Diabetes-associated periodontitis molecular features in infrared spectra of gingival crevicular fluid.

BACKGROUND: It has been established previously that infrared spectroscopy (IRS) can be used to identify periodontitis-specific molecular signatures in gingival crevicular fluid (GCF) and to confirm clinical diagnoses. This follow-up study is designed to assess whether this novel technique is also able to differentiate diseased from healthy sites in patients with diabetes mellitus (DM) by analyzing the molecular fingerprints embedded in the GCF. METHODS: A total of 65 patients with DM with moderate-to-severe chronic periodontitis (CP) was recruited, and 15 individuals without DM (65 sites) without periodontal diseases were used as control. Clinical examination and GCF samples were taken from a total of 351 sites, including periodontitis (109), gingivitis (115), and healthy (127) sites. Corresponding absorption spectra of GCF samples were acquired and processed, and the relative contributions of key functional groups in the infrared spectra were identified and analyzed. The qualitative assessment of clinical relevance of these GCF spectra was interpreted with multivariate statistical analysis: linear discriminant analysis (LDA). RESULTS: Spectral analysis revealed several molecular signatures representing vibrations in protein (amide I and II), lipid ester, and sugar moieties in the GCF of patients with DM with CP and non-DM controls. The diagnostic accuracy for distinction between healthy and CP sites in patients with DM determined by LDA of GCF spectra was 95.3% for the training set of samples and 87.5% for the validation set. Additional LDA of GCF spectra from healthy sites of non-DM controls and patients with DM revealed 100% diagnostic accuracy for the training set and 86.7% for the validation set. The regions robotically selected by LDA for the two analyses were slightly different in that first LDA identified major regions clustered with the side chain vibrations originating from protein and DNA contents, whereas the second was predominantly the glycation and protein components. CONCLUSION: IRS is a feasible method to differentiate disease-specific molecular signatures in GCF in the presence of DM and to generate a complex biochemical profile of GCF to identify DM-specific spectral features.

Surgical and non-surgical therapy with systemic antimicrobials for residual pockets in type 2 diabetics with chronic periodontitis: a pilot study.

AIM: This study evaluated the effects of surgical (SD) and non-surgical (NSD) debridements, associated with systemic antimicrobials, on clinical and immunological outcomes of residual pockets [RP; probing depth (PD) ≥5 mm with bleeding on probing] in type 2 diabetics. MATERIAL AND METHODS: A split-mouth, randomized controlled trial was conducted in 21 subjects presenting at least two RP per contralateral quadrant. Subjects received metronidazole plus amoxicillin for 10 days and, contralateral quadrants were assigned to receive SD or NSD. Clinical parameters and local levels of interferon-γ, interleukin (IL)-17, IL-23 and IL-4 were assessed at baseline, 3 and 6 months post-therapies. RESULTS: Overall, the mean number, PD and clinical attachment level (CAL) of RP improved significantly after therapies (p < 0.05), without differences between groups at any time-point (p > 0.05). At quadrant level, only SD produced significant reductions in the mean CAL. Also, SD promoted higher reduction in PD from baseline to 6 months than NSD (p < 0.05). Levels of all cytokines were increased after SD compared with NSD (p < 0.05). CONCLUSION: SD and NSD associated with systemic antimicrobials did not differ in terms of clinical benefits for RP in diabetics up to 6 months post-therapies. RP treated by SD presented increased levels of cytokines.

Relationship between glycemic subsets and generalized chronic periodontitis in type 2 diabetic Brazilian subjects.

OBJECTIVE: The aim of the present study was to evaluate the relationship between glycemic subsets and clinical periodontal conditions in type 2 diabetic Brazilians with generalized chronic periodontitis. DESIGN: Ninety-one Brazilians with type 2 DM and generalized chronic periodontitis were involved in this study. The clinical examination included full-mouth assessment of plaque index (PI), bleeding on probing (BoP), probing depth (PD), suppuration (SUP), clinical attachment level (CAL) and number of remaining teeth. Blood analyses were carried out for glycated haemoglobin (HbA1c) and fasting plasma glucose (FPG). The relationship between the extent of periodontitis, defined as the percentage of sites with PD and CAL≥5 mm, and glycemic parameters were also analysed. In addition, clinical parameters were compared amongst four (HbA1c levels ≤7.5%, 7.6-9%, 9.1-11% and >11%) and two (<9% and ≥9%) glycemic subsets. RESULTS: The frequency of uncontrolled diabetic subjects (HbA1c>7.5%) was higher than well-controlled subjects (HbA1c≤7.5%). Amongst the clinical parameters evaluated, only PI was positively correlated with the levels of HbA1c and FPG (p<0.05). The number of remaining teeth was negatively associated with the levels of HbA1c (p<0.05). In addition, PI was significantly higher in subjects presenting HbA1c levels >11% and ≥9% than those with HbA1c levels ≤7.5% and <9%, respectively (p<0.05). CONCLUSION: Although an increased frequency of the subjects with generalized chronic periodontitis included presented type 2 uncontrolled DM, a dose-response relationship between the severity and extension of periodontitis and the glycemic control was not established in these subjects.

