Salivary proteomics in bisphosphonate-related osteonecrosis of the jaw.

OBJECTIVE: The objective of this study was to identify differentially expressed salivary proteins in bisphosphonate-related osteonecrosis of the jaw (BRONJ) patients that could serve as biomarkers for BRONJ diagnosis. SUBJECTS AND METHODS: Whole saliva obtained from 20 BRONJ patients and 20 controls were pooled within groups. The samples were analyzed using iTRAQ-labeled two-dimensional liquid chromatography-tandem mass spectrometry. RESULTS: Overall, 1340 proteins were identified. Of these, biomarker candidates were selected based on P-value (<0.001), changes in protein expression (≥1.5-fold increase or decrease), and unique peptides identified (≥2). Three comparisons made between BRONJ and control patients identified 200 proteins to be differentially expressed in BRONJ patients. A majority of these proteins were predicted to have a role in drug metabolism and immunological and dermatological diseases. Of all the differentially expressed proteins, we selected metalloproteinase-9 and desmoplakin for further validation. Immunoassays confirmed increased expression of metalloproteinase-9 in individual saliva (P = 0.048) and serum samples (P = 0.05) of BRONJ patients. Desmoplakin was undetectable in saliva. However, desmoplakin levels tended to be lower in BRONJ serum than controls (P = 0.157). CONCLUSIONS: Multiple pathological reactions are involved in BRONJ development. One or more proteins identified by this study may prove to be useful biomarkers for BRONJ diagnosis. The role of metalloproteinase-9 and desmoplakin in BRONJ requires further investigation.

Design features of the Diabetes and Periodontal Therapy Trial (DPTT): a multicenter randomized single-masked clinical trial testing the effect of nonsurgical periodontal therapy on glycosylated hemoglobin (HbA1c) levels in subjects with type 2 diabetes and chronic periodontitis.

BACKGROUND: Evidence suggests that periodontitis is associated with prevalent and incident type 2 diabetes mellitus (T2DM), raising the question of whether periodontitis treatment may improve glycemic control in patients with T2DM. Meta-analyses of mostly small clinical trials suggest that periodontitis treatment results in a modest reduction in glycosylated hemoglobin (Hb) A1c. PURPOSE: The purpose of the Diabetes and Periodontal Therapy Trial (DPTT) was to determine if periodontal treatment reduces HbA1c in patients with T2DM and periodontitis. METHODS: DPTT was a phase-III, single-masked, multi-center, randomized trial with a planned enrollment of 600 participants. Participants were randomly assigned to receive periodontal treatment immediately (Treatment Group) or after 6 months (Control Group). HbA1c values and clinical periodontal measures were determined at baseline and 3 and 6 months following randomization. Medication usage and dosing were assessed at each visit. Periodontal treatment consisted of scaling and root planing for a minimum of two 90-minute sessions, plus the use of an antibacterial mouth rinse for at least 32 days afterwards. The primary outcome was change in HbA1c from baseline to 6 months and the trial was powered to detect a between-group difference of 0.6%. Secondary outcomes included changes in periodontal clinical measures, fasting plasma glucose, the Homeostasis Model Assessment (HOMA2) and the need for rescue diabetes or periodontal therapy. CONCLUSION: Dental and medical researchers collaborated to recruit, treat and monitor participants with two chronic diseases to determine if treatment of one condition affects the status of the other.

A pilot study of the association between cariogenic oral bacteria and preterm birth.

OBJECTIVES: We examined the associations between preterm birth and low birth weight and maternal caries history, maternal periodontal status, and salivary levels of mutans streptococci and Lactobacilli. DESIGN: This study was a matched case-control study in women during their pregnancy or up to 8 weeks after delivery. SUBJECTS AND METHODS: Thirty-four women delivering before 37 weeks gestation were recruited along with 73 term controls matched for age and race/ethnicity. Demographic and obstetric information was collected from questionnaires and medical records and oral examinations along with commercial salivary tests were completed within the study groups. MAIN OUTCOME MEASURES: The main outcome variables were the preterm birth and low birth weight status. The independent variables measured were the salivary levels of Lactobacilli and mutans streptococci and the caries and periodontal status of the subjects. RESULTS: The odds ratio comparing low levels of bacteria in preterm mothers and controls was statistically significant for Lactobacilli (odds ratio (OR) = 3.45, 95% confidence interval (CI) = 1.27 to 10.00) and almost significant for mutans streptococci (OR = 2.63, 95% CI = 0.95 to 8.33). Clinical caries and periodontal disease measures did not differ significantly between groups. CONCLUSION: Within the limitation of our study, low levels of Lactobacilli in saliva were found to be associated with preterm birth.

