Cardiovascular Disease Parameters in Periodontitis

Volume 80 • Number 3

Cardiovascular Disease Parameters in Periodontitis Andre´a M. Monteiro,* Maria A.N. Jardini,† Sarah Alves,‡ Viviana Giampaoli,§ ˆ nio M. Figueiredo Neto,‡ and Magnus Gidlund* Elisete C.Q. Aubin,§ Anto

Background: Recently, there has been an increasing in the impact of oral health on atherosclerosis and subsequent cardiovascular disease. The aim of this study is to investigate the association between chronic periodontitis and cardiovascular risk markers. Methods: Forty patients with periodontitis and 40 healthy gender-, body mass index–, and age-matched individuals were compared by measuring total cholesterol, high-density lipoprotein, low-density lipoprotein, triglycerides, levels of cytokines, antibodies against oxidized low-density lipoprotein, thiobarbituric acid reactive substances, total and differential white blood cell counts, and the non-linear index of refraction. Results: The levels of triglycerides and high-density lipoprotein in periodontitis patients were significantly higher and lower, respectively (P = 0.002 and P = 0.0126), compared to controls. Total cholesterol, low-density lipoprotein, and lipid peroxide levels were the same in both groups (P = 0.2943, P = 0.1284, and P = 0.067, respectively). Interleukin (IL)-6 and -8, antibodies against oxidized low-density lipoprotein, and leukocyte and neutrophil counts were significantly higher in periodontitis patients (P 5 mm. Inclusion criteria for controls were the absence of clinical and radiographic manifestations of periodontal disease, no history of periodontal disease, and ‡16 teeth present. Clinical examination. All clinical parameters were assessed by a single trained periodontitis expert, and a calibration exercise was performed to obtain acceptable interexaminer reproducibility. The evaluation included bleeding on probing as an indication of existing periodontal inflammation. This was done by measuring the PD and gingival recession at six sites (mesio-buccal, mid-buccal, disto-buccal, mesio-lingual, mid-lingual, and disto-lingual) of the teeth, with the exception of the third molars, by using a pressurecalibrated digital recording device.i Each individual was codified and randomized, and further analysis was done without previous knowledge of the state of the individual from which the sample was drawn. Plasma Collection Ten milliliters of blood was collected from each individual after 12 hours of fasting. The plasma was obtained after centrifugation of the blood at 1,000 · g at 4C for 15 minutes. Peripheral blood was also collected for the determination of the total and differential

i Florida Probe, Gainesville, FL.

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Risk Markers for Coronary Artery Disease in Periodontitis

white blood cell counts, which were performed in standard automated set-ups. Cytokine Assays The plasma was stored at -70C until analyzed. Interleukin (IL)-1b, -6, -8, and -10 and tumor necrosis factor-alpha (TNF-a) concentrations were tested using commercially available quantitative enzyme-linked immunosorbent assay kits.¶ Detection of Antibodies Against oxLDL Plates were coated with human oxLDL (7.5 mg/ml) in 0.1 M carbonate/bicarbonate buffer (pH 9.6) overnight at 4C. The plates were blocked with gelatin for 2 hours at room temperature. Serum samples (50 ml) diluted at 1:400 were added in duplicate and incubated for 2 hours. Then the plates were washed four times with phosphate buffered saline (PBS)polysorbate.# A peroxidase-conjugated mouse antihuman immunoglobulin G (dilution 1:1,000)** was added, and the plates were washed after 1 hour at room temperature. Substrate solution (3,39,5,59-tetramethylbenzidine in 0.1 M citrate-phosphate buffer, pH 5.5; plus H2O2) was added, and the reaction was interrupted 5 minutes later with the addition of 5 ml 2 M H2SO4.†† The optical density (OD) was measured using a microplate reader‡‡ at 450 nm, and the results were expressed as absorbance. Determination of TC, TGs, and HDL Plasma TC, TGs, and HDL were quantified by enzymatic methods.§§ LDL was estimated by using the Friedewald formula.26 To identify individuals with pathologic values, the following cutoff points were used according to the manufacturer’s recommendation: TC ‡200 mg/dl, LDL ‡130 mg/dl, TG ‡150 mg/dl, and low HDL 0.3 using the Wald test). Thus, within this model, we concluded that there is no significant association between age, gender, or BMI and periodontitis. In a second step, we established new groups for analyzed risk factors for cardiovascular disease using the following categorical criteria: gender (female versus male), age (