Proteome and Microbiome Mapping of Human Gingival Tissue in Health and Disease.

Efforts to map gingival tissue proteomes and microbiomes have been hampered by lack of sufficient tissue extraction methods. The pressure cycling technology (PCT) is an emerging platform for reproducible tissue homogenisation and improved sequence retrieval coverage. Therefore, we employed PCT to characterise the proteome and microbiome profiles in healthy and diseased gingival tissue. Healthy and diseased contralateral gingival tissue samples (total n = 10) were collected from five systemically healthy individuals (51.6 ± 4.3 years) with generalised chronic periodontitis. The tissues were then lysed and digested using a Barocycler, proteins were prepared and submitted for mass spectrometric analysis and microbiome DNA for 16S rRNA profiling analysis. Overall, 1,366 human proteins were quantified (false discovery rate 0.22%), of which 69 proteins were differentially expressed (≥2 peptides and p < 0.05, 62 up, 7 down) in periodontally diseased sites, compared to healthy sites. These were primarily extracellular or vesicle-associated proteins, with functions in molecular transport. On the microbiome level, 362 species-level operational taxonomic units were identified. Of those, 14 predominant species accounted for >80% of the total relative abundance, whereas 11 proved to be significantly different between healthy and diseased sites. Among them, Treponema sp. HMT253 and Fusobacterium naviforme and were associated with disease sites and strongly interacted (r > 0.7) with 30 and 6 up-regulated proteins, respectively. Healthy-site associated strains Streptococcus vestibularis, Veillonella dispar, Selenomonas sp. HMT478 and Leptotrichia sp. HMT417 showed strong negative interactions (r < -0.7) with 31, 21, 9, and 18 up-regulated proteins, respectively. In contrast the down-regulated proteins did not show strong interactions with the regulated bacteria. The present study identified the proteomic and intra-tissue microbiome profile of human gingiva by employing a PCT-assisted workflow. This is the first report demonstrating the feasibility to analyse full proteome profiles of gingival tissues in both healthy and disease sites, while deciphering the tissue site-specific microbiome signatures.

Coronally advanced flap with connective tissue graft or xenogeneic acellular dermal matrix in the treatment of multiple gingival recessions: A split-mouth randomized clinical trial.

OBJECTIVE: To evaluate the clinical efficacy of xenogeneic acellular dermal matrix (XADM) or connective tissue graft (CTG) combined with modified-coronally advanced flap (M-CAF) in the treatment of multiple gingival recessions. MATERIALS AND METHODS: Twelve participants with bilateral MGRs (multiple gingival recession) (82 gingival recessions) randomly received XADM (test group, 41 teeth) on one side and subepithelial CTG (control group, 41 teeth) on the other side in conjunction with M-CAF in the same session and completed the 18-months study period. Recession depth (RD), recession width (RW), keratinized tissue width (KTW), probing depth (PD), and clinical attachment level (CAL) were recorded at baseline, and 6-, 18-months postoperatively. RESULTS: PD was significantly higher in the test group at 18-months (P < .05). PD in the test group was also significantly higher at 6- and 18-months compared to baseline (P < .05). RD and RW were significantly lower at 6- and 18-months compared to baseline in both groups (P < .05) and both parameters were significantly higher in the test group at 18-months (P < .05). Percentage of teeth with complete root coverage in the test and control groups were similar at 6-months (78% and 70.7%, respectively) and at 18-months (both 87.8%) (P > .05). CONCLUSION: Within the limits of the study, M-CAF combined with XADM or CTG seems to be similarly effective in RD reduction of class I and II MGRs at least in the short term. Soft tissue shrinkage and increase in PD may be observed with XADM, while; CTG seems to provide stable clinical outcomes for 18-months follow-up. CLINICAL SIGNIFICANCE: Even though the CTG and XADM in conjunction with M-CAF may provide similar RD reduction in class I and II multiple gingival recessions in the short term. CTGs may be superior in terms of soft tissue shrinkage and PD values.

Annexin-1 as a salivary biomarker for gingivitis during pregnancy.

