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Background: Generalized aggressive periodontitis (GAgP) encompasses a distinct type of periodontal disease exhibiting much more rapid periodontal tissue destruction than chronic periodontitis. The best method for management of GAgP may include the use of both regenerative periodontal techniques and the administration of systemic antibiotics., Methods: The treatment of a case of GAgP over a period of 6.7 years is presented in this case report. Initial periodontal therapy (week 1- 32) consisted of supragingival plaque control and three appointments of scaling and root planing. Based on the periodontal pathogens isolated (5 species), the patient also received metronidazole plus amoxicillin for one week, followed 10 weeks later by metronidazole plus amoxicillin/clavulanate for one week. The patient was put on regular supportive periodontal therapy (SPT) thereafter. Orthodontic treatment was performed after completion of the initial therapy for 96 weeks. Measurements of clinical attachment level, bleeding on probing and plaque index were obtained at every examination., Results: Antimicrobial and mechanical treatment resulted in eradication of all periopathogens and significantly improved all clinical parameters. During orthodontic treatment and active maintenance, there was no relapse of GAgP. The patient participated in SPT for 194 weeks and thereafter decided to discontinue SPT. Twenty-four months later a relapse of GAgP was diagnosed and all teeth had to be extracted., Conclusions: These results indicate that a combined mechanical and antimicrobial treatment approach can lead to consistent resolution of GAgP. Further studies including a larger number of cases are warranted to validate these findings.

Background: This study determines the gingival crevicular fluid (GCF) levels of interleukin (IL)-1 beta, IL-2, IL-4, IL-8, interferon (IFN)-gamma and elastase activity in inflamed shallow and deep periodontal sites from patients with generalized chronic (GCP) and generalized aggressive periodontitis (GAgP), and to compare them to shallow sites from subjects with gingivitis. A secondary aim analyzes the microbiologic profile of these subjects., Methods: Cross-sectional clinical data were obtained from 20 GCP, 17 GAgP, and 10 gingivitis subjects. GCF samples were collected with paper strips and the levels of IL-1 beta, IL-2, IL-4, IL-8, and IFN-gamma were measured using a multiplexed bead immunoassay. Elastase activity was assessed by an enzymatic assay. Subgingival plaque samples were analyzed using checkerboard DNA-DNA hybridization. Significance of differences among groups for immunologic and microbiologic data was examined using Kruskal-Wallis adjusting for multiple comparisons., Results: Mean clinical parameters and GCF volumes were higher in patients with GCP and GAgP compared to the gingivitis group. Higher levels of IL-1 beta and higher elastase activity were found in deep sites compared to shallow sites in both periodontitis groups (P <0.05). The microbiologic data showed significantly higher levels of the red complex species in patients with GCP and GAgP compared to gingivitis (P <0.05). There were no statistically significant differences in levels of GCF biomarkers and in levels of subgingival bacterial species between subjects with GCP and GAgP., Conclusion: There were no statistically significant differences in the measured immunologic and microbiologic parameters between subjects with GCP and GAgP.

Background and Objective: Cytolethal distending toxin (CDT) is a genotoxin produced by Aggregatibacter actinomycetemcomitans. In spite of its association with pathogenesis, little is known about the humoral immune response against the CDT. This study aimed to test whether subgingival colonization and humoral response to A. actinomycetemcomitans would lead to a response against CDT., Material and Methods: Sera from periodontally healthy, localized and generalized aggressive periodontitis and chronic periodontitis subjects (n = 80) were assessed for immunoglobulin G titers to A. actinomycetemcomitans serotypes a/b/c and to each CDT subunit (CdtA, CdtB and CdtC) by ELISA. A. actinomycetemcomitans subgingival levels and neutralization of CDT activity were also analyzed., Results: Sera from 75.0% localized and 81.8% generalized aggressive periodontitis patients reacted to A. actinomycetemcomitans. A response to serotype b was detected in localized (66.7%) and generalized aggressive periodontitis (54.5%). Reactivity to A. actinomycetemcomitans correlated with subgingival colonization (R = 0.75, p < 0.05). There was no correlation between A. actinomycetemcomitans colonization or response to serotypes and the immunoglobulin G response to CDT subunits. Titers of immunoglobulin G to CdtA and CdtB did not differ among groups; however, sera of all generalized aggressive periodontitis patients reacted to CdtC. Neutralization of CDT was not correlated with levels of antibodies to CDT subunits., Conclusion: Response to CdtA and CdtB did not correlate with the periodontal status of the subject in the context of an A. actinomycetemcomitans infection. However, a response to CdtC was found in sera of generalized but not of localized aggressive periodontitis subjects. Differences in response to CdtC between generalized and localized aggressive periodontitis subjects indicate that CDT could be expressed differently by the infecting strains. Alternatively, the antibody response to CdtC could require the colonization of multiple sites.

Background: The aim of this study is to evaluate the effects of metronidazole-amoxicillin combination on clinical and microbiologic parameters in patients with generalized aggressive periodontitis., Methods: Twenty-eight patients were randomly included. The test group (n = 12) received amoxicillin-metronidazole combination and scaling and root planing; the control group (n = 16) received scaling and root planing alone. In addition to the clinical examinations, subgingival plaque samples were analyzed for total cultivable bacteria and the presence of Porphyromonas gingivalis, Tannerella forsythia (previously T. forsythensis), Treponema denticola, Prevotella intermedia, Prevotella nigrescens, Prevotella pallens, and Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans) using polymerase chain reaction., Results: All clinical parameters improved significantly compared to baseline (P <0.05) in both groups. There was a statistically significant reduction of pockets and clinical attachment gain in the combined group compared to the control group (P <0.05). Total counts of bacteria also decreased significantly at 3 and 6 months in both groups (P <0.05). T. denticola and T. forsythia were the most prevalent bacteria throughout the study. T. denticola showed a continuous decrease over 6 months in the test group, whereas no change was seen in the control group beyond 3 months. P. gingivalis decreased significantly at 3 months (P <0.05), whereas T. forsythia was the only pathogen decreased below detection limits by the combination therapy with a significant difference compared to the control group (P <0.05)., Conclusions: The results from this study suggest that combined amoxicillin and metronidazole use as an adjunct to scaling and root planing leads to better clinical healing compared to mechanical treatment alone. The polypharmaceutical approach used results in a significant and substantial decrease in T. forsythia and prevents its recolonization for 6 months, suggesting that T. forsythia may determine the long-term stability of periodontal treatment outcomes.

Background and Objective: Periodontitis is influenced by specific host-dependent immune responses. Periodontopathogens induce innate immune responses, amongst others, via toll-like receptor 2 (TLR2), resulting in activation of the nuclear transcription factor nuclear factor-kappaB (NF-kappaB). The aim of this case-control study was to evaluate links between genetic variants of these genes and chronic/aggressive periodontitis in a multivariate model., Material and Methods: A total of 141 patients with periodontitis (63 with chronic periodontitis and 78 with aggressive periodontitis) and 81 controls without periodontitis were included in the study. Polymorphisms in TLR2 (Arg677Trp, Arg753Gln) and in NF-kappaB (-94ins/delATTG) were determined by restriction fragment length polymorphism and fragment length analyses, respectively. Subgingival bacterial colonization was evaluated using a PCR/DNA probe test (micro-Ident)., Results: Although there was no association of the TLR2 polymorphism Arg753Gln with periodontitis, heterozygous carriers (Arg/Gln) were at a higher risk for colonization with bacteria of the 'red complex' (corrected p-value = 0.042). The del/del genotype of the NF-kappaB polymorphism was associated with aggressive periodontitis considering age, gender, smoking and approximal plaque index as potential confounders (odds ratio = 2.81, p = 0.035, 95% confidence interval: 1.08-7.33). del/del carriers had a higher risk for subgingival colonization with Aggregatibacter actinomycetemcomitans (odds ratio = 2.36, p = 0.030, 95% confidence interval: 1.09-5.1; adjusted for age, gender, smoking and pocket depth(bacteria))., Conclusions: The del/del genotype of NF-kappaB was shown to be associated with the occurrence of aggressive periodontitis.

