imunologia , Doenças Periapicais/imunologia , Ligante RANK/imunologia , Linfócitos T Reguladores/imunologia , Transferência Adotiva , Perda do Osso Alveolar/imunologia , Perda do Osso Alveolar/microbiologia , Animais , Anticorpos Monoclonais/farmacologia , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Expressão Gênica , Imunidade nas Mucosas , Inflamação/imunologia , Inflamação/microbiologia , Infliximab/farmacologia , Interleucina-10/imunologia , Ativação Linfocitária/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Osteólise/microbiologia , Doenças Periapicais/microbiologia , Ligante RANK/antagonistas & inibidores , Reação em Cadeia da Polimerase em Tempo Real , Fator de Crescimento Transformador beta/imunologiaRESUMO
OBJECTIVE: Nonsyndromic cleft lip with or without cleft palate (NSCL±P) is a common craniofacial anomaly with multifactorial etiology. Evidence suggests that variations in WNT pathway genes contribute to an increased susceptibility to NSCL±P. The aim of this study was to investigate the association of AXIN1, APC, CTNNB1, DVL2, and GSK3ß gene variants with NSCL±P in a case-control data set from Brazil. PATIENTS: 471 individuals with NSCL±P and 504 unrelated control individuals of Caucasian ethnicity. DESIGN: Twenty single-nucleotide polymorphisms (SNPs) in/nearby AXIN1, APC, CTNNB1, DVL2, and GSK3B genes were genotyped using Taqman chemistry in a Viia7 sequence detection instrument. Genotype, allele, and haplotype frequencies were compared among NSCL±P patients and controls using Fisher exact test, implemented in PLINK software. The level of significance was established at P ≤.002 under Bonferroni correction. In silico analysis of SNP function was assessed using MirSNP database. RESULTS: Significant association was found between GSK3B rs13314595 genotypes and NSCL±P ( P = .0006). Additionally, nominal associations were found between DVL2 (rs35594616) and APC (rs448475) with NSCL±P ( P = .02 and P = .03, respectively). SNP haplotypes for GSK3B and APC genes showed nominal associations with NSCL±P ( P < .05). In silico analysis predicted that APC rs448475 harbors a binding site for the microRNA miR-617 and that the switch from a G allele to C allele enhances binding, whereas DVL2 rs35594616 did not appear to harbor microRNA-binding sites. CONCLUSION: This study shows for the first time the association between GSK3B and NSCL±P and confirms the role of additional WNT pathway genes as candidates for NSCL±P.
Assuntos
Fenda Labial/genética , Fissura Palatina/genética , Glicogênio Sintase Quinase 3 beta/genética , Via de Sinalização Wnt/genética , Proteína da Polipose Adenomatosa do Colo/genética , Alelos , Proteína Axina/genética , Brasil , Estudos de Casos e Controles , Proteínas Desgrenhadas/genética , Feminino , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Haplótipos , Humanos , Masculino , Polimorfismo de Nucleotídeo Único , beta Catenina/genéticaRESUMO
Tooth agenesis is a common craniofacial abnormality in humans and represents failure to develop 1 or more permanent teeth. Tooth agenesis is complex, and variations in about a dozen genes have been reported as contributing to the etiology. Here, we combined whole-exome sequencing, array-based genotyping, and linkage analysis to identify putative pathogenic variants in candidate disease genes for tooth agenesis in 10 multiplex Turkish families. Novel homozygous and heterozygous variants in LRP6, DKK1, LAMA3, and COL17A1 genes, as well as known variants in WNT10A, were identified as likely pathogenic in isolated tooth agenesis. Novel variants in KREMEN1 were identified as likely pathogenic in 2 families with suspected syndromic tooth agenesis. Variants in more than 1 gene were identified segregating with tooth agenesis in 2 families, suggesting oligogenic inheritance. Structural modeling of missense variants suggests deleterious effects to the encoded proteins. Functional analysis of an indel variant (c.3607+3_6del) in LRP6 suggested that the predicted resulting mRNA is subject to nonsense-mediated decay. Our results support a major role for WNT pathways genes in the etiology of tooth agenesis while revealing new candidate genes. Moreover, oligogenic cosegregation was suggestive for complex inheritance and potentially complex gene product interactions during development, contributing to improved understanding of the genetic etiology of familial tooth agenesis.
Assuntos
Anodontia/genética , Feminino , Ligação Genética/genética , Variação Genética/genética , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Laminina/genética , Proteína-6 Relacionada a Receptor de Lipoproteína de Baixa Densidade/genética , Masculino , Proteínas de Membrana/genética , Mutação de Sentido Incorreto/genética , Linhagem , Reação em Cadeia da Polimerase em Tempo Real , Turquia , Sequenciamento do Exoma/métodos , Proteínas Wnt/genéticaRESUMO
Evidence from biological and human studies strongly supports a role for MMP and TIMP genes as candidate genes for non-syndromic cleft lip with or without cleft palate (NSCL/P). We previously showed the association of promoter polymorphisms in MMP3 (rs3025058 and rs522616) and TIMP2 (rs8179096) with NSCL/P. In this study, we examined the functional significance of these polymorphisms. A specific DNA-protein complex for MMP3 rs522616 A was detected, and this allele by itself showed greater promoter activity than the G allele. However, the effect of rs522616 was ultimately regulated by the rs3025058 allele on the background. For TIMP2 rs8179096, the T allele showed a 2.5-fold increase in promoter activity when compared with allele C, whereas both C and T alleles were found to bind to nuclear factor kappa B. Our results provide new evidence that promoter polymorphisms in MMP3 and TIMP2 are functional and may affect gene transcription with possible effects on craniofacial development leading to NSCL/P.