BACKGROUND: It is well established that there is higher susceptibility to gingival inflammation during pregnancy. Annexin-1 (ANXA1) is an anti-inflammatory protein which has been identified in gingival tissue exudates by discovery proteomics. This cross-sectional case-control study investigated the levels and association of ANXA1 and pro-inflammatory mediator interleukin (IL)-1ß in the saliva of pregnant and non-pregnant women. METHODS: Whole unstimulated saliva from 69 non-pregnant and 78 pregnant women was collected prior to measurement of probing depth, clinical attachment level, bleeding on probing, and plaque. Then, the women were split into 3 subgroups depending on their periodontal status (healthy, gingivitis, and periodontitis). The levels of ANXA1 and IL-1ß were measured with enzyme-linked immunosorbent assay and reported as pg/mg after normalizing against the total protein levels. RESULTS: Significantly higher ANXA1 levels were exhibited in pregnant women with gingivitis compared with health (P < 0.05) and in pregnant women with gingivitis compared with the respective non-pregnant group (P < 0.0001). There was a significantly higher level of IL-1ß in gingivitis than in health in pregnant women (P < 0.05) and significantly higher levels in periodontitis compared with health in non-pregnant women (P < 0.05). Looking at the IL-1 ß:ANXA1 ratio, the non-pregnant periodontitis group displayed a significantly higher ratio compared with the respective pregnant group (P < 0.05). In the non-pregnant subpopulation, the ratio was significantly higher in periodontitis compared with health (P < 0.01). CONCLUSION: Salivary ANXA1 levels are elevated in the presence of gingivitis only in pregnant, but not non-pregnant women, rendering this molecule as a potential salivary biomarker for non-invasive early screening for gingival inflammation during pregnancy.

Saliva, Serum Levels of Interleukin-21, -33 and Prostaglandin E2 in Patients with Generalised Aggressive or Chronic Periodontitis.

PURPOSE: This cross-sectional study aims to evaluate saliva, serum levels of interleukin-21 (IL-21), IL-33, and prostaglandin E2 (PGE2) in patients with generalised chronic periodontitis or aggressive periodontitis. MATERIALS AND METHODS: Before initiation of any periodontal treatment, saliva and serum samples were collected and clinical periodontal measurements were recorded from 94 participants (25 aggressive periodontitis patients, 25 chronic periodontitis patients, 44 periodontally healthy individuals). IL-21, IL-33 and PGE2 levels in serum and saliva samples were determined by ELISA. Data were tested statistically using Kruskal-Wallis, Mann-Whitney U-, and Spearman-rho rank tests. RESULTS: Saliva IL-33 levels were statistically significantly higher in the chronic than the aggressive group (p < 0.05). Serum IL-33, saliva and serum IL-21 and PGE2 levels were similar in the two periodontitis groups. Saliva IL-33 levels correlated with age in the chronic periodontitis group (p < 0.05). Statistically significant positive correlations were found between serum, saliva PGE2 levels and plaque index (p < 0.05). IL-33 and IL-21 levels in serum samples positively correlated in the periodontitis groups (p < 0.05). CONCLUSION: IL-21 and PGE2 analysis did not exhibit discriminating data between generalised chronic and aggressive periodontitis, but the present findings support the role of these cytokines in periodontitis. Statistically significantly higher saliva IL-33 levels in the chronic periodontitis group warrant further research.

Gingival Inflammation and Salivary or Serum Granulocyte-Secreted Enzymes in Patients With Polycystic Ovary Syndrome.

