Silvana Regina Perez Orrico

Effect of periodontal treatment on metabolic control, systemic inflammation and cytokines in patients with type 2 diabetes.

OBJECTIVE The aim of this study was to investigate the effect of periodontal therapy on the circulating concentration of high-sensitivity capsule-reactive protein (hs-CRP), fibrinogen (FIB), interleukin (IL)-4, IL-6, IL-8, IL-10 and tumour necrosis factor-alpha (TNF-alpha) and on the metabolic control in type 2 diabetes mellitus (T2DM) patients. MATERIAL AND METHODS Twenty-three T2DM patients with chronic periodontitis were enrolled in this study. Periodontal clinical parameters, namely visible plaque index, gingival bleeding index, bleeding on probing, probing depth and clinical attachment levels, were evaluated. Blood samples for plasma were collected and assessed for the levels of hs-CRP, FIB, IL-4, IL-6, IL-8, IL-10 and TNF-alpha. The glycated haemoglobin (HbA(1c)) and fasting plasma glucose were also measured. All parameters were evaluated before and 3 months after non-surgical periodontal therapy. RESULTS All clinical parameters were significantly improved 3 months after the periodontal therapy. A univariate comparison showed a tendency towards a decrease of the measured biomarkers, most pronounced for TNF-alpha and FIB, after therapy. Periodontal treatment also reduced HbA(1c) and hs-CRP levels, albeit not significantly. CONCLUSIONS The clinically successful non-surgical periodontal therapy tended to reduce systemic inflammation and the concentration of some circulating cytokines.

The Short-Term Effectiveness of Non-Surgical Treatment in Reducing Levels of Interleukin-1β and Proteases in Gingival Crevicular Fluid From Patients With Type 2 Diabetes Mellitus and Chronic Periodontitis

BACKGROUND This study aimed to compare the effectiveness of non-surgical periodontal treatment in improving periodontal status and reducing gingival crevicular fluid (GCF) levels of interleukin (IL)-1β and IL-18, elastase activity, and matrix metalloproteinase (MMP)--8 and --9 in periodontitis patients with and without type 2 diabetes mellitus (T2DM). METHODS Twenty-three patients with T2DM (diabetes group) and 26 systemically healthy subjects (control group) with chronic periodontitis participated in this study. The clinical examination included visible plaque index, gingival bleeding index, probing depth, clinical attachment level, and bleeding on probing. GCF samples were collected from five or six deep sites to evaluate the levels of IL-1β and -18, elastase, and MMP-8 and -9. Shallow sites were analyzed for IL-1β and elastase. The glycemic control was analyzed by the concentration of glycated hemoglobin (HbA1c). The subjects received non-surgical periodontal treatment and were reexamined 90 days later. RESULTS All clinical parameters showed a significant improvement after treatment, which was accompanied by a significant reduction in IL-1β, elastase activity, and MMP-8 and -9 levels in deep sites. The shallow sites also showed significant reductions in IL-1β and elastase activity levels. Treatment did not significantly reduce HbA1c concentrations in patients with T2DM. CONCLUSIONS Non-surgical periodontal treatment was effective in reducing the levels of IL-1β, elastase activity, and MMP-8 and -9 in GCF from diabetes and control groups. Patients with T2DM showed less reduction only in elastase activity in shallow sites compared to controls. This reduction was associated with improvement of the clinical periodontal status.

Lipid Peroxidation Is Associated with the Severity of Periodontal Disease and Local Inflammatory Markers in Patients with Type 2 Diabetes

CONTEXT Periodontitis is the most common lytic disease of bone and is recognized as a common complication of diabetes. Lipid peroxidation (LPO) is increased in diabetes and may be related to modulation of the inflammatory response. LPO levels in patients with diabetes and periodontal disease have not been evaluated. OBJECTIVE The aim of this study was to evaluate the levels of LPO and its correlation with periodontal status and inflammatory cytokines in type 2 diabetic and nondiabetic patients. DESIGN AND SETTING This is a cross-sectional study involving Brazilian patients recruited at the State University of São Paulo. PATIENTS The sample comprised 120 patients divided into four groups based upon diabetic and dyslipidemic status: poorly controlled diabetics with dyslipidemia, well-controlled diabetics with dyslipidemia, normoglycemic individuals with dyslipidemia, and healthy individuals. MAIN OUTCOME MEASURES Blood analyses were carried out for fasting plasma glucose, glycated hemoglobin, and lipid profile. Periodontal examinations were performed, and gingival crevicular fluid was collected. LPO levels were evaluated by measuring oxidized low-density lipoprotein (ELISA) and malondialdehyde (HPLC). Cytokines were evaluated by the multiplex bead technique. RESULTS LPO evaluated by malondialdehyde in plasma and gingival crevicular fluid was significantly increased in diabetes groups. Significant correlations between LPO markers and periodontal parameters indicate a direct relationship between these levels and the severity of inflammation and secretion of inflammatory cytokines, particularly in diabetic patients. CONCLUSION These findings suggest an important association for LPO with the severity of the local inflammatory response to bacteria and the susceptibility to periodontal disease in diabetic patients.

