The Association between Salivary Metabolites and Gingival Bleeding Score in Healthy Subjects: A Pilot Study.

Periodontal diseases, including gingivitis and periodontitis, are among the most prevalent diseases in humans. Gingivitis is the mildest form of periodontal disease, characterized by inflammation of the gingiva caused by the accumulation of dental plaque. Salivary diagnostics are becoming increasingly popular due to the variation in saliva composition in response to pathological processes. We used a metabolomics approach to investigate whether a specific saliva metabolic composition could indicate preclinical stage of gingivitis. 1H-NMR spectroscopy was used to obtain the salivary metabolite profiles of 20 healthy subjects. Univariate/multivariate statistical analysis evaluated the whole saliva metabolite composition, and the Full-Mouth Bleeding Score (FMBS) was employed as a classification parameter. Identifying a signature of specific salivary metabolites could distinguish the subjects with high FMBS scores but still within the normal range. This set of metabolites may be due to the enzymatic activities of oral bacteria and be associated with the early stages of gingival inflammation. Although this analysis is to be considered exploratory, it seems feasible to establish an FMBS threshold that distinguishes between the absence and presence of early inflammatory alterations at the salivary level.

Assessment of Myeloperoxidase and Nitric Levels around Dental Implants and Natural Teeth as a Marker of Inflammation: A Comparative Study.

INTRODUCTION: Dental implants form the mainstay of dental treatment involving rehabilitation of missing teeth. One of the major concerns for the clinicians doing dental implants is the postsurgical failure of dental implants. Success of dental implants is dependent upon the skills of the surgeon and the amount and quality of the bone remaining at the edentulous area where dental implant has to be placed. Myeloperoxidase (MPO) and nitrites are few of the enzymes and molecules which are said to be altered in inflammation. However, their exact role in the inflammatory processes around natural tooth and dental implant is still unclear. Hence we comparatively evaluated the levels of MPO and nitrites in the areas around the dental implants and natural teeth. MATERIALS AND METHODS: The present study comprises 42 patients who underwent prosthetic rehabilitation by dental implants from 2011 to 2014. Depth of probing value (DP), score of plaque index (SPI), gingival index (GI), and index of gingival bleeding time (GBT) were evaluated for the assessment of the periimplant soft tissue changes. Assessment of inflammation around the dental implant surface and around natural tooth was done based on the readings of these parameters. For the measurement of the MPO levels, spectrophotometric MPO assay was used. All the results were analyzed by Statistical Package for the Social Sciences (SPSS) software. RESULTS: The mean plaque index values were 1.56 and 0.97 in periodontitis cases of natural teeth and inflamed cases of dental implants respectively. While comparing mean plaque index, mean probing depth, and mean gingival bleeding index in between the two groups, significant difference was obtained. Mean MPO concentration in periodontitis and gingivitis cases in natural teeth were 0.683 and 0.875 U/µL, while in inflamed dental implant cases, the mean value was 0.622 U/µL. While comparing the total MPO levels, total nitrite levels, and total nitrite concentration in between two study groups, significant difference was obtained. On comparing the healthy and periodontitis cases in natural teeth, significant difference was obtained. CONCLUSION: In the inflammatory processes occurring around dental implant and natural teeth, MPO and NO make some amount of significant contribution. CLINICAL SIGNIFICANCE: The present study enforces on the role of MPO and nitrite as diagnostic and prognostic marker.

Response of chronic gingivitis to hygiene therapy and experimental gingivitis. Clinical, microbiological and metabonomic changes.

