Placental TLR recognition of salivary and subgingival microbiota is associated with pregnancy complications.

BACKGROUND: Pre-term birth, the leading cause of neonatal mortality, has been associated with maternal periodontal disease and the presence of oral pathogens in the placenta. However, the mechanisms that underpin this link are not known. This investigation aimed to identify the origins of placental microbiota and to interrogate the association between parturition complications and immune recognition of placental microbial motifs. Video Abstract METHODS: Saliva, plaque, serum, and placenta were collected during 130 full-term (FT), pre-term (PT), or pre-term complicated by pre-eclampsia (PTPE) deliveries and subjected to whole-genome shotgun sequencing. Real-time quantitative PCR was used to measure toll-like receptors (TLR) 1-10 expression in placental samples. Source tracking was employed to trace the origins of the placental microbiota. RESULTS: We discovered 10,007 functionally annotated genes representing 420 taxa in the placenta that could not be attributed to contamination. Placental microbial composition was the biggest discriminator of pregnancy complications, outweighing hypertension, BMI, smoking, and maternal age. A machine-learning algorithm trained on this microbial dataset predicted PTPE and PT with error rates of 4.05% and 8.6% (taxonomy) and 6.21% and 7.38% (function). Logistic regression revealed 32% higher odds of parturition complication (95% CI 2.8%, 81%) for every IQR increase in the Shannon diversity index after adjusting for maternal smoking status, maternal age, and gravida. We also discovered distinct expression patterns of TLRs that detect RNA- and DNA-containing antigens in the three groups, with significant upregulation of TLR9, and concomitant downregulation of TLR7 in PTPE and PT groups, and dense correlation networks between microbial genes and these TLRs. 70-82% of placental microbiota were traced to serum and thence to the salivary and subgingival microbiomes. The oral and serum microbiomes of PTPE and PT groups displayed significant enrichment of genes encoding iron transport, exosome, adhesion, quorum sensing, lipopolysaccharide, biofilm, and steroid degradation. CONCLUSIONS: Within the limits of cross-sectional analysis, we find evidence to suggest that oral bacteria might translocate to the placenta via serum and trigger immune signaling pathways capable of inducing placental vascular pathology. This might explain, in part, the higher incidence of obstetric syndromes in women with periodontal disease.

Examining awareness of tobacco's oral health effects: Dentists' role in smoking cessation among dental patients.

INTRODUCTION: Tobacco products are well-known as a major risk factor for systemic and oral diseases. Dentists may play an important role in the prevention and progression of oral problems related to smoking. The aim of this study was to evaluate the level of awareness about the poor oral health effects of tobacco products and the role of dentists in smoking cessation among dental patients. METHODS: A survey containing 40 questions was prepared, and patients seeking dental treatment between June and October 2019 at the School of Dentistry, Ege University, were asked to participate. The survey included demographic variables in the first part, habits of using tobacco products in the second part, relations between smoking and oral health, and the possible role of dentists in smoking cessation in the last part. Data were tested statistically by Mann Whitney U and chi-squared tests. RESULTS: A total of 501 patients participated in the survey; more than half of the participants were non-smokers (63.7%). Cigarettes (95.06%), hookah (7.69%), e-cigarettes (2.75%), and cigars (1.65%) were the most frequently consumed tobacco products. The biggest obstacle to quitting smoking was 'having smoker friends'. The rate of non-smokers (41.4%) agreeing that smoking is related to periodontal diseases was more than that of smokers (32.4%) (p<0.05). The most known side effect of tobacco products was halitosis (81.6%). Half of the respondents (46.7%) did not know about dentists' role in helping them quit smoking. The rate of participants previously recommended by a dentist to quit smoking was only 36%. CONCLUSIONS: The aesthetic and social consequences of using tobacco products are well known, but smokers are substantially less aware than non-smokers of the relationship between tobacco products and oral diseases. The present findings suggest that dentists should inform their patients about the detrimental effects of tobacco products and play an active role in advising them to quit.

Clinical, oral immunological and microbiological shifts during and after pregnancy.

OBJECTIVES: Physiological changes and shifts in the oral microbiota composition during pregnancy may affect the maternal immune system. Uncomplicated pregnancy is associated with a T-helper (Th) 2 predominant cytokine regulation (anti-inflammatory), while oral health deterioration during pregnancy is reflected by severe gingival inflammation, a primarily Th1 cytokine phenotype (pro-inflammatory), and oral microbiome alterations. This prospective observational study aimed to evaluate Th cytokine shifts and changes in the oral microbiota composition in saliva of women before and after birth. MATERIAL AND METHODS: Saliva (n = 96) was collected before and 6 months after birth, and medical, oral health, and periodontal status were assessed. In a multiplex immunoassay, 10 cytokines were simultaneously analyzed and cumulative Th1 and Th2 cytokine levels and Th1/Th2 ratio were calculated for all groups. Putative periodontal pathogens (n = 6) were evaluated by quantitative real-time polymerase chain reaction. RESULTS: Th2 cytokine levels were significantly lower (p = 0.014) while pro-inflammatory cytokine levels were significantly higher (p < 0.01) during pregnancy than postpartum. Similar Th1 levels were found between the groups (p = 0.143). Th1 and Th2 cytokines positively correlated with periodontal parameters (p < 0.001) and levels of studied bacteria during pregnancy (p < 0.05). CONCLUSIONS: This study identified a significantly increased Th1/Th2 cytokine ratio during pregnancy and a positive association with putative periodontal pathogens. This immunological and microbiological deregulation in the oral milieu during pregnancy is suggestive of a destructive inflammatory periodontal profile. STUDY REGISTRATION: Clinical Trials.gov (Record BAP-2015). CLINICAL RELEVANCE: Understanding altered oral immunological and microbiological regulation patterns during pregnancy may help improve the inflammatory periodontal profile in pregnant women.

