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Background: Previous research indicated that patients with localized aggressive periodontitis (LAgP) had minimal proximal decay. We speculated that differences in these two proximal dental diseases (LAgP and proximal decay) in LAgP could be due to the effect of saliva on the growth of key microorganisms related to these two infections. Carbon dioxide (CO(2)) is required for growth of Actinobacillus actinomycetemcomitans (Aa), the reputed cause of LAgP. Bicarbonate, a source of CO(2), buffers acid production by Streptococcus mutans (Sm), a key organism associated with caries. The purpose of this study was to determine whether the saliva of LAgP patients and subjects with Aa had higher levels of bicarbonate, or an elevated pH, and/or reduced survival of Sm., Methods: Eleven Aa-positive subjects (seven with LAgP) were matched with 11 Aa-negative controls. A total of 5 ml saliva obtained from each subject was tested for CO(2) levels, pH, and effects on survival of Aa and Sm. Saliva from 22 additional subjects was used for confirmatory data., Results: CO(2) levels in the test group (Aa-positive subjects) and controls (Aa-negatives) were similar. No clinically relevant differences were found in salivary pH. However, saliva from the test group killed Sm by more than two logs (P <0.05). No effect was seen on Aa. The saliva from the Aa-negative group killed Aa by two logs (P <0.05). No effect was seen on Sm., Conclusion: Aa-positive subjects had a salivary factor that significantly reduced survival of Sm, which may help to explain the fact that this group typically has minimal proximal decay.

Purpose: Periodontal disease is a commonly prevalent problem faced alike by both the developed and third world countries but showing wide variations in prevalence and severity across different geographical areas. The purpose was to identify Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Ekinella corrodens (Ec), Campylobacter rectus (Cr), Bacteroides forsythus (Bf), Treponema denticola (Td) and Fusobacterium nucleatum (Fn) in Indian adult periodontitis and rapidly progressive periodontitis patients., Materials and Methods: Paper points were used to collect the sample from 28 sites in both adult periodontitis and rapidly progressive periodontitis (8 healthy/20 diseased sites) patients and DNA analysis done. The categorical data was analysed by Fishers exact test and difference in the clinical parameters was tested by Mann-Whitney test., Results: In healthy sites of adult and rapidly progressive periodontitis patients, Aa, Ec, Bf and Aa, Pg, Pi, Td, Fn were detected respectively. However, when diseased and healthy sites were compared in both adult periodontitis and rapidly progressive periodontitis patients respectively, only Pg( P =0.004), Cr( P =0.04), Fn( P =0.014) and Pg( P =0.002), Cr( P =0.02), Fn( P =0.008) were statistically significant., Conclusion: The prevalence of the microorganisms correlate with the clinical parameters like probing depth and bleeding on probing as seen in the Japanese and Western periodontitis patients' population.

The highly leukotoxic JP2 clone of Actinobacillus actinomycetemcomitans is strongly associated with periodontitis in adolescents. Availability of the DNA sequence of the complete genome of A. actinomycetemcomitans strain HK1651, a representative strain of the JP2 clone (http://www.genome.ou.edu/act.html), has provided new possibilities in basic research regarding the understanding of the pathogenesis of A. actinomycetemcomitans in periodontitis. This case report describes the periodontal treatment of the original source of A. actinomycetemcomitans HK1651, a 16-year-old Ghanaian adolescent girl with aggressive periodontitis. The bacterial examination involved polymerase chain reaction analysis for presence of JP2 and non-JP2 types of A. actinomycetemcomitans. The treatment, including periodontal surgery supplemented by antibiotics, arrested the progression of periodontitis for more than 10 years. Initially, infection by A. actinomycetemcomitans, including the JP2 clone, was detected at various locations in the oral cavity and was not limited to the periodontal pockets. Post-therapy, the JP2 clone of A. actinomycetemcomitans disappeared, while the non-JP2 types of A. actinomycetemcomitans remained a part of the oral microflora.

Objective: To evaluate the prevalence of putative periodontal microorganism in Chinese patients with aggressive periodontitis (AgP)., Methods: A total of 72 subgingival plaque samples were collected from 55 AgP patients and 17 healthy subjects. Seven putative periodontal microorganisms including Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Tannerella forsythensis (Tf), Treponema denticola (Td), Campylobacter rectus (Cr), Prevotella intermedia (Pi) and Prevotella nigrescens (Pn) were detected by using 16S rRNA based polymerase chain reaction (PCR)., Results: The prevalence of Aa in AgP patients was very low (1.8%), while Pg, Tf, Td and Cr were more frequently detected in AgP patients (prevalence: 81.8%, 83.6%, 80% and 81.8%) than in healthy controls (prevalence: 17.6%, 11.8%, 5.9%, 29.4%, P < 0.01)., Conclusions: The prevalence of Pg, Tf, Td and Cr were very high in aggressive periodontitis patients, and the combination of these bacteria may play an important role in AgP.

Background: Patients with Kostmann syndrome (severe congenital neutropenia [SCN]) typically normalize their absolute neutrophil count (ANC) upon granulocyte colony-stimulating factor (G-CSF) therapy. However, although they no longer experience life-threatening bacterial infections, they frequently still have recurrent gingivitis and even severe periodontitis, often starting in early childhood., Methods: We studied the periodontal disease in the four surviving patients belonging to the family originally described by Kostmann. Their odontological records, x-rays, color photos, bacterial cultures, serum antibodies to oral bacteria, and histopathological examinations were reviewed. The data were also correlated to previous investigations on their antibacterial peptides and molecular biology., Results: Three patients had periodontal disease, despite normal ANC and professional dental care, and had neutrophils deficient in antibacterial peptides. One of these patients also had a heterozygous mutation in the neutrophil elastase gene, had severe periodontal disease and overgrowth of the periodontal pathogen Actinobacillus actinomycetemcomitans in the dental flora, and 15 permanent teeth had been extracted by the age of 27. One bone marrow-transplanted patient had no periodontal disease., Conclusions: Normalized ANC levels are not sufficient to maintain normal oral health in SCN patients, and because neutrophils are important for first-line defense and innate immunity, the deficiency of the antibacterial peptide LL-37 probably explains their chronic periodontal disease. Professional dental care is still important for SCN patients, despite treatment with G-CSF and normal ANC levels. Whether antibacterial peptides play a role in the pathogenesis of periodontitis in other patients remains to be elucidated.

The highly leukotoxic JP2 clone of Actinobacillus actinomycetemcomitans is strongly associated with aggressive periodontitis in adolescents of African descent. DNA fingerprinting using the frequently cutting restriction enzyme MspI and multilocus sequence typing (MLST) showed that five strains of this clone were genetically virtually identical, although ribotyping of the six rrn genes and EcoRI RFLP analysis of the seven IS150-like elements revealed differences. PCR analyses demonstrated that these multi-copy sequences are subject to intragenomic homologous recombination, resulting in translocations or large inversions. The genome rearrangements were reflected in differences among 25 strains representing the JP2 clone in DNA fingerprinting using the rare-cutting restriction enzyme XhoI and resolved by PFGE. XhoI DNA fingerprinting provides a tool for studying local epidemiology, including transmission of this particularly pathogenic clone of A. actinomycetemcomitans.

The diagnosis of localized aggressive periodontitis includes first molar attachment loss as an obligatory criterion. This tooth-specific based diagnosis has never been questioned or tested previously. We present a rare case of aggressive periodontitis that developed during orthodontic treatment, which included extraction of right lower first molar and bodily movement of the second molar to the original first molar site. At the end of the orthodontic therapy, localized periodontal disease was diagnosed at the site of the lower left first molar and the second lower right molar that was now occupying the site of the former first lower molar. The patient's periodontal condition was stabilized, and the bony defects were filled following periodontal treatment. This report shows that bacterial induced aggressive periodontitis developed during orthodontic treatment in a site-specific manner and suggest the hypothesis that localized aggressive periodontitis was targeted to a specific alveolar site rather than a tooth-specific site.