PURPOSE: To compare the clinical, microbiological and metabonomic profiles of subjects with high and low levels of chronic gingival bleeding during a controlled oral hygiene regimen intervention including sequential phases of rigorous therapeutic oral hygiene followed by experimental gingivitis (EG). METHODS: Two cohorts of qualified study subjects with differences in gingival bleeding on probing levels at their baseline clinical examination were entered into the study. These two cohorts were followed through three separate study phases including a 1-week baseline phase, a 2-week phase of rigorous oral hygiene including dental prophylaxis, and a 3-week EG phase of no oral hygiene to encourage relapse of gingivitis. The 58 subjects were assessed during each phase of the study for clinical presentation of gingivitis and concurrently had plaque sampled for real-time polymerase chain reaction (RTPCR) microbiological characterization and salivary lavage samples for 'systems biology' metabonomics assessment by 1H-NMR. RESULTS: Subjects presenting with different levels of gingival bleeding on probing when they entered the study responded differently to rigorous oral hygiene and EG. Specifically, the high bleeding cohort responded sluggishly to rigorous oral hygiene and exhibited markedly greater relapse to gingivitis during EG. RTPCR analysis showed changes in bacterial populations that were associated with study phases, particularly the increases in putative periodontal pathogens during EG. However, the microbiological profiles of high- and low-susceptibility gingival bleeding patients were largely similar. Metabonomic analysis likewise revealed significant changes in metabolite composition during study phases associated with differences in plaque toxicity, especially the short chain carboxylic acids propionate and n-butyrate, which tracked clinical changes in gingivitis severity. Systems analysis of metabonomic changes suggested differences between cohorts, although analysis to date has not elucidated whether these differences are causative (population predictive) or simply diagnostic of clinical status within populations.

Anti-inflammatory protein tumor necrosis factor-α-stimulated protein 6 (TSG-6) promotes early gingival wound healing: an in vivo study.

BACKGROUND: Human multipotent mesenchymal stromal cells (hMSCs) produce tumor necrosis factor (TNF)-α-stimulated protein 6 (TSG-6). TSG-6 modulates proinflammatory cytokine cascades and enhances tissue repair. This study tests the effects of recombinant human TSG-6 (rhTSG-6) on gingival wound healing within the first 2 days post-surgery. METHODS: After gingival resection in 120 Sprague-Dawley rats, 2 µg rhTSG-6 in 5-µL phosphate-buffered saline (PBS) or the same volume of only PBS solution was injected into gingival tissue approximating the surgical wound. Control animals did not receive injections. Tissue biopsies and blood were collected at 1 to 2, 6 to 8, 24, and 48 hours post-surgery (n = 10 per group). Specimens were analyzed via histologic analysis and enzyme-linked immunosorbent assay (ELISA) for quantification and comparison of inflammatory markers interleukin (IL)-1ß, IL-6, TNF-α, and myeloperoxidase (MPO). Wound photographs were taken for a double-masked clinical assessment at each time period. Weights were recorded for all animals pre- and post-surgery. RESULTS: Animals injected with rhTSG-6 had significantly less severe clinical inflammation at 6 to 8 (P = 0.01228), 24 (P = 0.01675), and 48 (P = 0.0186) hours. Sham and control animals had more weight loss at 24 and 48 hours. Sham and control animals had more pronounced cellular infiltrate. rhTSG-6-treated animals had significantly less MPO (P = 0.027) at 24 hours and IL-1ß (P = 0.027) at 24 and 48 hours. IL-6 showed a marginal significant difference at 6 to 8 hours, but there was no significant difference for TNF-α. CONCLUSION: rhTSG-6 reduced postoperative gingival inflammation by reducing levels of proinflammatory cytokines and cellular infiltrate and may offer significant promise as an anti-inflammatory agent for gingival surgery.

Sex differences in inflammatory and apoptotic signaling molecules in normal and diseased human gingiva.

BACKGROUND: The purpose of this study is to determine whether sex dimorphism exists in the expression of inflammatory and apoptotic mediators in gingiva obtained from normal and diseased sites of periodontal disease. METHODS: Gingival papillae were obtained from individuals (56 males and 62 females) who required extraction of adjacent teeth. Gingival samples were grouped by adjacent sulcus depth: 1 to 3 mm (normal), 3 mm with bleeding on probing (slight disease), 3 to 6 mm (moderate disease), and >6 mm (severe disease). The tissue concentrations of cysteine-requiring aspartate-directed protease 3 (caspase-3), interleukin-2, tumor necrosis factor-related apoptosis-inducing ligand, Fas ligand, p38α mitogen-activated protein kinase, extracellular signal-related kinase 1/2, and survivin were determined by enzyme-linked immunosorbent assay. These mediator concentrations, age of donor, sex of donor, and gingival sulcular depth were the outcome variables. Data were compared by factorial analysis of variance, post hoc Tukey, and Pearson correlation test. P <0.05 was used to indicate significant differences among the outcome variables. RESULTS: The mean gingival sulcular depth was significantly greater in male than in female groups (P <0.05). The majority of the tested mediators were significantly correlated with both sex and sulcular depth and with caspase-3 (P <0.05). The concentration of caspase-3 in female gingiva at all diseased sites was significantly greater than in gingiva derived from male sites (P <0.05). CONCLUSIONS: These data suggest sex dimorphism in the presence of gingival apoptosis at sites of periodontal disease, with females having the highest incidence of apoptosis. Because apoptosis clears inflammatory cells and promotes healing, this phenomenon could provide a mechanism for sex dimorphism for the incidence of periodontal disease.