BACKGROUND: The objective of this cross-sectional study is to investigate levels of salivary and serum matrix metalloproteinase (MMP)-9, myeloperoxidase (MPO), neutrophil elastase (NE), and MMP-9/tissue inhibitor of MMP-1 (TIMP)-1 ratio in patients with polycystic ovary syndrome (PCOS) and systemically healthy controls in the presence or absence of gingivitis. METHODS: Serum and salivary levels of these biomarkers were evaluated in the following: 1) periodontally healthy women with PCOS (n = 45); 2) women with PCOS and gingivitis (n = 35); 3) systemically and periodontally healthy women (n = 25); and 4) systemically healthy women with gingivitis (n = 20). Enzyme-linked immunosorbent assay was used to determine levels of these biomarkers. A full-mouth clinical periodontal evaluation was performed for each patient. RESULTS: Salivary MMP-9 and NE levels, as well as MMP-9/TIMP-1 ratios, were higher in the systemically healthy women with gingivitis compared with periodontally healthy women with PCOS (P <0.001; P <0.01; and P <0.0001, respectively). Serum MMP-9 and MPO levels were higher in women with PCOS and gingivitis compared with periodontally healthy women with PCOS (P <0.05). Serum MMP-9 levels were lower in healthy women with gingivitis than systemically and periodontally healthy women or women with PCOS and gingivitis (P <0.05). PCOS groups exhibited a positive correlation among clinical periodontal parameters and serum MMP-9 levels or salivary MPO, NE levels, and MMP-9/MMP-1 ratio. Correlation was negative among clinical periodontal parameters and serum MMP-9 levels and MMP-9/TIMP-1 ratio in systemically healthy patients (P <0.05). CONCLUSIONS: The present findings emphasize that PCOS and gingival inflammation are associated with each other, as evidenced by salivary and serum levels of neutrophilic enzymes. This interaction may contribute to the perturbation of ovarian remodeling in PCOS.

Biomarkers and Bacteria Around Implants and Natural Teeth in the Same Individuals.

BACKGROUND: This cross-sectional study assesses cytokine levels in peri-implant crevicular fluid (PICF)/gingival crevicular fluid (GCF) and a selection of subgingival/submucosal plaque bacteria from clinically healthy or diseased sites in the same individuals. METHODS: Samples from 97 implants/teeth (58 implants [19 healthy, 20 mucositis, 19 peri-implantitis] and 39 natural teeth [19 healthy, 12 gingivitis, eight periodontitis] in 15 systemically healthy patients were investigated by immunoassay and real-time polymerase chain reaction. Samples were obtained first, with probing depth, clinical attachment level, bleeding on probing, plaque index scores, and keratinized tissue width then recorded. Data were analyzed by Wilcoxon, Mann-Whitney U, and permutation tests on dependent, independent, and mixed dependent and independent samples and Spearman correlation. RESULTS: Interleukin (IL)-1ß levels were significantly higher in PICF samples of healthy implants than in GCF samples of healthy teeth (P = 0.003), and soluble receptor activator of nuclear factor-κB ligand (sRANKL) concentrations were significantly higher in the gingivitis than the mucositis group (P = 0.004). Biomarker levels were similar in peri-implantitis and periodontitis groups (P >0.05). Actinomyces naeslundi and Streptococcus oralis levels were significantly higher in the healthy implant group than in healthy teeth (P <0.05). Prevotella intermedia and Treponema denticola (Td) levels were lower in the mucositis group than the gingivitis group (P <0.05). Prevotella oralis and S. oralis levels were significantly higher in the periodontitis group (P <0.05), and Td levels were significantly higher in the peri-implantitis group (P <0.05). CONCLUSION: There were many similarities but, crucially, some differences in biomarker levels (IL-1ß and sRANKL) and bacterial species between peri-implant and periodontal sites in the same individuals, suggesting similar pathogenic mechanisms.

Proteolytic Mediators in Gestational Diabetes Mellitus and Gingivitis.