Generalized aggressive periodontitis (GAgP) is an inflammatory condition resulting in destruction of tooth-supporting tissues. We examined the production of IL-1beta, IL-6, tumour necrosis factor (TNF)-alpha, IL-12 and IL-10 in cultures of peripheral mononuclear cells (MNC) from 10 patients with GAgP and 10 controls stimulated with periodontal pathogens or a control antigen, tetanus toxoid (TT) in the presence of autologous serum. The pathogens used were Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum, either as type strains or bacteria isolated from the participants' inherent oral flora. The P. gingivalis -induced production of IL-6 was approximately 2.5-fold higher in patients with GAgP than in healthy controls (P < 0.05), while the corresponding TNF-alpha production was non-significantly elevated. IL-1beta production induced by P. gingivalis, as all cytokine responses induced by Pr. intermedia, F. nucleatum and TT was similar in the two groups. A reduced IL-12p70 response to Pr. intermedia and F. nucleatum was observed in smokers compared to non-smoking patients (P < 0.02). To assess the role of serum factors in the elevated IL-6 response to P. gingivalis, MNC from two donors free of disease were stimulated with this bacterium in the presence of the various patient and control sera. An elevated IL-6 and TNF-alpha response was observed in the presence of patient sera (P < 0.01 and P < 0.04, respectively). The data suggest that an exaggerated production of IL-6 occurs in GAgP, and that pro-inflammatory serum factors play an essential role in the response.

This study examined the distribution pattern of Aggregatibacter actinomycetemcomitans serotypes in the subgingival plaque of subjects residing in the United States. A. actinomycetemcomitans was identified in 256 subgingival plaque samples from 161 subjects. For 190 of the 256 samples, the total cultivable bacteria and selected periodontal pathogenic species were determined. A. actinomycetemcomitans isolates were confirmed by a16S rDNA-based PCR analysis, genotyped by arbitrarily-primed PCR, and serotyped by PCR analysis of serotype-specific gene clusters. A total of 82 distinct A. actinomycetemcomitans strains were identified. The serotype distribution pattern of the strains was 21 (25.6%) serotype a, 12 (14.6%) b, 41 (50%) c, 6 (7.3%) e, 1 (1.2%) f, and 1 (1.2%) non-typeable. For 14 subjects where multiple colonies of A. actinomycetemcomitans were identified, 11 subjects (78.6%) were each infected by a single serotype, while the remaining three subjects (21.3%) were each infected by two serotypes of A. actinomycetemcomitans. There was an inverse relationship between the level of cultivable A. actinomycetemcomitans and Porphyromonas gingivalis. Within subgingival plaque of study cohort A. actinomycetemcomitans serotype c was the dominant serotype and comprised 50% of all strains, followed by (in order of detection frequency) serotypes a and b. Serotypes d, e, and f strains were either not detected or less frequently found. Serotype distribution patterns of subgingival A. actinomycetemcomitans may vary among subjects of different race orethnicity.

Aggregatibacter actinomycetemcomitans is a Gram-negative bacterium that colonizes the human oral cavity and is the causative agent for localized aggressive periodontitis (LAP), an aggressive form of periodontal disease that occurs in adolescents. A. actinomycetemcomitans secretes a protein toxin, leukotoxin (LtxA), which helps the bacterium evade the host immune response during infection. LtxA is a membrane-active toxin that specifically targets white blood cells (WBCs). In this review, we discuss recent developments in this field, including the identification and characterization of genes and proteins involved in secretion, regulation of LtxA, biosynthesis, newly described activities of LtxA, and how LtxA may be used as a therapy for the treatment of diseases.

Aggressive periodontitis is characterized by a rapid and severe periodontal destruction in young systemically healthy subjects. A greater prevalence is reported in Africans and African descendent groups than in Caucasians and Hispanics. We first fine mapped the interval 1q24.2 to 1q31.3 suggested as containing an aggressive periodontitis locus. Three hundred and eighty-nine subjects from 55 pedigrees were studied. Saliva samples were collected from all subjects, and DNA was extracted. Twenty-one single nucleotide polymorphisms were selected and analyzed by standard polymerase chain reaction using TaqMan chemistry. Non-parametric linkage and transmission distortion analyses were performed. Although linkage results were negative, statistically significant association between two markers, rs1935881 and rs1342913, in the FAM5C gene and aggressive periodontitis (p = 0.03) was found. Haplotype analysis showed an association between aggressive periodontitis and the haplotype A-G (rs1935881-rs1342913; p = 0.009). Sequence analysis of FAM5C coding regions did not disclose any mutations, but two variants in conserved intronic regions of FAM5C, rs57694932 and rs10494634, were found. However, these two variants are not associated with aggressive periodontitis. Secondly, we investigated the pattern of FAM5C expression in aggressive periodontitis lesions and its possible correlations with inflammatory/immunological factors and pathogens commonly associated with periodontal diseases. FAM5C mRNA expression was significantly higher in diseased versus healthy sites, and was found to be correlated to the IL-1beta, IL-17A, IL-4 and RANKL mRNA levels. No correlations were found between FAM5C levels and the presence and load of red complex periodontopathogens or Aggregatibacter actinomycetemcomitans. This study provides evidence that FAM5C contributes to aggressive periodontitis.

Objectives: To investigate the short-term effects of nonsurgical therapy (scaling and root planing, SRP) on the subgingival microbiota in chronic (CP) and aggressive (AP) periodontal disease., Method and Materials: Ninety-seven CP and AP subjects underwent full-mouth SRP on 2 consecutive days. AP patients were randomly assigned to either receive systemic metronidazole plus amoxicillin (AP+AB) or were treated mechanically alone (AP). Pathogens were identified with 16S rRNA oligodeoxynucleotide probes and dot-blot hybridization before and at days 2, 3, 4, 7, 10, and 21 of healing. CP subjects were treated by scaling and root planing along with placebo tablets., Results: Initially, AP cell counts were 69.9- (Porphyromonas gingivalis), 10.2- (Aggregatibacter actinomycetemcomitans), 5.7- (Tannerella forsythia), and 3.3-fold (Prevotella intermedia) enhanced compared to CP cell counts. Following SRP, immediate elimination occurred in single individuals of all three treatment groups at day 2. After SRP plus antibiotic therapy (AP+AB), the prevalence scores dropped beyond the levels of AP and CP, beginning at day 7, and remained low until day 21 (P =or< .05). Clinical healing statistically benefited from SRP with no differences among the three treatment groups., Conclusion: Nonsurgical therapy resulted in both a suppression and early elimination of single taxa immediately after completion of active treatment. Systemic antibiotics significantly accelerate the suppression of the periodontal microflora, but have limited effect on the elimination of target isolates during healing.