Assessment of tissue oxygenation of periodontal inflammation in smokers using optical spectroscopy.

BACKGROUND: We have recently developed a periodontal diagnostic tool that was validated in non-smokers with periodontitis. Tobacco smoking is a recognized risk factor for periodontal diseases that can mask gingival bleeding and lead to a false negative diagnosis. Therefore, the purpose of current study is to further validate this instrument in smokers with periodontal diseases. METHODS: Using a portable optical near-infrared spectrometer, optical spectra were obtained, processed and evaluated from healthy (n = 108), gingivitis (n = 100), and periodontitis (n = 79) sites of 54 systemically healthy smokers. A modified Beer-Lambert unmixing model that incorporates a non-parametric scattering loss function was used to determine the relative contribution of deoxygenated haemoglobin (Hb) and oxygenated haemoglobin (HbO2 ) to the overall spectrum. The balance between tissue oxygen delivery and utilization in periodontal tissues was then assessed. RESULTS: Tissue oxygen saturation was significantly decreased in the gingivitis (p = 0.016) and periodontitis (p = 0.007) sites, compared to the healthy sites. There was a trend towards increased concentration of Hb and decreased concentration of HbO2 from healthy to diseased sites, without statistical significance (p > 0.05). CONCLUSIONS: Optical spectroscopy can determine tissue oxygenation profiles of healthy and diseased sites in smokers. The spectral profile of periodontal sites in smokers generally resembles those from non-smoking patients.

Profiling biomarkers in gingival crevicular fluid using multiplex bead immunoassay.

OBJECTIVE: Biomarkers in gingival crevicular fluid (GCF) have been investigated; however, measurements were limited by the small sample volume available. The aim of this study was to determine the levels of 40 different cytokines and chemokines in GCF samples. DESIGN: Eleven patients with generalised chronic periodontitis participating in a supportive periodontal therapy programme with remaining probing pocket depths (PDs) of >5mm were enrolled. One healthy and two diseased sites were sampled in each subject. Forty biomarkers in GCF were examined using a multiplex bead immunoassay. Porphyromonas gingivalis from the diseased sites was quantified by real-time polymerase chain reaction. RESULTS: Twenty-six biomarkers were detected in the GCF samples using the multiplex bead immunoassay. The levels of nine biomarkers were significantly different between the diseased and healthy sites after adjustment with Bonferroni's correction. The level of 26 biomarkers in diseased sites was compared between bleeding on probing (BOP)-positive and BOP-negative sites. Interleukin (IL)-1ß and interferon-inducible protein (IP)-10 levels were significantly higher in BOP-positive diseased sites than BOP-negative diseased sites after adjustment for multiple comparisons (IL-1ß, p=0.0007, IP-10; p=0.0009). In addition, the levels of IL-1ß in GCF were found to be strongly correlated with the P. gingivalis ratio (r=0.646, p=0.0012). CONCLUSION: IL-1ß levels in GCF correlate with the PDs, BOP and the presence of P. gingivalis in subgingival plaque. Multiplex bead assays can be useful in GCF studies. These findings can help in identifying new diagnostic methods in the diagnosis of periodontal disease.

Altered relationship between MMP-8 and TIMP-2 in gingival crevicular fluid in adolescents with Down's syndrome.

