Kinetics of neutrophil extracellular traps and cytokines in oral mucositis and Candida infection.

OBJECTIVE: This study investigated the concentrations of neutrophil extracellular traps (NET) and salivary cytokines (IL-1ß, IL-6, IL-8/CXCL8, TNF, and TGF-ß1) in patients undergoing chemotherapy and their associations with oral mucositis (OM) and Candida infection. MATERIALS AND METHODS: This prospective longitudinal study performed at a Brazilian service included 60 adults diagnosed with hematolymphoid diseases. Saliva samples were collected on days D0, D3, D10, and D15. Cytokines were analyzed by ELISA and NET formation by identification of the myeloperoxidase-DNA complex. Oral Candida spp. was cultured. RESULTS: OM occurred in 43.3% of patients and oral candidiasis in 20%. However, 66% of individuals had positive cultures for C. albicans. Higher concentrations of IL-6, IL-8/CXCL8, and TNF and lower concentrations of TGF-ß1 were observed in patients with OM. C. albicans infection contributed to the increase in IL-8/CXCL8, TGF-ß1, and TNF. Individuals with OM or with oral candidiasis had significant reductions in NET formation. In contrast, individuals with C. albicans and with concomitant C. albicans and OM exhibited higher NET formation. CONCLUSION: The kinetics of cytokine levels and NET formation in chemotherapy-induced OM appears to be altered by Candida infection, even in the absence of clinical signs of oral candidiasis.

Clinical, immunological, and microbiological analysis of the association between periodontitis and COVID-19: a case-control study.

PURPOSE: Periodontitis and coronavirus disease (COVID-19) share risk factors and activate similar immunopathological pathways, intensifying systemic inflammation. This study investigated the clinical, immunological and microbiological parameters in individuals with COVID-19 and controls, exploring whether periodontitis-driven inflammation contributes to worsening COVID-19 endpoints. METHODS: Case (positive RT-PCR for SARS-CoV-2) and control (negative RT-PCR) individuals underwent clinical and periodontal assessments. Salivary levels of TNF-α, IL-6, IL-1ß, IL-10, OPG, RANKL, neutrophil extracellular traps, and subgingival biofilm were analyzed at two timepoints. Data on COVID-19-related outcomes and comorbidity information were evaluated from medical records. RESULTS: Ninety-nine cases of COVID-19 and 182 controls were included for analysis. Periodontitis was associated with more hospitalization (p = 0.009), more days in the intensive care unit (ICU) (p = 0.042), admission to the semi-ICU (p = 0.047), and greater need for oxygen therapy (p = 0.042). After adjustment for confounders, periodontitis resulted in a 1.13-fold increase in the chance of hospitalization. Salivary IL-6 levels (p = 0.010) were increased in individuals with COVID-19 and periodontitis. Periodontitis was associated with increased RANKL and IL-1ß after COVID-19. No significant changes were observed in the bacterial loads of the periodontopathogens Porphyromona gingivalis, Aggregatibacter actinomycetemcomitans, Tanerella forsythia, and Treponema denticola. CONCLUSIONS: Periodontitis was associated with worse COVID-19 outcomes, suggesting the relevance of periodontal care to reduce the burden of overall inflammation. Understanding the crosstalk between SARS-CoV-2 infection and chronic conditions such as periodontitis that can influence disease outcome is important to potentially prevent complications of COVID-19.

Influence of anxiety/depression on chemotherapy-induced oral mucositis and related quality of life: A prospective cohort study.

OBJECTIVE: The impact of anxiety and depression on chemotherapy-induced oral mucositis has not been extensively explored in the literature. The aim of the present study was to evaluate anxiety/depressive symptoms, health-related quality of life, and oral health-related quality of life and their association with oral mucositis among individuals receiving chemotherapy. METHODS: This is a prospective longitudinal study carried out at a Brazilian referral service. The Hospital Anxiety and Depression Scale (HADS), World Health Organization Quality of Life-BREF (WHOQOL-BREF), and Oral Health Impact Profile questionnaire (OHIP-14) were applied at D0 (before chemotherapy) and D15 of chemotherapy. Clinicodemographic data and oral mucositis severity scores were evaluated. RESULTS: A total of 37 individuals (median age: 49 years) were included in the study. Nearly 38% of patients developed chemotherapy-induced oral mucositis and had higher anxiety/depression scores at baseline. Oral mucositis had a negative impact on oral health-related quality of life regarding functional limitation, physical pain, physical disability, and handicap. CONCLUSION: Anxiety/depressive symptoms are associated with oral mucositis that affects overall health and oral health-related quality of life.