Assuntos
Fenda Labial/enzimologia , Fissura Palatina/enzimologia , Metaloproteinase 3 da Matriz/genética , Polimorfismo Genético/genética , Inibidores de Proteases/análise , Inibidor Tecidual de Metaloproteinase-2/genética , Adenina , Alelos , Linhagem Celular Tumoral , Fenda Labial/genética , Fissura Palatina/genética , Citosina , Guanina , Haplótipos/genética , Humanos , NF-kappa B/genética , Regiões Promotoras Genéticas/genética , Timina , Transcrição Gênica/genéticaRESUMO
AIM: To evaluate the cytotoxic effects of triple antibiotic paste (TAP), double antibiotic paste (DAP), minocycline and calcium hydroxide and their influence on cytokine mRNA expression levels on human periodontal ligament (HPDL) fibroblasts. METHODOLOGY: Triple antibiotic paste, DAP and Ca(OH)2 test samples were immersed in culture medium and incubated at 37 °C for 24 and 48 h. HPDL cells were seeded at a density of 2 × 10(4) cells and exposed to either culture media (negative control), 0.1% SDS (positive control), 24- or 48-h elutes of each test material and incubated for 24 h. A multiparametric cytotoxicity assay kit (XTT, NR and CVDE) was used to evaluate the cytotoxic effects of each test material. Results were analysed using an ELISA plate reader and light absorbances of 450 and 530 nm as references. Cytokine mRNA expression levels in HPDL cells treated with the materials were also investigated using real-time PCR. Expression levels were calculated using the comparative 2(-ΔΔCt) method. Statistical analyses included anova followed by Bonferroni correction. RESULTS: Triple antibiotic paste and minocycline were the most cytotoxic materials when compared with DAP and Ca(OH)2 in all three (XTT, NR and CVDE) assays (P < 0.0001). No significant differences (P > 0.05) were found for cytokine gene expression levels after exposure to 24- or 48-h elutes of any of the materials except for IL6, which had significantly higher mRNA levels with the 24-h TAP elute (P < 0.01). CONCLUSION: Ca(OH)2 had a minimal effect on cell viability and cytokine production. The TAP showed deleterious effects on HPDL viability and increased expression of the pro-inflammatory cytokine IL6.
Assuntos
Antibacterianos/farmacologia , Ligamento Periodontal/efeitos dos fármacos , Células Cultivadas , Citocinas/genética , Citocinas/metabolismo , Fibroblastos/efeitos dos fármacos , Humanos , Ligamento Periodontal/citologia , Ligamento Periodontal/metabolismo , RNA Mensageiro/genéticaRESUMO
We have previously shown the association of AXIN2 with oral clefts in a US population. Here, we expanded our study to explore the association of 11 AXIN2 markers in 682 cleft families from multiple populations. Alleles for each AXIN2 marker were tested for transmission distortion with clefts by means of the Family-based Association Test. We observed an association with SNP rs7224837 and all clefts in the combined populations (p = 0.001), and with SNP rs3923086 and cleft lip and palate in Asian populations (p = 0.004). We confirmed our association findings in an additional 528 cleft families from the United States (p < 0.009). We tested for gene-gene interaction between AXIN2 and additional cleft susceptibility loci. We assessed and detected Axin2 mRNA and protein expression during murine palatogenesis. In addition, we also observed co-localization of Axin2 with Irf6 proteins, particularly in the epithelium. Our results continue to support a role for AXIN2 in the etiology of human clefting. Additional studies should be performed to improve our understanding of the biological mechanisms linking AXIN2 to oral clefts.
Assuntos
Proteína Axina/genética , Fenda Labial/genética , Fissura Palatina/genética , Animais , Povo Asiático/genética , Proteína Axina/biossíntese , China , Epistasia Genética , Europa (Continente) , Frequência do Gene , Estudo de Associação Genômica Ampla , Humanos , Índia , Fatores Reguladores de Interferon/biossíntese , Fatores Reguladores de Interferon/genética , América Latina , Desequilíbrio de Ligação , Camundongos , Palato Duro/embriologia , Polimorfismo de Nucleotídeo Único , Saliva/química , Turquia , Estados Unidos , População Branca/genéticaRESUMO
Oral squamous cell carcinoma (OSCC) accounts for more than 90% of the malignant neoplasms that arise in the mucosa of the upper aerodigestive tract. Recent studies of cleft lip/palate have shown the association of genes involved in cancer. WNT pathway genes have been associated with several types of cancer and recently with cleft lip/palate. To investigate if genes associated with cleft lip/palate were also associated with oral cancer, we genotyped 188 individuals with OSCC and 225 control individuals for markers in AXIN2, AXIN1, GSK3ß, WNT3A, WNT5A, WNT8A, WNT11, WNT3, and WNT9B. Statistical analysis was performed with PLINK 1.06 software to test for differences in allele frequencies of each polymorphism between cases and controls. We found association of SNPs in GSK3B (p = 0.0008) and WNT11 (p = 0.03) with OSCC. We also found overtransmission of GSK3B haplotypes in OSCC cases. Expression analyses showed up-regulation of WNT3A, GSK3B, and AXIN1 and down-regulation of WNT11 in OSCC in comparison with control tissues (P < 0.001). Additional studies should focus on the identification of potentially functional variants in these genes as contributors to human clefting and oral cancer.
