A quantitative bias analysis to assess the impact of unmeasured confounding on associations between diabetes and periodontitis.

AIM: To investigate unmeasured confounding in bidirectional associations between periodontitis and diabetes using quantitative bias analysis. METHODS: Subsamples from the Veterans Affairs Dental Longitudinal Study were selected. Adjusted for known confounders, we used Cox proportional hazards models to estimate associations between pre-existing clinical periodontitis and incident Type II Diabetes (n = 672), and between pre-existing diabetes and incident severe periodontitis (n = 521), respectively. Hypothetical confounders were simulated into the dataset using Bernoulli trials based on pre-specified distributions of confounders within categories of each exposure and outcome. We calculated corrected hazard ratios (HR) over 10,000 bootstrapped samples. RESULTS: In models using periodontitis as the exposure and incident diabetes as the outcome, adjusted HR = 1.21 (95% CI: 0.64-2.30). Further adjustment for simulated confounders positively associated with periodontitis and diabetes greatly attenuated the association or explained it away entirely (HR = 1). In models using diabetes as the exposure and incident periodontitis as the outcome, adjusted HR = 1.35 (95% CI: 0.79-2.32). After further adjustment for simulated confounders, the lower bound of the simulation interval never reached the null value (HR ≥ 1.03). CONCLUSIONS: Presence of unmeasured confounding does not explain observed associations between pre-existing diabetes and incident periodontitis. However, presence of weak unmeasured confounding eliminated observed associations between pre-existing periodontitis and incident diabetes. These results clarify the bidirectional periodontitis-diabetes association.

The impact of periodontitis exposure misclassification bias from partial-mouth measurements on association with diabetes and cardiovascular disease.

AIM: To quantify exposure misclassification bias arising from use of partial-mouth protocols in studies of periodontitis-systemic disease associations. MATERIALS AND METHODS: Using data from 10,134 adults participating in the National Health and Nutrition Examination Survey, we classified periodontal status based on full-mouth clinical examinations and three commonly used partial-mouth protocols. Associations between periodontitis and self-reported diabetes and cardiovascular disease were evaluated under each protocol using adjusted logistic regression. Percent relative bias was calculated to evaluate magnitude and direction of bias. RESULTS: Misclassification primarily resulted in underestimation of associations, the extent of which depended on both the outcome under study and exposure severity. Bias due to misclassification of severe periodontitis was negligible for cardiovascular disease (0%-4.1%) compared to diabetes (177.7%-234.1%). In contrast, bias in moderate periodontitis associations was comparable across each outcome-diabetes (28.4%-39.5%) and cardiovascular disease (8.9%-46.7%). Results did not meaningfully change based on the partial-mouth protocol implemented. Stratified analyses showed increased bias among those with ≤15 teeth. Use of mean attachment loss as a continuous exposure resulted in minimal-to-no bias. CONCLUSIONS: Exposure misclassification bias due to use of partial-mouth protocols can yield inaccurate conclusions about periodontitis-systemic disease associations, the extent of which may depend on periodontitis classification and the association under study.

Diminished bone formation during diabetic fracture healing is related to the premature resorption of cartilage associated with increased osteoclast activity.

UNLABELLED: Histological and molecular analysis of fracture healing in normal and diabetic animals showed significantly enhanced removal of cartilage in diabetic animals. Increased cartilage turnover was associated with elevated osteoclast numbers, a higher expression of genes that promote osteoclastogenesis, and diminished primary bone formation. INTRODUCTION: Diminished bone formation, an increased incidence of nonunions, and delayed fracture healing have been observed in animal models and in patients with diabetes. Fracture healing is characterized by the formation of a stabilizing callus in which cartilage is formed and then resorbed and replaced by bone. To gain insight into how diabetes affects fracture healing, studies were carried out focusing on the impact of diabetes on the transition from cartilage to bone. MATERIALS AND METHODS: A low-dose treatment protocol of streptozotocin in CD-1 mice was used to induce a type 1 diabetic condition. After mice were hyperglycemic for 3 weeks, controlled closed simple transverse fractures of the tibia were induced and fixed by intramedullary pins. Histomorphometric analysis of the tibias obtained 12, 16, and 22 days after fracture was performed across the fracture callus at 0.5 mm proximal and distal increments using computer-assisted image analysis. Another group of 16-day samples were examined by microCT. RNA was isolated from a separate set of animals, and the expression of genes that reflect the formation and removal of cartilage and bone was measured by real-time PCR. RESULTS: Molecular analysis of collagen types II and X mRNA expression showed that cartilage formation was the same during the initial period of callus formation. Histomorphometric analysis of day 12 fracture calluses showed that callus size and cartilage area were also similar in normoglycemic and diabetic mice. In contrast, on day 16, callus size, cartilage tissue, and new bone area were 2.0-, 4.4-, and 1.5-fold larger, respectively, in the normoglycemic compared with the diabetic group (p < 0.05). Analysis of microCT images indicated that the bone volume in the normoglycemic animals was 38% larger than in diabetic animals. There were 78% more osteoclasts in the diabetic group compared with the normoglycemic group (p < 0.05) on day 16, consistent with the reduction in cartilage. Real-time PCR showed significantly elevated levels of mRNA expression for TNF-alpha, macrophage-colony stimulating factor, RANKL, and vascular endothelial growth factor-A in the diabetic group. Similarly, the mRNA encoding ADAMTS 4 and 5, major aggrecanases that degrade cartilage, was also elevated in diabetic animals. CONCLUSIONS: These results suggest that impaired fracture healing in diabetes is characterized by increased rates of cartilage resorption. This premature loss of cartilage leads to a reduction in callus size and contributes to decreased bone formation and mechanical strength frequently reported in diabetic fracture healing.