BACKGROUND AND OBJECTIVE: Periodontitis is more frequently found in subjects with Down's syndrome. The aim was to investigate whether the relationship between MMPs and TIMPs) in the gingival crevicular fluid of subjects with Down's syndrome is altered compared with controls. MATERIAL AND METHODS: Twenty-one adolescents with Down's syndrome and gingivitis (DS-G), 12 subjects with Down's syndrome and periodontitis (DS-P), 26 controls with gingivitis (HC-G) and eight controls with periodontitis (HC-P) were clinically examined. All patients were between 11 and 20 years of age. Gingival crevicular fluid was collected from each subject and the concentrations of MMPs (2, 3, 8, 9 and 13) and TIMPs (1, 2 and 3) (expressed as pg/µL adjusted for volume of gingival crevicular fluid) were determined using multianalyte kits from R&D Systems. RESULTS: The concentrations of MMP-2, MMP-3, MMP-8, MMP-9 and TIMP-2 in gingival crevicular fluid were significantly higher (p < 0.005) in the DS-G group compared with the HC-G group. The correlation coefficient between MMP-8 and TIMP-2 differed significantly (p = 0.006) between the DS-G group and the HC-G group. On the contrary, the correlation coefficients between MMPs and TIMPs did not differ significantly between the DS-P group and the HC-P group. However, the DS-P group exhibited a significantly lower concentration of TIMP-2 in the gingival crevicular fluid compared with the HC-P group. CONCLUSION: Down's syndrome subjects with gingivitis exhibit higher concentrations of MMPs in gingival crevicular fluid with an altered relationship between MMP-8 and TIMP-2, which might impair the periodontal tissue turnover.

Elevated levels of salivary lactoferrin, a marker for chronic periodontitis?

BACKGROUND AND OBJECTIVE: Whole saliva is a complex mixture of fluids essential for the well-being of the oral hard and soft tissues. Saliva contains numerous antimicrobial proteins that help protect the oral ecosystem from infectious agents. Chronic periodontitis is an infectious chronic inflammatory condition that affects the tooth-supporting structures and leads to their destruction. The aim of the present study was to investigate differences in concentrations of salivary lactoferrin in subjects with and without periodontal disease and correlate these values with clinical variables associated with periodontal disease. MATERIAL AND METHODS: Stimulated whole saliva was collected from 17 subjects with chronic periodontitis and 17 periodontally healthy control subjects. Data relating to bleeding on probing, probing pocket depth and horizontal bone loss were registered. Concentrations of lactoferrin, lysozyme and IgA in stimulated whole saliva were quantified using ELISA. RESULTS: Subjects with chronic periodontits showed higher concentrations of lactoferrin in stimulated whole saliva compared with periodontally healthy control subjects (p < 0.05). Salivary concentrations of lactoferrin were positively correlated with bleeding on probing (p < 0.001) and the number of sites with probing pocket depth ≥ 6 mm (p < 0.001). CONCLUSION: Lactoferrin is raised in stimulated whole saliva in subjects with chronic periodontitis and is correlated with probing pocket depth ≥ 6 mm.

Relationship between salivary antioxidant capacity and phases of the menstrual cycle.

BACKGROUND AND OBJECTIVE: Reactive oxygen species and free radicals are involved in the pathogenesis of periodontal disease. Previous studies have shown that the stage of the menstrual cycle is associated with the levels of gingival inflammation and discomfort. This study examined changes in salivary antioxidant activities, clinical parameters and bacterial levels during the menstrual cycle. MATERIAL AND METHODS: The study group consisted of 16 women with periodontitis and 12 healthy women. Clinical and bacterial measurements were performed for all subjects during the ovulatory and follicular phases. RESULTS: Salivary antioxidant activity during the ovulatory phase was significantly lower than during the follicular phase in the women with periodontitis. The antioxidant activity in all subjects during the ovulatory phase was negatively correlated with Prevotella intermedia (r = -0.430; p = 0.023) and total bacterial counts (r = -0.496; p = 0.007); however, these correlations were not significant for subjects in the follicular phase. CONCLUSION: This study showed that salivary antioxidant capacity decreased, while bleeding on probing and P. intermedia increased, over the course of the menstrual cycle in women with periodontitis. Antioxidant capacity could be involved in the pathogenesis of periodontitis.