Regulation of matrix metalloproteinases-8, -9 and endogenous tissue inhibitor-1 in oral biofluids during pregnancy and postpartum.

OBJECTIVE: During pregnancy, mothers undergoe considerable physiological changes affecting the whole body including periodontal tissues. Susceptibility to gingival inflammation during pregnancy could be mediated by modulation of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs). Therefore, the aim of this study was to investigate salivary and gingival crevicular fluid (GCF) levels of MMPs and TIMPs during the second and third trimester of pregnancy and postpartum. DESIGN: Saliva and GCF samples were collected from 96 pregnant women (PW) before and after giving birth. The sixty matched non-pregnant women (N-PW) were recruited as a control group and full-mouth periodontal examination was performed. The levels of MMP-8, MMP-9 and TIMP-1 were determined by immunofluorometric and enzyme-linked immunosorbent assays. RESULTS: The PW group exhibited significantly higher levels of MMP-8 and MMP-9 in their saliva than the N-PW group while corresponding salivary TIMP-1 levels were significantly lower in NPW compared to the postpartum stage. This resulted in significantly higher MMP-8/TIMP-1 and MMP-9/TIMP-1ratio in the saliva from PW before and after birth than in that from N-PW. MMP-8, MMP-9 and TIMP-1 levels were higher in GCF from PW and postpartum than in that from N-PW. CONCLUSIONS: MMP-8 and MMP-9 levels in saliva and GCF reflect inflammatory burden during pregnancy. They could be used for monitoring the inflammatory state of gingival tissues during pregnancy.

Cholinergic signalling mechanisms and early implant healing phases in healthy versus generalized aggressive periodontitis patients: A prospective, case-control study.

AIMS: Periodontal diseases negatively affect implant osseointegration. Perturbations in non-neuronal cholinergic signalling mechanisms are associated with periodontitis; however, their role in generalized aggressive periodontitis (GAgP) is unknown. The aim of this prospective case-control study was to determine the relationship between non-neuronal cholinergic signalling mechanisms, secreted Ly-6/uPAR-related protein-1 (SLURP-1), interleukin-17 (IL-17) family cytokines and healing of dental implants in health and GAgP. MATERIAL AND METHODS: Thirteen GAgP patients and seven periodontally healthy individuals (PH) were recruited. Peri-implant crevicular fluid (PICF) was obtained at baseline and 1 month post-placement. Acetylcholine (ACh) levels and cholinesterase activity were determined biochemically. SLURP-1, IL-17A and IL-17E levels were determined by ELISA. Marginal bone loss (MBL) at 1 and 6 months post-placement was determined radiographically. RESULTS: The concentration of ACh, cholinesterase activity and IL-17A levels was elevated in PICF of patients with GAgP compared to PH individuals at baseline and 1 month post-placement. The concentration of ACh and cholinesterase activity levels in PICF correlated with levels of IL-17A and MBL around implants 1 month post-placement in patients with GAgP. CONCLUSIONS: Non-neuronal cholinergic mechanisms may play a role in the aetiopathogenesis of GAgP and may directly or indirectly, through modulation of IL-17A, influence early implant osseointegration and potential long-term implant survival.

Altered levels of inhibitory cytokines in patients with thalassemia major and gingival inflammation / Níveis alterados de citocinas inibitórias em pacientes com talassemia major e inflamação gengival

Objective: To evaluate local and systemic levels of interleukin-10 (IL-10), IL-33, and tumor necrosis factor alpha (TNF-α) in Thalassemia major (TM) in the presence of gingival inflammation. Material and Methods: 58 patients (TM, n=29 and systemically healthy controls, n=29) were included to the study. IL-10, IL-33, and TNF-α levels were evaluated in gingival crevicular fluid (GCF), saliva and serum. Clinical periodontal measurements were recorded. Results: GCF IL-33 total amounts in TM and gingivitis group were elevated compared to systemically and periodontally healthy group (p=0.01). GCF IL-10, IL33 and TNF-α concentrations were higher in TM and periodontally healthy group than the systemically healthy and gingivitis group (p=0.02, p=0.008, p=0.003). Serum IL-10 levels were elevated in TM and gingivitis compared to the systemically healthy and gingivitis (p=0.0009) and systemically and periodontally healthy (p=0.0007) groups. Serum IL-10 and TNF-α levels in TM and periodontally healthy group were higher than systemically and periodontally healthy group (p=0.01 and p=0.02). Conclusion: TM may potentially alter circulating levels of IL-33 and IL-10 and therefore, may affect the degree of periodontal inflammation locally or vice versa. Yet, the underlying mechanism linking the hematologic condition is not clear and deserves further investigation. (AU)