Background: Report of a combined periodontal and orthodontic treatment in a patient with Papillon-Lefevre Syndrome (PLS)., Methods: A patient with PLS was treated orthodontically 26 months after the start of a combined mechanical and antibiotic therapy. Clinical periodontal parameters were obtained 26 (t1), 60 (t2), and 79 (t3) months after anti-infective therapy. The deepest site of each tooth was sampled for microbiological analysis at 26 and 60 months. Periodontal maintenance therapy was provided every 6 weeks. After a stable periodontal situation was achieved, orthodontic treatment, consisting of space opening for the upper canines with a multibracket appliance and coil springs, was carried out. In the lower jaw, crowding was resolved by an orthodontic mesialization of the canines., Results: Twenty-six months (t1) after the beginning of the combined mechanical and antibiotic therapy, 6% of the sites exhibited 4 mm probing depth (PD) with bleeding on probing (BOP) or PD > or =5 mm. Sixty months (t2) after therapy the number of sites with 4 mm PD with BOP or PD > or =5 mm had increased to 17%, and 79 months after therapy (t3) 13% of all sites were similarly affected. From 26 to 60 months, a slight mean clinical attachment level (CAL) gain was observed, whereas the mean PD increased. From 60 to 79 months, there was a mean PD reduction. However, a significant mean attachment loss was also noted. After 26 months (t1), RNA probes failed to detect A. actinomycetemcomitans, P. gingivalis, or T. forsythensis from any site. Thirty-four months later (t2), subgingival recolonization was observed. A. actinomycetemcomitans was detected by RNA probes at three sites. At 26 and 60 months (t1, t2), trypticase-soy with serum, bacitracin, and vancomycin (TSBV) culture failed to detect A. actinomycetemcomitans at any of the sampled sites. Eighty-two months after the beginning of therapy (t4), none of the applied methods could detect A. actinomycetemcomitans from the pooled samples from the deepest pockets of each quadrant or the oral mucosa. In the present case, concomitant orthodontic treatment with a fixed appliance could be performed without further pronounced periodontal deterioration. Space for eruption of the canines and premolars was created, in addition to an alignment of the teeth., Conclusion: After a successful combined mechanical and antibiotic periodontal therapy of the PLS periodontitis, moderate orthodontic tooth movements may be possible within a complex interdisciplinary treatment regimen.

The JP2 clone of Actinobacillus actinomycetemcomitans has been implicated in the etiology of periodontitis in adolescents. The aim of this two-year longitudinal study was to describe clinical attachment loss (CAL) progression and to assess its association with baseline occurrence of the JP2 and non-JP2 types of A. actinomycetemcomitans. Clinical re-examination of 121 adolescents in Morocco was performed. Progression of CAL > or = 1 mm, > or = 2 mm, > or = 3 mm, and > or = 4 mm on at least one site was found in 58%, 48%, 22%, and 6% of the subjects, respectively. Subjects who, at baseline, harbored the JP2 clone had a significantly higher progression of CAL than did subjects harboring non-JP2 types of A. actinomycetemcomitans. Subjects harboring non-JP2 types displayed a marginally higher CAL progression than did subjects who were culture-negative for A. actinomycetemcomitans.

Objectives: The objectives of this study were to characterize the bacterial profile and to seek possible bacterial associations in the subgingival microbiota of early onset periodontitis/aggressive periodontitis patients by using two different techniques, culture and immunofluorescence., Material and Methods: The study group consisted of 66 systemically healthy individuals with evidence of early onset periodontitis - 41 females and 25 males aged 23-35 years (mean 31.1 +/- 3.1 years). Bacterial samples were collected from the deepest site in each quadrant, resulting in a total of 264 sites with a mean probing pocket depth of 6.6 +/- 1.5 mm. Samples were cultured anaerobically and in 10% CO(2) using selective and nonselective media, and isolates were characterized to species level. Indirect immunofluorescence using monoclonal antibodies was applied to detect Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia (Bacteroides forsythus, Tannerella forsythensis), Prevotella intermedia/Prevotella nigrescens, Campylobacter rectus, Peptostreptococcus micros and Actinomyces israelii., Results: 93.6% of sampled sites showed bleeding on probing and 23.5% were positive for suppuration. P. intermedia/P. nigrescens, P. gingivalis, and C. rectus were detected in 77.3-85.9% of samples using culture methods and in 85.6-91.3% using immunofluorescence. P. micros and A. actinomycetemcomitans were found, respectively, in 63.3% and 25.0% of all sites using culturing and in 58.7% and 27.7% sites using immunofluorescence. Significantly strong positive associations were observed between T. forsythia and C. rectus (odds ratio 109.46), and T. forsythia and P. gingivalis (odd ratio 90.26), whereas a negative association was seen between P. intermedia/P. nigrescens and A. actinomycetemcomitans (odds ratio 0.42). Coinfection by P. gingivalis, T. forsythia, P. intermedia/P. nigrescens and C. rectus was observed in 62.1% of the test sites, and in 89.4% of the studied subjects. The sensitivity of immunofluorescence for T. forsythia, C. rectus, P. intermedia/P. nigrescens and P. gingivalis was found to be very high (0.99-0.94) using culture as the reference detection method. The agreement between culture and immunofluorescence in detecting the presence or absence of the investigated species was 85.2-88.1% for P. gingivalis, P. intermedia/P. nigrescens, C. rectus, and T. forsythia, 75.9% for A. actinomycetemcomitans and 70.4% for P. micros., Conclusions: The microbial profile of the early onset/aggressive periodontitis population was complex. The agreement between the two detection methods was very high.

The purpose of this case report is to detail an initial periodontal manifestation of Crohn's disease in a 6-year-old boy. The first clinical diagnosis, localized aggressive periodontitis, was based on the microbiological isolation of Actinobacillus actinomycetemcomitans from subgingival sites. On examination, gingival lesions, together with bleeding on probing, edema, and erythema, were observed. Although an increased probing depth was detected, no radiographically visible alveolar bone loss was observed. According to these findings, periodontitis as a manifestation of a systemic disease was assumed. Furthermore, fissural ulcerations of the lips were noted. The patient also reported a swelling of the upper lip in the morning. Oral hygiene procedures, scaling, root planning, and the application of metronidazole and amoxicillin were not successful. Metabolic and several immunological tests, however, showed normal values. Two months after the first periodontal signs, the child suffered from severe malnutrition, accompanied by diarrhaea and abdominal pain. Active colitis with multiple granulomas was detected histopathologically from biopsies. Crohn's disease was then diagnosed by the internist. If in doubt, medical examinations in every case of childhood periodontitis are recommended to determine whether the findings speak for initial symptoms of a systemic disorder (eg, Crohn's disease).

The purpose of this report was to describe an approach to diagnose and effectively treat a pediatric patient with localized aggressive periodontitis. A 6-year-old female presented with clinical and radiographic evidence of severe attachment loss around several primary teeth. She had no history of systemic disease, periodontal disease, or caries prior to the periodontal abscess that prompted her referral. Routine immunological tests did not reveal any functional defects, but DNA testing for periodontal pathogens revealed the presence of all 8 aggressive periodontal pathogens assayed. Treatment consisted of the extraction of 2 severely affected primary teeth, increased frequency of recall appointments, and administration of systemic antibiotics. The patient's periodontal condition was stabilized 18 months post-treatment, and the 8 pathogens were no longer at detectable levels. With a treatment goal of preventing disease progression into the erupting permanent dentition, this treatment regimen provides an effective alternative to more aggressive strategies.

This pilot study examined gene expression signatures in pathological gingival tissues of subjects with chronic or aggressive periodontitis, and explored whether new subclasses of periodontitis can be identified based on gene expression profiles. A total of 14 patients, seven with chronic and seven with aggressive periodontitis, were examined with respect to clinical periodontal status, composition of subgingival bacterial plaque assessed by checkerboard hybridizations, and levels of serum IgG antibodies to periodontal bacteria assayed by checkerboard immunoblotting. In addition, at least two pathological pockets/patient were biopsied, processed for RNA extraction, amplification and labeling, and used to study gene expression using Affymetrix U-133 A arrays. Based on a total of 35 microarrays, no significantly different gene expression profiles appeared to emerge between chronic and aggressive periodontitis. However, a de novo grouping of the 14 subjects into two fairly robust clusters was possible based on similarities in gene expression. These two groups had similar clinical periodontal status and subgingival bacterial profiles, but differed significantly with respect to serum IgG levels against the important periodontal pathogens Porphyromonas gingivalis, Tannerella forsythensis and Campylobacter rectus. These early data point to the usefulness of gene expression profiling techniques in the identification of subclasses of periodontitis with common pathobiology.

