Generalized Aggressive Periodontitis Subjects Research Articles

Biomarker levels in gingival crevicular fluid of generalized aggressive periodontitis patients after non-surgical periodontal treatment.

The aim of this study was to assess the effects of non-surgical periodontal treatment on gingival crevicular fluid (GCF) cytokines in patients with generalized aggressive periodontitis (GAgP), in relation to clinical parameters. Data were obtained from 16 GAgP patients and 15 periodontally healthy controls. Periodontal parameters and GCF biomarker levels were evaluated at baseline and repeated 3 and 6months after treatment for GAgP subjects. Moderate and deep pocket sites were analyzed separately. The amount of interleukin (IL)-1β, IL-9, tumor necrosis factor (TNF)-α, platelet-derived growth factor (PDGF-bb), and vascular endothelial growth factor (VEGF) were measured using a highly specific and sensitive multiplex bead immunoassay. At baseline, cytokine levels in the moderate and deep pocket sites of GAgP patients were higher than those of the healthy control sites. In GAgP group, periodontal treatment led to improvement in all examined clinical parameters and resulted in a statistically significant reduction in the total amounts of IL-1β, VEGF, and TNF-α, in comparison to baseline, already 3months after therapy in both moderate and deep pocket sites and of PDGF-bb in deep sites (p<0.01). At the concentration level, only IL-1β and VEGF were affected. Non-surgical treatment of GAgP provided significant clinical benefits leading to a marked decrease in the GCF levels of some pro-inflammatory and pro-angiogenic cytokines, but not of IL-9 and PDGF-bb. Although the periodontal therapy successfully decreased clinical signs of inflammation, the GCF levels of some inflammatory cytokines were still elevated.

Synergistetes cluster A in saliva is associated with periodontitis

Synergistetes is a novel bacterial phylum consisting of gram-negative anaerobes. Increasing lines of evidence demonstrate that this phylum is associated with periodontal diseases. This study aimed to compare the presence and levels of Synergistetes clusters A and B, in saliva of patients with chronic periodontitis (CP), generalized aggressive periodontitis (G-AgP) and non-periodontitis subjects, and investigate their correlation with clinical parameters. Saliva was collected from patients with CP (n=20), G-AgP (n=21) and non-periodontitis subjects (n=18). Full mouth clinical periodontal measurements were recorded. The numbers of Synergistetes cluster A and cluster B or the associated species Jonquetella anthropi were quantified by fluorescent in situ hybridization and microscopy. Synergistetes cluster A bacteria were detected more frequently, and at higher numbers and proportions in the two periodontitis groups, than the non-periodontitis control group. The prevalence was 27.7% in the control group, 85% in CP and 86% in G-AgP. Compared to the control group, the numbers were significantly higher by 12.5-fold in CP and 26.5-fold in G-AgP, whereas the difference between the two forms of periodontitis was not statistically significant. Within the total bacterial population, the proportion of this cluster was increased in CP and G-AgP compared to the control group, with the difference between the two forms of periodontitis being also significant. There was a positive correlation between the levels of Synergistetes cluster A in saliva and all full mouth clinical periodontal parameters. Nevertheless, Synergistetes cluster B bacteria and J.anthropi species were detected infrequently and at low levels in all the three subject groups. Synergistetes cluster A, but not cluster B, bacteria are found at higher prevalence, numbers and proportions in saliva from patients with periodontitis, than non-periodontitis subjects. These findings support the association of this cluster with periodontitis.

Detection of Mogibacterium timidum in subgingival biofilm of aggressive and non-diabetic and diabetic chronic periodontitis patients.

The aim of the present study was to evaluate the frequency of detection of Mogibacterium timidum in subgingival samples of subjects with generalized aggressive periodontitis (GAgP) and uncontrolled diabetic and non-diabetic subjects with generalized chronic periodontitis (GChP). 48 patients with GAgP, 50 non-diabetic and 39 uncontrolled (glycated hemoglobin >7%) type 2 diabetic subjects with GChP were enrolled in this study. Subgingival biofilm were collected from deep pockets (probing depth > 7 mm). After DNA extraction, M. timidum was detected by Nested Polymerase Chain Reaction and chi-square test was used to data analysis (p>0.05). There were no differences in the frequency of detection of M. timidum between subjects with GAgP (35%) and non-diabetic subjects with GChP (40%) (p>0.05). The frequency of detection of M. timidum was significantly higher in deep pockets of diabetic subjects with GChP (56%) when compared to GAgP (p<0.05), but similar to non-diabetic subjects with GChP (p>0.05). The frequency of detection of M. timidum was higher in subjects GChP presenting uncontrolled type 2 diabetes mellitus, when compared to GAgP subjects.