Evidence of a substantial genetic basis for risk of adult periodontitis.

BACKGROUND: A few previous studies have suggested that risk for adult periodontitis (AP) has a genetic (heritable) component. We estimated genetic and environmental variances and heritability for gingivitis and adult periodontitis using data from twins reared together. METHODS: One hundred seventeen (117) pairs of adult twins (64 monozygotic [MZ] and 53 dizygotic [DZ] pairs) were recruited. Probing depth (PD), attachment loss (AL), plaque, and gingivitis (GI) were assessed on all teeth by two examiners. Measurements were averaged over all sites, teeth, and examiners. Extent of disease in subjects was defined at four thresholds: the percentage of teeth with AL > or = 2, AL > or = 3, PD > or = 4, or PD > or = 5 mm. Genetic and environmental variances and heritability were estimated using path models with maximum likelihood estimation techniques. RESULTS: MZ twins were more similar than DZ twins for all clinical measures. Statistically significant genetic variance was found for both the severity and extent of disease. AP was estimated to have approximately 50% heritability, which was unaltered following adjustments for behavioral variables including smoking. In contrast, while MZ twins were also more similar than DZ twins for gingivitis scores, there was no evidence of heritability for gingivitis after behavioral covariates such as utilization of dental care and smoking were incorporated into the analyses. CONCLUSIONS: These results confirm previous studies and indicate that approximately half of the variance in disease in the population is attributed to genetic variance. The basis for the heritability of periodontitis appears to be biological and not behavioral in nature.

No heritability of temporomandibular joint signs and symptoms.

The causes of temporomandibular joint (TMJ)-related signs and symptoms are largely unknown. We tested the hypotheses that these signs and symptoms, as well as oral parafunctional habits, are substantially heritable. Questionnaire and clinical data were collected from 494 twins, including pairs of reared-apart and reared-together monozygotic (MZ) and dizygotic (DZ) twins. A history of joint-area pain, joint noises, and clenching and grinding habits was scored as present or absent. Twenty-nine percent of the population experienced at least one sign or symptom. Nearly one-quarter of subjects clenched or ground their teeth, and 8.7% reported a history of joint-area pain. Pain was associated with clenching, grinding, and joint noises. MZ twins were no more similar than DZ twins for any outcome, suggesting that genetic factors do not influence these traits in the population. Reared-together MZ twins were no more similar than reared-apart MZ twins, suggesting a negligible effect of the family environment on these outcomes. Environmental factors unique to each twin appeared to be the major determinants of variation in this population.

Human herpesviruses and Porphyromonas gingivalis are associated with juvenile periodontitis.

BACKGROUND: Although herpesviruses have been associated with adult periodontitis, their relationship with juvenile periodontitis (JP) has not been established. This case-control study examined possible associations between JP and pathogenic bacteria, the human cytomegalovirus (HCMV), and the Epstein-Barr type 1 virus (EBV-1). METHODS: Subjects were participants in a larger survey of schoolchildren in North-Central Jamaica. Subgingival plaque samples from 15 subjects with JP, 20 with incipient periodontitis (IP), and 65 randomly-selected healthy controls were assayed for Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans using a 16S rRNA polymerase chain reaction (PCR) identification method, and for HCMV and EBV-1 using nested PCR identification. RESULTS: Strong bivariate associations were found between JP and P. gingivalis (odds ratio [OR] = 12.7; 95% CI = 2.6, 61.4), HCMV (OR = 10.0; 95% CI = 2.7, 36.3), and A. actinomycetemcomitans (OR = 8.0; 95% CI = 2.3, 27.5), but not EBV-1. In multivariate analyses, P. gingivalis remained a significant explanatory variable (OR = 7.8; 95% CI = 1.5, 40.9); however, the associations were marginal for HCMV (OR = 4.6; 95% CI = 0.9, 22.7), and non-significant for A. actinomycetemcomitans (OR = 2.0; 95% CI = 0.4, 9.7). The associations with JP and the extent of attachment loss were even stronger when both P. gingivalis and HCMV were detected together. P. gingivalis (OR = 3.9; 95% CI = 1.3, 12.0) and EBV-1 (OR = 3.3; 95% CI = 1.0, 10.3) were the only significant explanatory variables in the multivariate analysis of IP. CONCLUSIONS: P. gingivalis is the strongest and most stable indicator of periodontitis in Jamaican adolescents. Co-infection with P. gingivalis and HCMV appears to be particularly deleterious to periodontal health.