BACKGROUND: This study evaluates levels of matrix metalloproteinase (MMP)-8, MMP-9, and tissue inhibitor of MP-1 (TIMP-1) in biofluids of women with gestational diabetes mellitus (GDM) and systemically healthy counterparts with different statuses of periodontal health. METHODS: Seventy-one women with GDM and gingivitis (Gg), 30 women with GDM and healthy periodontium (Gh), 28 systemically and periodontally healthy women (Hh), and 37 systemically healthy women with gingivitis (Hg) were evaluated. MMP-8, MMP-9, and TIMP-1 levels were determined in gingival crevicular fluid (GCF), saliva, and serum by immunofluorometric and enzyme-linked immunosorbent assays. Full-mouth clinical periodontal parameters were recorded. RESULTS: GCF and serum MMP-8 concentrations, serum MMP-9 concentrations, and serum MMP-8/MMP-1 and MMP-9/MMP-1 molar ratios were significantly higher in Gg compared with Hg group (P <0.05). Serum MMP-8 levels and salivary TIMP-1 levels were higher in Gh compared with Hg group (P <0.05) whereas salivary MMP-8/TIMP-1 molar ratio was lower in Gh compared with Hg group (P <0.05). Elevated concentrations of GCF MMP-8 and MMP-9 were found in Gg compared with Gh group (P <0.05). Significant correlations were found between local levels of biomarkers and clinical periodontal parameters in only GDM group. CONCLUSION: GDM may modulate both local and circulating levels of MMP-8 especially when associated with gingivitis.

Clinical periodontal status and inflammatory cytokines in gestational diabetes mellitus.

OBJECTIVES: The aim of the present cross-sectional study was to compare clinical periodontal findings as well as gingival crevicular fluid (GCF) and serum levels of tumour necrosis factor-alpha (TNF-α), interleukin-10 (IL-10), and IL-33 between women with and without gestational diabetes mellitus (GDM). METHODS: Serum and GCF samples were collected, full-mouth recordings comprising plaque index, bleeding on probing and probing depth were performed in 96 females with GDM (cases) and 65 non-diabetic pregnant females (controls). Age, smoking status, pre-pregnancy body mass index, pregnancy outcomes were recorded. Serum and GCF IL-10, IL-33, TNF-α levels were determined. RESULTS: The GDM group was significantly older than the control group with an age difference of 3.27 years (mean ages were 32.05 and 28.78 years, respectively) (p<0.0001). Plaque Index (50.0 and 30.0 p=0.005), bleeding on probing (50.0 and 30.0 p=0.003) values were significantly higher in the GDM group. Serum TNF-α concentrations were significantly higher in the nonGDM group than the GDM group (p=0.001). GCF IL-10 concentrations and total amounts were significantly higher in the GDM group than the controls (p=0.004 and p<0.0001, respectively). CONCLUSION: Elevated GCF IL-10 levels may be a consequence of higher levels of inflammation as indicated by higher PI and BOP in the GDM group. However, the investigated clinical parameters may not have prominent effects on TNF-α and IL-33 levels. These findings provide further support for the importance of periodontal health during pregnancy.

Association of thalassemia major and gingival inflammation: A pilot study.

OBJECTIVES: This cross-sectional study aimed to investigate the relationship between thalassemia major (TM) and gingival inflammation through the salivary, serum, and gingival crevicular fluid (GCF) levels of matrix metalloproteinase (MMP)-8, MMP-9 and tissue inhibitor of MMP (TIMP)-1. METHODS: Biofluid samples and full-mouth clinical periodontal recordings were obtained from 29 otherwise healthy patients with TM and 25 systemically healthy (SH) individuals. Biofluid samples were evaluated by immunofluorometric assay (IFMA) and enzyme-linked immunoassays (ELISAs). Data were tested statistically by Kolmogorov Simirnov, Mann-Whitney U tests, Spearman correlation analysis. RESULTS: Age, smoking status, bleeding on probing, plaque index were similar in the study groups, but probing depth, gender data exhibited significant differences (p=0.037 for both). Salivary MMP-8, MMP-9, TIMP-1 concentrations were significantly higher in the TM than SH group (p=0.014; p<0.001; p=0.042, respectively). Serum TIMP-1 concentrations were significantly higher; MMP-8/TIMP-1, MMP-9/TIMP-1 molar ratios were significantly lower in the TM than SH group (p<0.001; p=0.005; p=0.022, respectively). Very few GCF samples revealed biochemical data above the detection limits. Numerous correlations were found between clinical periodontal parameters and biochemical data. CONCLUSIONS: It may be suggested that TM may exacerbate the local inflammatory response as manifested in salivary MMP-8, MMP-9, TIMP-1 levels.