Effects of periodontal therapy on GCF cytokines in generalized aggressive periodontitis subjects

To examine changes in levels of gingival crevicular fluid (GCF) cytokines, after periodontal therapy of generalized aggressive periodontitis (GAgP). Twenty-five periodontally healthy and 24 GAgP subjects had periodontal clinical parameters measured and gingival crevicular fluid (GCF) samples collected from up to 14 sites/subject. GCF samples were analysed using multiplex bead immunoassay for: GM-CSF, IFN-γ, IL-10, IL-1β, IL-2, IL-6 and TNF-α. Aggressive periodontitis subjects were randomly assigned to either scaling and root planing (SRP) alone or SRP plus systemic amoxicillin (500 mg) and metronidazole (400 mg) 3 times a day for 14 days. Clinical parameters and GCF cytokines were re-measured 6 months after treatment. Differences over time were analysed using the Wilcoxon test and between groups using the Mann-Whitney test. Significant reductions in GCF GM-CSF, IL-1β and the ratio IL-1β/IL-10 and increases in GCF IL-6 were detected after therapy. The mean change in GCF cytokines did not differ significantly between groups. Periodontal therapy improved GCF cytokine profiles by lowering IL-1β and increasing IL-10 levels. The reduction in GCF GM-CSF after therapy implicates this cytokine in the pathogenesis of GAgP. There was no difference between therapies in changes of GCF cytokines.

Low prevalence of subgingival viruses in periodontitis patients

Viruses such as Human Cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) have been proposed to be periodontal pathogens. The aim of this study was to analyse the presence of herpesvirus DNA in subgingival plaque samples of patients with different forms of periodontitis and in healthy periodontia. A total of 140 ethnically mixed (prevalently Caucasian) subjects took part in the study. Sixteen were affected by localized aggressive periodontitis (LAgP), 64 by generalized aggressive periodontitis (GAgP), 20 by chronic periodontitis (CP) and 40 were periodontally healthy. Polymerase chain reaction (PCR) analyses were performed to detect HCMV and EBV. Sera were tested for anti-HCMV and EBV IgG antibodies. PCRs for herpes simplex (HSV) and varicella zoster virus (VZV) were performed in subgingival samples from a subset of 20 AgP subjects. HCMV DNA was not detected in any plaque samples. EBV DNA was detected in four LAgP (25%), two GAgP (3%) subjects and four healthy individuals (10%). HSV DNA and VZV DNA were not detected in the subset of studied individuals. This study challenges the previously reported high prevalence of herpesvirus DNA in subgingival samples from periodontitis patients and so questions whether they act as pathogens in such patients.

Initial comparison of proteomic profiles of whole unstimulated saliva obtained from generalized aggressive periodontitis patients and healthy control subjects

Salivary proteomics technology can be used to evaluate the disease progression of periodontitis and the systemic screening of proteomes of saliva from subjects with aggressive periodontitis has not been available. The objective of this preliminary study was to compare the proteomic profile of whole unstimulated saliva of subjects with generalized aggressive periodontitis (GAgP) with that of healthy volunteers to identify proteins, the levels of which were significantly altered between the two groups. Whole unstimulated saliva was obtained from five subjects with GAgP and five healthy subjects, and proteins were separated using two-dimensional gel electrophoresis. Proteins, the levels of which were significantly different between the two groups, were identified by computer image analyses and subsequent electrospray ionization tandem mass spectrometry. Eleven proteins that exhibited a different level in the GAgP group vs. the control group were identified. Compared with whole saliva of healthy control subjects, the levels of serum albumin, immunoglobulin (Ig) gamma2 chain C region, Ig alpha2 chain C region, vitamin D-binding protein, salivary alpha-amylase and zinc-alpha2 glycoprotein were increased in whole unstimulated saliva of GAgP subjects, while those of lactotransferrin, elongation factor 2, 14-3-3 sigma, short palate, lung and nasal epithelium carcinoma-associated protein 2 precursor and carbonic anhydrase 6 were decreased. Comparison of the proteomic profile of whole unstimulated saliva of GAgP subjects with that of healthy control subjects revealed at least 11 differential proteins. The approach applied herein might be helpful to aid understanding of the etiology of GAgP.