Immunization enhances inflammation and tissue destruction in response to Porphyromonas gingivalis.

It is well established that host-bacterium interactions play a critical role in the initiation and progression of periodontal diseases. By the use of inhibitors, it has been shown that mediators associated with the innate immune response significantly contribute to the disease process. Less is known regarding the role of the acquired immune response. To investigate mechanisms by which the acquired immune response to Porphyromonas gingivalis could affect connective tissue, we used a well-documented calvarial model to study host-bacterium interactions. Injection of P. gingivalis stimulated gamma interferon, interleukin 6, macrophage inflammatory protein 2, and monocyte chemoattractant protein 1 expression as determined by real-time PCR. Prior immunization against P. gingivalis significantly enhanced the mRNA levels of these cytokines and chemokines. Similarly, immunization significantly increased and prolonged the formation of a polymorphonuclear leukocyte and mononuclear cell infiltrate (P < 0.05). In addition, the area of connective tissue destruction, osteoclastogenesis, bone loss, mRNA expression of proapoptotic genes, and degree of fibroblast apoptosis were increased in immunized mice (P < 0.05). These results indicate that activation of the acquired immunity by P. gingivalis increases the inflammatory and destructive responses which occur in part through up-regulating the innate immune response and enhancing osteoclastogenesis and fibroblast apoptosis.

Morphometric analysis of the furcation anatomy of mandibular molars.

BACKGROUND: Successful treatment of molar furcation defects remains a challenge in clinical practice. Knowledge of anatomic factors facilitates predictable management of furcation involvement lesions. The degree of success in managing furcation involvement is inversely related to the horizontal probing depth. The depth of the horizontal component of attachment loss can vary depending on the external tooth-surface reference points used. However, the anatomical factors affecting horizontal component of attachment loss have not been previously assessed. Therefore, this study determined the bucco-lingual measurements of the cemento-enamel junction and the mesial and distal roots and at the level of root separation. METHODS: One hundred extracted permanent human mandibular first (N = 50) and second (N = 50) molars were studied. Four horizontal bucco-lingual widths were measured with calibrated calipers: 1) furcation entrance/roof (FE); 2) cemento-enamel junction level (CEJ); 3) mesial root width (MRW); and 4) distal root width (DRW). RESULTS: The mean widths at FE, CEJ, MRW, and DRW were, respectively, 5.53 +/- 0.45 mm, 8.71 +/- 0.54 mm, 8.57 +/- 0.54 mm, and 7.97 +/- 0.65 mm in the first molars and 5.61 +/- 0.65 mm, 8.40 +/- 0.65 mm, 7.95 +/- 0.88 mm, and 7.16 +/- 0.84 mm in the second molars. Analysis of variance revealed significant differences between FE and the other variables tested. The results showed that the bucco-lingual width of the furcation roof is considerably shorter than the MRW and DRW. The difference in the mean bucco-lingual dimension between FE and the other measurements occurred in all teeth evaluated and varied between 0.7 and 4.30 mm. CONCLUSIONS: Our findings demonstrate that clinical measurements of horizontal probing depth that use the external surfaces of roots as reference points overestimate the true anatomical component of furcation involvement in mandibular molars. Conversely, positive treatment outcomes in these teeth may be underestimated. This has implications not only for clinical practice but also for clinical research studies evaluating treatment outcomes.