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

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

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

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

Periodontite associada à artrite reumatoide: o papel das redes extracelulares de neutrófilos (NETs) / Periodontitis associated with rheumatoid arthritis: the role of neutrophil extracellular traps (NETs)

Introdução: As redes extracelulares de neutrófilos (NETs) são apontadas como um dos mecanismos relevantes na patogênese da periodontite e artrite reumatoide (AR). No entanto, permanece pouco compreendido a participação das NETs como mecanismo de ligação entre as duas doenças. Objetivos: 1) Investigar a concentração das NETs na saliva, no plasma e in vitro em indivíduos com AR e controles saudáveis e a associação com a periodontite e atividade da AR; 2) Avaliar o impacto do tratamento periodontal não cirúrgico na concentração das NETs na saliva e no plasma; 3) Investigar a associação entre a presença de polimorfismos de nucleotídeo único no gene codificador da enzima peptidil arginina deaminase 4 (PAD4) com a AR, a produção de NETs in vitro e a periodontite; 4) Sistematizar as evidências disponíveis na literatura sobre o efeito do tratamento periodontal não cirúrgico sobre os principais parâmetros clínicos e laboratoriais da AR e score de atividade da doença 28 (DAS28). Material e Métodos: Para atender aos objetivos 1 e 2, a concentração de NETs na saliva, no plasma e na cultura de neutrófilos isolados do sangue periférico foi determinada por meio da identificação do complexo mieloperoxidase (MPO)-DNA com o uso do kit PicoGreen®. Para atender ao objetivo 3 foi realizada a extração do DNA genômico das células mononucleares do sangue periférico de indivíduos com AR e controles e foi realizada a genotipagem para os polimorfismos de nucleotídeo único PADI4_89, PADI4_90, PADI4_92 e PADI_104. Para atender ao objetivo 4 foi realizada uma overview incluindo revisões sistemáticas que avaliaram o efeito do tratamento periodontal não cirúrgico sobre os parâmetros da AR. A busca foi realizada nas principais bases de dados, sem restrição de idioma ou data de publicação. Foi realizada ainda uma meta-análise incluindo dados dos estudos primários identificados nas revisões sistemáticas analisadas. Resultados: 1) e 2) Indivíduos com AR e com periodontite apresentaram maior concentração de NETs na saliva, no plasma e in vitro. O tratamento periodontal não cirúrgico reduziu a concentração de NETs na saliva e plasma de indivíduos com AR. 3) Não foi observada associação entre a presença de genótipos polimórficos e a AR. A presença de um haplótipo homozigoto para o polimorfismo foi associada a uma maior produção de NETs in vitro e piores parâmetros periodontais. 4) Foram incluídas na overview nove revisões sistemáticas. Os principais desfechos avaliados foram DAS28; proteína C-Reativa e/ou velocidade de hemossedimentação. A meta-análise mostrou que o tratamento periodontal não cirúrgico resultou em diminuição significativa do DAS28. Conclusão: A concentração das NETs na saliva, no plasma e na cultura de neutrófilos de sangue periférico está associada a AR e a periodontite, podendo representar o elo entre as duas doenças. O tratamento periodontal não cirúrgico leva à redução da atividade da AR. Polimorfismos no gene PADI4 estão associados a produção de NETs in vitro e à presença de periodontite.

Introduction: Neutrophil extracellular traps (NETs) are recognized as one of the relevant mechanisms in the pathogenesis of periodontitis and rheumatoid arthritis (RA). However, the participation of NETs as a linking mechanism between the two diseases remains poorly understood. Objectives: 1) To investigate the concentration of NETs in saliva, plasma, and in vitro in individuals with RA and healthy controls and the association of NETs with periodontitis and RA activity; 2) To evaluate the impact of non-surgical periodontal treatment on the concentration of NETs in saliva and plasma; 3) To investigate the association between the presence of single nucleotide polymorphisms in the gene coding for the enzyme peptidyl arginine deaminase 4 (PAD4) with RA, in vitro production of NETs, and periodontitis; 4) To systematize the evidence available in the literature on the effect of non-surgical periodontal treatment on the main clinical and laboratory parameters of RA and disease activity score 28 (DAS28). Material and Methods: To accomplish Objectives 1 and 2, the concentration of NETs in saliva, plasma, and culture of neutrophils isolated from peripheral blood was determined by identifying the myeloperoxidase (MPO)-DNA complex using the PicoGreen kit®. To accomplish Objective 3, genomic DNA was extracted from peripheral blood mononuclear cells of individuals with RA and healthy controls, and genotyping was performed for single nucleotide polymorphisms PADI4_89, PADI4_90, PADI4_92, and PADI_104. To accomplish the Objective 4, an overview, including systematic reviews that evaluated the effect of non-surgical periodontal treatment on RA parameters, was performed. The search was carried out in the main databases, with no restriction on language or date of publication. A meta- analysis, including data from the primary studies identified in the analyzed systematic reviews, was also performed. Results: For Objectives 1 and 2, individuals with RA and periodontitis showed a higher concentration of NETs in saliva, plasma, and in vitro. Non-surgical periodontal treatment reduced the concentration of NETs in saliva and plasma of individuals with RA. For Objective 3, no association between the presence of polymorphic genotypes and RA was observed. The presence of a homozygous haplotype for the polymorphism was associated with a higher production of NETs in vitro and worse periodontal parameters. For Objective 4, nine systematic reviews were included in the overview. The main outcomes evaluated were DAS28, C-Reactive protein, and/or erythrocyte sedimentation rate. The meta- analysis showed that non-surgical periodontal treatment resulted in a significant decrease in DAS28. Conclusion: The concentration of NETs in saliva, plasma and culture of peripheral blood neutrophils is associated with RA and periodontitis and may represent the link between the two diseases. Non-surgical periodontal treatment leads to reduced RA activity. Polymorphisms in the PADI4 gene are associated with the in vitro production of NETs and with presence of periodontitis.