Relationship of neutrophil phagocytosis and oxidative burst with the subgingival microbiota of generalized aggressive periodontitis

Polymorphonuclear neutrophil (PMN) dysfunctions have been associated with severe forms of periodontitis. This study evaluated the correlation between PMN phagocytosis and oxidative burst with the subgingival microbiota of patients with generalized aggressive periodontitis (GAgP). Heparinized peripheral blood samples were obtained from 18 GAgP patients and 11 periodontally healthy (PH) subjects, and PMNs were isolated on a Ficoll-Hypaque gradient. For phagocytosis analysis, PMNs were incubated with fluorescein-labeled Staphylococcus aureus. The oxidative burst was evaluated by incubation of PMNs with dihydroethidium and activation by S. aureus. The assays were examined using flow cytometry. Subgingival biofilm samples were obtained from periodontal sites with and without periodontitis and 24 species were detected by checkerboard. A significantly lower phagocytosis rate was observed for patients with GAgP compared with PH subjects over time (P < 0.05). No differences between groups were found for superoxide production. GAgP patients presented significantly higher prevalence and levels of Porphyromonas gingivalis, Tannerella forsythia, and Aggregatibacter actinomycetemcomitans serotype b than controls (P < 0.05). Significant negative correlations between T. forsythia and P. gingivalis and PMN functions were observed. GAgP subjects presented diminished phagocytic activity of peripheral PMNs and high prevalence and levels of classical periodontal pathogens.

Angiotensin-converting enzyme (ACE), angiotensinogen (AGT), and angiotensin II type 1 receptor (AT1R) gene polymorphisms in generalized aggressive periodontitis

Aim Host response to periodontopathic microorganisms can be modulated by genetic factors. Accumulated evidence highlighted the role of renin–angiotensin system (RAS) in inflammatory response thus potential implication of this molecular system in the pathogenesis of periodontitis can be suggested. The present study investigated common genetic variants of molecules within the RAS family namely angiotensin-converting enzyme (ACE), angiotensinogen (AGT) and angiotensin II type 1 receptor (AT1R) in relation to generalized aggressive periodontitis (G-AgP). Methods DNA was obtained from peripheral blood of 103 G-AgP patients and 100 periodontally healthy subjects. ACE I/D, AGT M235T and AT1R A1166C polymorphisms were genotyped by polymerase chain reaction and restriction fragment length polymorphism method. Chi-square, ANOVA and logistic regression were used in statistical analyses. Results Both ACE I/D and AT1R polymorphisms were similar in G-AgP and healthy groups ( p > 0.05). G-AgP subjects exhibited decreased AGT TT genotype and T allele frequency as compared to healthy subjects ( p < 0.05). The same trend was also observed in the nonsmoker subgroup regarding investigated RAS polymorphisms. Conclusions Present findings suggest that AGT M235T TT genotype and T allele might be associated with decreased risk for G-AgP in Turkish population.

TNF‐α and IL‐4 levels in generalized aggressive periodontitis subjects

The aim of this study was to evaluate tumor necrosis factor (TNF)-alpha and interleukin (IL)-4 levels in healthy sites and sites exhibiting signs of moderate and advanced generalized aggressive periodontitis (GAgP) in the same subject. The following sites were selected for crevicular fluid sampling in the same AgP subject (n = 14): Healthy sites (HS): no marginal bleeding or bleeding on probing (BOP) and probing depth (PD) <or= 3 mm; Moderate sites (MS): BOP and PD between 4 and 6 mm; Advanced sites (AS): BOP and PD >or= 7 mm. One site from periodontally healthy subjects (n = 13) was sampled for use as a control. TNF-alpha and IL-4 levels were measured using ELISA. The total amount of TNF-alpha was lower for control sites, while there were no differences among healthy and diseased sites from GAgP subjects (P < 0.05). The concentration of TNF-alpha was higher in HS, in relation to the other sites (P < 0.05). There were no significant differences among the groups regarding total amounts of IL-4 (P > 0.05), while IL-4 concentration was significantly higher in control sites, when compared with sites from GAgP subjects (P < 0.05). In conclusion, high levels of TNF-alpha and low levels of IL-4 were observed in both healthy and diseased sites within the same generalized AgP individuals.