Expression of immune-inflammatory markers in sites of chronic periodontitis in patients with type 2 diabetes.

BACKGROUND: The aim of this study is to evaluate the gene expression of immune-inflammatory markers in gingival biopsies of patients with type 2 diabetes with chronic periodontitis (CP). METHODS: Gingival biopsies were harvested from systemically and periodontally healthy patients (SPH), systemically healthy patients with CP (SHCP), and patients with better-controlled and poorly controlled diabetes and CP. The levels of mRNA of interleukin (IL)-17, IL-6, IL-23, IL-10, IL-4, interferon-γ, toll-like receptor (TLR)-2, TLR-4, osteoprotegerin, receptor activator of nuclear factor-kappa B ligand (RANKL), tumor necrosis factor-α, transforming growth factor-ß, transcription factor forkhead box p3, transcription factor orphan nuclear receptor C2 (RORC2), and receptor of advanced glycation end products (RAGE) were evaluated by quantitative real-time polymerase chain reaction. RESULTS: All CP groups presented higher levels of mRNA of TLR-2, TLR-4, IL-17, RANKL, and RAGE and a higher frequency of IL-17 and TLR-2 mRNA-positive biopsies when compared to SPH (P <0.05). There was a higher frequency of detection of RORC2 in the biopsies from both groups with diabetes compared to the other groups (P <0.05). The frequency of IL-4 mRNA-positive tissues was lower in patients with diabetes compared to SHCP (P <0.05). CONCLUSION: CP, but not type 2 diabetes mellitus, significantly affected the expressions of the evaluated genes related to the innate and adaptive immune responses.

Changes in the subgingival biofilm composition after coronally positioned flap.

OBJECTIVES: This study evaluated the effects of coronally positioned flap (CPF) on the subgingival biofilm composition. MATERIAL AND METHODS: Twenty-two subjects with gingival recessions were treated with CPF. Clinical parameters were assessed before and at 6 months after surgery. Subgingival biofilms were analyzed by checkerboard DNA-DNA hybridization technique for 40 bacterial species. RESULTS: Recession height, clinical attachment level and bleeding on probing improved significantly (p<0.05) at 6 months post-CPF. The proportions of 10 periodontal pathogens and the proportions of red and orange complexes decreased at 6 months. CONCLUSION: In conclusion, CPF can induce beneficial effects on the composition of the subgingival microbiota after 6 months.

Changes in the subgingival biofilm composition after coronally positioned flap

OBJECTIVES: This study evaluated the effects of coronally positioned flap (CPF) on the subgingival biofilm composition. MATERIAL AND METHODS: Twenty-two subjects with gingival recessions were treated with CPF. Clinical parameters were assessed before and at 6 months after surgery. Subgingival biofilms were analyzed by checkerboard DNA-DNA hybridization technique for 40 bacterial species. RESULTS: Recession height, clinical attachment level and bleeding on probing improved significantly (p<0.05) at 6 months post-CPF. The proportions of 10 periodontal pathogens and the proportions of red and orange complexes decreased at 6 months. CONCLUSION: In conclusion, CPF can induce beneficial effects on the composition of the subgingival microbiota after 6 months.

Cytokine levels in sites of chronic periodontitis of poorly controlled and well-controlled type 2 diabetic subjects.

AIM: This study compared the levels of tumour necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-4, IL-17 and IL-23 in the gingival crevicular fluid (GCF) from well-controlled and poorly controlled type 2 diabetic subjects with chronic periodontitis, before and after periodontal therapy. MATERIAL AND METHODS: Eighteen well-controlled (glycated haemoglobin levels ≤8%) and 20 poorly controlled (glycated haemoglobin levels >8%) diabetic subjects were enrolled in this study. All subjects were submitted to non-surgical periodontal therapy. GCF sampling and clinical periodontal parameters were assessed before, 3 and 6 months post-therapy. Total amounts and concentrations of TNF-α, IFN-γ, IL-4, IL-17 and IL-23 in the GCF were analysed by enzyme-linked immunosorbent assay (ELISA). RESULTS: The levels of IL-17 were higher in poorly than in well-controlled subjects (p<0.05), whereas the levels of IFN-γ were increased in well- compared with poorly controlled subjects at all experimental groups (p<0.05). In addition, IL-4 levels were lower in well- than poorly controlled diabetic subjects at baseline (p<0.05). There were no differences between groups for TNF-α and IL-23 at any time points (p>0.05). CONCLUSION: These results indicate a predominance of pro-inflammatory T-helper type 1 (Th1)- or Th17-cytokines in sites of chronic periodontitis from type 2 diabetic subjects, according to their glycaemic control.