Polymorphisms and Haplotypes in the Interleukin-4 Gene are Associated With Chronic Periodontitis in a Brazilian Population

BACKGROUND Some haplotypes in the interleukin-4 (IL4) gene were reported to influence IL-4 cytokine production and were associated with inflammatory diseases. Association studies focusing on IL4 gene polymorphisms and periodontal disease provided conflicting results. The aim of this study is to investigate whether IL4 gene polymorphisms and haplotypes were related to chronic periodontitis in a Brazilian population. METHODS The polymorphisms -590(C/T) and +33(C/T) in the IL4 gene were identified by polymerase chain reaction (PCR) and restriction fragment-length polymorphism methods; the variable number of tandem repeats (VNTR) was identified by PCR. To assess the differences between the periodontitis group (n = 125) and control group (n = 125), the chi(2) test was used to assess genotype and allele distributions of individual polymorphisms. For haplotypes reconstructed by an expectation-maximization algorithm, the CLUMP program and Fisher exact test were used. Multivariate logistic regression modeling was used to assess the association of age, gender, smoking status, and polymorphism/haplotype with periodontitis. RESULTS The -590(T), +33(C), and insertion (I) of 70-base pair (bp) alleles and genotypes were more prevalent in the periodontitis group, even after adjusting for covariates. The -590, +33, and insertion (TCI) haplotype was associated with a susceptibility to periodontitis (adjusted odds ratio [OR(adjusted)] = 2.68; 95% confidence interval [CI] = 1.50 to 4.80) as was the genotype TCI/CCI (OR(adjusted) = 5.27; 95% CI = 2.28 to 12.18), whereas the TTD (OR(adjusted) = 0.48; 95% CI = 0.26 to 0.91), CTI (OR(adjusted) = 0.28, 95% CI = 0.11 to 0.70), and TTD/CTI (OR(adjusted) = 0.29; 95% CI = 0.08 to 1.13) genotypes were a associated with protection against the development of chronic periodontitis. CONCLUSION Significant associations between alleles, genotypes, and haplotypes of polymorphisms in the IL4 gene and chronic periodontitis were verified in Brazilian individuals.

Haplotypes in the Interleukin 8 Gene and Their Association with Chronic Periodontitis Susceptibility

Interleukin-8 (IL-8), which is responsible for the migration and activation of neutrophils, is an important inflammatory mediator involved in the initiation and amplification of acute inflammatory reactions and chronic inflammatory processes. IL-8 plays an important role in periodontitis, an inflammatory disease characterized by the loss of connective tissue and alveolar bone. The aim of this study was to investigate whether the SNPs rs2227307 (+396) and rs2227306 (+781), and the haplotypes they formed together with the previously investigated rs4073 (−251), were associated with chronic periodontitis susceptibility. Clinical periodontal exams were performed and DNA samples were collected from 493 individuals (223 with periodontitis and 270 controls). Associations between SNPs, haplotypes, and subject phenotypes were analyzed using the χ2 test followed by multivariate logistic regression modeling. We conclude that the +396TT genotype and the haplotypes ATC/TTC and AGT/TGC were significantly associated with chronic periodontitis susceptibility in Brazilians.

The effect of non-surgical periodontal therapy on peroxidase activity in diabetic patients: a case-control pilot study.

OBJECTIVE To evaluate the effect of periodontal therapy on clinical parameters as well as on total salivary peroxidase (TSP) activity and myeloperoxidase (MPO) activity in the gingival crevicular fluid (GCF) of patients with type 2 diabetes mellitus (DM2) and of systemically healthy individuals. MATERIAL AND METHODS Twenty DM2 subjects with inadequate metabolic control (test group) and 20 systemically healthy individuals (control group), both groups with chronic periodontitis, were enrolled. Periodontal clinical parameters, namely periodontal probing depth (PD), clinical attachment level (CAL), visible plaque index (VPI), bleeding on probing (BOP), gingival bleeding index (GBI) and presence of suppuration (SUP), as well as TSP activity and GCF MPO activity, were assessed before and 3 months after non-surgical periodontal therapy. RESULTS At baseline and 3 months post-treatment, the test group presented a higher percentage of sites with VPI and BOP (p<0.01). MPO activity in the GCF presented lower values (p<0.05) for the test group at both baseline and the post-treatment period. The periodontal treatment resulted in a significant improvement of most clinical and enzymatic parameters for both groups (p<0.05). CONCLUSIONS In both groups, the periodontal therapy was effective in improving most clinical parameters and in reducing salivary and GCF enzymatic activity. The diabetic individuals presented lower MPO activity in the GCF.