Osseointegrated Implants in Subjects Treated for Generalized Aggressive Periodontitis: 10‐Year Results of a Prospective, Long‐Term Cohort Study

The aim of this prospective 10-year study of partially edentulous subjects treated for generalized aggressive periodontitis and periodontally healthy subjects was a clinical, microbiologic, and radiographic comparison of teeth and implants and assessment of the implants' success rate. Five subjects treated for generalized aggressive periodontitis (GAgP) and five periodontally healthy subjects who were orally rehabilitated with osseointegrated implants participated in the study. First, they were examined 2 to 4 weeks before extraction of the non-retainable teeth (baseline) and 3 weeks after insertion of the final abutments. All further examinations were performed during a 3-month recall schedule over a 10-year period. At every session, clinical parameters were recorded, and the composition of the subgingival microflora was determined. Radiographs were taken at baseline after insertion of the superstructure and 1, 3, 5, 8, and 10 years later. Throughout the follow-up period, the peri-implant gingival index of GAgP subjects was significantly higher than in periodontally healthy subjects. There was no difference in plaque index between teeth and implants or between the two groups. The peri-implant probing depths were comparable in the two groups and remained < or =4 mm throughout the follow-up. The probing depth was significantly higher around the teeth of the GAgP subjects compared to periodontally healthy subjects. Implants of GAgP subjects showed a significantly higher attachment loss (Ø 2.4 mm). The attachment level at teeth and implants of the periodontally healthy subjects and at teeth of the GAgP subjects was almost unchanged. Microbiologically, GAgP subjects had fewer cocci and more motile rods and filaments at teeth and implants than periodontally healthy subjects. GAgP subjects showed significantly more peri-implant bone loss in the first year (Ø 2.07 mm) and in the subsequent 9 years (total 1.3 mm). Bone loss at teeth also was significantly higher at baseline (Ø 26.39%) and in the following years (total 9.3%). Implant survival rates were 100% in periodontally healthy subjects versus 83.33% in GAgP subjects. This 10-year study showed that partially edentulous subjects treated for GAgP can be rehabilitated successfully with osseointegrated implants. However, the bone and attachment loss at the implants were higher than in periodontally healthy subjects.

Matrix Metalloproteinase‐2, ‐9, and ‐12 Gene Polymorphisms in Generalized Aggressive Periodontitis

Matrix metalloproteinases (MMPs) are involved in periodontal tissue remodeling and degradation. Polymorphisms in the promoter region of the MMP-2 and -9 genes and in the coding region of the MMP-12 gene could affect transcription and the function of these enzymes. The aim of the present study was to determine the association between the aforementioned MMP polymorphisms and generalized aggressive periodontitis (GAgP). Genomic DNA was obtained from the peripheral blood of 92 subjects with GAgP and 157 periodontally healthy subjects. MMP-2 -735C/T, MMP-9 -1562C/T, and MMP-12 357Asn/Ser polymorphisms were genotyped by polymerase chain reaction and restriction fragment length polymorphism. Probing depth, clinical attachment loss, supragingival plaque accumulation, and bleeding on probing were recorded. The data were analyzed by chi(2), logistic regression, and Mann-Whitney U tests. The genotype distributions, allele frequencies, and rare allele carriage of MMP-2 and MMP-12 genes were similar in GAgP and healthy subjects (P >0.05). T allele frequency and T allele carriage of the MMP-9 -1562 C/T polymorphism were significantly lower in the GAgP group than in the healthy group (P <0.05). In addition, logistic regression analysis revealed a protective effect for MMP-9 -1562 T allele carriers (odds ratio = 0.52; P = 0.04). MMP-2 -735C/T and MMP-12 357Asn/Ser polymorphisms are not related to GAgP. Conversely, the MMP-9 -1562 gene T allele might be associated with a decreased risk for GAgP in the Turkish population.