Assuntos
Carcinoma de Células Escamosas/genética , Fenda Labial/genética , Fissura Palatina/genética , Neoplasias Bucais/genética , Proteínas Wnt/genética , Proteínas Adaptadoras de Transdução de Sinal/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteína Axina , Estudos de Casos e Controles , Feminino , Frequência do Gene , Quinase 3 da Glicogênio Sintase/genética , Glicogênio Sintase Quinase 3 beta , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas de Neoplasias/genética , Polimorfismo de Nucleotídeo Único , Proteínas Repressoras/genética , Transdução de Sinais/genética , Proteína Wnt3 , Proteína Wnt3ARESUMO
Cleft lip/palate is a defect of craniofacial development. In previous reports, chromosome 6q has been suggested as a candidate region for cleft lip/palate. A multipoint posterior probability of linkage analysis of multiplex families from the Philippines attributed an 88% probability of harboring a cleft-susceptibility gene to a narrower region on bands 6q14.2-14.3. We genotyped 2732 individuals from families and unrelated individuals with and without clefts to investigate the existence of possible cleft-susceptibility genes in this region. We found association of PRSS35 and SNAP91 genes with cleft lip/palate in the case-control cohort and in Caucasian families. Haplotype analyses support the individual associations with PRSS35. We found Prss35 expression in the head and palate of mouse embryos at critical stages for palatogenesis, whereas Snap91 was expressed in the adult brain. We provide further evidence of the involvement of chromosome 6q in cleft lip/palate and suggest PRSS35 as a novel candidate gene.
Assuntos
Cromossomos Humanos Par 6 , Fenda Labial/genética , Fissura Palatina/genética , Predisposição Genética para Doença , Serina Proteases/genética , Animais , Brasil , Estudos de Casos e Controles , Distribuição de Qui-Quadrado , Bases de Dados Genéticas , Frequência do Gene , Loci Gênicos , Haplótipos , Humanos , Desequilíbrio de Ligação , Camundongos , Proteínas Monoméricas de Montagem de Clatrina/genética , Palato Duro/embriologia , Polimorfismo de Nucleotídeo Único , Serina Endopeptidases/genética , População Branca/genéticaRESUMO
There is evidence for a genetic component in caries susceptibility, but the disease is greatly influenced by environmental factors, which are extremely difficult to control in humans. For the present study, we used DNA samples collected from 110 unrelated, non-cleft individuals older than 12 years of age from Tiquisate, Guatemala: a population with similar cultural, dietary and hygiene habits, similar access to the dentist and fluoride exposure. Forty-four individuals were designated 'very low caries experience' (DMFT < or = 2), and 66 were designated 'higher caries experience' (DMFT > or = 3). Single-nucleotide polymorphism markers were genotyped in selected candidate genes (ameloblastin, amelogenin, enamelin, tuftelin-1, and tuftelin interacting protein 11) that influence enamel formation. Having at least one copy of the rare amelogenin marker allele was associated with increased age-adjusted caries experience. This association was stronger in individuals with higher DMFT (DMFT > or = 20; p = 0.0000001). Our results suggest that variation in amelogenin may contribute to caries susceptibility in the population studied. The approach of comparing individuals with extremely distinct caries experiences could be valuable for decreasing the potential influence of environmental factors on genetic studies of caries.
Assuntos
Amelogenina/genética , Suscetibilidade à Cárie Dentária/genética , Cárie Dentária/genética , Adolescente , Adulto , Cromossomos Humanos X , Índice CPO , Feminino , Frequência do Gene , Guatemala , Humanos , Indígenas Centro-Americanos/genética , Modelos Lineares , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo ÚnicoRESUMO
Individuals with clefts present considerably more dental anomalies than do individuals without clefts. We used dental development to subphenotype clefts with the goal of identifying cleft subgroups that could have specific genetic contributions. We examined 1000 individuals, 500 with clefts and 500 without. We used several clinical features, such as cleft completeness or incompleteness, laterality, and the presence of dental anomalies to assess each individual's cleft status. We performed chi-square and Fisher's exact tests to compare the frequencies of observed anomalies between individuals with and individuals without clefts, and among individuals with different cleft subphenotypes. Agenesis of the lateral incisor on the non-cleft side was the most remarkable observation, and may suggest that such cases could be considered incomplete forms of bilateral clefts of the lip.