Background: To our knowledge, the association of the five serotypes of Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) to the new diagnostic classification scheme defined by the American Academy of Periodontology in 1999 has not yet been described. The goal of this study was to characterize the frequencies of the five serotypes of A. actinomycetemcomitans in A. actinomycetemcomitans isolates from various forms of periodontitis using both old and new diagnostic classifications and to determine the relationships between serotype and age and clinical diagnosis., Methods: A total of 345 A. actinomycetemcomitans isolates from 115 A. actinomycetemcomitans culture-positive subjects (mean age 38.0 +/- 18.3 years, 59% female) were collected. Based on the new classifications, 33 subjects had aggressive periodontitis and 82 chronic periodontitis. According to old classifications, there were six prepubertal periodontitis (PPP), 12 localized juvenile periodontitis (LJP), 15 post-localized juvenile periodontitis (PLJP), 28 refractory periodontitis (Ref-P), and 54 adult periodontitis (AP) cases. Serotypes of A. actinomycetemcomitans were determined by an indirect immunofluorescence assay using serotype-specific polyclonal antisera to A. actinomycetemcomitans strains ATCC 29523, ATCC 43728, ATCC 33384, IDH 781 and IDH 1705 (serotype a, b, c, d, and e, respectively). Proportions of serotype b were examined between different diagnostic and age groups with a Z-test for proportions., Results: Most subjects (n = 100, 86.96%) were infected with a single serotype (22 serotype a, 44 serotype b, 30 serotype c, 1 serotype d, and 3 serotype e). There were 11 subjects (9.57%) with two serotypes and two subjects (1.74%) with 3 serotypes. Two individuals had isolates lacking any detectable serotype antigen. Serotype b was the predominant serotype in children under 18 years of age and young adults between 19 to 35 years, although serotype b status was not significantly associated with age. Serotypes d and e were not found in patients under 35 years old. In 62 adult patients, one subject had serotype d and three had serotype e. Serotype b was the most common serotype in aggressive periodontitis (60.61%). The proportion of cases with serotype b was significantly higher in aggressive periodontitis compared to chronic periodontitis (P = 0.031). Other serotypes were not significantly associated with new diagnostic categories. Serotypes d and e were not detected in aggressive periodontitis., Conclusion: The results of this study show that proportions of serotype b of A. actinomycetemcomitans are significantly greater in culture-positive patients with aggressive periodontitis than those with chronic periodontitis.

The iron-binding protein lactoferrin is a ubiquitous and abundant constituent of human exocrine secretions. Lactoferrin inhibits bacterial growth by sequestering essential iron and also exhibits non-iron-dependent antibacterial, antifungal, antiviral, antitumor, anti-inflammatory, and immunoregulatory activities. All of these non-iron-dependent activities are mediated by the highly charged N terminus of lactoferrin. In this study we characterized a Lys/Arg polymorphism at position 29 in the N-terminal region of human lactoferrin that results from a single nucleotide polymorphism in exon 1 of the human lactoferrin gene. We expressed cDNAs encoding both lactoferrin variants in insect cells and purified the two proteins by ion exchange chromatography. The two lactoferrin variants exhibited nearly identical iron-binding and iron-releasing activities and equivalent bactericidal activities against a strain of the gram-negative bacterium Actinobacillus actinomycetemcomitans. When tested against the gram-positive species Streptococcus mutans and Streptococcus mitis, however, lactoferrin containing Lys at position 29 exhibited significantly greater bactericidal activity than did lactoferrin containing Arg. In addition, the Lys-containing lactoferrin stimulated bovine tracheal epithelial cells to synthesize much higher levels of tracheal antimicrobial peptide mRNA than did the Arg-containing variant. A genotyping assay that distinguished between the two alleles based on a polymorphic EarI restriction site showed that the Lys and Arg alleles had frequencies of 24% and 76%, respectively, among 17 healthy human subjects, and 72% and 28%, respectively, among nine patients with localized juvenile periodontitis. Our findings suggest that these two lactoferrin variants are functionally different and that these differences may contribute to the pathogenesis of localized juvenile periodontitis.

The JP2 clone of Actinobacillus actinomycetemcomitans is associated with early-onset periodontitis in certain ethnic populations of African origin. Here, we describe and evaluate a set of primers for PCR to assay for the presence of A. actinomycetemcomitans and to discriminate between JP2-like strains and other genotypes in subgingival plaque samples.

The term periodontal disease refers to all diseases that involve the supportive structures of the periodontium. Peridontal diseases commonly begin as a gingivitis and progress to periodontitis. Necrotizing ulcerative gingivitis (NUG) is the most fulminate form of gingivitis. The two main forms of periodontitis are chronic periodontitis (also known as adult periodontitis) and aggressive periodontitis (also known as early onset periodontitis, destructive periodontitis, and juvenile periodontitis). Gingivitis treatment involves removing dental plaques and maintaining good oral hygiene. Periodontitis therapy should include root debriding, draining the infected root, and surgically resecting inflamed periodontal tissues. Systemic antimicrobials often are indicated in NUG, chronic periodontitis, and aggressive periodontitis. When possible, antimicrobial selection should be based upon culture and susceptibility testing of the subgingival flora.

Background: Microbial associations in aggressive periodontitis versus different ethnic origins are substantially unknown. We undertook this study to determine the prevalence of seven putative periodontopathogens in Korean patients and to evaluate microbial differences in localized and generalized aggressive periodontitis patients., Methods: Thirty-nine aggressive periodontitis patients between 20 and 35 years old (24 males and 15 females; mean age 29.6 years) were selected according to clinical criteria. The patients were subclassified into 17 localized and 22 generalized aggressive periodontitis patients. In each of the 39 individuals, subgingival plaque samples were collected from four diseased teeth (> or = 6 mm probing depth, 156 sites) and one healthy site (< or = 3 mm probing depth, 39 sites). Polymerase chain reaction (PCR) of the 16S ribosomal RNA gene fragments (about 530 bp) of plaque bacteria and their subsequent detection by dot-blot hybridization using specific oligonucleotide probes were performed to determine the presence of seven periodontopathogens., Results: The prevalences were 75% for Actinobacillus actinomycetemcomitans, 94.2% for Tannerella forsythensis (formerly Bacteroides forsythus), 99.4% for Fusobacterium sp., 85.9% for Micromonas micros (formerly Peptostreptococcus micros), 96.8% for Porphyromonas gingivalis, 78.8% for Prevotella intermedia, and 96.8% for Treponema sp. The prevalences of these bacteria were significantly higher in diseased sites than in healthy sites. Logistic regression analysis showed that P. intermedia was more significantly associated with generalized aggressive periodontitis than the localized form, with an odds ratio of 3.28 (95% confidence interval 1.26-8.56, P = 0.015)., Conclusions: Our results demonstrate that the seven periodontal pathogens analyzed are strongly associated with Korean aggressive periodontitis. In particular, P. intermedia are more significantly associated with generalized aggressive periodontitis, a more severe and progressive form, than with localized aggressive periodontitis.

