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.

Salivary secretory leukocyte protease inhibitor levels in patients with stage 3 grade C periodontitis: a comparative cross-sectional study.

Secretory leukocyte protease inhibitor (SLPI) is an anti-protease that protects mucosal tissue integrity owing to its anti-microbial and immunomodulatory properties. This study aimed to investigate SLPI levels in periodontal diseases, and analyze the potential correlation with clinical periodontal parameters. Whole saliva samples were obtained from healthy (n = 24), gingivitis (n = 24) and patients with stage 3 grade C periodontitis (n = 24). SLPI was measured by ELISA and normalized by total protein. Receiver operating characteristics (ROC) curve was used for estimating the area under the curve (AUC). The normalized SLPI levels were significantly reduced in periodontitis compared with gingivitis (4.84-fold) or health (1.83-fold) and negatively correlated with periodontal parameters. The ROC curves showed a good predictor value of the SLPI for differentiation of periodontitis versus health or gingivitis (AUC ≥ 0.80). This study demonstrates that the levels of SLPI are high in periodontal health, further elevated in gingivitis, but eventually decreased in severe periodontitis beyond the former two states. This observation may have broader implications in the context of inflammatory diseases affecting the oral mucosa, as it shows that the bacterial burden is disturbing the homeostatic balances of anti-microbial and anti-protease factors in the oral cavity.

Salivary Fingerprinting of Periodontal Disease by Infrared-ATR Spectroscopy.

PURPOSE: Periodontal diseases, the most common chronic inflammatory diseases in humans, do not only affect tooth-supporting tissues but also other body parts by contributing to the development of life-threatening conditions. Since currently available diagnostic methods in periodontics lack the ability to identify patients at high risk for periodontal disease progression, development of innovative, non-invasive, rapid detection methods for diagnosing periodontal diseases is needed. This study aims to assess the potential of infrared attenuated total reflection (IR-ATR) spectroscopy to detect differences in composition of saliva supernatant in non-periodontitis individuals (control) and patients with generalized aggressive periodontitis (G-AgP). EXPERIMENTAL DESIGN: IR-ATR is performed with a wavelength interval from 1230 to 1180 cm-1 , analyzed with a simple subtraction in absorbance data. RESULTS: Ten samples show in the analysis of variance of the two data sets a true difference (99.8%). A principal component analysis (PCA) is able to discriminate between G-AgP and control groups. CONCLUSION AND CLINICAL RELEVANCE: This study demonstrates for the first time that IR-ATR spectroscopy is a promising tool for the analysis of saliva supernatant for the diagnosis of periodontitis, and potentially other periodontal conditions. IR-ATR spectroscopy holds the potential to be miniaturized and utilized as a non-invasive screening test.

Effect of orthodontic force magnitude on cytokine networks in gingival crevicular fluid: a longitudinal randomized split-mouth study.

OBJECTIVE: To compare effect of two different orthodontic forces on maxillary canine distalization via evaluation of 30 analytes including cytokines, growth factors, and chemokines in gingival crevicular fluid (GCF) obtained from tension and compression sites. DESIGN: Longitudinal, split-mouth, randomized controlled trial. METHODS: The upper right and left canines were randomly distalized by a continuous force of either 75 or 150 g, in 15 individuals with Class II division 1 malocclusion. GCF samples were obtained from the tension and the pressure sides of each canine at appliance placement (baseline) and after force application at 24 hours and 28 days without reactivation of the coil spring. The protein content of GCF was analysed by a multiplexed immunoassay. The effects of force, side, and time on the analyte levels were assessed by the Brunner-Langer method. OUTCOME: The changes of GCF analyte levels from baseline to 24 hours and 28 days. RANDOMIZATION: Coin flipping was used for allocation of two forces. BLINDING: The participants and periodontist who performed clinical measurements and GCF sampling were blinded to group assignment and interventions (double-blinded trial). RESULTS: All patients completed the study. No harm was observed. When compared to baseline, both forces caused significant up-regulation of tumour necrosis factor-α and interleukin (IL)-1RA in the tension and the pressure sides at 28 days (P < 0.05), but not at 24 hours. Although GCF volume was similar between the two force groups over time (P > 0.05), IL-8 and MCP-1 levels in GCF were significantly lower at the pressure sites receiving higher force (150 g) at 24 hours (P < 0.05). LIMITATIONS: Although sample size (15 patients, 30 teeth) was adequate according to the initial power calculation, borderline significances may indicate lack of power or large variability among the samples. CONCLUSIONS: Although a higher force of 150 g did not result in increased cumulative canine movement or GCF production, selective host mediators were differentially regulated by the magnitude and duration of the force. REGISTRATION AND TRIAL PROTOCOL: The trial was registered retrospectively in the U.S. National Institutes of Health Clinical Trials Registry. Full details of trial protocol NCT03555747 are available on request.