Frequency of 530-bp deletion in Actinobacillus actinomycetemcomitans leukotoxin promoter region.

Actinobacillus actinomycetemcomitans strains showing a 530-bp deletion in the promoter region of the leukotoxin gene operon elaborate high amounts of leukotoxin that may play a role in the pathogenesis of periodontal disease. This study used polymerase chain reaction detection to determine the occurrence of the 530-bp deletion in 94 A. actinomycetemcomitans strains from individuals of various ethnic backgrounds. Eleven blacks and one Hispanic subject but no Caucasian or Asian subjects showed the 530-bp deletion in the leukotoxin promoter region, suggesting that the deletion is mainly a characteristic of individuals of African descent. A. actinomycetemcomitans strains exhibiting a deletion in the leukotoxin promoter region occurred both in individuals having severe periodontitis and in adolescents revealing no evidence of destructive periodontal disease.

Periodontal bacteria in adult twins.

BACKGROUND: Both environmental and genetic factors are known to influence clinical measures of periodontal disease. The purpose of this study was to determine whether genetic factors similarly influence the presence of specific periodontal bacteria in subgingival plaque. METHODS: Reared-together and reared-apart monozygous (MZ) and dizygous (DZ) adult twins were examined clinically. Demographic and behavioral information was obtained from each subject by questionnaire. Subgingival plaque samples were obtained from the index teeth, and the presence of P. intermedia, P. gingivalis, A. actinomycetemcomitans, E. corrodens, and F. nucleatum was determined using an immunoassay. RESULTS: Microbiological and clinical data were available for 169 twin pairs. The subject-based prevalences of the bacteria in the twin groups ranged from 11% for Porphyromonas gingivalis to 40% for F. nucleatum. For all species examined, the concordance rates were not significantly different (P > 0.05) between MZ and DZ twin groups. These findings were apparent despite similar smoking histories, self-reported oral hygiene practices, and antibiotic use in the twin groups. Furthermore, MZ twins reared together were not more similar than MZ reared-apart twins with respect to any bacterial species examined. CONCLUSIONS: These findings suggest that in a population with access to routine dental care, any effects that host genes and the early family environment have on the presence of specific bacteria in subgingival plaque are not apparent in adulthood. Most twins with disease in this study had early periodontitis. Results from this study may not necessarily be extrapolated to more advanced disease states.

Cell-mediated immune system regulation in periodontal diseases.

The adaptive immune system consists of humoral and cell-mediated immunity. T-lymphocytes are the key components of cell-mediated immunity. CD4+ helper T-lymphocytes facilitate B-cells to differentiate and produce specific antibodies, whereas CD8+ cytotoxic T-lymphocytes kill virally infected cells. Periodontal diseases have been associated with a variety of imbalances in the regulation of immune responses. Changes in the ratios of peripheral blood CD4+ and CD8+ T-lymphocytes, depressed proliferative responses of peripheral blood lymphocytes, and increased frequency of CD45RO+ memory T-lymphocytes in diseased tissues have been reported in individuals with various forms of periodontal disease. While some studies have shown an increased frequency of gamma delta + T-cells in periodontal lesions, the role of gamma delta + T-cells in periodontal disease remains controversial. The ability of putative periodontopathic bacteria selectively to stimulate certain V beta-expressing T-cells is intriguing and could determine whether a CD4+ Th1 or a CD4+ Th2 cell response is elicited. The prominence of a particular subset of helper T-cells within the periodontal lesion could be a reflection of the stage and activity of the disease, or the types of bacteria present. Regardless, longitudinal studies of the involvement of T-cell subsets and cytokines in periodontal disease are clearly needed.