Objectives: Denaturing gradient gel electrophoresis (DGGE) was applied to the microbiologic examination of subgingival plaque., Materials and Methods: The PCR primers were designed from conserved nucleotide sequences on 16S ribosomal RNA gene (16SrDNA) with GC rich clamp at the 5'-end. Polymerase chain reaction (PCR) was performed using the primers and genomic DNAs of typical periodontal bacteria. The generated 16SrDNA fragments were separated by denaturing gel., Results: Although the sizes of the amplified DNA fragments were almost the same among the species, 16SrDNAs of the periodontal bacteria were distinguished according to their specific sequences. The microflora of clinical plaque samples were profiled by the PCR-DGGE method, and the dominant 16SrDNA bands were cloned and sequenced. Simultaneously, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia were detected by an ordinary PCR method. In the deep periodontal pockets, the bacterial community structures were complicated and P. gingivalis was the most dominant species, whereas the DGGE profiles were simple and Streptococcus or Neisseria species were dominant in the shallow pockets. The species-specific PCR method revealed the presence of A. actinomycetemcomitans, P. gingivalis and P. intermedia in the clinical samples. However, corresponding bands were not always observed in the DGGE profiles, indicating a lower sensitivity of the DGGE method., Conclusion: Although the DGGE method may have a lower sensitivity than the ordinary PCR methods, it could visualize the bacterial qualitative compositions and reveal the major species of the plaque. The DGGE analysis and following sequencing may have the potential to be a promising bacterial examination procedure in periodontal diseases.

Objectives: The purpose of this study was to evaluate the clinical and microbiological status of patients with early-onset or aggressive periodontitis (EOP) who had received supportive periodontal care (SPC) every 3-6 months for a period of 5 years, following active periodontal treatment., Material and Methods: The study population consisted of 25 individuals with early-onset periodontitis. Clinical examination and recordings of probing pocket depth (PPD) and clinical attachment level (CAL) were performed at baseline prior to treatment (T0), 3 months following the termination of active periodontal treatment (T1) and annually at the SPC appointments (T2,T3,T4,T5). Microbiological samples were obtained at the 5-year SPC (T5). Subgingival plaque samples for each individual were collected from one deep pocket (>5 mm), based on pretreatment measurements, randomly selected in each quadrant. The levels of Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Porphyromonas gingivalis and Treponema denticola were determined using oligonucleotide probe hybridization., Results: During the 5-year period, the mean of SPC/patient was 12.7 sessions. A significant improvement was observed in PPD, CAL, gingival bleeding index and suppuration following treatment. However, between T1 and T5, 134 sites in 20 patients deteriorated with a CAL loss of> or =2 mm. Out of these 134 sites showing disease progression, microbial samples were randomly obtained in 13 sites (9.7%) from 8 patients. Among other factors, smoking and stress were found to have significant predictive value on the future attachment loss. P. gingivalis, T. denticola and total bacterial load were statistically significantly higher in patients who experienced disease progression during the 5-year maintenance period., Conclusions: For most EOP patients, regular SPC was effective in maintaining clinical and microbiological improvements attained after active periodontal therapy. However, a small percentage of sites was identified as progressive in 20 patients. Variables found to be related to periodontal progression were the presence of as well as the high bacterial counts of P. gingivalis, T. denticola and total bacterial load, number of acute episodes, number of teeth lost, smoking and stress.

Objective: The aim of this study was to investigate effects of immunodeficiency on the periodontal infection characters of the specific pathogens of juvenile periodontitis., Methods: A total of 36 immunodeficient guinea pigs produced by twice whole-body irradiation with 60Co were divided randomly into four groups, in which Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and A. actinomycetemcomitans with P. gingivalis were inoculated into the gingival sulcus of two mandibular incisors respectively. The pigs in the control group did not receive any inoculation. At 2, 3 and 6 weeks after inoculation, three animals in each group were sacrificed successively. Clinical and histological examinations were used to examine the changes in the periodontal tissues. The other 36 normal guinea pigs were divided into four groups and treated in a similar way described above., Results: Significant periodontal damages were noted in immunodeficient pigs inoculated with A. actinomycetemcomitans, P. gingivalis or A. actinomycetemcomitans and P. gingivalis in 2 and 3 weeks after bacterial inoculation. The damages were more severe than in the normal groups. The immunodeficient groups demonstrated larger numbers of osteoclasts than the normal groups (P < 0.05)., Conclusion: The loss of periodontal tissue in immunodeficient hosts is much serious than those with normal defence system, after they are infected with A. actinomycetemcomitans and P. gingivalis. Abnormal defence system in hosts may play an important role in onset and development of juvenile periodontitis.

Cells of the periodontal pathogen Actinobacillus actinomycetemcomitans exhibit tight adherence to surfaces such as glass, plastic and hydroxyapatite, a property that probably plays an important role in the ability of this bacterium to colonize teeth and other surfaces. Tight adherence is mediated by long fibrils of bundled pili (fimbriae) that form on the surface of the cell. The flp-1 gene encodes the major pilin protein component of A. actinomycetemcomitans fimbriae. In this study we compared flp-1 DNA sequences from 43 strains of A. actinomycetemcomitans isolated in Europe, Japan and the United States and identified seven distinct flp-1 allelic classes. DNA and predicted protein sequences were almost completely conserved within each flp-1 class but were highly divergent between classes. Most amino acid substitutions occurred in the C-terminus of the pilin protein, a region that has been shown to be important for the bundling and adhesive properties of the pili. flp-1 classes correlated with serotypes and 16S rRNA genotypes in most strains. At least five strains showed evidence of horizontal transfer of flp-1 between strains of different serotypes and 16S rRNA genotypes. Four of the seven flp-1 classes were present in geographically diverse isolates. Strains representing all seven flp-1 classes, but not a strain carrying a transposon insertion in flp-1, bound avidly to polystyrene in an in vitro adherence assay. Strains representing six of the seven flp-1 classes were isolated from localized juvenile periodontitis patients, suggesting that phylogenetically diverse strains carry pathogenic potential. Our findings provide a framework for future biochemical, immunological and genetic studies of A. actinomycetemcomitans fimbriae.

The local salivary immunoglobulin A (IgA) response in patients with aggressive periodontitis to oral microorganisms and its role for the pathogenesis has not been determined. This study investigated the hypothesis that aggressive periodontitis patients have impaired oral secretory immunity. Our test group was made-up of 19 aggressive periodontitis patients and 19 age- and gender-matched periodontally healthy controls. Total IgA, IgA subclass 1, IgA subclass 2 and IgA reactive to Actinobacillus actinomycetemcomitans Y4, Treponema denticola ATCC 35404 and Candida albicans DSM 3454 were determined by enzyme-linked immunosorbent assay in whole unstimulated and stimulated saliva. A statistically significantly lower concentration and secretion rate of total salivary IgA (P < 0.01) and IgA1 (P < 0.001) was found in the aggressive periodontitis group in resting and stimulated saliva. A decrease of IgA2 (P < 0.05) was seen in resting saliva. Although only minor differences were detected in the concentration and secretion of bacteria-reactive IgA in both groups, the proportion of bacteria-reactive IgA from the total IgA was significantly higher (P < 0.01) in the aggressive periodontitis group in all three microorganisms tested. Our results indicate an inhibition of total secretory IgA. In particular an IgA subclass 1-specific decrease in aggressive periodontitis was noted, while the bacteria-reactive humoral immune system in saliva was activated. The role of the decrease of IgA1 immunoglobulins in aggressive periodontitis with respect to susceptibility for periodontal diseases has to be elucidated.

The terms 'early onset periodontitis' (EOP) and 'juvenile periodontitis' (JP) were replaced by that of 'aggressive periodontitis' in a recent international workshop for the classification of periodontal diseases and conditions. The chief etiologic agent for aggressive periodontitis is considered to be Actinobacillus actinomycetemcomitans in localized juvenile periodontitis. Porphyromonas gingivalis is also mentioned as the etiologic agent of the aggressive periodontitis, although to date its role remains questionable. This communication describes three cases of aggressive periodontitis found to be associated with P. gingivalis but not A. actinomycetemcomitans by polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Our findings clarify the role of P. gingivalis as an etiologic agent in this type of periodontitis and confirm its inclusion in the current definition of aggressive periodontitis.