Aim: To examine relationships between subgingival biofilm composition and levels of gingival crevicular fluid (GCF) cytokines in periodontal health and generalized aggressive periodontitis (GAP)., Materials and Methods: Periodontal parameters were measured in 25 periodontally healthy and 31 GAP subjects. Subgingival plaque and GCF samples were obtained from 14 sites from each subject. Forty subgingival taxa were quantified using checkerboard DNA-DNA hybridization and the concentrations of eight GCF cytokines were measured using Luminex. Cluster analysis was used to define sites with similar subgingival microbiotas in each clinical group. Significance of differences in clinical, microbiological and immunological parameters among clusters was determined using the Kruskal-Wallis test., Results: GAP subjects had statistically significantly higher GCF levels of interleukin-1beta (IL-1beta) (p<0.001), granulocyte-macrophage colony-stimulating factor (GM-CSF) (p<0.01) and IL-1beta/IL-10 ratio (p<0.001) and higher proportions of Red and Orange complex species than periodontally healthy subjects. There were no statistically significant differences in the mean proportion of cytokines among clusters in the periodontally healthy subjects, while the ratio IL-1beta/IL-10 (p<0.05) differed significantly among clusters in the aggressive periodontitis group., Conclusions: Different subgingival biofilm profiles are associated with distinct patterns of GCF cytokine expression. Aggressive periodontitis subjects were characterized by a higher IL-1beta/IL-10 ratio than periodontally healthy subjects, suggesting an imbalance between pro- and anti-inflammatory cytokines in aggressive periodontitis.

Background: Genetic polymorphisms that influence neutrophil function were proposed as possible risk factors for aggressive periodontitis (AgP). The aim of this study is to test whether a clinical diagnosis and specific genetic variants are associated with neutrophil activity in subjects with AgP and healthy subjects., Methods: This study describes the results of neutrophil analyses performed in 40 subjects (20 subjects with AgP and 20 healthy control subjects). Neutrophils were extracted from peripheral blood and analyzed in a masked manner for phagocytosis and oxidative burst by flow cytometry in response to Escherichia coli, Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), and Porphyromonas gingivalis. Genomic DNA was extracted and genotyped for the nicotinamide adenine dinucleotide phosphate p22 phagocytic oxidase subunit (p22(phox)) cytochrome b alpha gene (CYBA) C242T, crystallizable gamma fragment (Fcgamma)IIa H/R, and FcgammaIIIb neutrophil antigen (NA)1/NA2 polymorphisms., Results: Analyses in pairs of white subjects (n = 24) revealed that patients with AgP exhibited a higher oxidative burst in response to E. coli (P = 0.002) and P. gingivalis (P = 0.002) compared to healthy control subjects. The p22(phox) CYBA 242 T allele was associated with an oxidative burst in response to the challenge by two strains of A. actinomycetemcomitans (P = 0.018 and P = 0.046). The FcgammaIIa polymorphism was associated with the phagocytic index of E. coli (P = 0.024)., Conclusion: This study confirms previous reports of a higher oxidative burst associated with AgP and presented preliminary evidence that genetic factors may influence neutrophil function in patients with AgP and healthy individuals.

Aim: The aim of this study was to evaluate the clinical and microbiological effects of scaling and root planing (SRP) alone or combined with metronidazole (MTZ) and amoxicillin (AMX) in the treatment of subjects with generalized aggressive periodontitis (GAgP)., Materials and Methods: A double-blind, placebo-controlled, randomized clinical trial was conducted in 30 subjects receiving SRP alone or combined with MTZ (400 mg 3 x per day) and AMX (500 mg 3 x per day) for 14 days. Clinical and microbiological examinations were performed at baseline and 3 months post-SRP. Nine subgingival plaque samples per subject were analysed using checkerboard DNA-DNA hybridization., Results: Subjects receiving MTZ and AMX showed the greatest improvements in the mean full-mouth probing depth and clinical attachment level and at initially intermediate and deep sites. The most beneficial changes in the microbial profile were also observed in the MTZ+AMX group, which showed the lowest proportions of the red complex as well as a significant decrease in the proportions of the orange complex after treatment. The antibiotic therapy also reduced the levels of Aggregatibacter actinomycetemcomitans at initially deep sites., Conclusion: Subjects with GAgP significantly benefit from the adjunctive use of MTZ and AMX. The short-term advantages are observed in the clinical and microbiological parameters.

Objective: The aim of this study was to compare the outcome of two different microbiological tests for detection of Aggregatibacter actinomycetemcomitans (A.a.), Porphyromonas gingivalis (P.g.), Tannerella forsythia (T.f.) and Treponema denticola (T.d.)., Material and Methods: A total of 69 adult patients with severe chronic (sCP) or aggressive periodontitis (AgP) participated in the study. Microbiological samples were examined for A.a., P.g., T.f. and T.d. using an RNA probe test (PADO) and a real-time polymerase chain reaction test (MERI)., Results: For all periodontal pathogens under investigation, the median bacterial counts detected with PADO were smaller compared to those detected with MERI. P.g., T.f. and T.d. could be found in the majority of all patients with both tests. With MERI, A.a. was detected more often (24.6%) than with PADO (18.8%). Only 10.1% of the patients tested positive for A.a. with both tests., Conclusion: Both tests showed a high percentage of agreement for P.g., T.f. and T.d., but exhibited marked differences in the detection of A.a.

Background: Bacteria in periodontal pockets develop complex sessile communities that attach to the tooth surface. These highly dynamic microfloral environments challenge both clinicians and researchers alike. The exploration of structural organisation and bacterial interactions within these biofilms is critically important for a thorough understanding of periodontal disease. In recent years, Filifactor alocis, a fastidious, Gram-positive, obligately anaerobic rod was repeatedly identified in periodontal lesions using DNA-based methods. It has been suggested to be a marker for periodontal deterioration. The present study investigated the epidemiology of F. alocis in periodontal pockets and analysed the spatial arrangement and architectural role of the organism in in vivo grown subgingival biofilms., Results: A species-specific oligonucleotide probe, FIAL, was designed and evaluated. A total of 490 subgingival plaque samples were submitted to PCR and subsequent dot blot hybridization to compare the prevalence of F. alocis in patients suffering from generalized aggressive periodontitis (GAP), chronic periodontitis (CP), and control subjects resistant to periodontitis. Moreover, a specially designed carrier system was used to collect in vivo grown subgingival biofilms from GAP patients. Subsequent topographic analysis was performed using fluorescence in situ hybridization.While the majority of patients suffering from GAP or CP harboured F. alocis, it was rarely detected in the control group. In the examined carrier-borne biofilms the organism predominantly colonized apical parts of the pocket in close proximity to the soft tissues and was involved in numerous structures that constitute characteristic architectural features of subgingival periodontal biofilms., Conclusions: F. alocis is likely to make a relevant contribution to the pathogenetic structure of biofilms accounting for periodontal inflammation and can be considered an excellent marker organism for periodontal disease.

Introduction: Very little is known of the diversity and expression of virulence factors of serotypes of Aggregatibacter actinomycetemcomitans. Toxic activity on Chinese hamster ovary (CHO) cells and cdt and ltx genotyping were evaluated in A. actinomycetemcomitans serotypes., Methods: Forty-one A. actinomycetemcomitans isolates were analysed for CHO cell growth inhibition. Genotyping was performed by polymerase chain reactions specific to the ltx promoter region, serotype-specific and cdt region and by sequencing of cdtB., Results: cdtABC was detected in 40 strains. Analysis of the cdtA upstream region revealed 10 cdt genotypes. Toxicity to CHO cells was detected for 92.7% of the isolates; however, no correlation between the toxic activity and the cdt genotype was detected. Serotype c was more prevalent among Brazilian samples (68.0%). Four serotype b isolates from subjects with aggressive periodontitis were associated with high leukotoxin production and exhibited moderate to strong toxic activity in CHO cells, but were classified in different cdt genotypes. High levels of toxicity in CHO cells were not associated with a particular serotype; 57.1% of serotype a isolates presented low toxicity to CHO cells whereas the highly toxic strains belonged to serotypes b and c. Sequencing of cdtB revealed a single nucleotide polymorphism of amino acid 281 but this was not related to the toxic activity in CHO cells., Conclusion: Differences in prevalence of the low and highly cytotoxic strains among serotypes reinforce the hypothesis that serotype b and c isolates of A. actinomycetemcomitans are more virulent than serotype a strains.