Detection of local and systemic cytokines in adult periodontitis.

Periodontitis is a chronic inflammatory disease of the soft and hard supporting tissues of the teeth and is a major cause of tooth loss in adults. The local host response to periodontopathic bacteria results in the release of inflammatory mediators and cytokines. Since cytokines are indicative of effector functions, we compared the pattern of cytokine production in periodontal patients and healthy controls. Specifically, we investigated the simultaneous presence of cytokines produced by T helper (Th)1, Th 2, and inflammatory cells which could be involved in periodontitis. We also compared the expression of these cytokine mRNAs in healthy and diseased tissues. No significant differences were detected at the protein or mRNA levels of the cytokines in the systemic circulation of patients and controls. The surface markers CD16 and CD56 were expressed on significantly fewer peripheral mononuclear cells of patients when compared to controls. gamma delta + T cells were found in half of the diseased tissues, but in none of the healthy tissues of either patients or controls. Finally, significant differences were observed between healthy and inflamed gingival tissues in the cytokine mRNA profile. Expression of IL-6 and IFN-alpha mRNA was significantly higher in diseased tissues compared to healthy tissues in patients.

Crevicular fluid enzymes from endosseous dental implants and natural teeth.

Several neutrophil-derived enzymes that are present in the gingival crevicular fluid have been evaluated for use as risk markers for periodontal disease progression. However, very little information is available about the presence of these enzymes in peri-implant tissues. The purpose of this cross-sectional study was to compare levels of enzymes in gingival crevicular fluid between natural teeth and endosseous dental implants and between well-integrated and failing implants. Scores of plaque and gingivitis were recorded for 68 integrated implants, five failing implants, and 34 natural teeth in 12 completely edentulous and 18 partially edentulous subjects. Samples of gingival crevicular fluid were obtained from these sites using filter paper strips and were assayed for levels of neutral protease, neutrophil elastase, myeloperoxidase, and beta-glucuronidase. Neutral protease levels were higher (P = .066) at moderately to severely inflamed implant sites (Gingival Index of 2, 3) compared to mildly or noninflamed sites (Gingival Index of = 0, 1). Despite the small number (n = 5) of failing implants evaluated in this study, levels of neutrophil elastase, myeloperoxidase, and beta-glucuronidase were significantly higher (P < or = .001) around failing implants compared to successful implants. Neutral protease levels were also elevated around failing implants, but the difference was not statistically significant. Results of this study indicate that neutrophil elastase, myeloperoxidase, and beta-glucuronidase levels in GCF appear to be good candidates for study as risk markers of implant failure.

Evaluation of periodontal treatments using controlled-release tetracycline fibers: clinical response.

The purpose of this investigation was to evaluate the clinical efficacy of controlled-release tetracycline fiber therapy in adult periodontitis patients. One hundred-twenty-two (122) adult patients from 3 dental centers were enrolled at baseline for this study. each patient provided at least one site in each of four quadrants that was > or = 5 mm and bled on probing. One or two such sites were selected as test sites and were randomly assigned to receive one of four treatments: scaling and root planing (S), scaling and root planing plus tetracycline fiber for 10 days (SF), fiber therapy alone for 10 days (F), or two 10-day serial fiber applications (FF). After treatment, no periodontal maintenance or supportive care was provided until the end of this 12-month study. Probing depth (PD), clinical attachment level (CAL), plaque, and bleeding on probing (BOP) were measured at baseline and at 1, 3, 6, 9, and 12 months following treatment. Repeated PD and CAL measurements were taken at three locations within each site and averaged for each site. One hundred-sixteen (116) subjects completed the study. All treatments resulted in similar improvements in clinical parameters compared to baseline and were equally effective in the treatment of periodontitis as measured by probing depth reduction, clinical attachment level gain, and reduction of bleeding on probing. The clinical response, established primarily by 3 months following therapy, was generally sustained in all treatment groups for 12 months without the benefit of supportive maintenance therapy.

Evaluation of periodontal treatments using controlled-release tetracycline fibers: microbiological response.