The purpose of this review is to provide the clinician with some practical rationale for the selection and use of antibiotics in the treatment of destructive periodontal diseases. We have attempted to integrate approximately 20 years of periodontal literature describing antibiotic therapy with personal experience and 21st century ideas. This article addresses antibiotic use during treatment of aggressive periodontitis with emphasis on juvenile disease and adult refractory diseases. The literature review revealed few large, controlled studies that compared efficacy of adjunctive antibiotic use to mechanical therapy alone. Even fewer studies evaluated the efficacy of one antibiotic relative to another. However, based on the evidence available, certain conclusions were drawn. Adjunctive use of an antibiotic along with mechanical debridement is recommended for the treatment of Actinobacillus actinomycetemcomitans-associated periodontitis as an acceptable therapeutic regimen. Due to the emergence of tetracycline-resistant A. actinomycetemcomitans, the combination of metronidazole and amoxicillin may be preferable. In aggressive refractory periodontitis, compelling evidence exists that the use of an appropriate adjunctive antibiotic frequently gives a more favorable clinical response than mechanical therapy alone. Unfortunately, the selection of antibiotic is not as clear and is probably case-dependent. Positive responses have been reported with amoxicillin/clavulanic acid, clindamycin, metronidazole, and the combination therapy metronidazole plus amoxicillin. The introduction of local delivery antibiotics specifically for the treatment of periodontitis offers a novel concept for the treatment of localized disease. The latter, in particular, may prove useful in the treatment of recurrent disease activity or where only a few individual sites are involved.

We investigated the variable Valpha and Vbeta gene usage of Actinobacillus actinomycetemcomitans-reactive periodontal CD4+ T cell receptors (TCR) from: (i) four A. actinomycetemcomitans-infected localized juvenile periodontitis (LJP) patients, (ii) four groups of A. actinomycetemcomitans-inoculated NOD/SCID mice engrafted with individual LJP-derived HuPBL and (iii) HuPBL samples of four LJP patients and two healthy control subjects, by quantitative PCR analyses. The results show that: (i) the majority of the TCR genes (82.5% of Valpha and 91.1% of Vbeta) used by periodontal CD4+ T cells in A. actinomycetemcomitans-inoculated HuPBL-engrafted NOD/SCID mice overlap with those used by local periodontal T cells in LJP patients, (ii) although A. actinomycetemcomitans-reactive periodontal CD4+ TCR repertoire is relatively widespread, there are a few dominant genes shared by the LJP patients, suggesting a limited number of antigens or epitopes commonly recognized and (iii) A. actinomycetemcomitans likely lacks superantigenic characteristics. These results suggest A. actinomycetemcomitans-associated human CD4+ T cell repertoire established in HuPBL-NOD/SCID mice provides a useful approach to study specific aspects of immune-parasite interactions in the periodontium.

The microflora, immunological profiles of host defence functions, and human leukocyte antigen (HLA) findings are reported for a mother, son and daughter who were diagnosed as having 'periodontitis as a manifestation of systemic diseases, associated with hematological disorders'. Examinations were made of the bacterial flora from the periodontal pocket, neutrophil chemotaxis, neutrophil phagocytosis, and the genotypes (DQB1) and serotypes (DR locus) of HLA class II antigens. Phenotypic analyses of the peripheral lymphocytes were also conducted. The subgingival microflora from the mother was dominated by Gram-negative rods, especially Porphyromonas endodontalis, Prevotella intermedia/Prevotella nigrescens and Fusobacterium nucleatum. Subgingival microflora samples from the son and daughter were dominated by Gram-positive cocci and Gram-positive rods. Through the use of polymerase chain reaction, Campylobacter rectus and Capnocytophaga gingivalis were detected in all subjects, whereas Porphyromonas gingivalis, P. intermedia, and Treponema denticola were not detected in any subjects. All three subjects showed a remarkable level of depressed neutrophil chemotaxis to N-formyl-methionyl-leucyl-phenylalanine, although their phagocyte function levels were normal, in comparison to healthy control subjects. Each subject had the same genotype, HLA-DQB1*0601, while the mother had HLA-DR2 and HLA-DR8, and the son and daughter had HLA-DR2 only. In summary, the members of this family showed a similar predisposition to periodontitis with regard to certain host defence functions. It is suggested that the depressed neutrophil chemotaxis that was identified here could be a significant risk factor for periodontitis in this family.

Background: The purpose of this study was to determine the prevalence and distribution of Eikenella corrodens (E. corrodens) in subgingival plaque in different age and periodontitis groups and to examine whether its presence is related to periodontal diseases., Methods: A total of 273 subgingival plaque samples from 213 periodontitis patients and 60 healthy subjects were assessed. Smears from each plaque sample were made and E. corrodens was detected by means of indirect immunofluorescent technique. Mean percentage of E. corrodens per total bacteria (distribution) was calculated in each sample. The prevalence (% of positive samples) and distribution of E. corrodens were statistically analyzed based on age or diagnosis by means of Fisher's exact test and analysis of variance (ANOVA)., Results: Prevalence of E. corrodens decreased by age in the healthy control group; however, prevalence did not change in periodontitis groups. Distribution of E. corrodens was highest in juvenile periodontitis (JP) (2.3 +/- 1.5%) followed by post-JP (1.7 +/- 2.1%), prepubertal periodontitis (1.4 +/- 1.1%), rapidly progressive periodontitis (0.8 +/- 0.7%), adult periodontitis (0.7 +/- 0.6%), and healthy subjects (0.3 +/- 0.3%) (ANOVA, P<0.0001). The <20-year-old age group with periodontitis showed the highest distribution of E. corrodens (2.2 +/- 1.6%) compared to the older age groups who were either healthy or had periodontitis (ANOVA, P<0.0001)., Conclusions: Since the distribution of E. corrodens is significantly higher in JP, post-JP, and PP, E. corrodens might play an important role in the occurrence or progression of periodontitis in young patients.

Young, systemically healthy subjects may suffer from early-onset forms of periodontitis characterized by the presence of localized deep vertical bony defects. The aim of this study was to compare the healing response after guided tissue regeneration (GTR) treatment of similar intrabony defects in patients affected by early-onset and chronic adult periodontitis. Twenty systemically healthy, nonsmoking subjects were enrolled in the study; 10 were affected by early-onset periodontitis (EOP) and 10 by chronic adult periodontitis (CAP). In each subject, only one deep vertical bony defect (intrabony component > 4 mm, probing attachment level > or = 8 mm) was treated according to the principles of GTR therapy with titanium-reinforced e-PTFE membranes. At the time of the surgery and at the 1-year follow-up, a microbiologic test for the identification of the main periodontopathogens was performed in each of the treated sites. There was no statistically significant difference at 1 year in the amount of clinical attachment gain (P = .4), reduction of probing pocket depth (P = .3), or increase in gingival recession (P = 1.0) between EOP and CAP patients. The 1-year microbiologic results demonstrated the complete disappearance of the putative periodontopathogens from all surgically treated sites in both patient groups. The results of the study demonstrated that deep intrabony defects in patients with EOP can be successfully treated by means of GTR procedures and that the suppression of periodontopathogens under threshold values can be maintained for at least 1 year, provided that the patient is enrolled in a maintenance program consisting of recalls for professional tooth cleaning and reinforcement of self-performed oral hygiene measures at 1-month intervals.

Background: A clone of Actinobacillus actinomycetemcomitans (JP2) with increased leukotoxin production and characterized by a 530-bp deletion in the leukotoxin gene operon is endemically present in Morocco and strongly associated with the presence of early onset periodontitis (EOP)., Objectives: To compare patterns of attachment loss among EOP-patients with or without JP2-type of A. actinomycetemcomitans in dental plaque., Material and Methods: Among 45 Moroccan adolescents with EOP (i.e. one or more teeth with attachment loss >/= 3 mm) 39 had cultivable plaque samples. Fifteen (38.5%) were culture-positive for A. actinomycetemcomitans of the JP2-type as determined by PCR, and 24 (61.5%) were not (mean age 16.5 years in both groups)., Results: EOP-patients culture-positive for A. actinomycetemcomitans of the JP2-type had significantly more teeth with attachment loss (mean 5.1, median 4.0) than EOP-patients not culture-positive for A. actinomycetemcomitans of the JP2-type (mean 2.8 teeth, median 1.0) (p = 0.02), and higher attachment loss (mean 4.3 mm vs. 3.4 mm; median 4.0 mm vs. 3.0 mm) (p = 0.01). No major differences could be detected between the two groups in the pattern of affected teeth in the dentition., Conclusions: The study demonstrates increased periodontal destruction among EOP-patients culture-positive for A. actinomycetemcomitans of the JP2-type compared with EOP-patients without the JP2-clone.