Objetivo: Avaliar os níveis locais e sistêmicos de interleucina-10 (IL-10), IL-33 e fator de necrose tumoral alfa (TNF-α) na Talassemia Major (TM) na presença de inflamação gengival. Material e Métodos: 58 pacientes (TM, n = 29 e controles sistemicamente saudáveis, n = 29) foram incluídos no estudo. Os níveis de IL-10, IL-33 e TNF-α foram avaliados em fluido crevicular gengival (FCG), saliva e soro. As medições periodontais clínicas foram registradas. Resultados: As quantidades totais de IL-33 no FCG do grupo de TM e gengivite foram elevadas em comparação com o grupo sistemicamente e periodontalmente saudável (p = 0,01). As concentrações de IL-10 , IL-33 e TNF-α do FCG foram maiores no grupo TM e periodontalmente saudáveis do que no grupo sistemicamente saudável e gengivite (p = 0,02, p = 0,008, p = 0,003). Os níveis séricos de IL-10 estavam elevados na TM e gengivite em comparação com os grupos sistemicamente saudável e gengivite (p = 0,0009) e sistemicamente e periodontalmente saudáveis (p = 0,0007). Os níveis séricos de IL-10 e TNF-α no grupo TM e periodontalmente saudáveis foram maiores do que os grupos sistemicamente e periodontalmente saudáveis (p = 0,01 ep = 0,02). Conclusão: A TM pode alterar potencialmente os níveis circulantes de IL-33 e IL-10 e, portanto, pode afetar o grau de inflamação periodontal localmente ou vice-versa. No entanto, o mecanismo subjacente que liga a condição hematológica não é claro e merece uma investigação mais aprofundada. (AU)

Salivary Adiponectin and Leptin Levels are Increased in Women with Gestational Diabetes Mellitus and Gingival Inflammation.

PURPOSE: Gestational diabetes mellitus (GDM) is defined as glucose intolerance with first onset or diagnosis in pregnancy. This study evaluated clinical and biochemical parameters in a possible association between GDM and gingival inflammation. MATERIALS AND METHODS: A total of 87 pregnant women - 44 with GDM and 43 without (NGDM) - were included. Subgroups were created according to gingival inflammation. Plaque index (PI), bleeding on probing (BOP), and probing depth (PD) were recorded. RESULTS: Age, anthropometric variables and baby weight (g) were all statistically significantly higher in the GDM group (p < 0.0001). Systolic and diastolic blood pressure (mmHg), saliva, serum leptin and adiponectin levels were similar in the GDM and NGDM groups (p = 0.605, p = 0.662, p = 0.737, and p = 0.596, respectively). Salivary adiponectin levels were statistically significantly higher in the two subgroups with gingivitis compared to those with clinically healthy periodontium (p < 0.01). Serum adiponectin levels were statistically significantly higher in the NGDM subgroup with gingivitis than the NGDM group with clinically healthy periodontium (p < 0.05). Statistically significant positive correlations were found between PD, PI, BOP and saliva adiponectin levels in the GDM group (p < 0.05). Positive correlations were also found between clinical periodontal parameters and saliva, serum levels of adiponectin in the control group without GDM (p < 0.05). CONCLUSION: The higher salivary adiponectin levels in the gingivitis groups suggest that gingival inflammation is more likely to influence local inflammatory parameters both in the presence and absence of GDM. Further larger-scale studies are required to better clarify the possible interactions between gingival inflammation and GDM.

Diabetes-related molecular signatures in infrared spectra of human saliva.

BACKGROUND: There is an ongoing need for improvements in non-invasive, point-of-care tools for the diagnosis and prognosis of diabetes mellitus. Ideally, such technologies would allow for community screening. METHODS: In this study, we employed infrared spectroscopy as a novel diagnostic tool in the prediction of diabetic status by analyzing the molecular and sub-molecular spectral signatures of saliva collected from subjects with diabetes (n = 39) and healthy controls (n = 22). RESULTS: Spectral analysis revealed differences in several major metabolic components - lipid, proteins, glucose, thiocyanate and carboxylate - that clearly demarcate healthy and diseased saliva. The overall accuracy for the diagnosis of diabetes based on infrared spectroscopy was 100% on the training set and 88.2% on the validation set. Therefore, we have established that infrared spectroscopy can be used to generate complex biochemical profiles in saliva and identify several potential diabetes-associated spectral features. CONCLUSIONS: Infrared spectroscopy may represent an appropriate tool with which to identify novel diseases mechanisms, risk factors for diabetic complications and markers of therapeutic efficacy. Further study into the potential utility of infrared spectroscopy as diagnostic and prognostic tool for diabetes is warranted.