Oral and endotracheal tubes colonization by periodontal bacteria: a case-control ICU study.

Periodontal infection is a possible risk factor for respiratory disorders; however, no studies have assessed the colonization of periodontal pathogens in endotracheal tubes (ET). This case-control study analyzed whether periodontal pathogens are able to colonize ET of dentate and edentulous patients in intensive care units (ICU) and whether oral and ET periodontal pathogen profiles have any correlation between these patients. We selected 18 dentate and 18 edentulous patients from 78 eligible ICU patients. Oral clinical examination including probing depth, clinical attachment level, gingival index , and plaque index was performed by a single examiner, followed by oral and ET sampling and processing by quantitative polymerase chain reaction (total bacterial load, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythia). Data were statistically analyzed by Mann-Whitney U, two-way analysis of variance (p < 0.05). Among dentate, there was no correlation between clinical parameters and ET bacterial levels. Both dentate and edentulous patients showed similar ET bacterial levels. Dentate patients showed no correlation between oral and ET bacterial levels, while edentulous patients showed positive correlations between oral and ET levels of A. actinomycetemcomitans, P. gingivalis, and T. forsythia. Periodontal pathogens can colonize ET and the oral cavity of ICU patients. Periodontal pathogen profiles tend to be similar between dentate and edentulous ICU patients. In ICU patients, oral cavity represents a source of ET contamination. Although accompanied by higher oral bacterial levels, teeth do not seem to influence ET bacterial profiles.

Tooth loss in individuals under periodontal maintenance therapy: 5-year prospective study.

BACKGROUND AND OBJECTIVE: Tooth loss (TL), one of the most visible results of the evolution of periodontitis, causes physiological and psychological impacts on a patient's life. This prospective study aimed to evaluate the incidence, underlying reasons and influence of risk predictors for the occurrence of TL in a program of periodontal maintenance therapy (PMT) over 5 years. METHODS: The sample comprised 212 individuals diagnosed with chronic moderate-severe periodontitis, who had finished active periodontal treatment, were incorporated in a PMT program. Individuals were divided in to two groups: 96 regular compliers (RC) and 116 irregular compliers (IC). Full-mouth periodontal examination was performed. Social, demographic, behavioral and biological variables of interest were collected at all PMT visits. The effect of risk predictors and confounders for TL, as well as the underlying reasons of TL, were assessed by univariate and multivariate analysis. RESULTS: TL was significantly lower among RC (0.12 teeth lost/year) in comparison to IC (0.36 teeth lost/year; p < 0.01). Individuals that were > 55 years old, males and smokers lost significantly more teeth in both groups (with IC > RC). The number of teeth lost due to periodontal reasons was significantly higher than TL for other reasons in both groups (p < 0.01). The final linear and logistic model for TL included: male gender, smoking, probing depth 4-6 mm in up to 10% of sites and irregular compliance. CONCLUSION: IC individuals undergoing PMT presented higher rates of TL when compared to RC individuals. Findings demonstrated the influence of irregular compliance and the importance of monitoring other risk predictors for TL such as smoking, male gender and severity of probing depth during PMT.

Catecholamines promote the expression of virulence and oxidative stress genes in Porphyromonas gingivalis.