Objectives: Microbiological laboratory procedures are involved in diagnosis and therapy control of progressive and refractory forms of periodontitis. In recent years techniques have been developed based on the detection of nucleic acids. The purpose of this study was to validate the commercially available micro-Dent(R) test which employs probes for A. actinomycetemcomitans, P. gingivalis, P. intermedia, B. forsythus and T. denticola., Methods: 122 plaque samples obtained from periodontal pockets with various depths from 33 early onset periodontitis (EOP) patients and 15 periodontally healthy subjects were analysed by cultivation and the microDent(R) kit., Results: Both cultivation and the nucleic acid based assay showed a positive correlation of pocket depth with the frequency and quantity of periodontopathogenic species. T. denticola was found only in pockets > 4 mm in EOP patients. Comparison of the two methods revealed that the microDent(R) kit identified both P. gingivalis and B. forsythus more often than did the cultivation method., Conclusions: Nucleic acid techniques should replace cultivation methods as gold standard in microbiological diagnosis of progressive periodontitis. The micro-Dent(R) kit can be recommended for microbiological laboratories analysing subgingival plaque samples.

Background: Prior reports have suggested that the iron-binding protein lactoferrin (LF) may either kill Actinobacillus actinomycetemcomitans (Aa) or interfere with its binding to host cells. Other studies have indicated that the degree of iron saturation of LF might play a role in these interactions. However, these studies utilized strains that had lost critical attachment characteristics found in well-preserved clinical isolates of Aa. The purpose of this work was to study the effect of LF iron levels on survival and attachment of well-preserved clinical isolates of Aa., Methods: LF containing 0%, 30%, and 100% iron saturation was tested for its ability to kill clinical isolates of Aa and to inhibit their binding to buccal epithelial cells (BECs)., Results: Neither iron-free LF (apo-LF) nor iron-saturated LF killed Aa clinical isolates. Increasing the iron saturation of LF resulted in an increased inhibition of Aa binding to BECs (P < or =0.005). This effect was consistent for the 3 clinical isolates tested. Pretreatment of Aa with iron-saturated LF reduced binding to BECs by 58%, 61.8%, and 64.2%, respectively, for each of the 3 clinical strains tested (P < or =0.005). Pretreatment of Aa strains with apo-LF, iron alone, or bovine serum albumin had no effect on binding. Pretreatment of BECs with LF (either apo-LF or iron-containing LF) had no influence on Aa binding., Conclusions: These results indicate that reduction in binding of Aa to epithelial cells is maximized by pretreatment of Aa cells with iron-saturated lactoferrin. These in vitro results suggest that patients with lactoferrin containing lowered levels of iron would be more susceptible to Aa colonization.

A 10-year-old Japanese girl with severe tooth mobility in her lower permanent incisors was examined clinically, as well as radiographic and microbiological means. The incisors had severe alveolar bond loss and pocket depths exceeding 7 mm at the first visit, however, 10 periodontal bacteria were not detected in subgingival plaque samples taken from the lower central incisors using a 16S rRNA-based polymerase chain reaction method. Periodontal treatment consisting of mechanical debridement and antibiltic medication resulted in a significant improvement of the clinical parameters. Three months after the first examination, dental radiographs showed refilling of alveolar bone in the region. Further, microbiological examinations after remission detected only oral microflora commonly found in health children including A. actinomycetemcomitans. Based on the clinical, readiographic, and microbiological findings, the present case was diagnosed as acute periodontitis.

Background: Periodontal diseases are among the most frequent diseases affecting children and adolescents. These include gingivitis, localized or generalized aggressive periodontitis (a.k.a., early onset periodontitis which includes generalized or localized prepubertal periodontitis and juvenile periodontitis) and periodontal diseases associated with systemic disorders. The best approach to managing periodontal diseases is prevention, followed by early detection and treatment., Methods: This paper reviews the current literature concerning the most common periodontal diseases affecting children: chronic gingivitis (or dental plaque-induced gingival diseases) and early onset periodontitis (or aggressive periodontitis), including prepubertal and juvenile periodontitis. In addition, systemic diseases that affect the periodontium and oral lesions commonly found in young children are addressed. The prevalence, diagnostic characteristics, microbiology, host-related factors, and therapeutic management of each of these disease entities are thoroughly discussed.

The phylogeny of 20 Actinobacillus actinomycetemcomitans strains isolated from patients with localized juvenile periodontitis (LJP) was investigated by using partial sequence analysis of 16S rRNA genes, arbitrarily primed PCR (AP-PCR), and four additional PCR assays that amplified polymorphic regions in the leukotoxin (lkt), cytolethal distending toxin (cdt), major fimbrial subunit (flp-1), and serotype-specific O polysaccharide gene clusters. Our analysis also included four strains isolated from healthy subjects and nine reference strains. We found that A. actinomycetemcomitans strains comprised three major phylogenetic lineages. One lineage consisted of serotype b strains, a second lineage consisted of serotype c strains, and a third lineage consisted of serotype a, d, e, and f strains. 16S rRNA sequences within each lineage were highly conserved (<1% base substitutions), whereas sequences between lineages were exceptionally divergent (1.9 to 5.0% substitutions). Two strains exhibited 16S rRNA sequences that were even more distantly related to those of the three major lineages (2.7 to 6.7% substitutions), indicating that additional minor lineages or variants exist. The distribution of 16S rRNA sequences and lkt, cdt, flp-1, and AP-PCR genotypes was consistent with a clonal population structure, with little evidence of assortative recombination between strains of different serotypes. Strains from all three major lineages were recovered from LJP patients, suggesting that phylogenetically diverse strains of A. actinomycetemcomitans carry pathogenic potential.

Background: Limited information is available as to whether local cellular immunity in early-onset periodontitis (EOP) subjects harboring Actinobacillus actinomycetemcomitans (Aa) differs from that in patients without Aa. In addition, the effect of scaling and root planing on various lymphocyte subsets is described rather sparsely., Methods: In 10 subjects with early-onset periodontitis harboring Aa (EOP-Aa) and in 10 subjects without Aa (EOP-nonAa), clinical measurements were recorded and gingival biopsies were performed before and after scaling and root planing. The specimens were cut into serial sections; using the alkaline phosphatase-antialkaline phosphatase technique, monoclonal antibodies to CD20 (B cells), CD30 (plasma cells), and CD45RO (T-memory cells) were applied as well as polyclonal antibodies to alpha, gamma, and mu chains (Ig A, G, and M). Cells were counted from an area of 0.25 mm2 in areas showing the largest infiltration., Results: Before therapy, mean counts of all cell phenotypes were found to be markedly enhanced in the EOP-Aa group compared to EOP-nonAa subjects. Following scaling and root planing, the numbers of all phenotypes decreased in both groups. However, comparing the data before and after therapy in the EOP-Aa group, the P value was <0.05 only for CD30-positive cells. In the EOP-nonAa group, the differences before and after therapy reached statistical significance (P<0.05) for all cell counts, except for IgM-positive cells. Furthermore, reduction of probing depth and gain of clinical attachment reached the 0.05 level of statistical significance only in EOP-nonAa subjects., Conclusions: In EOP subjects harboring Aa, inflammatory cell subsets were detected in 2- to 3-fold higher numbers compared to patients without Aa. Scaling and root planing resulted in a decrease of all cell phenotypes studied in individuals without Aa, whereas in subjects with Aa, the only significant decrease that was seen occurred in plasma cells.