Aim: To monitor the intra-oral microbiological changes after full-mouth extraction using quantitative polymerase chain reaction (qPCR)., Material and Methods: Nine patients with severe, aggressive periodontitis, for whom a full-mouth tooth extraction was the only remaining treatment option were recruited. Before and 6 months after extraction, microbial samples were obtained (tongue, saliva and subgingival plaque) and analysed by qPCR., Results: The elimination of subgingival niches, by extraction of all natural teeth, resulted in a 3-log reduction of Porphyromonas gingivalis and Tannerella forsythia, and more modest reductions of Aggregatibacter actinomycetemcomitans and Prevotella intermedia. However, the detection frequencies of these periodontopathogens in saliva and on the tongue remained unchanged after full-mouth tooth extraction., Conclusion: In contrast to what has been believed so far, full-mouth tooth extraction does not result in eradication of all periodontopathogens but only in a significant reduction. The clinical consequences of this observation remain speculative.

Background: The identification of Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans) serotypes may add some important information to the understanding of the pathogenetic background of severe periodontal infections. This study compared serotypes of A. actinomycetemcomitans in two groups of periodontal patients with different ethnic backgrounds., Methods: A total of 194 patients (96 Germans and 98 Koreans) with aggressive or severe chronic periodontitis participated in the study. Microbiologic analysis of pooled samples from subgingival plaque was performed by using a real-time polymerase chain reaction (PCR) test for A. actinomycetemcomitans. In patients who tested positive for A. actinomycetemcomitans, serotypes (a through f) were determined by nucleic acid-based methods., Results: The prevalence of patients who tested positive for A. actinomycetemcomitans with the real-time PCR was comparable in both groups (Germans: 27.0%; Koreans: 22.2%). In German patients, the serotypes detected most frequently were b (33.3%), c (25.0%), and a (20.8%), whereas in Korean patients, the serotype distribution was different, with serotypes c (61.9%) and d (19.0%) accounting for >80% of the complete serotype spectrum., Conclusion: Even if the percentage of patients who tested positive for A. actinomycetemcomitans was identical in patients with generalized aggressive and severe chronic periodontitis and different ethnic backgrounds, the distribution of A. actinomycetemcomitans serotypes may exhibit marked differences.

Aim: To look for clinical signs of periodontal disease in young adults who exhibited radiographic bone loss and detectable numbers of Aggregatibacter actinomycetemcomitans in their primary dentition., Material and Methods: Periodontal status and radiographic bone loss were examined in each of the subjects 16 years after the baseline observations. Techniques for anaerobic and selective culture, and checkerboard, were used to detect periodontitis-associated bacterial species. The isolated A. actinomycetemcomitans strains were characterized by polymerase chain reaction., Results: Signs of localized attachment loss were found in three out of the 13 examined subjects. A. actinomycetemcomitans was recovered from six of these subjects and two of these samples were from sites with deepened probing depths and attachment loss. Among the isolated A. actinomycetemcomitans strains, serotypes a-c and e, but not d or f, were found. None of the isolated strains belonged to the highly leucotoxic JP2 clone, and one strain lacked genes for the cytolethal distending toxin., Conclusions: This study indicates that the presence of A. actinomycetemcomitans and early bone loss in the primary dentition does not necessarily predispose the individual to periodontal attachment loss in the permanent dentition.

Background and Objective: As a pro-inflammatory cytokine, interleukin-2 mediates the activation, growth and differentiation of T and B lymphocytes and natural killer cells. Promoter polymorphisms of the interleukin-2 gene have been associated with altered interleukin-2 production or identified as prognostic markers for various infectious diseases. Therefore, the aim of this study was to evaluate two polymorphisms at positions -330 T/G and 166 G/T in patients with generalized chronic periodontitis (n = 58) or generalized aggressive periodontitis (n = 73) in comparison with periodontitis-free controls (n = 69)., Material and Methods: Both interleukin-2 polymorphisms were analyzed using the polymerase chain reaction with sequence-specific primers. Distributions of single alleles, genotypes and haplotypes were calculated using the chi-square test. Risk factor analyses were carried out by logistic regression with respect to established cofactors for periodontitis. The presence of subgingival bacteria in an individual were analyzed using a molecular biological method (the micro-Ident test)., Results: The interleukin-2 genotype -330 TG occurred less frequently in patients with chronic periodontitis (25.9% vs. 49.3%). Moreover, this genotype decreased the adjusted odds ratio for chronic periodontitis (odds ratio = 0.394), whereas the interleukin-2 genotype 166 TT and the haplotype combination interleukin-2 -330,166 TT : TT were associated with an increased adjusted odds ratio (odds ratio = 2.82 or 2.97). For the latter interleukin-2 combination, a positive association for the subgingival presence of Porphyromonas gingivalis (81.3% vs. 59.5%) and bacteria of the 'red complex' (78.1% vs. 56.0%) was shown., Conclusion: The interleukin-2 genotypes -330 TG and 166 TT, as well as the combination genotype interleukin-2 TT : TT, could be putative prognostic factors for chronic periodontitis.

The JP2 clone of Aggregatibacter actinomycetemcomitans is strongly associated with aggressive periodontitis. To obtain information about colonization dynamics of the JP2 clone, we used PCR to examine its presence in 365 Moroccan juveniles from whom periodontal plaque samples were collected at baseline and after one and two years. Periodontal attachment loss was measured at baseline and at the two-year follow-up. At baseline, 43 (12%) carriers of the JP2 clone were found. Nearly half (44 %) of these were persistently colonized with the clone. The relative risk for the development of aggressive periodontitis, adjusted for the concomitant presence of other genotypes of A. actinomycetemcomitans, was highest for individuals continuously infected by the JP2 clone (RR = 13.9; 95% CI, 9.0 to 21.4), indicating a relationship between infectious dose and disease, which further substantiates the evidence for the JP2 clone as a causal factor in aggressive periodontitis.

Aim: The microbial profile of localized aggressive periodontitis (LAgP) has not yet been determined. Therefore, the aim of this study was to evaluate the subgingival microbial composition of LAgP., Material and Methods: One hundred and twenty subjects with LAgP (n=15), generalized aggressive periodontitis (GAgP, n=25), chronic periodontitis (ChP, n=30) or periodontal health (PH, n=50) underwent clinical and microbiological assessment. Nine subgingival plaque samples were collected from each subject and analysed for their content of 38 bacterial species using checkerboard DNA-DNA hybridization., Results: Red complex and some orange complex species are the most numerous and prevalent periodontal pathogens in LAgP. The proportions of Aggregatibacter actinomycetemcomitans were elevated in shallow and intermediate pockets of LAgP subjects in comparison with those with GAgP or ChP, but not in deep sites. This species also showed a negative correlation with age and with the proportions of red complex pathogens. The host-compatible Actinomyces species were reduced in LAgP., Conclusion: A. actinomycetemcomitans seems to be associated with the onset of LAgP, and Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Campylobacter gracilis, Eubacterium nodatum and Prevotella intermedia play an important role in disease progression. Successful treatment of LAgP would involve a reduction in these pathogens and an increase in the Actinomyces species.