BACKGROUND AND OBJECTIVE: Stress has been identified as an important risk factor in the development of many infectious diseases, including periodontitis. Porphyromonas gingivalis, a gram-negative oral anaerobic bacterium, is considered an important pathogen in chronic periodontitis. Microorganisms, including P. gingivalis, that participate in infectious diseases have been shown to respond to catecholamines released during stress processes by modifying their growth and virulence. Therefore, the purpose of this study was to evaluate the effects of adrenaline and noradrenaline on the growth, antimicrobial susceptibility and gene expression in P. gingivalis. MATERIAL AND METHODS: P. gingivalis was incubated in the presence of adrenaline and noradrenaline (100 µm) for different time-periods in rich (Tryptic soy broth supplemented with 0.2% yeast extract, 5 µg/mL of hemin and 1 µg/mL of menadione) and poor (serum-SAPI minimal medium and serum-SAPI minimal medium supplemented with 5 µg/mL of hemin and 1 µg/mL of menadione) media, and growth was evaluated based on absorbance at 660 nm. Bacterial susceptibility to metronidazole was examined after exposure to adrenaline and noradrenaline. The expression of genes involved in iron acquisition, stress oxidative protection and virulence were also evaluated using RT-quantitative PCR. RESULTS: Catecholamines did not interfere with the growth of P. gingivalis, regardless of nutritional or hemin conditions. In addition, bacterial susceptibility to metronidazole was not modified by exposure to adrenaline or noradrenaline. However, the expression of genes related to iron acquisition (hmuR), oxidative stress (tpx, oxyR, dps, sodB and aphC) and pathogenesis (hem, hagA and ragA) were stimulated upon exposure to adrenaline and/or noradrenaline. CONCLUSION: Adrenaline and noradrenaline can induce changes in gene expression related to oxidative stress and virulence factors in P. gingivalis. The present study is, in part, a step toward understanding the stress-pathogen interactions that may occur in periodontal disease.

Aggregatibacter actinomycetemcomitans serotypes infections and periodontal conditions: a two-way assessment.

This study investigated a large population of individuals positive for A. actinomycetemcomitans and performed a two way analysis assessing the relation between the different serotypes of the bacterium and periodontal conditions. The serotypes analysis (serotypes a, b, c, d, e, f) showed that out of the 204 selected individuals positive for A. actinomycetemcomitans, 152 were positive for a single serotype, 27 showed a variable mixed infection and 25 individuals were not positive for any of the serotypes tested. Serotypes a, b and c were largely found (98%), and serotype c was the most prevalent. Serotypes d, e, and f were either not detected or relatively rare. It was also verified that in non-periodontitis individuals, serotypes a and c were more prevalent (p<0.05); in individuals with mild or moderate/severe chronic periodontitis serotype c was also more common (p<0.05); and aggressive periodontitis subjects showed high prevalence of both serotypes b and c (p<0.05). In conclusion, our study showed that serotype c was the most prevalent in both diseased and healthy subjects. Aggressive periodontitis subjects were not exclusively associated with A. actinomycetemcomitans serotype b. Non-typeable strains were either not detected or were relatively infrequent, and serotypes d and f were not detected in the examined Brazilian population.

Detection of periodontal pathogens in newborns and children with mixed dentition.

We report the age-related prevalence of red complex periodontal pathogens, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia, along with four strains of orange complex pathogens. The bacteria present in samples isolated from tongue, cheek, and subgingival sulcus in edentulous newborns and children with mixed dentition were monitored by polymerase chain reaction (PCR). P. gingivalis was not detected in any site of any subject in the two groups tested. However, T. denticola was not only found in the 6-13 years age group, but also in edentulous newborns at a relatively high prevalence, indicating non-dentition-related colonization by T. denticola. Campylobacter rectus, Prevotella intermedia, T. forsythia, Eikenella corrodens, and Parvimonas micra were found in the oral cavity of most subjects belonging to the 6-13 years age group compared to newborns. This suggested a pronounced association between these colonizing bacteria and the presence of teeth. There was also a strong relation between T. denticola and T. forsythia for their prevalence in the subgingival sulcus of the 6-13 years age group (p < 0.0001), but not in the other sites tested, suggesting that the colonization of dentition-related T. forsythia may be associated with the increased prevalence of non-dentition-related T. denticola in the subgingival sulcus. Overall, these results suggest that dentition is a key determinant of bacterial colonization, especially orange complex bacteria and the red complex bacterium T. forsythia.

Prevalence of periodontopathogens in a black Brazilian secluded community matched with a black urban population.