Gingival crevicular fluid and salivary HIF-1α, VEGF, and TNF-α levels in periodontal health and disease.

BACKGROUND: Hypoxia-inducible factor-1 alpha (HIF-1α) is expressed as an adaptive response to hypoxia, mediates angiogenesis through the expression of vascular endothelial growth factor (VEGF) and can be induced by tumor necrosis factor-alpha (TNF-α). This study aimed to investigate the gingival crevicular fluid (GCF) and salivary HIF-1α, VEGF, and TNF-α levels in periodontal health and disease. METHODS: A total of 87 individuals, 20 generalized aggressive periodontitis (G-AgP), 20 chronic periodontitis (CP), 26 gingivitis patients, and 21 periodontally healthy individuals, were included. Clinical periodontal parameters were recorded; GCF and salivary samples were collected; and HIF-1α, VEGF, and TNF-α levels were measured by enzyme-linked immunosorbent assay. Nonparametric tests were used for the statistical analyses. RESULTS: G-AgP and CP groups had significantly higher GCF HIF-1α, VEGF, and TNF-α total amounts than gingivitis and healthy groups (P < 0.05). GCF HIF-1α and TNF-α total amounts in gingivitis group were significantly higher than the healthy group (P < 0.05). GCF and salivary concentrations of biomarkers were similar in both periodontitis groups (P > 0.05). Salivary HIF-1α concentrations in gingivitis group were significantly higher than G-AgP and healthy groups (P < 0.05). GCF HIF-1α, VEGF, and TNF-α total amounts were positively correlated with the site-specific clinical periodontal parameters and with each other (P < 0.05). CONCLUSIONS: HIF-1α is detectable in GCF and saliva of periodontally diseased and healthy individuals, and the GCF levels of the biomarker can be affected by disease status. Increased GCF HIF-1α, VEGF, and TNF-α levels in both chronic and aggressive form of periodontitis might suggest the role of TNF-α/HIF-1α/VEGF pathway in the pathogenesis of periodontal diseases.

Targeted Proteomics Guided by Label-free Quantitative Proteome Analysis in Saliva Reveal Transition Signatures from Health to Periodontal Disease.

Periodontal diseases are among the most prevalent worldwide, but largely silent, chronic diseases. They affect the tooth-supporting tissues with multiple ramifications on life quality. Their early diagnosis is still challenging, due to lack of appropriate molecular diagnostic methods. Saliva offers a non-invasively collectable reservoir of clinically relevant biomarkers, which, if utilized efficiently, could facilitate early diagnosis and monitoring of ongoing disease. Despite several novel protein markers being recently enlisted by discovery proteomics, their routine diagnostic application is hampered by the lack of validation platforms that allow for rapid, accurate and simultaneous quantification of multiple proteins in large cohorts. Here we carried out a pipeline of two proteomic platforms; firstly, we applied open ended label-free quantitative (LFQ) proteomics for discovery in saliva (n = 67, including individuals with health, gingivitis, and periodontitis), followed by selected-reaction monitoring (SRM)-targeted proteomics for validation in an independent cohort (n = 82). The LFQ platform led to the discovery of 119 proteins with at least 2-fold significant difference between health and disease. The 65 proteins chosen for the subsequent SRM platform included 50 functionally related proteins derived from the significantly enriched processes of the LFQ data, 11 from literature-mining, and four house-keeping ones. Among those, 60 were reproducibly quantifiable proteins (92% success rate), represented by a total of 143 peptides. Machine-learning modeling led to a narrowed-down panel of five proteins of high predictive value for periodontal diseases with maximum area under the receiver operating curve >0.97 (higher in disease: Matrix metalloproteinase-9, Ras-related protein-1, Actin-related protein 2/3 complex subunit 5; lower in disease: Clusterin, Deleted in Malignant Brain Tumors 1). This panel enriches the pool of credible clinical biomarker candidates for diagnostic assay development. Yet, the quantum leap brought into the field of periodontal diagnostics by this study is the application of the biomarker discovery-through-verification pipeline, which can be used for validation in further cohorts.