Influence of smoking on interleukin-1beta level, oxidant status and antioxidant status in gingival crevicular fluid from chronic periodontitis patients before and after periodontal treatment.

BACKGROUND AND OBJECTIVE: The aim of this study was to evaluate the impact of smoking on the relationship between interleukin-1 (IL-1ß) and oxidation in patients with periodontitis and response to nonsurgical periodontal therapy. MATERIAL AND METHODS: Data were obtained from 30 patients with generalized chronic periodontitis (15 smokers and 15 nonsmokers) and from 10 periodontally healthy controls. IL-1ß level, total oxidant status (TOS) and total antioxidant status (TAS) were recorded in gingival crevicular fluid. Probing depth, clinical attachment level, gingival and plaque indices and bleeding on probing were also measured. The gingival crevicular fluid and clinical parameters were recorded at baseline and 6 wk after periodontal treatment. RESULTS: The study showed statistically significant improvement of clinical parameters in both smokers and nonsmokers after periodontal treatment. Moreover, the baseline IL-1ß levels were significantly higher in smokers compared with nonsmokers (p < 0.05). After periodontal treatment, the IL-1ß levels were significantly reduced in both smokers and nonsmokers (p < 0.05). There were no significant differences in TOS and TAS between periodontitis patients and healthy controls at baseline and 6 wk after periodontal treatment. The level of IL-1ß in gingival crevicular fluid was positively correlated with TOS in both smokers and nonsmokers. CONCLUSIONS: Periodontal treatment improved the clinical parameters in both smokers and nonsmokers. The results confirm that periodontal therapy has an effect on IL-1ß levels in gingival crevicular fluid, but not on TOS and TAS.

Smoking influences salivary histamine levels in periodontal disease.

OBJECTIVES: Histamine, a potent vasoactive amine, is increased in saliva of periodontitis patients. The present study aimed to further investigate the diagnostic potential of histamine for periodontal disease and assessed smoking, a major risk factor of periodontitis, as a possible influencing factor. METHODS: Salivary and serum samples of 106 participants (60 periodontitis patients, 46 controls) were collected. Salivary histamine was determined by a commercially available ELISA kit, and serum C-reactive protein was measured by a routine laboratory test. Cigarettes per day and packyears were assessed as smoking exposure parameters. RESULTS: Statistically significantly increased levels of salivary histamine and serum C-reactive protein were detected between the patient and control group (P = 0.022 and P = 0.001). Salivary histamine levels were significantly higher in smoking compared with non-smoking patients (P < 0.001), and salivary histamine as well as serum C-reactive protein correlated significantly positively with smoking exposure parameters (P < 0.05). CONCLUSIONS: Smoking, an established and common risk factor of periodontitis, was assessed as a possible influencing factor for salivary histamine. Most interestingly, salivary histamine differed highly significantly between smoking and non-smoking periodontitis patients. Our results suggest a possible involvement of histamine in tobacco-exacerbated periodontal disease, but do not suggest salivary histamine as a reliable diagnostic marker for periodontitis.

Changes of four proinflammatory proteins in whole saliva during experimental gingivitis.

OBJECTIVE: To examine changes of four proinflammatory proteins in whole saliva in the early stage of plaque-induced experimental gingivitis. METHODS: Eleven young male volunteers were recruited following the cessation of all oral hygiene measures for a period of 21 days. The levels of Interleukin-6 (IL-6), Interleukin-1ß (IL-1ß), calprotectin in saliva were determined with enzyme linked immunosorbent assay. The activity of elastase in saliva was examined. RESULTS: IL-1ß, IL-6 and calprotectin in saliva increased gradually as plaque accumulated and peaked on the 14th and 21st day respectively. Moreover, the three proinflammatory proteins showed good correlations with clinical parameters, with IL-1ß correlating with clinical parameters more closely in particular. The activity of elastase in saliva elevated rapidly and peaked on the second day (P < 0.01). However, after the seventh day, elastase activity declined slowly continuously. The change of IL-6 and IL-1ß in saliva showed a similar tendency throughout the experiment, the correlation coefficient was r = 0.687 (P < 0.01), but there was no obvious correlation between calprotectin and elastase, even though both mainly come from neutrophils. CONCLUSION: Our study demonstrated that IL-6, IL-1ß and calprotectin concentrations in saliva could reflect the degree of gingival inflammation. The longitudinal change of elastase activity in saliva during the experimental gingivitis period was quite different from that of other pro-inflammatory proteins; reasons for the decrease of elastase activity in the late gingivitis period need further study.