Subgingival Plaque in Periodontal Health Antagonizes at Toll-Like Receptor 4 and Inhibits E-Selectin Expression on Endothelial Cells.

The ability of the subgingival microbial community to induce an inappropriate inflammatory response ultimately results in the destruction of bone and gingival tissue. In this study, subgingival plaque samples from both healthy and diseased sites in the same individual were obtained from adults with chronic periodontitis and screened for their ability to either activate Toll-like receptor 2 (TLR2) or TLR4 and to antagonize TLR4-specific activation by agonist, Fusobacterium nucleatum LPS. Subgingival plaque from diseased sites strongly activated TLR4, whereas matched plaque samples obtained from healthy sites were significantly more variable, with some samples displaying strong TLR4 antagonism, while others were strong TLR4 agonists when combined with F. nucleatum LPS. Similar results were observed when TLR4 dependent E-selectin expression by endothelial cells was determined. These results are the first to demonstrate TLR4 antagonism from human plaque samples and demonstrate that healthy but not diseased sites display a wide variation in TLR4 agonist and antagonist behavior. The results have identified a novel characteristic of clinically healthy sites and warrant further study on the contribution of TLR4 antagonism in the progression of a healthy periodontal site to a diseased one.

Evaluation of the gingival inflammation in pregnancy and postpartum via 25-hydroxy-vitamin D3, prostaglandin E2 and TNF-α levels in saliva.

BACKGROUND: Physiological changes and immunological modifications occur during pregnancy. The clinical and biological features of periodontal infections are affected by pregnancy. The aim of the present study was to evaluate saliva levels of 25-hydroxy-vitamin D3 (25(OH)D3), prostaglandin E2 (PGE2) and TNF-alpha (TNF-α) in pregnancy, postpartum and non-pregnant controls. METHODS: Whole saliva samples together with full-mouth clinical periodontal recordings were obtained from 59 pregnant, 47 post partum and 70 systemically healthy non-pregnant women. Groups were also evaluated according to the periodontal health status. 25(OH)D3, PGE2 and TNF-α levels in the saliva samples were determined by enzyme-linked immunoassays. Data were statistically tested by nonparametrical tests. RESULTS: Saliva TNF-α and PGE2 levels were significantly lower and 25(OH)D3 levels were significantly higher in the pregnant group than postpartum group (p<0.0001). Saliva TNF-α and 25(OH)D3 levels were significantly higher and PGE2 levels were significantly lower in the control group than postpartum group (p<0.0001). In the pregnant healthy, gingivitis and periodontitis groups saliva TNF-α levels were significantly lower than postpartum and control counterparts (p<0.0001, p=0.032, p=0.003 and p=0.013; p=0.027; p=0.007, respectively). In control healthy, gingivitis and periodontitis groups saliva 25(OH)D3 levels were significantly higher than the postpartum counterparts (p<0.0001, p<0.0001, p=0.002, respectively). In the control healthy and gingivitis groups saliva 25(OH)D3 levels were significantly higher than pregnant healthy and gingivitis (p<0.0001). CONCLUSIONS: In conclusion, within the limits of the present study it seems that pregnancy have an effect on parameters in saliva in relation to the periodontal status of the women. Further studies are required for better understanding of the impact of periodontal diseases on pregnancy or otherwise.

Oxidative stress markers in saliva and periodontal disease status: modulation during pregnancy and postpartum.