Objectives: The objective of this study was to determine the relationship between the activity of the enzyme aspartate aminotransferase (AST) in gingival crevicular fluid (GCF) using the colorimetric PerioGard (PTM) test and the subgingival microflora in early onset periodontitis lesions., Material and Methods: The study population consisted of 25 otherwise healthy individuals exhibiting early onset periodontitis (EOP). In each patient four experimental sites were identified comprising one deep periodontal pocket (PD >5 mm) randomly chosen in each quadrant. Bacterial samples were obtained from the experimental sites, consecutively cultured anaerobically and in 10% CO(2) using selective and nonselective media. Isolates were characterized to species level by conventional biochemical tests and various identification kits. Clinical measurements as well as AST activity, assessed either as positive or negative using the PTM, were recorded at the same sites., Results: Sixty-two sites exhibited AST positive and 38 AST negative activity. Analysis of bacterial counts using the ANOVA (Mann Whitney U-test) showed that Streptococcus intermedius, Peptostreptococcus micros, Campylobacter concisus, Bacteroides forsythus, Camplobacter gracilis, Campylobacter rectus and Selenomonas sputigena were significantly higher in sites with AST-positive activity. The odds ratio of having high prevalence of S. intermedius, P. micros, C. concisus, B. forsythus, C. gracilis, C. rectus and S. sputigena in the presence of a positive AST site was very high (range: 3.5-17.0). Streptococcus sanguis, Actinomyces naeslundii, Gemella morbillorum, Capnocytophaga gingivalis, Veillonella parvula, Fusobacterium varium, Eubacterium lentum and Prevotella oralis were detected in significantly higher proportions in sites with AST negative activity and manifested a negative odds ratio in the presence of AST positive sites. The logistic regression analysis revealed that smoking and bleeding upon probing showed a significant association with AST activity, while plaque and suppuration were not found to be significant predictors of AST activity. The co-infection of Porphyromonas gingivalis, B. forsythus and P. micros, or P. gingivalis, B. forsythus and C. rectus were found to be significantly associated with the AST activity (p<0.001). AST positive sites revealed significantly higher occurrence of co-infections by P. gingivalis, B. forsythus, S. sputigena or by P. gingivalis, B. forsythus, S. intermedius than AST negative sites (p<0.001). P. gingivalis, B. forsythus, A. naeslundii co-infection was found significantly higher in the AST negative sites (p<0.001)., Conclusions: The present study found a high level of agreement between the presence of putative periodontal pathogens and positive AST scores at periodontal sites that clinically were considered to be potentially disease active. Prospective studies should be performed to confirm the findings.

Background: The role of antibodies to periodontal microorganisms in the development of periodontal tissue destruction is still unclear. The aim of this study was to investigate the association between serum levels of IgG, IgA, and IgM antibodies to 6 periodontal microorganisms and clinical subtypes of varying severity of early-onset periodontitis (EOP) in young African American adults., Methods: The study group consisted of 159 African Americans aged 19 to 25 years (mean 22 years) and included 97 cases with EOP and 62 controls with no clinical signs of EOP. These subjects were selected from a nationally representative sample of adolescents who received an oral examination as part of the National Survey of Oral Health of United States Children in 1986-1987. The group was examined clinically a second time 6 years later and blood samples were collected. Serum levels of IgG, IgA, and IgM reactive to Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Campylobacter rectus, Eikenella corrodens, and Fusobacterium nucleatum were assessed., Results: Serum levels of IgG and IgA antibody reactive to P. gingivalis and A. actinomycetemcomitans and IgA antibody to P. intermedia were significantly higher in generalized EOP cases compared to healthy controls. IgM antibody levels did not show any significant associations with EOP for any of the 6 bacterial species tested. There were no significant differences in antibody levels between controls and the 13 subjects in our study who were classified with localized EOP., Conclusions: The findings suggest that antibodies to P. gingivalis, P. intermedia, and A. actinomycetemcomitans may play a significant role in the pathogenesis of EOP. Substantial longitudinal studies that monitor antibody levels and avidity prior to disease onset, during progression, and following clinical intervention will be necessary to fully understand the role of this component of the immune response in protection versus tissue destruction and the potential use in EOP risk assessment and disease management.

Sixty-five patients with generalised early-onset periodontitis (G-EOP) (age range 16-42 years, 32 smokers and 33 non-smokers) were assessed for antibody titres and avidity to a panel of five suspected periodontal pathogens (Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Prevotella intermedia, Treponema denticola and Bacteroides forsythus). Thirty-four of these patients were untreated (17 smokers and 17 non-smokers), and thirty-one were in the maintenance phase of periodontal therapy (15 smokers and 16 non-smokers). Previous studies have investigated the effect of smoking on IgG levels in periodontitis patients in the context of the more extensive periodontal destruction seen in smokers. Based on this literature our hypothesis was that smokers would have depressed serum IgG levels directed against recognised periodontal pathogens compared with non-smokers. Antibody titres were measured by ELISA deploying fixed whole cells as coating. The IgG response was detected with biotin-anti-human IgG and avidin-peroxidase; avidity was determined by elution with ammonium thiocyanate. Median titres to A. actinomycetemcomitans, P. intermedia and T. denticola were significantly lower in maintenance patient smokers (p= 0.02, 0.02 and 0.002 respectively) but not in untreated patients. Avidity to P. gingivalis was also lower in smoking maintenance patients (p = 0.003) but not in untreated patients. These findings may imply some interruption of immune maturation in smokers following periodontal treatment.

Background: The successful use of osseointegrated implants in periodontally healthy patients has been documented in numerous longitudinal studies in recent years. However, the extent to which these positive results apply to periodontally diseased patients remains unclear. The aim of the present prospective longitudinal study of partially edentulous patients treated for generalized chronic periodontitis and generalized aggressive periodontitis was a clinical, microbiological, and radiographic comparison of teeth and implants and assessment of the implant success rate., Methods: Five partially edentulous patients treated for generalized aggressive periodontitis (GAgP) and 5 treated for generalized chronic periodontitis (GCP) were enrolled in this study. The GAgP patients received 36 implants, and the GCP patients 12 implants. The teeth were examined 2 to 4 weeks before extraction of the non-retainable teeth (baseline), and 3 weeks after insertion of the final abutments (second examination). All further examinations were performed during a 3-month recall schedule over a 5-year period for the GAgP patients and over a 3-year period for the GCP patients. At each session clinical parameters were recorded at teeth and implants and the composition of the subgingival microflora was determined by dark-field microscopy and DNA analysis. Intraoral radiographs of the teeth and implants were taken for control purposes at baseline; after insertion of the superstructure; and 1, 3, and 5 years later., Results: The clinical findings indicated healthy periodontal and peri-implant conditions in both patient groups throughout the study. However, an increased probing depth and an attachment loss were recorded in the GAgP patients after the third year (P<0.001). The distribution of the microorganisms revealed no significant differences between the patient groups or between implants and teeth. Moderate bone loss at teeth and implants was registered in both groups. The success rates recorded were 100% in the GCP patients and 88.8% (maxilla: 85.7%; mandible: 93.3%) in the GAgP patients., Conclusions: The 3-year and 5-year follow-ups show that osseointegrated implants may be successful in oral rehabilitation of partially edentulous patients treated for generalized aggressive periodontitis and generalized chronic periodontitis. However, as no significant differences were recorded between conditions at teeth and at implants, progression of the disease cannot be ruled out.

Background: Recent studies provide strong evidence implicating Peptostreptococcus micros in the pathogenesis of various oral infections, including oropharyngeal abscesses and periodontal disease. To date, very little is known regarding the role of P. micros in periodontal disease. Therefore, a genetic analysis was initiated to differentiate among strains of P. micros infecting periodontal patients., Methods: Sixty DNA samples of P. micros isolated from 15 patients with periodontal disease were evaluated. Arbitrarily primed polymerase chain reactions (AP-PCR) were performed using primer 3 (AGTCAGCCAC) and primer 13 (CAGCACCCAC). The PCR products were analyzed by gel electrophoresis., Results: The primers produced several unique patterns among the strains tested. Primer 3 resulted in 30 different patterns, whereas primer 13 resulted in 31 different patterns, which were distinct from those seen with primer 3. In 8 of 15 patients, the PCR profile was identical for all isolates cultured from that patient, indicating a clonal infection. In 4 of 15 patients, 2 different genotypes were identified. In the remaining 3 patients, all isolates cultured from these patients exhibited a unique genotype., Conclusions: While P. micros appears to be heterogeneous throughout a population of periodontal patients, each patient is, for the most part, infected with a limited number of genotypes. These results demonstrate the genetic diversity of P. micros and the usefulness of AP-PCR for future epidemiological studies in understanding the role P. micros plays in periodontal disease pathogenesis.