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.

Alarm anti-protease trappin-2 negatively correlates with proinflammatory cytokines in patients with periodontitis.

BACKGROUND: Trappin-2 is a potent biologically active serine protease inhibitor with anti-inflammatory properties that has also been characterized as an "alarm anti-protease." Although the importance of trappin-2 in several chronic infections has been demonstrated, its potential involvement in periodontitis remains undefined. This study aims to investigate salivary levels of trappin-2 and interleukin (IL)-1ß in periodontally healthy individuals and patients with gingivitis or generalized chronic periodontitis (CP) or aggressive periodontitis (GAgP). METHODS: Whole unstimulated saliva samples were collected from 80 systemically healthy and non-smoking individuals before full-mouth periodontal examination. Trappin-2 and IL-1ß were analyzed by enzyme-linked immunosorbent assay and reported as nanograms per milligram after calibration for total protein levels. RESULTS: Correlation analysis revealed negative association between trappin-2 and IL-1ß levels. Trappin-2 also showed strong negative correlation with clinical periodontal parameters, in contrast to IL-1ß, which showed positive correlation. Trappin-2 levels were significantly lower in individuals with CP and GAgP, but not gingivitis, compared with healthy individuals. Reduced salivary concentrations of trappin-2 had high sensitivity and specificity to distinguish health from periodontitis. CONCLUSIONS: Trappin-2 is abundant in the saliva of individuals with healthy periodontium in line with its role as an "anti-alarm" protease. Decreased salivary trappin-2 and increased IL-1ß levels in individuals with periodontitis, compared with healthy individuals, may implicate a potential antiprotease/proinflammatory cytokine imbalance, resulting in impaired host protective capacity.

Salivary and Serum Markers Related to Innate Immunity in Generalized Aggressive Periodontitis.

BACKGROUND: Host inflammatory and immune responses play an important role in aggressive periodontitis (AgP). Thus, this study aims to evaluate levels of the innate immunity-related markers calprotectin, colony-stimulating factor (CSF)-1, macrophage migration inhibitory factor (MIF), monokine induced by interferon-γ (MIG), and matrix metalloproteinase (MMP)-8 in serum and saliva from patients with generalized AgP and those with gingivitis or a healthy periodontium. METHODS: This study enrolled 40 individuals (17 males and 23 females; mean age 33.30 ± 9.31 years), 15 with generalized AgP, 15 with gingivitis, and 10 who were periodontally healthy. Full-mouth periodontal examinations were performed, and serum and saliva were collected. Levels of calprotectin, CSF-1, MIF, MIG, and MMP-8 were measured using enzyme-linked immunosorbent assays. RESULTS: In serum, mean levels of calprotectin were 2.06-fold higher in patients with AgP than in healthy patients (P = 0.01). Serum levels of MMP-8 were significantly elevated in patients with AgP compared with both healthy patients and those with gingivitis, by 2.60-fold and 2.77-fold, respectively (P = 0.03 and P = 0.009, respectively). In saliva, levels of MMP-8 were 5.66-fold higher in patients with AgP than in healthy patients (P = 0.02). CSF-1, MIF, and MIG levels in both serum and saliva did not differ significantly among the groups. CONCLUSIONS: Serum levels of calprotectin and MMP-8 are elevated in patients with AgP. MMP-8 levels are also increased in saliva from patients with AgP. These results support involvement of innate immune response in the pathogenesis of AgP.

Gingival crevicular fluid and plasma oxidative stress markers and TGM-2 levels in chronic periodontitis.