BACKGROUND: Periodontal diseases may affect local and systemic inflammation, and reactive oxygen species (ROS) levels. This systemic health burden could compromise the outcome of pregnancy in expectant mothers. The aim of the present study was to evaluate oxidative stress markers, including glutathione peroxidase (GPx), thiobarbituric acid-reactive substances (TBARS) and 8-hydroxy-2'-deoxyguanosine (8-OHdG), and total bacterial loads in the saliva of pregnant and postpartum women, and to investigate their association with periodontal disease severity. METHODS: A total of 187 women were originally recruited for this case-control study, assigned to the following groups a) pregnant group, b) postpartum group: the pregnant group re-evaluated 6 months after giving birth, c) control group: systemically healthy and non-pregnant women. The levels of the studied oxidative stress markers in saliva were measured by commercially available kits. RESULTS: The levels of salivary 8-OHdG were significantly elevated in the pregnant, compared with the control group. Although salivary 8-OHdG levels slightly decreased after giving birth (postpartum group), the difference did not reach significance. In contrast, the activity of antioxidant enzyme GPx in saliva was significantly lower in the pregnant than the control group. Although no differences in lipid peroxidation (represented by TBARS) were observed between the pregnant and control groups, after giving birth TBARS levels were significantly lowered. Only in the postpartum and control groups did clinical measurements of periodontal disease severity correlate with oxidative stress markers. Interestingly, there were no such correlations with TBARS in the pregnant and postpartum groups. CONCLUSIONS: The present study shows changes in the oxidant/antioxidant balance in saliva during pregnancy and after birth, which may be affected by periodontal health status in the latter case. Whether this is associated with adverse pregnancy outcomes, or not, remains to be elucidated. Early identification of ROS markers in saliva may be of clinical value in the periodontal management of pregnant women.

Increased infection with key periodontal pathogens during gestational diabetes mellitus.

AIM: Gestational diabetes mellitus (GDM), gingivitis, infection with specific periodontal pathogens and systemic inflammation each increase the risk for poor pregnancy outcome. We set out to monitor the interactions of gingivitis and GDM with respect to oral infection and the systemic inflammatory burden. MATERIALS AND METHODS: Four case-control groups (n = 117) were recruited, (1) No gingivitis, No GDM (n = 27); (2) Gingivitis, No GDM (n = 31); (3) No gingivitis, GDM (n = 21); and (4) Gingivitis, GDM (n = 38). Oral infection with three key periodontal pathogens was determined by PCR. Systemic inflammation was determined by quantification of CRP by EIA. RESULTS: Gingivitis during pregnancy was associated with oral infection with Porphyromonas gingivalis, Filifactor alocis and Treponema denticola and combinations thereof (all p < 0.01). GDM was also associated with increased infection with individual and multiple oral pathogens (all p < 0.05). Gingivitis during pregnancy led to a 325% increase in systemic CRP (mean, 2495 versus 8116 ng/ml, p < 0.01). CONCLUSIONS: Diabetes and gingivitis act in concert to increase risk biomarkers for poor pregnancy outcome.

The Association Between Thalassemia Major and Periodontal Health.

BACKGROUND: The aim of this cross-sectional study is to compare the local and systemic levels of soluble receptor activator of nuclear factor-κB ligand (sRANKL), osteoprotegerin (OPG), a proliferation-inducing ligand (APRIL), B-cell activating factor (BAFF), interleukin (IL)-6, and IL-8 in biofluids of patients with thalassemia major (TM) with or without gingivitis. METHODS: Seventy-seven patients are included in this study (TM, n = 29; systemically healthy, n = 48). Gingival crevicular fluid (GCF), saliva, and serum levels of IL-6, IL-8, sRANKL, OPG, BAFF, and APRIL were determined by enzyme-linked immunosorbent assay. Data were analyzed by appropriate non-parametric or parametric statistical tests. RESULTS: Median GCF, serum, and saliva levels for BAFF (P <0.001) and IL-6 and IL-8 (P <0.005) were higher in TM gingivitis than in systemically healthy gingivitis (P <0.001). GCF, serum, and saliva levels for APRIL, sRANKL, IL-6, and IL-8 were higher in TM than in systemically and periodontally healthy comparison groups (P <0.05). Positive correlations were found between bleeding on probing (BOP), plaque index (PI) scores, and GCF APRIL, serum sRANKL, serum OPG, and sRANKL concentrations in TM groups (P <0.05). Several significant positive correlations were found between BOP, PI scores, and biofluid parameters also in systemically healthy groups. CONCLUSION: TM may have a role in the underlying systemic hematologic condition and potentially affect gingival inflammation via dysregulation of lymphocytes and increased activation of osteoclasts.