Lack of Association of a Functional Polymorphism in the Interleukin 8 Gene with Susceptibility to Periodontitis

The important inflammatory mediator interleukin-8 (IL-8) is responsible for the migration and activation of neutrophils. The IL8 gene contains a functional single-nucleotide polymorphism (SNP) (rs4073) in its promoter region that may influence the expression of IL-8, and which has been associated with inflammatory diseases. The purpose of this study was to investigate the association of the SNP (rs4073) in the IL8 gene with susceptibility to periodontitis. DNA was extracted from the buccal epithelial cells of 500 individuals (control n = 224 and periodontitis n = 276). Individuals were genotyped for the SNP (rs4073) using sequence-specific primer polymerase chain reaction. Associations between the SNP (rs4073) and subject phenotypes were analyzed using the chi-squared test, followed by univariate and multivariate logistic regression modeling. The genotype distributions in both groups were consistent with Hardy-Weinberg equilibrium. Univariate and multivariate analysis showed that age, skin color, and smoking status were associated with periodontitis. No significant differences were found for sex and frequencies of alleles and genotypes between the control and periodontitis groups in the univariate analysis. These findings were replicated in the multivariate analysis. The SNP (rs4073) in the IL8 gene is not associated with susceptibility to periodontitis in Brazilian individuals, even after controlling for covariates.

Modulation of cell proliferation, survival and gene expression by RAGE and TLR signaling in cells of the innate and adaptive immune response: role of p38 MAPK and NF-KB

Objective The aim of this study was to evaluate a possible synergism between AGE-RAGE and TLR4 signaling and the role of p38 MAPK and NF-kB signaling pathways on the modulation of the expression of inflammatory cytokines and proliferation of cells from the innate and adaptive immune response. Material and Methods T lymphocyte (JM) and monocyte (U937) cell lines were stimulated with LPS and AGE-BSA independently and associated, both in the presence and absence of p38 MAPK and NF-kB inhibitors. Proliferation was assessed by direct counting and viability was assessed by a biochemical assay of mitochondrial function. Cytokine gene expression for RAGe, CCL3, CCR5, IL-6 and TNF-α was studied by RT-PCR and RT-qPCR. Results RAGE mRNA expression was detected in both cell lines. LPS and AGE-BSA did not influence cell proliferation and viability of either cell line up to 72 hours. LPS and LPS associated with AGE induced expression of IL-6 and TNF-α in monocytes and T cells, respectively. Conclusions There is no synergistic effect between RAGE and TLR signaling on the expression of IL-6, TNF-α , RAGE, CCR5 and CCL3 by monocytes and lymphocytes. Activation of RAGE associated or not with TLR signaling also had no effect on cell proliferation and survival of these cell types.

Diabetes mellitus and oral mucosa alterations: Prevalence and risk factors

AIMS To investigate the prevalence of oral mucosa alterations in patients with type 2 diabetes and to identify possible risk factors related to oral mucosa alterations. METHODS 146 patients with type 2 diabetes and 111 age- and gender-matched healthy controls subjects were consecutively recruited from Araraquara School of Dentistry to answer a structured questionnaire designed to collect demographic data as well as current and former history of diabetes. Clinical examination of the oral mucosa was carried out by a stomatologist. RESULTS A higher prevalence of oral mucosa alterations was found in patients with diabetes than in patients without diabetes (p<0.001), with significant difference to development conditions (p<0.0001), potentially malignant disorders (p<0.0001) and fungal infections (p<0.05). In the multiple logistic regression, diabetes (odds ratio 9.9 IC 5.11-19.16) and smoking habit (odds ratio 3.17 IC 1.42-7.12) increased the odds of oral mucosa alterations significantly. CONCLUSIONS Patients with diabetes mellitus not only showed an increased prevalence of oral mucosa alterations but also a significant percentage of potentially malignant disorders. These findings elucidate the necessity of regular clinical examination to ensure early diagnosis and prompt management of oral mucosa lesions in patients with diabetes.

Effects of magnesium intake deficiency on bone metabolism and bone tissue around osseointegrated implants.

OBJECTIVES This study evaluated the effect of magnesium dietary deficiency on bone metabolism and bone tissue around implants with established osseointegration. MATERIALS AND METHODS For this, 30 rats received an implant in the right tibial metaphysis. After 60 days for healing of the implants, the animals were divided into groups according to the diet received. Control group (CTL) received a standard diet with adequate magnesium content, while test group (Mg) received the same diet except for a 90% reduction of magnesium. The animals were sacrificed after 90 days for evaluation of calcium, magnesium, osteocalcin and parathyroid hormone (PTH) serum levels and the deoxypyridinoline (DPD) level in the urine. The effect of magnesium deficiency on skeletal bone tissue was evaluated by densitometry of the lumbar vertebrae, while the effect of bone tissue around titanium implants was evaluated by radiographic measurement of cortical bone thickness and bone density. The effect on biomechanical characteristics was verified by implant removal torque testing. RESULTS Magnesium dietary deficiency resulted in a decrease of the magnesium serum level and an increase of PTH and DPD levels (P ≤ 0.05). The Mg group also presented a loss of systemic bone mass, decreased cortical bone thickness and lower values of removal torque of the implants (P ≤ 0.01). CONCLUSIONS The present study concluded that magnesium-deficient diet had a negative influence on bone metabolism as well as on the bone tissue around the implants.