Gene polymorphisms of tissue plasminogen activator and plasminogen activator inhibitor‐1 in Turkish patients with generalized aggressive periodontitis

Tissue plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) have important roles in proteolytic events in periodontitis. The aim of this study was to investigate TPA and PAI-1 gene polymorphisms in relation to susceptibility to generalized aggressive periodontitis (G-AgP). Genomic DNA was obtained from peripheral blood of 90 G-AgP patients and 154 periodontally healthy subjects. 4G/5G polymorphism in the promoter region of the PAI-1 gene and Alu-repeat insertion/deletion (I/D) polymorphism in intron 8 of the TPA gene were genotyped by polymerase chain reaction and endonuclease digestion. The genotype distributions of TPA and PAI-1 genes were similar between G-AgP and healthy subjects (p>0.05). The distribution of TPA genotypes in G-AgP patients was 33.4% D/D, 44.4% I/D, and 22.2% I/I and was 26.3% D/D, 40.4% I/D, and 33.3% I/I in healthy subjects. The D allele was 55.6% in G-AgP and 46.6% in healthy subjects. There was a significant difference among study groups in D allele frequencies (p=0.044). The PAI-1 genotype distribution in G-AgP was 29.1% 4G/4G, 43.0% 4G/5G, and 27.9% 5G/5G, while it was 35.7% 4G/4G, 43.8% 4G/5G, and 20.5% 5G/5G in healthy subjects. These data suggest that the D polymorphic allele of TPA gene polymorphism could be associated with susceptibility to G-AgP in Turkish subjects.

Subgingival microbiota of periodontally untreated Mexican subjects with generalized aggressive periodontitis

Specific microbial profiles that may distinguish between generalized aggressive-periodontitis (GAgP) and generalized chronic-periodontitis (GCP) have, to date, not been described. The purpose of the present study was to describe the subgingival microbial composition of Mexican subjects with GAgP and compare it with that of individuals with GCP and periodontal health (PH). Seventy-seven subjects with GAgP (n=19), GCP (n=39) and PH (n=19) were selected. Clinical measurements included plaque accumulation, gingival erythema, bleeding on probing, suppuration, pocket depth and attachment level. Up to 28 subgingival plaque samples were obtained from each subject and analysed using the checkerboard DNA-DNA hybridization technique. GAgP and GCP subjects harboured significantly higher levels and/or proportion of Porphyromonas gingivalis, Tannerella forsythia (levels: p<0.001, proportion: p<0.01), Prevotella nigrescens (p<0.05 levels) and "red" complex species (p<0.001 proportion) than PH subjects. All GAgP subjects were carriers of P. gingivalis and P. nigrescens. No significant differences in any of the 40 microbial species tested were detected between GAgP and GCP subjects. Our results revealed that the microbial differences between GAgP and GCP subjects were only discrete and none of the bacterial species tested seemed to specifically differentiate the subgingival microbial profile of either periodontitis group.

Distribution of FcγRIIa and FcγRIIIb Genotypes in Patients With Generalized Aggressive Periodontitis.

Polymorphisms in FcγR have been associated with different forms of periodontitis. This study determined the frequency of FcγRIIa and FcγRIIIb alleles/genotypes in patients with generalized aggressive periodontitis (GAgP). Thirty-one GAgP and 49 periodontally healthy Brazilian subjects participated in the study. Full-mouth periodontal examinations were carried out, and mouthwash samples were collected for human DNA isolation. FcγR genotyping was performed by polymerase chain reaction and hybridization with allele-specific oligonucleotide probes. Significant differences between groups were sought by Mann-Whitney, χ2 , and Fisher exact tests and configural frequency analysis. FcγRIIa-H131 (53.8%) and FcγRIIIb-NA1 (75%) were the most prevalent alleles in this sample population. A significant overrepresentation of FcγRIIIb-NA2 was observed in the GAgP group, whereas FcγRIIIb-NA1 was detected more often in healthy individuals (odds ratio, 32.5; 95% confidence interval [CI], 10.6 to 99.8; P <0.001). No significant differences in the distribution of the FcγRIIa genotypes were observed between the groups. The prevalence of FcγRIIIb-NA2/NA2 was higher in GAgP patients, whereas FcγRIIIb-NA1/NA1 was predominant in the healthy group (χ2 = 45.1; P <0.001). The combination of the genotypes FcγRIIIb-NA2/NA2 plus FcγRIIa-H/H131 was observed more frequently in GAgP subjects than expected from marginal frequencies (χ2 = 12.5; P <0.001). The data suggest that the FcγRIIIb-NA2 allele and/or FcγRIIIb-NA2/NA2 genotype and the composite genotype FcγRIIIb-NA2/NA2 plus FcγRIIa-H/H131 may be associated with GAgP, whereas FcγRIIIb-NA1 and/or FcγRIIIb-NA1/NA1 may be related to periodontal health in this sample of the Brazilian population.