Influence of glycemic control on Epstein-Bar and Cytomegalovirus infection in periodontal pocket of type 2 diabetic subjects.

OBJECTIVE: The aim of the present study was to evaluate the influence of glycemic control on the frequency of Epstein-Bar (EBV) and Cytomegalovirus (CMV) in periodontal pockets of type 2 diabetic subjects with chronic periodontitis. DESIGN: Forty-six subjects presenting generalized chronic periodontitis and type 2 diabetes mellitus (DM) were selected for this study. Polymerase chain reaction (PCR) was used to determine the presence of EBV and CMV in shallow [Probing Depth (PD)≤3mm], moderate (PD=4-6mm) and deep (PD>7mm) pockets. HbA1c levels ≤7%, >7 to <10%, and ≥10% defined good, moderate and poor glycemic control, respectively. RESULTS: Higher frequency of EBV was found in the shallow pockets of the subjects with poor glycemic control (p<0.05; chi-square test). Moreover, EBV-free subjects presented moderate or good glycemic control. Glycemic control did not influence the frequency of CMV in all pocket categories. CONCLUSION: Poor glycemic control in type 2 diabetic subjects can increase the occurrence of EBV in shallow periodontal pockets.

Receptor activator of nuclear factor-kappa B ligand/osteoprotegerin ratio in sites of chronic periodontitis of subjects with poorly and well-controlled type 2 diabetes.

BACKGROUND: The aim of this study is to evaluate the levels of osteoclastogenesis-related factors (soluble receptor activator of nuclear factor-kappa B ligand [sRANKL] and osteoprotegerin [OPG]) in gingival crevicular fluid (GCF) from subjects with poorly and well-controlled type 2 diabetes and chronic periodontitis before and after periodontal therapy. METHODS: Eighteen subjects with well-controlled diabetes (glycated hemoglobin [HbA1c] levels ≤ 8%) and 20 subjects with poorly controlled diabetes (HbA1c levels >8%) were enrolled in this study. All subjects were submitted to non-surgical periodontal therapy. GCF sampling and clinical periodontal parameters were assessed at baseline and 3 and 6 months post-therapy. Total amounts and concentrations of sRANKL and OPG in GCF were analyzed by enzyme-linked immunosorbent assay (ELISA). RESULTS: Total amounts and concentrations of sRANKL and RANKL/OPG ratios were higher in poorly controlled subjects than in well-controlled subjects at baseline and 3 and 6 months post-therapy (P <0.05). In addition, RANKL/OPG ratios decreased in well-controlled subjects (P <0.05) but not in poorly controlled subjects (P >0.05) at 3 months post-therapy. Almost all clinical parameters improved significantly for both groups post-treatment (P <0.05). CONCLUSION: RANKL/OPG ratios in untreated and treated periodontitis sites may be negatively influenced by poor glycemic control in subjects with type 2 diabetes.

Effectiveness of full-mouth and partial-mouth scaling and root planing in treating chronic periodontitis in subjects with type 2 diabetes.

BACKGROUND: This study evaluated the clinical and metabolic effects of full-mouth scaling and root planing (FMSRP) compared to partial-mouth scaling and root planing (PMSRP) in patients with type 2 diabetes and chronic periodontitis, and it assessed the impact of the glycemic status on the clinical and metabolic response to periodontal therapy. METHODS: In this clinical trial, 18 subjects with diabetes received FMSRP in a maximum of 24 hours, and 18 subjects received PMSRP in a maximum of 21 days. Visible plaque accumulation, bleeding on probing, suppuration, probing depth, clinical attachment level (CAL), and glycosylated hemoglobin (HbA1c) levels were obtained at baseline and at 3 and 6 months post-therapy. Baseline HbA1c values > or =9% and <9% defined subjects with poorly and better-controlled diabetes, respectively. RESULTS: All clinical parameters improved after therapy (P <0.05). No significant differences were observed between treatment groups for clinical and metabolic parameters at any time (P >0.05). There were no changes in the HbA1c levels after therapy (P >0.05). No subject reported any adverse effects during the study. Individuals with better-controlled diabetes achieved a lower mean CAL at 6 months post-therapy, when FMSRP and PMSRP were evaluated together (P <0.05). CONCLUSIONS: FMSRP and PMSRP were equally effective in treating chronic periodontitis in subjects with type 2 diabetes, without significant improvements in the glycemic control at 3 and 6 months. Considering the periodontal therapy as a whole (FMSRP plus PMSRP), subjects with better-controlled diabetes exhibited a benefit in CAL at 6 months compared to subjects with poorly controlled disease.