Elevated matrix metalloproteinase-8 in saliva and serum in polycystic ovary syndrome and association with gingival inflammation.

This study aimed to investigate the levels of matrix metalloproteinase-8 (MMP-8) and tissue inhibitors of MMP-1 (TIMP-1) in saliva and serum samples of women with polycystic ovary syndrome (PCOS; n = 80) and matched systemically healthy controls (n = 45), with varying degrees of gingival inflammation. Salivary levels of MMP-8 and the MMP-8/TIMP-1 ratio were significantly elevated in women with PCOS, who also exhibited more gingivitis than systemically healthy women. No major changes were observed in salivary TIMP-1 levels with regard to PCOS. Serum levels of MMP-8 and the MMP-8/TIMP-1 ratio were significantly higher in women with PCOS, irrespective of the presence of gingivitis, while there were no differences in TIMP-1 levels. A positive correlation was indicated between probing depth, bleeding on probing, plaque index and salivary or serum MMP-8 levels or MMP-8/TIMP-1 ratio in the case of PCOS, while a negative such correlation was revealed for TIMP-1 in systemically healthy women. Increased levels of MMP-8 in saliva and serum seem to be more pronounced in women with PCOS, and potentiated in the presence of gingival inflammation. Alterations in MMP/TIMP system triggered by local and systemic inflammation may be implicated in the pathogenesis of PCOS, or the deterioration of its clinical presentation.

Evaluation of biochemical parameters and local and systemic levels of osteoactive and B-cell stimulatory factors in gestational diabetes in the presence or absence of gingivitis.

BACKGROUND: Gestational diabetes mellitus (GDM) is defined as varying glucose intolerance, with first onset or recognition in pregnancy. This study evaluates clinical and biochemical parameters in a possible association between GDM and gingivitis. METHODS: A total of 167 pregnant females was included in the study. There were 101 females with GDM and 66 females without GDM. Subgroups were created according to the presence or absence of gingival inflammation. Plaque index, bleeding on probing, and probing depth were recorded at four sites per tooth. Serum, saliva, and gingival crevicular fluid (GCF) levels of interleukin (IL)-6, IL-8, soluble receptor activator of nuclear factor-kappa B ligand (sRANKL), osteoprotegerin (OPG), B-cell activating factor (BAFF), and a proliferation-inducing ligand (APRIL) were determined by enzyme-linked immunosorbent assay. Data were analyzed by Kruskal-Wallis and Mann-Whitney U tests and Spearman correlation analysis. RESULTS: Age and anthropometric indices were higher in the GDM than non-GDM group (P <0.0001). Clinical periodontal recordings, serum BAFF, IL-8, and saliva sRANKL levels were higher in the GDM group (P <0.05). Saliva IL-6 level was higher in the GDM with gingivitis group than non-GDM with gingivitis group (P = 0.044). Serum and GCF BAFF (P <0.0001), serum, saliva, and GCF APRIL (P <0.0001; P <0.0001; P = 0.032, respectively), GCF OPG (P = 0.036), and serum and saliva sRANKL (P <0.0001) were higher in the GDM with gingivitis group than GDM without gingivitis group. CONCLUSIONS: The inflammatory response seems to be more pronounced in females with GDM. The observed increase in both local and systemic levels of inflammatory cytokines may suggest an interaction between gingivitis and GDM.

Association between polycystic ovary syndrome, oral microbiota and systemic antibody responses.