OBJECTIVE: To evaluate the prevalence of periodontopathogens according to periodontal profile in a black Brazilian secluded community matched with an urban black population. PARTICIPANTS: A total of 84 subjects were selected, 42 (mean age 25.7 sd 18.0 years) from a secluded community called Santo Antonio do Guapore (SAG) and 42 (mean age 25.4 sd 18.1 years) from an urban area of Sao Paulo State (SPT). METHODS: Participants received clinical examinations as follows: periodontal pocket depth; clinical attachment loss; plaque and gingival indexes. After examination, the secluded population was classified as periodontal health (13), gingivitis (15) or periodontitis (14). Then, 182 urban volunteers were screened and 42 subjects were selected matched for the variables: periodontal diagnosis, age (+/- 2 years) and gender. Samples were taken for microbial analysis. Genomic DNA for Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Campylobacter rectus, Tannerella forsythia and Prevotella intermedia was provided by polymerase chain reaction. RESULTS: Except for C. rectus, all pathogens were present in both groups with no statistically significant difference. In particular, C. rectus was more prevalent only in gingivitis subjects from the SPT group (p<0.05). A high frequency of periodontopathogens was related to the severity of periodontal disease. CONCLUSION: In general, the prevalence of the examined periodontopathogens in this study did not differ between a secluded black Brazilian population and an urban black population.

Protease-activated receptor-2 (PAR(2)) in human periodontitis.

No evidence for the role of protease-activated receptor-2 (PAR(2)) in human periodontal disease has been demonstrated so far. Thus, we sought to investigate the expression of PAR(2) mRNA in chronic periodontitis, and to examine whether its expression is related to the presence of PAR(2) potential activators. Microbiological and gingival crevicular fluid samples were collected from individuals with chronic periodontitis and control individuals, and the presence of neutrophil serine proteinase 3 (P3) and Porphyromonas gingivalis was evaluated. PAR(2) mRNA expression was higher (p < 0.001) in those with chronic periodontitis compared with control individuals, and it was statistically decreased (p = 0.0006) after periodontal treatment. Furthermore, those with chronic periodontitis presented higher (p < 0.05) levels of IL-1alpha, IL-6, IL-8, and TNF-alpha, total proteolytic activity, P. gingivalis prevalence, and P3mRNA expression compared with control individuals. We conclude that PAR(2) mRNA expression and its potential activators are elevated in human chronic periodontitis, therefore suggesting that PAR(2) may play a role in periodontal inflammation.

Smoking increases salivary arginase activity in patients with dental implants.

It is believed that an increased arginase activity may lead to less nitric oxide production, which consequently increases the susceptibility to bacterial infection. Considering the hypothesis that smoking may alter the arginase activity and that smoking is considered a risk factor to dental implant survival, the present study aimed at evaluating the effect of smoking on the salivary arginase activity of patients with dental implants. Salivary samples of 41 subjects were collected: ten non-smoking and with no dental implants (group A), ten non-smoking subjects with dental implants (group B), ten smoking subjects with implants (group C), and 11 smoking subjects with no dental implants (group D). The levels of salivary arginase activity were determined by the measurement of L-ornithine and expressed as mIU/mg of protein. A significant increase in the salivary arginase activity was verified in groups C (64.26 +/- 16.95) and D (49.55 +/- 10.01) compared to groups A (10.04 +/- 1.95, p = 0.00001 and p = 0.0110, groups C and D, respectively) and B (11.77 +/- 1.45, p = 0.00001 and p = 0.0147, groups C and D, respectively). No significant difference was found between groups C and D (p = 0.32). Within the limits of the present study, it can be concluded that salivary arginase activity is increased in smoking subjects with dental implants in contrast to non-smoking subjects with dental implants, therefore suggesting a possible mechanism by which cigarette smoking may lead to implant failure. The analysis of salivary arginase activity may represent an important tool to prevent implant failure in the near future.

Periodontal therapy reduces arginase activity in saliva of patients with chronic periodontitis.