OBJECTIVE: This study was aimed to evaluate the gingival crevicular fluid (GCF) and plasma transglutaminase-2 (TGM-2), total antioxidant capacity (TAC), total oxidant status (TOS), ferric reducing antioxidant power (FRAP) and thiobarbituric acid reactive substances (TBARS) in patients with chronic periodontal disease. MATERIALS AND METHODS: Twenty patients with chronic periodontitis (CP), 20 patients with gingivitis and 20 healthy subjects were enrolled in the study. Clinical periodontal parameters including probing depth, clinical attachment level, plaque index and papillary bleeding index were recorded. GCF and plasma levels of TGM-2, TAC, TOS, TBARS and FRAP were analyzed. RESULTS: GCF TGM-2 was significantly lower in CP group than in gingivitis patients (P=0.006). GCF FRAP in CP and gingivitis groups was significantly lower than in healthy subjects (P<0.001). Plasma FRAP level was lower in gingivitis group when compared to healthy subjects (P=0.003). There was no significant difference in GCF and plasma TAC, TOS, TBARS and plasma TGM-2 levels among the study groups (P>0.05). GCF TGM-2 level was positively correlated with GCF TAC and negatively correlated with CAL. CONCLUSIONS: Decreased FRAP in GCF and plasma indicating lower antioxidant status of CP patients might suggest the role of oxidative stress in periodontitis. GCF TGM-2 data might suggest that TGM2 is associated with stabilization of the extracellular matrix and wound healing in periodontium rather than gingival inflammation.

Impact of implant-abutment connection on osteoimmunological and microbiological parameters in short implants: a randomized controlled clinical trial.

OBJECTIVES: The study aimed to determine the levels of soluble receptor activator of nuclear factor-кB ligand (sRANKL) and osteoprotegerin (OPG) as well as their relative calculated ratio in peri-implant crevicular fluid (PICF) obtained around two different types of implant-abutment connection on short implants following a 12-month monitoring period. Moreover, the levels of a number of oral bacterial species were investigated in the corresponding submucosal biofilm samples. MATERIALS AND METHODS: Thirty short implants were randomly placed in posterior maxillary edentulous sites using a split-mouth design in 15 periodontally healthy subjects. Tapered interference fit (TIF) and taper-integrated screwed-in (TIS) types of implant-abutment connections were selected for investigation. PICF and submucosal biofilm samples were collected 1 month after surgery and repeated 12 months after prosthetic loading. Clinical parameters, including probing depth, dichotomous presence of bleeding on probing, and plaque index, were recorded and digital periapical radiographs were taken at each time point. sRANKL and OPG levels in PICF were analyzed using an enzyme-linked immunosorbent assay. Total bacterial levels, as well as levels of Fusobacterium nucleatum, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, Prevotella intermedia, and Streptococcus oralis, were analyzed in the corresponding submucosal biofilm samples using quantitative real-time polymerase chain reaction. RESULTS: The total amount of sRANKL in TIF implants was 2.64-fold lower than that in TIS implants at baseline (P < 0.001), whereas similar levels were found after 12 months (P > 0.05). Accordingly, OPG and RANKL/OPG ratio were similar between the groups at each time point (P > 0.05). Microbiological results were similar in both groups at each time point (P > 0.05). CONCLUSION: The results of this longitudinal study suggested that sRANKL and OPG in PICF, as well as microbiological parameters in submucosal biofilms, were similar between TIF and TIS implants, after a 12-month monitoring period, despite early differences in the former. Therefore, the type of implant-abutment connection does not appear to influence longitudinally the levels of osteoimmunological and microbiological markers in the peri-implant tissues of short implants.

Impact of aging on TREM-1 responses in the periodontium: a cross-sectional study in an elderly population.