Effects of high-dose bisphenol A on the mouse oral mucosa: A possible link with oral cancers.

Bisphenol A (BPA) is an endocrine disrupting chemical able to promote hormone-responsive tumors. The major route of BPA contamination being oral, the aim of the present study was to investigate BPA effects on oral cells. Here, we evaluated the impact of sub-chronic in vivo exposure to BPA and its in vitro effects on neoplastic and non-neoplastic oral cells. We evaluated the oral mucosa of mice chronically exposed to BPA (200 mg/L). The response of keratinocytes (NOK-SI) and Head and Neck (HN) Squamous Cell Carcinoma (SCC), HN12 and HN13 cell lines to BPA was examined. In vivo, BPA accumulated in oral tissues and caused an increase in epithelial proliferative activity. BPA disrupted the function of keratinocytes by altering pro-survival and proliferative pathways and the secretion of cytokines and growth factors. In tumor cells, BPA induced proliferative, invasive, pro-angiogenic, and epigenetic paths. Our data highlight the harmful effects of BPA on oral mucosa and, tumorigenic and non-tumorigenic cells. Additionally, BPA may be a modifier of oral cancer cell behavior by prompting a functional shift to a more aggressive phenotype.

Association of oral mucosa hyperpigmentation with imatinib mesylate use: a cross-sectional study and a systematic literature review.

OBJECTIVES: To assess the association between oral mucosa hyperpigmentation in patients with leukemia and imatinib mesylate use. Additionally, we compared our data to those obtained from a systematic review. MATERIALS AND METHODS: A cross-sectional study was conducted with 74 patients undergoing treatment with imatinib mesylate. Sociodemographic characteristics, oral mucosa alterations, and medical history were evaluated. Oral hyperpigmentation was scored. The use of imatinib mesylate and hydroxyurea was evaluated. Association between oral hyperpigmentation and imatinib mesylate was assessed. A systematic review was also conducted to retrieve case reports or case series of patients with oral hyperpigmentation associated with imatinib mesylate. RESULTS: Among the 74 participants, 41 were male (55.4%) and 33 were female (44.6%). Participants' mean age was 49.3 years. Sixty-six (89.2%) patients developed hyperpigmented lesions in the hard palate mucosa. In multivariate analysis, patients who had used imatinib mesylate for > 72 months had a hyperpigmentation score 1.62 times higher than those who had used this medication during a shorter period. Patients who had used hydroxyurea for > 30 days had a hyperpigmentation score 1.43 times higher than those who had used this medication during a shorter period. The systematic review retrieved 20 clinical cases of patients undergoing imatinib mesylate treatment and exhibiting oral hyperpigmentation. CONCLUSIONS: The development of oral hyperpigmentation is associated with imatinib mesylate use. Hydroxyurea seems to increment such an association. CLINICAL RELEVANCE: To assist providers in the differential diagnosis of hyperpigmented lesions associated with imatinib mesylate, as well as in the clinical management of such lesions.

Bleeding assessment in oral surgery: A cohort study comparing individuals on anticoagulant therapy and a non-anticoagulated group.

Some prospective studies have been designed specifically to investigate perioperative bleeding in dental surgery. The quantitative assessment of intraoperative blood loss can be useful for indicating the real risk of bleeding complications, especially in medically compromised individuals. The aim of this study was to evaluate the pattern of bleeding in individuals under vitamin K antagonist (VKA) therapy and non-anticoagulated individuals submitted to dental extractions. Perioperative bleeding was evaluated by using a total collected bleeding corrected by absorbance reading (dental bleeding score). 138 procedures were performed. When the perioperative dental bleeding score was correlated with the number of extracted teeth, the quantity of bleeding was found to be directly proportional to the procedure. Extractions of two or more teeth presented higher scores than single extractions (p = 0.003). In a comparative analysis between the VKA and non-anticoagulated groups, no significant difference in the scores was found. The previous history of complications in dental procedures (p = 0.001) and the use of additional hemostatic measures were higher in the VKA group (p = 0.017). VKA therapy did not impact significantly the volume of blood lost during dental extractions. Perioperative bleeding assessment might be a useful parameter for evaluating patients under antithrombotic treatment.