Background and Objective: Mucosal inflammatory responses are orchestrated largely by pro-inflammatory chemokines. The chemokine granulocyte chemotactic protein 2 (CXCL6) is involved in neutrophil recruitment and migration. Previous studies have shown that granulocyte chemotactic protein 2 is up-regulated during mucosal inflammation (e.g. in inflammatory bowel disease), similarly to the functionally and structurally related chemokine interleukin-8. Nevertheless, unlike interleukin-8, a role of granulocyte chemotactic protein 2 in gingival inflammation has not been yet demonstrated. In this study we aimed to evaluate the expression of the chemokine granulocyte chemotactic protein 2 in clinically healthy vs. diseased gingival tissues and to explore possible correlations with clinical and microbiological markers of periodontitis., Material and Methods: Gene expression in 184 'diseased' and 63 'healthy' gingival tissue specimens from 90 patients with periodontitis was analyzed using Affymetrix U133Plus2.0 arrays. The expression of granulocyte chemotactic protein 2 was further confirmed by real-time reverse transcription-polymerase chain reaction, western blotting and enzyme-linked immunosorbent assay, while the localization of granulocyte chemotactic protein 2 in gingival tissues was analyzed by immunohistochemistry. Plaque samples from the adjacent periodontal pockets were collected and evaluated for 11 species of periodontal bacteria using checkerboard DNA-DNA hybridizations., Results: Among all known chemokines, GCP-2 expression was the most up-regulated (3.8-fold, p < 1.1 x 10(-16)), in 'diseased' vs. 'healthy' tissue as compared to a 2.6-fold increased expression of interleukin-8 mRNA (p < 1.2 x 10(-15)). Increased expression of granulocyte chemotactic protein 2 correlated with higher levels of 'red' and 'orange' complex pathogens and with increased probing depth, but not with attachment loss. Immunohistochemistry showed that granulocyte chemotactic protein 2 was expressed in gingival vascular endothelium., Conclusion: The level of expression of granulocyte chemotactic protein 2 correlates with the severity of periodontitis and appears to act as a hitherto unrecognized functional adjunct to interleukin-8 in diseased gingival tissues.

The Papillon-Lefevre syndrome (PLS) was first described by Papillon and Lefevre in 1924. It is an autosomal recessive disorder characterized by a diffuse palmoplanter hyperkeratosis and rapidly progressive and devastating periodontitis, affecting the primary as well as the permanent dentition, attributed to a point mutation of the cathepsin C gene. This paper presents a clinical presentation and a brief review of its etiology and treatment modalities.

Background and Objective: This study analyzed the interaction of Porphyromonas gingivalis ATCC 33277 and Aggregatibacter actinomycetemcomitans Y4 with peripheral blood polymorphonuclear neutrophils taken from patients with aggressive periodontitis and chronic periodontitis., Material and Methods: Peripheral blood polymorphonuclear neutrophils obtained from 12 patients with chronic periodontitis, six patients with aggressive periodontitis and 12 healthy controls were exposed to P. gingivalis and A. actinomycetemcomitans following opsonization of the bacteria using the patient's own serum. Serum immunoglobulin G (IgG) levels against both periodontopathogens were measured. Phagocytosis and killing of the bacteria, as well as the extracellular human neutrophil elastase activity, were quantified. The total amount and the extracellular release of reactive oxygen species were measured using luminol-dependent and isoluminol-dependent chemiluminescence., Results: Polymorphonuclear neutrophils from patients with chronic (62.16 +/- 19.39%) and aggressive (43.26 +/- 26.63%) periodontitis phagocytosed more P. gingivalis than the healthy controls (24.43 +/- 19.87%) at the 30-min time point after exposure to the bacteria (p < 0.05). High serum IgG levels against P. gingivalis and A. actinomycetemcomitans were detected in subjects with periodontitis. Polymorphonuclear neutrophils from subjects with chronic and aggressive periodontitis released significantly more reactive oxygen species and demonstrated greater human neutrophil elastase activity in the absence of any stimulus than polymorphonuclear neutrophils from healthy controls (p < 0.05). Polymorphonuclear neutrophils in chronic periodontitis released significantly more reactive oxygen species when exposed to P. gingivalis and A. actinomycetemcomitans than polymorphonuclear neutrophils in aggressive periodontitis., Conclusion: High serum IgG levels against P. gingivalis and A. actinomycetemcomitans promote phagocytosis in periodontitis. The extracellular release of reactive oxygen species and neutrophil elastase by polymorphonuclear neutrophils may also contribute to damage of the surrounding periodontal tissues.

Implants showing signs of peri-implantitis harbor a microbiota similar to that of periodontitis-affected teeth. This case report describes the subgingival microbiota of a 45-year-old female with advanced periodontitis before and after complete edentulation and reconstruction with dental implants. A 3-month healing period post extraction passed before implants were placed using a two-stage submerged implant protocol. At 4- to 6-month recall visits after definitive prosthetic reconstruction, some implant sites showed bleeding on probing and localized mucositis. Microbiological culture of three inflamed peri-implant sites showed an almost identical spectrum of pathogens, including Porphyromonas gingivalis, Tannerella forsythia, and other major pathogenic bacteria characteristic of aggressive periodontitis. As natural teeth were absent for 8 months, this case report suggests that periodontal pathogens can be retained for a prolonged period of time in nondental sites, from where they can later colonize and compromise the health of dental implants. The therapeutic implications of this finding are discussed.

Background: Compromised T-cell responses to periodontal pathogens may contribute to the pathogenesis of generalized aggressive periodontitis (GAgP). In this study, we attempted to characterize T-helper cell (Th1, Th2, and Th17) responses in patients with GAgP and healthy controls upon stimulation with disease-relevant pathogens., Methods: Mononuclear cells (MNCs) from 10 white patients with GAgP and 10 white controls were stimulated with Porphyromonas gingivalis American Type Culture Collection (ATCC) 33277 (Pg), Prevotella intermedia ATCC 25611, Fusobacterium nucleatum ATCC 49256 (Fn), and similar bacteria isolated from the participants' inherent oral flora. Tetanus toxoid (TT) was used as control antigen. The resulting production of interferon-gamma (IFN-gamma) and interleukin (IL)-2, -4, -5 and -17 and the induced proliferation of CD4+ T cells were measured., Results: MNCs from patients with GAgP exhibited decreased IL-2 responses to Pg and Fn. No difference was observed between patients with GAgP and controls with regard to CD4+ T-cell proliferation or the production of IFN-gamma and IL-4, -5, and -17, irrespective of whether type strains or bacteria isolated from the participants' oral cavity were used for stimulation. Moreover, similar proliferative and cytokine responses to TT were observed. Notably, smoking patients with GAgP exhibited significantly lower IFN-gamma responses to the bacteria and to TT than non-smoking patients or controls., Conclusions: The decreased IL-2 responses of patients with GAgP to Pg and Fn combined with adequate IL-2 responses to TT suggest an impaired antigen-specific T-cell reactivity with periodontal pathogens in GAgP. The decreased IFN-gamma responses of smokers within the patient group suggest that smoking may aggravate this impairment.

A case of a 31-year-old female with aggressive periodontitis over 14 years is presented. From 1993 to 2000, no periodontal therapy occurred; disease development and progression could be reconstructed upon radiographic findings. In 2000, full-mouth disinfection therapy and antibiotic therapy was performed, as well as regenerative surgical treatments. Seven years after surgical treatment, stable periodontal conditions and clear bone regeneration in the surgical areas was evident.