BACKGROUND: Aging is associated with altered immune response, which increases susceptibility to infections. sTREM-1 is involved in the amplification of the inflammatory response to bacterial infection. The present cross-sectional study aims to investigate local sTREM-1 levels in gingival crevicular fluid (GCF) as well as key periodontal pathogen levels in the subgingival plaque in an elderly cohort with periodontal health, gingivitis, and chronic periodontitis (CP). METHODS: Subjects were 51 systemically healthy, elderly individuals (mean age, 68 ± 4.5 years) who had undergone full-mouth periodontal examinations. Subgingival plaque and GCF samples were collected from the healthy sites of participants without periodontal disease (n = 17), the sites with gingival inflammation from patients with gingivitis (n = 19), and the periodontitis sites of patients with CP (n = 15). GCF volumes were measured by an electronic impedance device, and total protein levels were assessed by a flouremetric assay. sTREM-1 levels in GCF were measured by enzyme-linked immunosorbent assay. The subgingival plaque total bacteria, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Fusobacterium nucleatum, and Prevotella intermedia levels were determined by quantitative real-time polymerase chain reaction. Statistical analysis was performed using nonparametric methods. RESULTS: GCF volume, total protein concentrations, and sTREM-1 levels in GCF were similar among the groups (p > 0.05). Significantly higher T. forsythia levels were observed in subgingival plaque samples harvested from patients with gingivitis and CP, than in those from healthy participants (p < 0.05). However, the subgingival levels of the other four periodontal pathogens and total bacteria were not statistically different among the groups (p > 0.05). CONCLUSIONS: Our findings suggest that there are no differences in GCF volume, total protein, and sTREM-1 levels between healthy and periodontally diseased elderly adults. We found only limited differences in the studied subgingival microbial profile. This finding indicates an already deregulated, local inflammatory response in this elderly cohort, on which bacterial biofilm challenge may have a limited further impact.

Gingival Crevicular Fluid and Plasma Levels of Transglutaminase-2 and Oxidative Stress Markers in Cyclosporin A-Induced Gingival Overgrowth.

BACKGROUND: Transglutaminase (TGM)-2 has been shown to contribute to fibrosis by extracellular matrix accumulation in some organs and is activated by intracellular reactive oxygen species. The aim of this study is to investigate levels of gingival crevicular fluid (GCF) and plasma TGM-2 and oxidative stress markers (OSMs) in cyclosporin A (CsA)-induced gingival overgrowth (GO). METHODS: The study enrolled 20 healthy (H) individuals; 20 patients with gingivitis (G); 20 CsA-medicated patients with GO (CsA GO+); and 20 CsA-medicated patients without GO (CsA GO-). GCF and plasma levels of TGM-2 were analyzed by enzyme-linked immunosorbent assay. Spectrofluorometry was used to analyze thiobarbituric acid reactive substance (TBARS); ferric-reducing antioxidant power (FRAP); total oxidant status (TOS); and total antioxidant capacity (TAC). RESULTS: GCF TGM-2 level was elevated in CsA GO+ compared with G (P = 0.048) and H (P = 0.001) groups. GCF TBARS level was elevated in CsA GO+ compared with other groups (CsA GO- group: P = 0.003; G group: P <0.001; and H group: P <0.001) and was higher in CsA GO- than in H (P = 0.048). GCF FRAP level was lower in CsA GO- than in H (P = 0.04). Both CsA GO+ and CsA GO- groups had lower GCF TOS levels than H (P <0.001 and P = 0.002) and G (P = 0.003 and P = 0.04). GCF TAC was higher in CsA GO+ than in H (P = 0.02). Plasma TGM-2 level was elevated in CsA GO+ compared with G (P = 0.048) and H (P = 0.002). Plasma FRAP level was higher in H and CsA GO- than in CsA GO+ (P = 0.008 and P = 0.02). CONCLUSIONS: CsA use significantly alters GCF and plasma levels of TGM-2 and OSMs. TGM-2 may contribute to CsA-induced GO in CsA-treated patients by changing GCF and plasma levels of OSMs. Further studies are needed to prove causality and its direction.

Treatment of lateral open bite with vertical dentoalveolar distraction osteogenesis.

The aim of this article is to describe the surgical, orthodontic, and periodontal treatment of an adult patient with a lateral open bite, anterior crowding, and gingival recession on the mandibular right lateral incisor. The lateral open bite, which resisted conventional mechanics, was successfully corrected by the combination of dento-osseous osteotomies and vertical alveolar distraction using orthodontic multibracket appliances in conjunction with nickel-titanium archwires and intermaxillary elastics. After the orthodontic treatment, the denuded root surface of the mandibular right lateral incisor was closed using a coronally advanced flap technique with platelet-rich fibrin. The results at the 2-year posttreatment follow-up were satisfactory from both the occlusal and the periodontal standpoints.