This present study evaluated the salivary arginase activity (SAA) in patients with chronic periodontitis and the effect of periodontal therapy on the activity of such enzyme. Thirty-six patients (mean age, 45.97 +/- 14.52), 18 chronic periodontitis subjects (test group), and 18 periodontally healthy individuals (control group) participated in the study. Clinical periodontal examinations included measurements of probing pocket depth (PD), clinical attachment level (CAL), plaque (PI), and gingival (GI) indexes. The test group received periodontal therapy according to individual needs. The saliva sample was collected from all study population at baseline (both groups) and 30 days after periodontal therapy (test group). SAA was determined by measuring the L: -ornithine formation from L-arginine and was expressed as mU/ml. The results showed that the mean values of SAA were statistically different between control and test groups. SAA was about 2.5 times higher in test than control groups. Thirty days after periodontal therapy, enzyme levels were 1.56 times lower than before periodontal therapy. We concluded that SAA is increased in chronic periodontitis subjects when compared to periodontally healthy individuals and that periodontal therapy significantly reduced SAA levels. It was suggested that in the near future, SAA may be used as a salivary marker of periodontal status.

Prevalence and risk variables for peri-implant disease in Brazilian subjects.

OBJECTIVES: The aim of this study was to verify the prevalence of peri-implant disease and analyse possible risk variables associated with peri-implant mucositis and peri-implantitis. The study group consisted of 212 partially edentulous subjects rehabilitated with osseointegrated implants. MATERIAL AND METHODS: The implants placed were examined clinically and radiographically to assess the peri-implant status. The degree of association between peri-implant disease and various independent variables was investigated using a multinomial regression analysis. RESULTS: The prevalence of peri-implant mucositis and peri-implantitis were 64.6% and 8.9%, respectively. In univariate modelling, healthy peri-implant subjects presented lower plaque scores, less periodontal bleeding on probing, and less time elapsed since placement of supra-structures. In multivariate analyses, the risk variables associated with increased odds for having peri-implant disease included: gender, plaque scores, and periodontal bleeding on probing. Presence of periodontitis and diabetes were statistically associated with increased risk of peri-implantitis. The only two factors, which did not contribute to the presence of the disease, were the time elapsed since placement of supra-structures and the frequency of visits for maintenance care. CONCLUSION: Our data suggest that subjects with periodontitis, diabetes, and poor oral hygiene were more prone to develop peri-implantitis.

Evaluation of human leukocyte N-formylpeptide receptor (FPR1) SNPs in aggressive periodontitis patients.

Polymorphonuclear neutrophils (PMNs) are attracted to sites of infection by N-formylpeptide (fMLP) chemoattractants. The high-affinity fMLP receptor (FPR1) of phagocytic cells interacts with bacterial fMLP and mediates chemotaxis, degranulation, and superoxide production. These cellular functions are disrupted in PMN from aggressive periodontitis (AP) patients. Two FPR1 gene single nucleotide polymorphisms (SNPs), c.329T>C and c.378C>G, have been associated with a localized form of AP in African-American patients. To evaluate the generality of these SNPs in AP patients, we sequenced a 363 bp interval of the FPR1 gene in an ethnically diverse group of patients (n=111) and controls (n=115). Neither c.329T>C nor c.378C>G were detected in the 452 alleles sequenced. Six SNPs were identified including two located in the FPR1 second extracellular loop that were significantly associated with the AP phenotype in African-American patients (p.R190W, P=0.0033; and p.N192K, P=0.0018). These two SNPs show three predominant haplotypes, each associated with a different disease risk in African-Americans. These data do not support the hypothesis that the FPR1 SNPs c.329T>C and c.378C>G play an etiologic role in aggressive periodontitis, but do suggest that SNPs in the second extracellular loop may be etiologically important.

Novel COL1A1 mutation (G559C) [correction of G599C] associated with mild osteogenesis imperfecta and dentinogenesis imperfecta.