Introduction: Cytolethal distending toxin (Cdt) is potentially one of several virulence factors of Aggregatibacter actinomycetemcomitans, the prime etiological agent of localized aggressive periodontitis (LAP). Little is known regarding the Cdt-specific antibody response in humans. The current study is a quantitative and qualitative evaluation of the toxin-specific antibody response in a cohort of LAP patients and age-, race- and sex-matched controls., Methods: Ninety-five subjects provided a total of 692 serum samples. Sera were analysed by enzyme-linked immunosorbent assays to determine the titers of antibody against the intact Cdt holotoxin as well as the individual subunit proteins (CdtA, CdtB, and CdtC). Neutralization of growth inhibition mediated by Cdt was evaluated in a modified colony-forming assay using Chinese hamster ovary cells., Results: Fourteen of the 95 subjects exhibited significant serum Cdt-binding activity. There were no differences in the percentages of seropositive individuals or in the mean antibody titers between the control and LAP groups. Binding activity was detected against each of the three Cdt subunit proteins in all of the positive samples. Neutralization of Cdt-mediated growth inhibition was detected in samples from all of the seropositive subjects (range 20-75%)., Conclusions: Cdt, a recently identified A. actinomycetemcomitans virulence factor, is capable of inducing a neutralizing antibody response indicating that the toxin is produced during natural infection of humans. The failure of a vast majority (20 of 23) of the LAP subjects to mount a significant anti-Cdt response may in part explain their relative susceptibility to the disease.

Introduction: 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)., Methods: 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., Results: 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., Conclusions: GAgP subjects presented diminished phagocytic activity of peripheral PMNs and high prevalence and levels of classical periodontal pathogens.

Objective: The objective of this study was to evaluate the effect of a dental water jet on plaque biofilm removal using scanning electron microscopy (SEM)., Methodology: Eight teeth with advanced aggressive periodontal disease were extracted. Ten thin slices were cut from four teeth. Two slices were used as the control. Eight were inoculated with saliva and incubated for 4 days. Four slices were treated using a standard jet tip, and four slices were treated using an orthodontic jet tip. The remaining four teeth were treated with the orthodontic jet tip but were not inoculated with saliva to grow new plaque biofilm. All experimental teeth were treated using a dental water jet for 3 seconds on medium pressure., Results: The standard jet tip removed 99.99% of the salivary (ex vivo) biofilm, and the orthodontic jet tip removed 99.84% of the salivary biofilm. Observation of the remaining four teeth by the naked eye indicated that the orthodontic jet tip removed significant amounts of calcified (in vivo) plaque biofilm. This was confirmed by SEM evaluations., Conclusion: The Waterpik dental water jet (Water Pik, Inc, Fort Collins, CO) can remove both ex vivo and in vivo plaque biofilm significantly.

Objective: To measure the anti-Pg (porphyromonas gingivalis, Pg) IgG antibody titers in serum in patients with aggressive periodontitis (AgP); to study the association of IgG titers with periodontal status (alveolar bone loss); to analyze the age, gender or smoking status on IgG levels in patients with aggressive periodontitis., Methods: (1) Ninety-one AgP subjects with an average age of 27.6 years (females 56, males 35, and 14 male-smokers) were collected. Thirty-five healthy subjects with an average age of 29.4 years (males 10 and females 25) were also collected as control group. (2) Questionnaire, clinical evaluations and assessments of alveolar bone loss were made. (3) Patients were categorized into two groups: severe AgP group (58 subjects, with >14 teeth of moderate-severe alveolar bone loss); light-moderate group (33 subjects). (4) To collect serum sample of patients and determine the anti-Pg IgG antibody titers in serum using ELISA., Results: (1) The mean IgG antibody titers to Pg in AgP group (8.55 +/-3.24) were significantly higher than that in control group (4.78+/-2.58, P=0.0004). (2) No significant difference [(8.38+/-3.73) and (8.67+/-2.90), respectively] of mean anti-Pg IgG titers was found between male and female. (3) The mean of anti-Pg IgG titers (9.26+/-2.58) in elder group (aged>25) was significantly higher than that in younger patients (6.96+/-3.98, P=0.0014). (4) Smokers had significantly lower anti-Pg IgG titers (7.33+/- 3.72) than non-smokers (8.78+/-3.12, P=0.016). (5) Anti-Pg IgG titers in severe AgP group was significantly higher than that in light-moderate AgP group. (6)The results of multi-factor regression analysis showed that smoking, age and status of alveolar bone loss were the factors affecting anti-Pg IgG titers., Conclusion: The titers of IgG antibody to Pg in serum were related to age, smoking and periodontal status in AgP patients.

Localized aggressive periodontitis is a distinct entity of periodontal disease and is characterized by deep vertical bony defects that typically affect the first molars and incisors of young patients. Therapy is usually aimed at reducing the pathogenic microflora through scaling and root planing and the administration of systemic antibiotics. However, conservative periodontal therapy may result in reparative wound healing with limited regeneration of the lost tissues. Periodontal surgery combined with enamel matrix derivative has been introduced as a method to promote regeneration of the lost periodontium and has been studied extensively in the treatment of chronic periodontitis. This case report describes the treatment of a 27-year-old patient displaying severe localized aggressive periodontitis with documented disease progression. After initial therapy consisting of scaling and root planing and systemic administration of amoxicillin and metronidazole, the vertical defects were treated by minimally invasive access flaps combined with application of enamel matrix derivative. Clinical, microbiologic, and radiographic findings are reported for up to 1.5 years after initial therapy, indicating good efficacy of the therapeutic strategy and stability of the treatment outcome.

Objective: The aim was to compare the effect of scaling and root planing (SRP) alone, diode laser treatment (LAS) alone, and SRP combined with LAS (SRP + LAS) on clinical and microbial parameters in patients with aggressive periodontitis., Materials and Methods: Thirty patients with aggressive periodontitis were assessed for plaque, bleeding on probing (BOP), probing pocket depth (PPD), and clinical attachment level (CAL). Four plaque samples were randomly obtained, one from each quadrant that was randomly assigned to SRP alone, SRP + LAS, LAS alone, and control (CRL). A 980-nm diode laser was used in continuous mode at 2 W power. Plaque samples were collected 2 wk, 12 wk, and 6 mo post-treatment. The levels of Porphyromonas gingivalis, Tannerella forsythia, Aggregatibacter actinomycetemcomitans, Treponema denticola, and total bacterial load (TBL) were evaluated using ssrRNA probes., Results: Bacterial counts were decreased with all three treatment modalities and they did not reach baseline levels at 6 mo post-treatment. The SRP + LAS group showed statistically significantly lower TBL and bacterial levels of P. gingivalis and T. denticola at 6 mo post-treatment compared to SRP or LAS treatments alone. At the end of the observation period significant differences were observed for PPD and CAL between the SRP + LAS group and both the SRP alone and LAS alone groups. No differences were detected for percentage of plaque and percentage of BOP between any of the treatment groups at 6 mo post-treatment., Conclusions: Within the limits of this study, diode laser-assisted treatment with SRP showed a superior effect over SRP or LAS alone for certain microbial and clinical parameters in patients with aggressive periodontitis over the 6-mo monitoring period.

Aim: This study was designed to investigate the utility of two "new" definitions for assessment of bone loss in a rodent model of periodontitis., Material and Methods: Eighteen rats were divided into three groups. Group 1 was infected by Aggregatibacter actinomycetemcomitans (Aa), group 2 was infected with an Aa leukotoxin knock-out, and group 3 received no Aa (controls). Microbial sampling and antibody titres were determined. Initially, two examiners measured the distance from the cemento-enamel-junction to alveolar bone crest using the three following methods; (1) total area of bone loss by radiograph, (2) linear bone loss by radiograph, (3) a direct visual measurement (DVM) of horizontal bone loss. Two "new" definitions were adopted; (1) any site in infected animals showing bone loss >2 standard deviations above the mean seen at that site in control animals was recorded as bone loss, (2) any animal with two or more sites in any quadrant affected by bone loss was considered as diseased., Results: Using the "new" definitions both evaluators independently found that infected animals had significantly more disease than controls (DVM system; p<0.05)., Conclusions: The DVM method provides a simple, cost effective, and reproducible method for studying periodontal disease in rodents.