In a 12-month multi-center study of 116 adult periodontitis subjects, six putative periodontal pathogens were monitored by DNA probe methods in a subset of 31 subjects. Monitored species included Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Fusobacterium nucleatum (Fn), Eikenella corrodens (Ec), Campylobacter rectus (Cr), and Actinobacillus actinomycetemcomitans (Aa) with an average detection limit of 1.8 x 10(4) bacterial colony forming units/sample. The microbiological response to four periodontal treatments was studied, one treatment in each quadrant; scaling and root planing (S), scaling and root planing with tetracycline (TC) fiber (SF), a single application of TC fiber (F) and two serial applications of TC fiber (FF). Generally two sites were sampled in each quadrant, however, in some quadrants only one site was selected. These treatments were evaluated at baseline; immediately following therapy; and post-treatment at 1, 3, 6, and 12 months. The study was conducted with a split-mouth design with no maintenance therapy over a 12-month period. At baseline, 70.8% of sites had detectable Fn; 42.9% Pg; 63.5% Pi; 29.7% Ec; 28.3% Cr; and 5.5% Aa. No significant differences were seen in baseline proportions of these species between centers. Numbers and proportions of detectable pathogens (with the exception of Pg) exhibited a triphasic temporal response: a precipitous initial decrease immediately following therapy; a rise in proportions in the 1- to 3-month post-therapy period; and a spontaneous decline in the absence of therapy over the 3- to 12-month period.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of periodontal treatments using controlled-release tetracycline fibers: maintenance response.

The purpose of this investigation was to examine periodontal disease recurrence from 3 to 12 months following various treatments with scaling and root planing and controlled-release tetracycline fibers. One-hundred-twenty-two (122) adult volunteers with at least one bleeding pocket > or = 5 mm in each of four quadrants were enrolled in this study. One or two such sites in each quadrant were selected as test sites. Quadrants were randomly assigned to receive one of four treatments: scaling and root planing (S); scaling and root planing plus tetracycline fiber for 10 days (SF); fiber therapy alone for 10 days (F); or fiber therapy alone for 20 days (FF). After treatment, no supportive care was provided during the 12-month study period. Probing depth (PD), attachment loss (AL), plaque, and bleeding on probing were measured at baseline, and at 1, 3, 6, 9, and 12 months after treatment. PD and AL measures were taken at three locations within each site and averaged for each site. Disease recurrence was defined as > or = 1 mm mean attachment loss at a site during the 3- to 12-month period. One-hundred-sixteen (116) subjects completed the study. Sites treated with SF experienced significantly (P < 0.05) less disease recurrence (4%) than S, F, or FF (9%, 10%, and 12%, respectively). Results of this study suggest that, compared to S, F, or FF, scaling and root planing in conjunction with tetracycline fiber therapy for 10 days can significantly reduce disease recurrence 3 to 12 months following treatment in the absence of supportive care.

Genetic contributions to saliva protein concentrations in adult human twins.

The heritability of saliva protein concentrations was investigated in stored samples of clarified stimulated whole saliva from adult twins participating in a study of periodontal disease genetics. Saliva was obtained from 29 monozygous and 20 dizygous twin pairs. Visits were scheduled so that both twins in a pair donated saliva at the same time of day. Flow rate was determined, and frozen samples later assayed for lactoferrin, lysozyme, secretory IgA, total peroxidase, myeloperoxidase and total protein. Pairs were always assayed together. Within- and between-pair variances were used to estimate twin intraclass correlations. Pearson correlations were used to estimate associations between saliva variables and clinical indices of gingivitis, dental plaque, periodontal attachment loss, and probing depth. Significant genetic contributions to variance were seen for total protein, lactoferrin, and total peroxidase. Total protein showed a significant positive correlation with gingivitis. There were no other correlations with clinical indices, and intraclass correlations for saliva variables did not change after adjustment for gingivitis. Dizygous twin correlations were higher than monozygous twin correlations for flow rate, lysozyme, and secretory IgA. That may be an artefact due to small numbers of pairs. It seems unlikely that a common environmental factor would strongly affect saliva in twins living apart as adults. Present findings, taken as sib correlations, support a genetic contribution to saliva protein concentrations. Problems with the twin model in saliva might be resolved by longitudinal studies of large numbers of twins.