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.

Interleukin-6 Family of Cytokines in Crevicular Fluid of Renal Transplant Recipients With and Without Cyclosporine A-Induced Gingival Overgrowth.

BACKGROUND: Interleukin (IL)-6 family of cytokines, including IL-6, oncostatin M (OSM), leukemia inhibitory factor (LIF), and IL-11, have fibrogenic features. The current study determines gingival crevicular fluid (GCF) levels of fibrosis-related IL-6-type cytokines in cyclosporine A (CsA)-induced gingival overgrowth (GO). METHODS: Eighty non-smokers were included (40 CsA-medicated renal transplant patients with GO [GO+; n = 20] or without GO [GO-; n = 20], 20 individuals with gingivitis, and 20 healthy participants). Probing depth and plaque, papilla bleeding, and hyperplastic index scores were recorded. GCF samples were obtained from the mesio-buccal aspects of two teeth. GCF IL-6, IL-1ß, OSM, LIF, and IL-11 levels were analyzed by enzyme-linked immunosorbent assay. RESULTS: The GO+ and GO- groups had higher IL-6 total amounts than the healthy group (P <0.008). IL-1ß total amounts in the GO+ group were significantly higher than in both the healthy and GO- groups (P <0.008). OSM total amount was elevated in the GO+ and GO- groups compared with both the gingivitis and healthy groups (P <0.008). All groups had similar LIF and IL-11 total amounts (P >0.008). Moderate positive correlations were detected among IL-6, IL-1ß, OSM, and IL-11 total amount in GCF and clinical parameters (P <0.05). CONCLUSIONS: IL-6 and OSM increases in GCF as a result of CsA usage or an immunosuppressed state irrespective of the severity of inflammation and the presence of GO. The IL-6 family of cytokines might not be directly involved in biologic mechanisms associated with CsA-induced GO. Lack of an association between assessed IL-6 cytokines and CsA-induced GO might indicate distinct effects of these cytokines on fibrotic changes of different tissues.

Gingival crevicular fluid interleukin-36ß (-1F8), interleukin-36γ (-1F9) and interleukin-33 (-1F11) levels in different periodontal disease.

BACKGROUND: Periodontal inflammation is driven by the coordinated action of a number of factors, including the IL-1 family. Our study aimed to examine the levels of interleukin (IL)-36ß, IL-36γ and IL-33 levels in gingival crevicular fluid (GCF) from patients with different periodontal diseases. MATERIALS AND METHODS: A total of 80 subjects, 20 patients with generalized aggressive periodontitis (G-AgP), 20 patients with chronic periodontitis (CP), 20 with gingivitis and 20 periodontally healthy subjects were included. Periodontal status was evaluated by measuring probing depth, clinical attachment loss, papillary bleeding index and plaque index. GCF cytokine levels were analysed by ELISA. RESULTS: CP, gingivitis and healthy groups had similar GCF IL-36ß total amount (p>0.008). G-AgP group had elevated IL-36ß total amount compared to CP group (p<0.008). G-AgP group had similar GCF IL-36ß total amount to gingivitis and healthy groups (p>0.008). GCF IL-36γ and IL-33 total amounts of the study groups were similar (p>0.05). CONCLUSIONS: The present study demonstrated for the first time the presence of IL-36ß, IL-36γ and IL-33 GCF levels with different periodontal diseases. High levels of IL-36-ß in the AgP group in comparison to CP group might suggest that periodontitis in the aggressive form could be related to the increase in GCF IL-36ß.

Spectrophotometric and computerized evaluation of tooth bleaching employing 10 different home-bleaching procedures: In-vitro study.