Periodontal disease is a multiple factor disease caused by genetic factors, environmental factors, and periodontal bacteria (periodontal pathogens). The present study aimed to elucidate the risk factors for periodontal disease in Japanese adolescents. Subjects (11-16 years old) were classified into three groups: localized aggressive periodontitis (LAP), periodontal attachment loss (PAL), and periodontally healthy (PH) groups. Genomic DNA isolated from the buccal mucosa was used for single-nucleotide polymorphism analyses of the candidate genes (interleukin-1alpha-889; interleukin-1alpha +4845; interleukin-1beta +3954; an immunoglobulin G Fc gamma receptor, FcgammaRIIa-R/H131; and a human leukocyte antigen class II allele, HLA-DQB1) of aggressive periodontitis. Subgingival plaque samples obtained from the same subjects were used for 16S rRNAbased polymerase chain reaction analysis of five important periodontal pathogens (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Treponema denticola, and Tannerella forsythia). Tannerella forsythia was detected in the deepest periodontal pockets in all subjects in the LAP and PAL groups. The prevalence of an atypical BamHI restriction site in HLA-DQB1 of the LAP group was significantly higher than that in the PH and PAL groups. Furthermore, all subjects who had the atypical BamHI restriction site in HLA-DQB1 had T. forsythia infection. These results suggested that T. forsythia is associated with periodontal disease in Japanese adolescents and also suggested that HLA-DQB1 is related to LAP and is associated with T. forsythia infection.

Background: Periodontitis develops in a time-dependent manner. Cross-sectional studies document one moment in time but fail to capture the progressive nature of disease. Radiographic measures of bone loss are relatively insensitive but are reliable markers of irreversible disease. Longitudinal studies are needed to identify biomarkers that can precede radiographic evidence of bone loss and, thus, mark the period prior to clinical evidence of irreversible disease. A longitudinal study of students susceptible to localized aggressive periodontitis (LAgP) was conducted to evaluate chemokines/cytokines found in saliva derived from periodontally healthy children who subsequently developed alveolar bone loss., Methods: Students were screened, sampled for Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans [Aa]), and divided into a cohort of Aa+ and Aa- students. Ninety-six periodontally healthy Aa+ and Aa- students were recalled every 6 to 9 months following screening. Examinations, saliva collections, and radiographs were performed. After seven students developed bone loss, the levels of 21 cytokines were assessed and matched to saliva from seven Aa+ and seven Aa- students who remained healthy for > or =1 year. Subsequently, saliva from an additional 27 students who remained healthy was analyzed., Results: Nineteen cytokines were not detected or were detected at low levels. Macrophage inflammatory protein (MIP)-1alpha was elevated 50-fold in seven Aa+ students who developed disease 6 to 9 months prior to radiographic detection of bone loss compared to levels in 21 Aa+ and 20 Aa- students who remained healthy (P <0.001). Interleukin (IL)-1beta was also elevated (P = 0.01). MIP-1alpha had a specificity of 96.8% and a sensitivity of 100%, whereas IL-1beta showed 90.3% specificity and 85.7% sensitivity relative to bone loss. MIP-1alpha levels were also related to increasing probing depth and the number of pockets >6 mm., Conclusion: The superior sensitivity and specificity of MIP-1alpha, which correlated well with probing depths and the onset of bone loss, suggested that it could be used as an early biomarker for LAgP.

Background: Herpesviruses may be related to the etiology of aggressive periodontitis (AgP) and chronic periodontitis (CP) by triggering periodontal destruction or by increasing the risk for bacterial infection. This case-control study evaluated the presence of herpes simplex virus type I (HSV-1), Epstein-Barr virus type I (EBV-1), human cytomegalovirus (HCMV), Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), Porphyromonas gingivalis, Prevotella intermedia, and Tannerella forsythia (previously T. forsythensis) in patients with generalized AgP (AgP group), CP (CP group), or gingivitis (G group) and in healthy individuals (C group)., Methods: Subgingival plaque samples were collected with paper points from 30 patients in each group. The nested polymerase chain reaction (PCR) method was used to detect HSV-1, EBV-1, and HCMV. Bacteria were identified by 16S rRNA-based PCR., Results: HSV-1, HCMV, and EBV-1 were detected in 86.7%, 46.7%, and 33.3% of the AgP group, respectively; in 40.0%, 50.0%, and 46.7% of the CP group, respectively; in 53.3%, 40.0%, and 20.0% of the G group, respectively; and in 20.0%, 56.7%, and 0.0% of the C group, respectively. A. actinomycetemcomitans was detected significantly more often in the AgP group compared to the other groups (P <0.005). P. gingivalis and T. forsythia were identified more frequently in AgP and CP groups, and AgP, CP, and G groups had higher frequencies of P. intermedia compared to the C group., Conclusion: In Brazilian patients, HSV-1 and EBV-1, rather than HCMV, were more frequently associated with CP and AgP.

The term periodontitis encompasses several polymicrobial infectious diseases, of multifactorial etiology, with chronic and aggressive forms. In spite of the etiopathogenic differences between these two forms of the disease, few studies have analyzed the subgingival colonization by yeast. The objective of this investigation was to analyze the composition of the yeast microbiota present in the mucosa and subgingival sites of healthy individuals and patients with aggressive and chronic periodontitis. For this, samples were recovered from these two locations and the yeast recovered identified by phenotypic and genotypic methods. Patients with chronic periodontitis showed significant differences in relation to the other groups with respect to carrier status (69.2% versus 35.7% of healthy individuals; [chi(i)(2) test; p=0.014]), the total number of isolated colony forming units or CFU (mean and ranges 281.6 (0-6048) [K-W(2)=6.998; p=0.03]), the Simpson diversity index (I) in site b (I(b)=0.344 versus healthy subjet and aggresive periodontitis where I=0 [multiple t-test comparisons with the Bonferronni correction, p<0.05]), and the species profile. Interestingly, in spite of the varied profiles of the species present in the mucosa of the three groups analyzed we noted that only C. albicans and C. dubliniensis were capable of colonizing the periodontal pockets in patients with chronic periodontitis, while only C. albicans was identified in the subgingiva of healthy individuals and patients with aggressive periodontitis.

Background: The purpose of this study was to test the hypothesis that periodontal pathogens associated with aggressive periodontitis persist in extracrevicular locations following scaling and root planing, systemic antibiotics, and antimicrobial rinses., Methods: Eighteen patients with aggressive periodontitis received a clinical examination during which samples of subgingival plaque and buccal epithelial cells were obtained. Treatment consisted of full-mouth root planing, systemic antibiotics, and chlorhexidine rinses. Clinical measurements and sampling were repeated at 3 and 6 months. Quantitative polymerase chain reaction determined the number of Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), Prevotella intermedia, Porphyromonas gingivalis, Tannerella forsythia (previously T. forsythensis), and Treponema denticola in the plaque. Fluorescence in situ hybridization and confocal microscopy determined the extent of intracellular invasion in epithelial cells., Results: Clinical measurements improved significantly following treatment. All bacterial species except P. gingivalis were significantly reduced in plaque between baseline and 3 months. However, all species showed a trend to repopulate between 3 and 6 months. This increase was statistically significant for log T. denticola counts. All species were detected intracellularly. The percentage of cells infected intracellularly was not affected by therapy., Conclusions: The 6-month increasing trend in the levels of plaque bacteria suggests that subgingival recolonization was occurring. Because the presence of these species within epithelial cells was not altered after treatment, it is plausible that recolonization may occur from the oral mucosa. Systemic antibiotics and topical chlorhexidine did not reduce the percentage of invaded epithelial cells. These data support the hypothesis that extracrevicular reservoirs of bacteria exist, which might contribute to recurrent or refractory disease in some patients.