Genetic and heritable risk factors in periodontal disease.

The purpose of this paper is to review current knowledge of genetic risk factors for the periodontal diseases and to present updated and additional data from the Minnesota Twin Periodontal Study. Family studies suggest that susceptibility to the early onset forms of disease, particularly prepubertal and juvenile periodontitis, is, at least in part, influenced by host genotype. Inherited phagocytic cell deficiencies appear to confer risk for prepubertal periodontitis. The prevalence and distribution of juvenile periodontitis in affected families are most consistent with an autosomal recessive mode of inheritance. However, considerable etiologic as well as genetic heterogeneity within these clinically-defined diseases is evident. Whether or not genetic factors influence the more common adult chronic periodontitis is less clear. Although results from family studies suggest that environmental factors appear to be the major determinants of variance in adult periodontitis, data from our twin studies indicate that both genetic and environmental factors influence disease. Furthermore, comparisons between reared-together and reared-apart adult monozygous twins indicate that early family environment has no appreciable influence on probing depth and attachment loss measures in adults.

Genetic and inheritance considerations in periodontal disease.

Considerable evidence suggests that there is some genetic basis for the early onset forms of disease. The molecular abnormality and its genetic inheritance has been established in some cases of generalized prepubertal periodontitis. Family studies of juvenile periodontitis indicate that this disorder is transmitted by autosomal recessive genes, although additional data indicate genetic heterogeneity in this clinically defined disease. Recent evidence also suggests that susceptibility to periodontal disease may be related in part to genetically determined immune responsiveness to bacterial lipopolysaccharides. Although specific genetic risk factors have not been identified for the more common adult chronic periodontitis, recent studies indicate that there is significant genetic variance in the population for clinical and radiographic measures of disease. More precise definitions of disease phenotypes will facilitate future genetic epidemiologic studies of the periodontal diseases.

A twin study of genetic variation in proportional radiographic alveolar bone height.

The purpose of this study was to estimate the genetic variance for alveolar bone height by means of the classic twin method and the study of monozygous twins reared apart. Panoramic radiographs were obtained from 120 pairs of adult twins (mean age = 40.4 years, S.D. = 10.4 years), for comparison of 62 pairs of monozygous twins reared together (MZT), 25 pairs of like-sexed dizygous twins reared together (DZT), and 33 pairs of monozygous twins reared apart (MZA). Mesial and distal bone heights were determined as a proportion of tooth length. A full-mouth bone score was computed for each twin by averaging these proportions from all measurable teeth. Between-pair (B) and within-pair (W) variances were computed for each twin group. The population variances (B + W) of the MZT and DZT twin groups were similar, which validated a basic assumption of the twin model. Intraclass correlations and heritability estimates were also computed for the reared-together and reared-apart twin groups. Boot-strap sampling was used to provide estimates and confidence limits for these values. The intraclass correlations for the twin groups were: MZT = 0.70, DZT = 0.52, and MZA = 0.55. The results of this study suggest that there is significant genetic variance in the population for proportional alveolar bone height.

Periodontal findings in adult twins.

Gingivitis and periodontitis are among the most common diseases known to man. Although bacterial plaque is generally accepted as the primary etiologic agent, little information is available concerning the influence that host genetic factors have on these diseases. The purpose of the present study was to examine the relative contribution of environmental and host genetic factors to clinical measures of periodontal disease through the study of both reared-together twins and monozygous twins reared apart. Probing depth, clinical attachment loss, gingivitis, and plaque were assessed from the Ramfjord teeth in 110 pairs of adult twins (mean age 40.3 years), including 63 monozygous and 33 dizygous twin pairs reared together and 14 monozygous twin pairs reared apart. Bootstrap sampling was used to estimate and provide confidence limits of between-pair and within-pair variances, intraclass correlations and heritability. Based on ratios of within-pair variances or heritability estimates, a significant (P less than 0.05) genetic component was identified for gingivitis, probing depth, attachment loss and plaque. Heritability estimates indicated that between 38% to 82% of the population variance for these periodontal measures of disease may be attributed to genetic factors. While there is general agreement that bacteria are important in the pathogenesis of the periodontal diseases, future etiologic studies should consider the role of host genetic influences.