OBJECTIVE: The aim of this in-vitro study was to evaluate the efficacy of bleaching products, determine the applicability and validation of the measurement methods. MATERIALS AND METHODS: Freshly extracted 110 human incisor teeth were stained with whole blood and hemolysate solution prior to the application of 10 different home-bleaching products. Spectrophotometric measurements of the tooth shades were performed for each specimen before and after bleaching at the 1(st), 3(rd), 7(th), and 14 days. Differences in lightness (Δl), chroma (Δc), hue (Δh) values and shade changes were measured to evaluate process. Computerized digital imaging analyses to determine the color changes were performed with Photoshop CS4 software (Adobe, San Jose, CA, USA). Statistical analyses were performed with analysis of variance, Scheffe and Tukey tests. RESULTS: In all of the test groups regardless of the material used, a significant increase in lightness and hue, and decrease of chroma were observed, as compared to the control group. After recommended bleaching applications, Δl and Δh values respectively increased in group Zaris White and Brite (ZWB) and group Pola Night and Δc values showed significant decrease in groups ZWB and Rembrandt REM3 (P < 0.05). At the end of the procedure both spectrophotometric and digital imaging analysis showed ZWB was the most effective product among the others while Yotuel and Happy Smile were the least (P < 0.05). CONCLUSIONS: Home-bleaching systems showed slower but almost permanent bleaching effect likewise office-based methods. Both software and spectrophotometric analyses have advantages such as evaluating the results objectively and numerically, also treatment outcomes could be preserved.

Soluble triggering receptor expressed on myeloid cells 1 (sTREM-1) in gingival crevicular fluid: association with clinical and microbiologic parameters.

BACKGROUND: Soluble triggering receptor expressed on myeloid cells 1 (sTREM-1) belongs to the immunoglobulin superfamily and is involved in amplification of the inflammatory response to bacterial infection. This cross-sectional study aims to investigate the levels of sTREM-1 in gingival crevicular fluid (GCF) of individuals without periodontitis and with chronic periodontitis (CP) or generalized aggressive periodontitis (GAgP) and their association with the levels of key periodontal pathogens in subgingival plaque. METHODS: GCF and subgingival plaque samples were obtained from healthy sites of participants without periodontitis (n = 20) and periodontitis sites of patients with CP (n = 22) and GAgP (n = 20). sTREM-1 levels in GCF were measured by enzyme-linked immunosorbent assay. Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Aggregatibacter actinomycetemcomitans levels in subgingival plaque were analyzed by quantitative real-time polymerase chain reaction. RESULTS: sTREM-1 levels in GCF were higher in CP and GAgP than healthy sites by 3.6- and 4.4-fold, respectively, with no significant differences between the two forms of periodontitis. Moreover, sTREM-1 levels in GCF were positively correlated with site-specific clinical periodontal parameters and levels of P. gingivalis, T. denticola, and T. forsythia, but not A. actinomycetemcomitans, in subgingival plaque. CONCLUSION: Increased GCF levels of sTREM-1 at diseased sites and their positive correlation with clinical and microbiologic parameters strengthen the association of this inflammatory marker with periodontitis.

Gingival crevicular fluid and plasma acute-phase cytokine levels in different periodontal diseases.

BACKGROUND: The aim of the present study is to investigate gingival crevicular fluid (GCF) and plasma acute-phase cytokines, interleukin-1ß (IL-1ß), interleukin-6 (IL-6), interleukin-11 (IL-11), oncostatin M (OSM), and leukemia inhibitory factor (LIF) levels in patients with different periodontal diseases. METHODS: Eighty individuals were included in this study; 20 with chronic periodontitis (CP), 20 with generalized aggressive periodontitis (GAgP), 20 with gingivitis, and 20 classified as healthy (H). Probing depth, clinical attachment level, plaque index, and papilla bleeding index were recorded. Plasma and GCF IL-1ß, IL-6, IL-11, OSM, and LIF levels were analyzed by enzyme-linked immunosorbent assay. RESULTS: CP and GAgP groups had significantly higher GCF IL-1ß, IL-6, and IL-11 levels when compared with the H group (P <0.05). Conversely, GCF LIF levels of the CP and GAgP groups were lower than those of the H group (P <0.05). GCF OSM levels did not differ significantly among study groups. Plasma levels of all the cytokines studied were not significantly different among the study groups. CONCLUSIONS: Based on the present data, elevated IL-1ß, IL-6, and IL-11 GCF levels, but not plasma levels, are suggested as reliable inflammatory biomarkers in periodontal diseases. Decreased LIF levels in diseased groups might reflect the possible beneficial effects of LIF in the modulation of inflammatory response in gingiva.