Objective: To make qualitative and quantitative analysis of Archaea in subgingival plaque sample and to investigate the relationship between periodontal disease and Archaea., Methods: Subgingival plaque was collected from 23 patients with aggressive periodontitis, 29 with chronic periodontitis, 35 with plaque-induced gingivitis and 38 healthy controls. Qualitative and quantitative analysis of methanogenic archaea was performed by amplification of the 16S rRNA genes in the DNA extracted from the plaque samples., Results: Archaea were found in 65% of aggressive periodontitis patients, 72% of chronic periodontitis, 26% of gingivitis and zero of healthy subjects. Quantitative analysis showed the average abundance of archaeal 16S rRNA gene in Archaea-positive patients was different among the three groups. The average 16S rRNA gene copy number from per microg wet plaque was 6.66 x 10(6) in aggressive periodontitis sufferers, 4.47 x 10(6) in chronic periodontitis and 1.78 x 10(6) in gingivitis groups. The prevalence of Archaea and the average Archaea 16S rRNA gene numbers in periodontitis groups were higher than those in gingivitis group (P < 0.05)., Conclusions: This suggests that Archaea may be implicated as causative agents for periodontitis.

Objective: To examine Porphyromonas gingivalis (Pg) in subgingival plaque of the patients with periodontitis and to find out the rules of Pg colonization after periodontal initial treatment., Methods: A total of 1620 subgingival plaque samples were collected from 180 subjects including chronic periodontitis (CP) patients (n = 90), and aggressive periodontitis (AgP) patients (n = 90) in different periods of periodontal initial therapy-the baseline, 6 weeks, and 12 weeks after treatment. The following periodontal clinical parameters were recorded with Florida probe at sampled sites: probing depth (PD), clinical attachment loss (CAL), and bleeding on probing (BOP). Quantities of Pg were examined by AmpliFluor endpoint quantitative polymerase chain reaction., Results: At the 6th week of periodontal initial therapy, there were 61 (22.6%) and 66 (24.4%) Pg increased sites respectively, in which no significant difference was detected (P > 0.05). At baseline of periodontal initial therapy, more severe periodontal clinical parameters of Pg increased sites were observed than those of Pg stationary sites. At the 12th week, however, there were 96 (35.6%) and 18 (6.7%) Pg increased sites respectively, significant difference detected (P < 0.05). At 6th week of periodontal initial therapy, more severe periodontal clinical parameters of Pg increased sites were observed than those of Pg stationary sites., Conclusions: Pg colonization in AgP and CP patients started 6 weeks after periodontal initial therapy, but the recolonization pattern was different between these two groups of patients. Severe periodontitis sites in baseline seemed to place them at risk of Pg colonization after periodontal initial therapy.

Objective: To evaluate the prevalence rates of human cytomegalovirus(HCMV) and Epstein-Barr virus-1(EBV-1) in subgingival plaque and analyze the relationship between herpesviruses, periodontal pathogenic bacteria and periodontal clinical parameters in Chinese patients with aggressive periodontitis(AgP)., Methods: A total of one hundred and twenty subgingival plaque samples were collected from 89 AgP patients and 31 healthy subjects. HCMV and EBV-1 were detected using nested polymerase chain reaction(PCR).Contemporaneously, 8 periodontal pathogenic bacteria including Actinobacillus actinomycetemcomitans(Aa), Porphyromonas gingivalis(Pg), Tannerella forsythensis(Tf), Prevotella intermedia(Pi), Campylobacter rectus(Cr),Fusobacterium nucleatum(Fn), Treponema denticola(Td), Prevotella nigrescens(Pn) were detected by 16S rRNA based PCR., Results: HCMV was more frequently detected in AgP patients (43.8%) than in healthy controls (12.9%, P<0.01). The prevalence rates of HCMV and EBV-1 in AgP patients with 6-8 kinds of bacteria detected were 54.4% and 17.4%, respectively, significantly higher than those with 3-5 kinds of bacteria detected (P<0.05)., Conclusion: The prevalence rates of HCMV and EBV was higher in AgP patients than in healthy controls. Herpesviruses and periodontal pathogenic bacteria may cooperate synergistically in the development of AgP, which could be considered as a pathogenetic consortium in future investigation of periodontaltitis.

We developed a loop-mediated isothermal amplification method that detects the JP2 clone of Aggregatibacter actinomycetemcomitans, which induces aggressive periodontitis in adolescents of North and West African descents. Being independent of special equipment, this specific and sensitive method offers significant advantages for screening of patients on a population basis and in clinical settings.

Background: Periodontitis is a loss of supporting connective tissue and alveolar bone around teeth, and if it occurs in an aggressive form it can lead to tooth loss before the age of 20 years. Although the cause of periodontitis in general remains elusive, a particular clone (JP2) of the gram-negative rod Aggregatibacter (Actinobacillus) actinomycetemcomitans is considered a possible aetiological agent of the aggressive form in adolescents living in or originating from north and west Africa, where the disease is highly prevalent. We did a population-based longitudinal study of adolescents to assess the role of the JP2 clone in the initiation of aggressive periodontitis., Methods: A total of 700 adolescents from public schools in Rabat, Morocco, were enrolled in the study. We used PCR to detect A actinomycetemcomitans in plaque samples (taken from molar and incisor sites) and to differentiate between the JP2 clone and other non-JP2 genotypes of the bacterium. 18 individuals were found to already have periodontitis and were excluded. The 682 periodontally healthy adolescents (mean age 12.5 years; SD 1.0) were classified according to their A actinomycetemcomitans carrier status at baseline. After 2 years, 428 (62.8%) individuals returned for re-examination, which included recording of periodontal attachment loss measured from the cemento-enamel junction to the bottom of the periodontal pockets of all teeth present., Findings: Individuals who carried the JP2 clone of A actinomycetemcomitans alone (relative risk 18.0; 95% CI 7.8-41.2, p<0.0001) or together with non-JP2 clones of A actinomycetemcomitans (12.4; 5.2-29.9, p<0.0001) had a significantly increased risk of periodontal attachment loss. A much less pronounced disease risk was found in those carrying non-JP2 clones only (3.0; 1.3-7.1, p=0.012)., Interpretation: The JP2 clone of A actinomycetemcomitans is likely to be an important aetiological agent in initiation of periodontal attachment loss in children and adolescents. Co-occurrence of non-JP2 clones of A actinomycetemcomitans reduces the risk of development of periodontitis, suggesting competition for the ecological niche between the JP2 and non-JP2 clones of this species.

Leukocyte adhesion deficiency type 1 (LAD-1) is a rare, inherited immunodeficiency that affects 1 in 1 million people yearly and usually presents with recurrent, indolent bacterial infections of the skin, mouth, and respiratory tract and impaired pus formation and wound healing. Features of this disease result from mutations in the region of the CD18 gene, which is encoded on chromosome 21q22.3. This gene codes for the common subunit of the leukocyte integrins LFA-1, Mac 1, and p150,95. Failure to produce a functional subunit results in the defective expression of all 3 leukocyte integrins, and the leukocytes of LAD have subnormal adhesion properties. We present a case of the moderate-to-severe form of LAD in a 3-year-old girl who initially presented with generalized swelling and erythema of the gingiva, with slight tooth mobility and a nonhealing labial ulceration. Her medical history was significant for recurrent urinary tract infections. Periodontal pathogens, including Capnocytophaga, Eikenella corrodens, and Candida albicans, were cultured. The patient had a significantly elevated white blood cell count and absolute neutrophil count. The diagnosis of LAD was confirmed with flow cytometry, which revealed significantly decreased subunits. Twenty-four months after the diagnosis was made and after a series of granulocyte transfusions and bone marrow transplantations, she expired as the result of respiratory failure.