The gingiva contains leptin and leptin receptor in health and disease.

The presence of leptin (OB) and soluble OB receptor (s-OB-R) in gingival tissue extract and gingival crevicular fluid has led the studies investigating the relationship between OB and periodontal diseases. This study aims to investigate the levels of OB and s-OB-R in serum and their presence in gingiva of healthy controls (HC), gingivitis (G), aggressive periodontitis (AP), and chronic periodontitis (CP) patients; and whether correlations exist between clinical and serum parameters, OB and s-OB-R. Seventy-seven subjects [HC (n = 20), G (n = 20), CP (n = 21), and AP (n = 16)] were included in this study. After the clinical periodontal parameter recordings and venous blood sampling, gingival tissues obtained. Serum parameters' levels determined with enzyme linked immune sorbent assay; and OB and OB-R in gingiva immunohistochemically. No significant differences were observed regarding the serum parameters [high sensitivity C-reactive protein (hs-CRP), lipids, OB, and s-OB-R] when the groups were compared (P > 0.0125). The serum OB has positive correlations with hs-CRP in the G group (P < 0.05), and s-OB-R has presented significant negative correlations with BOP in HC group (P < 0.05), with hs-CRP in G (P < 0.05) and AP groups (P < 0.05). The positive correlations were observed between the serum OB and HDL and body mass index in the CP group (P < 0.05). In all of the tissue samples of all groups, there was positive OB and OB-R immunoreactivity in the gingival epithelium. The gingival tissues contain both OB and OB-R. The serum levels of OB and s-OB-R do not vary between patients and with different periodontal conditions.

Correlations between pentraxin 3 or cytokine levels in gingival crevicular fluid and clinical parameters of chronic periodontitis.

The gingival crevicular fluid (GCF) contains various biomarkers, such as interleukin (IL)-1ß, IL-6, IL-8, tumor necrosis factor-α (TNF-α), and IL-10, among others. These cytokines have been reported to correlate with gingival inflammation and periodontal status. Therefore, the analysis of GCF may be useful for the diagnosis of periodontal status. Pentraxin 3 (PTX3) is the first identified long pentraxin, and is released by several cell types in response to proinflammatory signals. The aim of this study was to determine the levels of IL-1ß, IL-6, IL-8, TNF-α, IL-10 and PTX3 in GCF from diseased and healthy sites in patients with chronic periodontitis. Cross-sectional clinical data were obtained from 50 patients with chronic periodontitis. GCF samples were collected with paper strips from one periodontal diseased site and one periodontally healthy site per subject. The levels of IL-1ß, IL-6, IL-8, IL-10 and TNF-α were determined using a multiplexed bead immunoassay, and the PTX3 level was measured using an enzyme-linked immunosorbent assay. Mean clinical parameters were significantly higher at diseased sites (P < 0.01) as compared to healthy sites, and the mean levels of PTX3, IL-1ß, IL-6, IL-8, IL-10 and TNF-α were higher in diseased sites (P < 0.01) than in healthy sites. There were strong correlations between PTX3 or IL-1ß and periodontal status. These results suggest that GCF PTX3 levels might be useful as a diagnostic marker for periodontal disease.

Trefoil factors in saliva and gingival tissues of patients with chronic periodontitis.