A genotype-phenotype analysis of a three-generation family segregating for an autosomal-dominant osteogenesis imperfecta (OI) variant is reported here. The family was ascertained through the presentation of a proband concerned about discoloration of her teeth, found to be dentinogenesis imperfecta (DGI). Examination of 36 family members identified 15 individuals with DGI. Linkage studies were performed for genetic markers from candidate intervals known to contain genes responsible for DGI on chromosomes 4q, 7q, and 17q. Conclusive evidence for linkage of DGI was obtained to genetic markers on chromosome 17q21-q22 (DLX-3, Z(max) = 5.34, theta = 0.00). All DGI-affected family members shared a common haplotype, which was not present in individuals without DGI. Haplotype analysis sublocalized the gene to a 5-cM genetic interval that contained the collagen 1 alpha 1 (COL1A1) gene. More than 150 different COL1A1 gene mutations have been associated with various forms of OI, and five of these have been associated with DGI and type IV OI. After excluding these five mutations, mutational analysis was performed on the remaining exons including intron--exon boundaries, which resulted in identification of a Gly559Cys mutation in exon 32, present in all DGI-affected family members. Clinical features segregating with this G559C mutation included hyperextensible joints, joint pain and an increased propensity for bone fractures with moderate trauma. This is the first report of joint pain associated with a COL1A1 mutation and DGI. The mild skeletal features and reduced penetrance of the non-dental findings illustrate the importance of genetic evaluations for families with a history of DGI.

Evidence of genetic heterogeneity for hereditary gingival fibromatosis.

Hereditary Gingival Fibromatosis (HGF) is the most common genetic form of gingival fibromatosis. The condition is most frequently reported to be transmitted as an autosomal-dominant trait, but autosomal-recessive inheritance has also been reported. The clinical presentation of HGF is variable, both in the distribution (number of teeth involved) and in the degree (severity) of expression. It is unknown if the variable clinical expression of HGF in different families is due to variable expression of a common gene mutation, allelic mutations, or non-allelic mutations. The apparently different modes of Mendelian inheritance of HGF suggest genetic heterogeneity. A gene locus for HGF has been localized to a 37-cM genetic interval on chromosome 2p21-p22 (D2S1352, Zmax = 5.10, theta = 0.00) flanked by D2S1788 and D2S441. To evaluate the generality of this linkage, we tested linkage with 9 markers from this candidate region in another large family, segregating for an autosomal-dominant form of generalized HGF, and found no support for linkage with any of these markers. Furthermore, statistical tests of this apparent heterogeneity were highly significant. Analysis of these data provides direct evidence that at least two genetically distinct loci are responsible for autosomal-dominant hereditary gingival fibromatosis.

Evidence of genetic heterogeneity for hereditary gingival fibromatosis / Evidência da heterogenidade genética para fibromatose gengival hereditária

Hereditary gingival fibromatosis (HGF) is the most common genetic form of gingival fibromatosis. The condition is most frequently reported to be transmitted as an autosomal-dominant trait, but autosomal-recessive inheritance has also been report. The clinical presentation of HGF is variable, both in the distribution (number of teeth involved) and in the degree (severity) of expression of HGF in different families is due to variable expression of a common gene mutation, allelic mutations, or non-allelic mutation, allelic mutations. The apparently different modes of Mendelian inheritance of HGF suggest genetic heterogeneity. A gene locus for HGF has been localized to a 37-cM genetic interval on chromosome 2p21-p22 (D2S1352, Zmax=5.10, O = 0.00) flanked by D2S1788 and D2S441. To evaluate the generality of this candidate region in another large family, segregating for an autosomal-dominant form of generalized HGF, and found no support for linkage with any of these markers. Furthermore, statistical tests of this apparent heterogeneity were highly significant. Analysis of these data provides direct evidence that at least two genetically distinct loci are responsible for autosomal-dominant hereditary gingival fibromatosis

Evidence for the role of highly leukotoxic Actinobacillus actinomycetemcomitans in the pathogenesis of localized juvenile and other forms of early-onset periodontitis.