Polycystic ovary syndrome (PCOS) is a hormonal disorder of women that not only is the leading cause of infertility but also shows a reciprocal link with oral health. This study aimed to investigate the hypothesis that the levels of putative periodontal pathogens in saliva and their antibody response in serum are elevated in PCOS, compared to systemic health. A total of 125 women were included in four groups; 45 women with PCOS and healthy periodontium, 35 women with PCOS and gingivitis, 25 systemically and periodontally healthy women, 20 systemically healthy women with gingivitis. Salivary levels of seven putative periodontal pathogens were analyzed by quantitative real-time polymerase chain reaction and serum antibody levels were analyzed by ELISA. In women with PCOS, salivary Porphyromonas gingivalis, Fusobacterium nucleatum, Streptococcus oralis and Tannerella forsythia levels were higher than matched systemically healthy women, particularly in the case of gingivitis. Aggregatibacter actinomycetemcomitans and Treponema denticola levels were similar among study groups. The presence of PCOS also enhanced P. gingivalis, Prevotella intermedia and S. oralis serum antibody levels, when gingivitis was also present. Gingival inflammation correlated positively with levels of the studied taxa in saliva, particularly in PCOS. The presence of P. gingivalis and F. nucleatum in saliva also exhibited a strong positive correlation with the corresponding serum antibody levels. In conclusion, as an underlying systemic endocrine condition, PCOS may quantitatively affect the composition of oral microbiota and the raised systemic response to selective members of this microbial community, exerting a confounding role in resultant gingival inflammation and periodontal health. The most consistent effect appeared to be exerted on P. gingivalis.

Serum and salivary matrix metalloproteinases, neutrophil elastase, myeloperoxidase in patients with chronic or aggressive periodontitis.

Salivary, serum matrix metalloproteinase-8 (MMP-8), tissue inhibitor of matrix metalloproteinases-1 (TIMP-1), neutrophil elastase (NE), and myeloperoxidase (MPO) levels were investigated in generalized chronic periodontitis (GCP), generalized aggressive periodontitis (GAgP), and healthy groups. Whole-mouth clinical periodontal measurements were recorded. Salivary, serum concentrations of MMP-8, MPO, TIMP-1, and NE were determined by immunofluorometric assay or ELISA in 18 patients with GCP, 23 patients with GAgP, 18 individuals with healthy periodontium. Patients in the GAgP group were younger than the other groups (p<0.05). The study groups were similar in gender, smoking status. Plaque index was higher in GCP than GAgP group (p<0.05). Biochemical data were similar in periodontitis groups. Salivary, serum MPO, and salivary NE concentrations were higher; TIMP-1 concentrations were lower in the periodontitis groups than the controls (p<0.05). The present data support a close relationship between salivary, serum protease content and clinical periodontal parameters in patients with generalized periodontitis.

Metatranscriptomics of the human oral microbiome during health and disease.

The human microbiome plays important roles in health, but when disrupted, these same indigenous microbes can cause disease. The composition of the microbiome changes during the transition from health to disease; however, these changes are often not conserved among patients. Since microbiome-associated diseases like periodontitis cause similar patient symptoms despite interpatient variability in microbial community composition, we hypothesized that human-associated microbial communities undergo conserved changes in metabolism during disease. Here, we used patient-matched healthy and diseased samples to compare gene expression of 160,000 genes in healthy and diseased periodontal communities. We show that health- and disease-associated communities exhibit defined differences in metabolism that are conserved between patients. In contrast, the metabolic gene expression of individual species was highly variable between patients. These results demonstrate that despite high interpatient variability in microbial composition, disease-associated communities display conserved metabolic profiles that are generally accomplished by a patient-specific cohort of microbes. IMPORTANCE The human microbiome project has shown that shifts in our microbiota are associated with many diseases, including obesity, Crohn's disease, diabetes, and periodontitis. While changes in microbial populations are apparent during these diseases, the species associated with each disease can vary from patient to patient. Taking into account this interpatient variability, we hypothesized that specific microbiota-associated diseases would be marked by conserved microbial community behaviors. Here, we use gene expression analyses of patient-matched healthy and diseased human periodontal plaque to show that microbial communities have highly conserved metabolic gene expression profiles, whereas individual species within the community do not. Furthermore, disease-associated communities exhibit conserved changes in metabolic and virulence gene expression.