BACKGROUND: Trefoil factors (TFFs) are secreted molecules that are involved in cytoprotection against tissue damage and the immune response. TFFs have been detected in saliva and oral tissues, but their clinical significance has never been investigated in patients with chronic periodontitis. The objective of this study is to determine whether TFF expression in saliva and gingival tissues is associated with periodontal pathology. METHODS: Saliva and gingival tissue samples were collected from 25 non-periodontitis individuals and 25 patients with chronic periodontitis (CP). Enzyme-linked immunosorbent assay and immunohistochemical methods were used to evaluate the expression of TFFs (TFF1, TFF2, and TFF3) in saliva and gingival tissues, respectively. Periodontopathic bacteria were quantified by real-time polymerase chain reaction. RESULTS: Reduced salivary TFF1 and TFF3 concentrations were observed in patients with CP (P = 0.003 and P <0.001, respectively). Decreased TFF3 expression in gingival tissues of patients with CP was demonstrated (P = 0.041). Levels of salivary TFF3 concentrations were negatively correlated with periodontal pathology and number of Porphyromonas gingivalis and Tannerella forsythia (formerly known as Bacteroides forsythus). CONCLUSIONS: Altered expression of TFFs in saliva and gingival tissues was detected in patients with CP. The results suggest that TFF3 may be involved in the pathogenesis of periodontal disease.

Bone remodeling-associated salivary biomarker MIP-1α distinguishes periodontal disease from health.

BACKGROUND AND OBJECTIVE: The field of salivary diagnostics lacks an accepted and validated biomarker of alveolar bone remodeling. To address this, we examined levels of salivary biomolecules specifically associated with biological aspects of bone remodeling in subjects with chronic periodontitis in a case-control study. MATERIAL AND METHODS: Levels of macrophage inflammatory protein-1α (MIP-1α), osteoprotegerin, C-telopeptide pyridinoline cross-links of type I collagen and ß-C-terminal type I collagen telopeptide in unstimulated whole saliva of 80 subjects (40 subjects with moderate to severe chronic periodontitis and 40 sex- and age-matched healthy control subjects) were measured using enzyme immunosorbent assays. Saliva was collected before clinical examination, which included probing depth, clinical attachment loss and bleeding on probing. RESULTS: The mean level of MIP-1α in subjects with periodontitis was 18-fold higher than in healthy subjects (p < 0.0001). Clinical periodontal indices correlated significantly with MIP-1α levels (p < 0.0001). Of the biomolecules examined, MIP-1α demonstrated the greatest ability to discriminate between periodontal disease and health as determined by the area under the curve (0.94) and classification and regression tree analysis (sensitivity 94% and specificity 92.7%). Osteoprotegerin levels were elevated 1.6-fold (p = 0.055), whereas C-telopeptide pyridinoline cross-links of type I collagen and ß-C-terminal type I collagen telopeptide levels were below the level of detection in the majority of subjects. CONCLUSION: These findings suggest that the chemokine MIP-1α may aid in identifying periodontitis. Future longitudinal studies are warranted to determine whether this biomarker can help in ascertaining the progression of bone loss in subjects with periodontal disease.

Correlation between leptin and the health of the gingiva: a predictor of medical risk.

CONTEXT: Over the past decade, a growing body of scientific evidence has suggested an exquisite association between oral infection and systemic diseases (e.g. atherosclerosis, cardiovascular diseases, premature or low birth weight babies, pulmonary diseases, etc.) and also between systemic diseases (e.g. arthritis, diabetes, HIV infection and osteoporosis) and oral and craniofacial diseases and disorders. Leptin is a hormone secreted by the adipocytes in varying quantities and regulates the body weight. The present study was undertaken in the context of knowing the role of leptin in the inflammatory process occurring in the gingiva as the disease progressed from gingivitis to periodontitis. AIMS: The present study was done to correlate the concentrations of leptin and interleukin (IL)-6 within the gingiva in healthy, gingivitis and periodontitis groups of patients and to correlate gingival leptin and IL-6 concentrations with plasma leptin and IL-6 concentrations in the healthy, gingivitis and periodontitis groups of patients. SETTINGS AND DESIGN: This was a cross-sectional study and was carried out on the patients from the out-patient department of Periodontics in A B Shetty Memorial Institute of Dental Sciences. PATIENTS AND METHODS: Seventy-five patients in the age group of 18-60 years were selected and grouped based on the gingival index (Loe and Sillness) and their clinical attachment levels into healthy, gingivitis and periodontitis groups. Leptin and IL-6 levels were estimated within gingiva and the plasma of each subject using an enzyme-linked immunosorbent assay kit. The results of this study were tabulated and subjected to statistical analysis. Mean and the standard deviation were calculated using analysis of variance Fisher's F-test and then the results were subjected to Tukey's Honest significance difference method for multiple comparison among the three groups. Correlation among the three groups was estimated using Pearson's correlation analysis. RESULTS: Results showed a statistically significant decrease in the concentration of gingival leptin and a statistically significant increase in the concentration of plasma leptin as the gingival disease progressed. CONCLUSION: It was concluded that as the gingival disease progressed, the gingival leptin concentration decreased, whereas the plasma leptin concentration increased, indicating a possible correlation between leptin concentration in the gingiva and the risk of developing systemic disease like the cardiovascular disease.