A particular clone (JP2) of Actinobacillus actinomycetemcomitans with increased leukotoxin production has been isolated from individuals with early-onset periodontitis (EOP). The aim of this study was to determine the frequency of carriers of this clone and its association with EOP in Moroccan schoolchildren. Of 217 plaque samples, 131 (60.4%) were culture-positive for A. actinomycetemcomitans. A total of 19 of these isolates had a 530-bp deletion in the leukotoxin promoter region characteristic of the JP2 clone. A strong association between the presence of A. actinomycetemcomitans with the 530-bp deletion and EOP was found (adjusted OR = 29.4; 95% Cl = 8.3 - 104.4; p < 0.0005), while no association could be demonstrated between the presence of A. actinomycetemcomitans without the deletion and EOP (adjusted OR = 1.3; 95% CI = 0.5 -2.9; p = 0.750). The study demonstrates that the endemic presence, in a human population, of the highly leukotoxic JP2 clone may result in an unusually high prevalence of EOP.

Background: Membrane-bound CD14 (mCD14) is expressed mainly on circulating monocytes and tissue macrophages. It is one of the receptors, which act at the recognition of lipopolysaccharides by host cells. Periodontopathic bacteria result in activation of cellular and humoral immune responses., Aim: The aim of the present study was to analyze the peripheral blood mCD14 concentrations as well as cell surface markers of lymphocyte subsets in periodontitis patients of various categories., Materials and Methods: Peripheral blood samples were obtained from 22 early onset periodontitis (EOP), 10 adult periodontitis (AP) patients and 13 systemically and periodontally healthy control subjects. Three-color flow cytometry and a panel of relevant monoclonal antibodies were used to determine the percent expression of various cell surface markers on peripheral blood mononuclear cells (PBMCs). The results were tested statistically by one-way variance analysis and Newman Keuls test., Results: No significant difference was observed between the study groups with regard to the relative counts of B-cells, T-cells, T-helper, T-cytotoxic/suppressor, activated T-cells and natural killer cells. EOP patients expressed significantly lower level of interleukin-2 receptor (IL-2R) when compared with AP patients (6.08% and 19.3% respectively) (p<0.05). The level of mCD14 in EOP patients (7.18%) was lower than that of AP patients (9.3%) and the control subjects (9.2%), but the differences were not statistically significant., Conclusions: The low level of IL-2R in the EOP group may be interpreted as an insufficient responsiveness to the periodontopathogens, which may be ultimately related with the more severe tissue destruction. Though not significant, the reduced expression of mCD14 in EOP group may also be related with the immune system deficiencies in these patients.

Objective: To describe the prevalence of the cultivable subgingival microbiota in periodontal diseases and to draw attention to the polymicrobial nature of periodontic infections., Methods: The study population consisted of 95 patients, 51 females and 44 males, aged 14-62 years. Twenty-nine patients exhibited adult periodontitis (AP), six localized juvenile periodontitis (LJP), and 60 rapidly progressive periodontitis (RPP). Two to four pooled bacterial samples were obtained from each patient. Samples were collected with sterile paper points from the deepest periodontal pockets. The samples were cultured under anaerobic and microaerophilic conditions using selective and non-selective media. Isolates were characterized to species level by conventional biochemical tests and by a commercial rapid test system., Results: Prevotella intermedia and Capnocytophaga spp. were the most frequently detected microorganisms in all diagnostic groups. Porphyromonas gingivalis and Peptostreptococcus micros were found more frequently in AP and RPP patients, while Actinobacillus actinomycetemcomitans and Eikenella corrodens were associated with AP, LJP and RPP patients. The other bacterial species, including Actinomyces spp., Streptococcus spp. and Eubacterium spp., were detected at different levels in the three disease groups., Conclusions: The data show the complexity of the subgingival microbiota associated with different periodontal disease groups, indicating that the detection frequency and levels of recovery of some periodontal pathogens are different in teeth affected by different forms of periodontal disease.

Background: Papillon Lefèvre syndrome (PLS) is a rare entity and, as such, it is almost impossible to evaluate an effective therapy in a randomized controlled study. The amount of success reported after therapy for prepubertal periodontitis (PP) in PLS is highly variable from case to case. The goal of this case report is to evaluate the effects of a combined mechanical and antibiotic periodontal therapy regimen in the management of PLS., Methods: A male patient was diagnosed as suffering from PP associated with PLS at the age of 7 years. He showed hyperkeratosis of the palms and soles, as well as advanced periodontal disease already affecting permanent teeth with maximal probing depth and vertical attachment loss of 12 mm and 11 mm, respectively. Subgingival debridement was performed with simultaneous administration of oral 250 mg amoxicillin 3 times daily and 250 mg metronidazole twice daily for one week. Clinical parameters were assessed and subgingival plaque was collected from all teeth prior to therapy and 7 and 26 months after treatment. Selective cultures for A. actinomycetemcomitans were incubated for each individual tooth and DNA probe analysis was performed for various periodontal pathogens., Results: Prior to combined mechanical and antibiotic treatment, all teeth but one harbored Actinobacillus actinomycetemcomitans subgingivally. However, at 7 and 26 months after therapy A. actinomycetemcomitans could be detected neither by culture nor by DNA probes. Clinical parameters improved markedly and teeth erupting after therapy did not exhibit attachment loss of more than 1.5 mm during the observation period., Conclusions: Eradication (suppression beneath detection levels) of A. actinomycetemcomitans seems to play a significant role in the successful treatment of localized prepubertal periodontitis in PLS.

Background, Aims: Actinobacillus actinomycetemcomitans (Aa) has been implicated as the putative micro-organism for localized juvenile periodontitis (LJP). The most distinct clinical features of LJP include severe angular bony defects of the mesial sides of permanent first molars and the onset of disease during puberty. Currently, no large-scale studies have been performed which address the change in detection rates of Aa on the mesial sides of permanent 1st molars following eruption and up to puberty., Method: In this study, subgingival plaque samples were taken from the mesial pockets of 2 randomly selected permanent 1st molars from 328 primary school children and 50 adult staff, and analyzed by polymerase chain reaction (PCR) to detect Aa., Results: The results showed a 5.5% prevalence rate of Aa which increased after the eruption of 1st molars and peaked near puberty. There were no significant differences in the detection rates of Aa among different groups in terms of gender, plaque index (PII), and gingival index (GI); however, the higher detection rates of Aa were significantly associated with increased probing depths at p<0.05., Conclusion: PCR analysis of the subgingival plaques demonstrated a prevalence of Aa which peaked near puberty, suggesting that Aa may be important for LJP in Taiwan.

A 10-year-old boy presented with generalized gingival inflammation, extensive alveolar bone loss, and tooth mobility. Clinical and radiographic examination supplemented by microbiologic investigation led to a diagnosis of classically termed prepubertal periodontitis (now known as generalized aggressive periodontitis). Other than severe periodontitis, the child was systemically healthy. Neither unusual infections nor abnormalities in neutrophil functions were detected. Microbiologic examinations by culture revealed the presence of the periodontal pathogen Actinobacillus actinomycetemcomitans. Treatment consisted of extraction of mobile teeth, supragingival and subgingival debridement, subgingival curettage, and root planing combined with a 1-week prescription of a combination of metronidazole and amoxicillin. Scanning electron microscopy of extracted teeth revealed hypoplastic and aplastic cementum at the periodontally exposed and intact surfaces. Clinical and microbiologic follow-up was continued over a 1-year period. No periodontal lesions have been detected, and A actinomycetemcomitans could not be isolated from the subgingival areas of the remaining teeth at the end of the first year. Since A actinomycetemcomitans was the main pathogen present in the subgingival microflora of the patient, it might play a key role in the etiology of prepubertal periodontitis.