C‐Reactive Protein Levels in Patients With Aggressive Periodontitis

Sera from patients with periodontal infections contain elevated levels of C-reactive protein (CRP) compared to periodontally healthy individuals. Most studies to date have included patients with chronic periodontitis, and few investigators have studied CRP levels in subjects with aggressive periodontitis (AgP). The purpose of this study was to determine the relative levels of serum CRP in AgP patients and periodontally healthy subjects and to examine patients' characteristics that might account for intergroup differences. Serum samples were collected from 93 patients with generalized AgP (GAgP), from 97 patients with localized AgP (LAgP), and from 91 healthy controls (non-periodontitis [NP]). Periodontal examination consisted of plaque index, gingival index, probing depth, bleeding index, and attachment loss measurements. Current smoking was assessed by determination of serum cotinine levels by enzyme-linked immunosorbent assay (ELISA), and serum CRP levels were determined using a high-sensitivity ELISA assay. The three groups were significantly different from one another (P <0.0001). The 95% confidence interval for serum CRP concentrations were as follows: NP, 0.65919 (0.4901 to 0.8869); LAgP, 1.10138 (0.8265 to 1.468); and GAgP, 2.05318 (1.5313 to 2.7538) mg/l. CRP levels in both LAgP and GAgP subjects were significantly greater than those in NP subjects, and levels in GAgP were significantly greater than those in LAgP. Following adjustment of the data for periodontal and demographic variables and current smoking, both mean probing depth and periodontal diagnosis remained correlated with CRP levels. Patients with AgP have statistically significant elevations in serum CRP levels compared to subjects without periodontitis. Elevated CRP in these subjects might represent a contribution of periodontal infections to systemic inflammation in relatively young individuals.

Altered lipoprotein subclass distribution and PAF-AH activity in subjects with generalized aggressive periodontitis

In this study, we examined whether the documented increase of plasma triglycerides in patients with generalized aggressive periodontitis (GAgP) is associated with changes in lipoprotein subclass distribution and/or LDL-associated platelet-activating factor acetylhydrolase (PAF-AH) activity. Lipoprotein subclasses were analyzed in whole plasma samples using nuclear magnetic resonance methods. Compared with subjects without periodontitis (NP subjects; n = 12), GAgP subjects (n = 12) had higher plasma levels of large, medium, and small VLDL (35.0 +/- 6.7 vs. 63.1 +/- 9.6 nmol/l; P = 0.025), higher levels of intermediate density lipoprotein (24.8 +/- 11.6 vs. 87.2 +/- 16.6 nmol/l; P = 0.006), lower levels of large LDL (448.3 +/- 48.5 vs. 315.8 +/- 59.4 nmol/l; P = 0.098), and higher levels of small LDL (488.2 +/- 104.2 vs. 946.7 +/- 151.6 nmol/l; P = 0.021). The average size of LDL from NP and GAgP subjects was 21.4 +/- 0.2 and 20.6 +/- 0.3 nm, respectively (P = 0.031). Compared with NP subjects, GAgP subjects had a greater number of circulating LDL particles (961.3 +/- 105.3 vs. 1,349.0 +/- 133.2 nmol/l; P = 0.032). Differences in the plasma levels of large, medium, and small HDL were not statistically significant. NP and GAgP subjects had similar plasma levels of total LDL-associated PAF-AH activity; however, LDL of GAgP subjects contained less PAF-AH activity per microgram of LDL protein (1,458.0 +/- 171.0 and 865.2 +/- 134 pmol/min/microg; P = 0.014). These results indicate that, in general, GAgP subjects have a more atherogenic lipoprotein profile and lower LDL-associated PAF-AH activity than NP subjects. These differences may help explain the increased risk of GAgP subjects for cardiovascular disease.