Expression of HMGB1 and HMGN2 in gingival tissues, GCF and PICF of periodontitis patients and peri-implantitis.

BACKGROUND AND OBJECTIVE: High mobility group chromosomal protein B1 (HMGB1) and N2 (HMGN2), two members of high mobility group (HMG) family, play important role in inflammation. The purpose of this study was to investigate the expression of HMGB1 and HMGN2 in periodontistis. MATERIALS AND METHODS: The expression of HMGB1 and HMGN2 mRNA in gingival tissues and gingival crevicular fluid (GCF) in chronic periodontitis (CP), generalised aggressive periodontitis (G-AgP) patients and healthy subjects was detected by real-time PCR. The protein level of HMGB1 and HMGN2 in peri-implant crevicular fluid (PICF), peri-implant crevicular fluid of peri-implantitis (PI-PICF) and normal patients was determined by Western blotting. Furthermore, IL-1ß, IL-6, IL-8, TNF-α and HMGB1 levels in GCF, PI-PICF and healthy-PICF samples from different groups were determined by ELISA. RESULTS: HMGN2 expression was increased in inflamed gingival tissues and GCF from CP and G-ApG groups compared to control group. HMGB1 expression was the highest in the gingival tissues and GCF from CP patients and was accompanied by increased concentrations of IL-1ß, IL-6, IL-8 proinflammaory cytokines. CONCLUSION: To our knowledge, this is the first study reporting that the expression of HMGB1 and HMGN2 was increased in the gingival tissues and GCF in CP and G-AgP and the PICF in PICF. Our data suggest that HMGB1 may be a potential target for the therapy of periodontitis and PI.

Expression of transient receptor potential vanilloid receptor 1 and toll-like receptor 4 in aggressive periodontitis and in chronic periodontitis.

BACKGROUND AND OBJECTIVE: The objective of the present study was to evaluate the expression and the distribution of the transient receptor potential vanilloid receptor 1 (TRPV1) and of toll-like receptor 4 (TLR4) in tissue samples from patients with periodontal disease (aggressive periodontitis and chronic periodontitis) and from healthy controls. MATERIAL AND METHODS: Ten tissue samples from each disease group (aggressive periodontitis and chronic periodontitis) and from healthy subjects were obtained during routine oral surgical procedures. Subgingival specimens were collected from sites with advanced loss of support (probing depth>5mm) and specimens from the corresponding healthy controls were obtained during tooth extraction for orthodontic reasons or following surgical extraction of an impacted third molar. The distribution of TRPV1 and TLR4 receptors in human gingival tissue was studied by immunohistochemistry. RESULTS: Both TLR4 and TRPV1 were detected in gingival tissues from healthy subjects, and from patients with chronic periodontitis and aggressive periodontitis, particularly in gingival keratinocytes, fibroblasts, inflammatory cells and the endothelial lining of capillaries in connective tissues. Histologic examination of the samples from healthy controls disclosed that clinically healthy gingiva does not correspond to histologically healthy gingiva. Subsequently, these samples were redesignated as gingivitis samples. TRPV1 was down-regulated in all cell types in samples obtained from patients with chronic periodontitis compared to samples obtained from patients with gingivitis, whereas TLR4 was down-regulated only in the epithelium and in gingival fibroblasts. In contrast, the levels of these markers in patients with aggressive periodontitis were similar to those in healthy patients. CONCLUSION: Local expression of TRPV1 and TLR4 in gingival tissues may contribute to both physiological and pathological processes in the periodontium. Our data suggest that TRPV1 and TLR4 may play a role specifically in the pathophysiology of chronic periodontitis.