BACKGROUND: Actinobacillus actinomycetemcomitans leukotoxin is thought to be an important virulence factor in the pathogenesis of localized juvenile and other forms of early-onset periodontitis. Some highly leukotoxic A. actinomycetemcomitans strains produce 10 to 20 times more leukotoxin than other minimally leukotoxic strains. The distribution, clonality, and intrafamilial transmission of highly leukotoxic A. actinomycetemcomitans were examined in order to determine the importance of leukotoxin in the pathogenesis of periodontitis. METHODS: The polymerase chain reaction (PCR) was used to differentiate highly leukotoxic from minimally leukotoxic strains in examining 1,023 fresh A. actinomycetemcomitans isolates and strains from our culture collection. These were obtained from 146 subjects including 71 with localized juvenile periodontitis (LJP), 4 with early-onset periodontitis, 11 with post-localized juvenile periodontitis, 41 with adult periodontitis, and 19 periodontally normal subjects. The arbitrarily primed polymerase chain reaction (AP-PCR) analysis of 30 oral isolates from each of 25 subjects was used to determine the intraoral distribution of A. actinomycetemcomitans clones. AP-PCR was also used to examine the transmission of A. actinomycetemcomitans in 30 members of 6 families. The clonality of 41 highly leukotoxic A. actinomycetemcomitans strains was evaluated by both AP-PCR and ribotyping. RESULTS: Highly leukotoxic A. actinomycetemcomitans was found only in subjects with localized juvenile and early-onset periodontitis. Fifty-five percent of the LJP subjects harbored highly leukotoxic A. actinomycetemcomitans isolates. Seventy-three percent of the A. actinomycetemcomitans isolates in these subjects were highly leukotoxic. Highly leukotoxic A. actinomycetemcomitans infected younger subjects (mean age 13.95 years, range 5 to 28 years) than minimally leukotoxic (mean age 35.47 years, range 6 to 65 years). Most subjects were infected with only one A. actinomycetemcomitans genotype. However, PCR of whole dental plaques and subsequent analysis of up to 130 individual oral isolates suggested a possible shift in A. actinomycetemcomitans over time in that a few subjects harbored both highly leukotoxic and minimally leukotoxic strains. AP-PCR analysis was consistent with intrafamilial A. actinomycetemcomitans transmission. Ribotyping and AP-PCR analysis confirmed a previous report that highly leukotoxic A. actinomycetemcomitans consists of a single clonal type. CONCLUSIONS: This study suggests that localized juvenile and other forms of Actinobacillus-associated periodontitis are primarily associated with the highly leukotoxic clone of A. actinomycetemcomitans.

Genetic heterogeneity of gingival fibromatosis on chromosome 2p.

Gingival fibromatosis (GF) occurs in several genetic forms as a simple Mendelian trait, in malformation syndromes, and in some chromosomal disorders. Specific genes responsible for GF have not been identified. An autosomal dominant form of hereditary gingival fibromatosis (HGF, MIM 135300) was recently mapped to chromosome 2p21 in a large Brazilian family and there was an earlier report of GF in a boy with a cytogenetic duplication involving 2p13-->p21. We thus hypothesised that a common gene locus may be responsible for GF in both the Brazilian family and the boy with the chromosome 2p duplication. We performed additional genetic linkage studies on the Brazilian family and molecular cytogenetic studies on the patient with the cytogenetic duplication to correlate more precisely the genetic interval of the HGF phenotype with the duplicated 2p interval. Additional linkage analysis of new family members resulted in refinement of the candidate region for HGF to an 8 Mb region. Molecular cytogenetic analysis of the 2p13-->p21 duplication associated with GF showed that the duplicated region was proximal to the candidate interval for HGF. Thus, our results support the presence of two different gene loci on chromosome 2p that are involved in GF.

Genetic linkage of hereditary gingival fibromatosis to chromosome 2p21.

Gingival fibromatosis is characterized by a slowly progressive benign enlargement of the oral gingival tissues. The condition results in the teeth being partially or totally engulfed by keratinized gingiva, causing aesthetic and functional problems. Both genetic and pharmacologically induced forms of gingival fibromatosis are known. The most common genetic form, hereditary gingival fibromatosis (HGF), is usually transmitted as an autosomal dominant trait, although sporadic cases are common and autosomal recessive inheritance has been reported. The genetic basis of gingival fibromatosis is unknown. We identified an extended family (n=32) segregating an autosomal dominant form of isolated gingival fibromatosis. Using a genomewide search strategy, we identified genetic linkage (Zmax=5.05, straight theta=.00) for the HGF phenotype to polymorphic markers in the genetic region of chromosome 2p21 bounded by the loci D2S1788 and D2S441. This is the first report of linkage for isolated HGF, and the findings have implications for identification of the underlying genetic basis of gingival fibromatosis.