Prevalencia de Porphyromonas gingivalis en un grupo de pacientes con aparatología fija de ortodoncia / Prevalence of Porphyromonas gingivalis in a group of patients with fixed orthodontic appliances

Introducción: existen diversos patógenos que pueden afectar no sólo la salud periodontal, sino también la salud general de los pacientes. Objetivo: determinar la Porphyromonas gingivalis (PG) en el primer molar superior derecho de adolescentes, de entre 12 y 18 años, con al menos un mes de tratamiento de ortodoncia con aparatología fija. Material y métodos: se realizó un estudio observacional, descriptivo, transversal de casos en un grupo de 26 adolescentes con tratamiento de ortodoncia, compuesto de brackets metálicos, tubos o bandas, arcos NiTi termoactivos, módulos, cadenas o ligaduras; sin importar sexo, edad, tiempo de tratamiento o maloclusión. Se formaron dos pares de grupos 1 y 2 (15 mujeres y 11 hombres), A y B (13 mujeres y 13 hom- bres) comparando los resultados obtenidos entre los grupos. Resulta- dos: dentro del grupo 1 y 2 la detección molecular de microorganismos arroja que 80% fueron positivas a la PG, 58.33% presenta maloclusión y en promedio 89% de las pacientes son positivas a PG. La detección molecular del grupo A y B indica que 54.54% fueron positivos a PG, mientras que 83.3% presenta maloclusión y en promedio 47% son positivos a PG. Conclusión: la explicación de los eventos moleculares que se desencadenan en la cavidad oral y los sistemas afectados por PG contribuyen a la prevención de complicaciones al tener una mejor comprensión de los fenómenos infecciosos (AU)

Introduction: there are various pathogens that can affect not only periodontal health, but also the general health of patients. Objective: to determine Porphyromonas gingivalis (PG) in the upper right first molar of adolescents, between 12 and 18 years old, with at least one month of orthodontic treatment with fixed appliances. Material and methods: a cross-sectional descriptive observational study of cases was carried out in a group of 26 adolescents with orthodontic treatment, consisting of metal brackets, tubes or bands, thermoactive NiTi archwires, modules, chains or ligatures; regardless of sex, age, treatment time or malocclusion. Two pairs of groups 1 and 2 (15 women and 11 men), A and B (13 women and 13 men) were formed, comparing the results obtained between the groups. Results: within group 1 and 2, the molecular detection of microorganisms shows that 80% were positive for PG, 58.33% presented malocclusion and an average of 89% of patients were positive for PG. The molecular detection of group A and B indicates that 54.54% were positive for PG while 83.3% presented malocclusion and on average 47% were positive for PG. Conclusion: the explanation of the molecular events that are triggered in the oral cavity and the systems affected by PG contribute to the prevention of complications by having a better understanding of the infectious phenomena (AU)

Microbial Screening Reveals Oral Site-Specific Locations of the Periodontal Pathogen Selenomonas noxia.

INTRODUCTION: Selenomonas noxia (SN) is an important periodontal pathogen, associated with gingivitis and periodontitis. Many studies have found associations between SN and indicators of poor health outcomes, such as smoking, low socioeconomic status and obesity. However, less is known about the prevalence of this organism and more specifically about other oral site-specific locations that may harbor this organism. METHODS: Using an existing patient repository (n = 47) of DNA isolated from saliva and other oral sites (n = 235), including the dorsum of the tongue, lower lingual incisor, upper buccal molar and gingival crevicular fluid (GCF), molecular screening for SN was performed. Screening results were analyzed for associations between demographic variables (age, sex, race/ethnicity) and clinical information (body mass index or BMI, presence of orthodontic brackets, primary/mixed/permanent dentition). RESULTS: qPCR screening revealed a total of n = 62/235 sites or 26.3% harboring SN with saliva and GCF (either alone or in combination with one or more sites) most often observed (Saliva, n = 23/27 or 85.18%, GCF, n = 14/27 or 51%). Analysis of site-specific data revealed most positive results were found among saliva and GCF alone or in combination, with fewer positive results observed among the tongue (33.3%), lower lingual incisor (29.6%), and upper buccal molar (25.9%). No significant associations were found between demographic or clinical variables and presence of SN at any site. CONCLUSIONS: These results may be among the first to describe site-specific locations of S. noxia among various additional oral biofilm sites. These data may represent a significant advancement in our understanding of the sites and locations that harbor this organism, which may be important for our understanding of the prevalence and distribution of these organisms among patients of different ages undergoing different types of oral treatments, such as orthodontic treatment or therapy.

Treatment of Porphyromonas gulae infection and downstream pathology in the aged dog by lysine-gingipain inhibitor COR388.

COR388, a small-molecule lysine-gingipain inhibitor, is currently being investigated in a Phase 2/3 clinical trial for Alzheimer's disease (AD) with exploratory endpoints in periodontal disease. Gingipains are produced by two species of bacteria, Porphyromonas gingivalis and Porphyromonas gulae, typically associated with periodontal disease and systemic infections in humans and dogs, respectively. P. gulae infection in dogs is associated with periodontal disease, which provides a physiologically relevant model to investigate the pharmacology of COR388. In the current study, aged dogs with a natural oral infection of P. gulae and periodontal disease were treated with COR388 by oral administration for up to 90 days to assess lysine-gingipain target engagement and reduction of bacterial load and downstream pathology. In a 28-day dose-response study, COR388 inhibited the lysine-gingipain target and reduced P. gulae load in saliva, buccal cells, and gingival crevicular fluid. The lowest effective dose was continued for 90 days and was efficacious in continuous reduction of bacterial load and downstream periodontal disease pathology. In a separate histology study, dog brain tissue showed evidence of P. gulae DNA and neuronal lysine-gingipain, demonstrating that P. gulae infection is systemic and spreads beyond its oral reservoir, similar to recent observations of P. gingivalis in humans. Together, the pharmacokinetics and pharmacodynamics of COR388 lysine-gingipain inhibition, along with reduction of bacterial load and periodontal disease in naturally occurring P. gulae infection in the dog, support the use of COR388 in targeting lysine-gingipain and eliminating P. gingivalis infection in humans.

Levels of pro- and anti-inflammatory cytokines in cystic fibrosis patients with or without gingivitis.

BACKGROUND: Inflammatory periodontal diseases are caused by interaction between gram negative, anaerobic bacteria and host response. Persistent infection of Pseudomonas aeruginosa in cystic fibrosis (CF) patients also cause increased pro-inflammatory response and the imbalance of pro- and anti-inflammatory response in brochoalveolar lavage fluid which leads to destruction of lungs. The aim of this study is to evaluate periodontal status of CF patients, to measure level of cytokines and biochemical molecules in gingival crevicular fluid (GCF), and to detect presence of P. aeruginosa in dental plaque samples. MATERIALS AND METHODS: GCF samples were collected from 41 CF patients and 39 healthy (non-CF) subjects. Interleukin (IL)-1ß, IL-17, IL-10, human neutrophil elastase (HNE), cystic fibrosis transmembrane regulator (CFTR) protein, and human ß-defensin-1 (HBD1) in GCF were evaluated by ELISA method. Dental plaque samples were collected from 18 CF patients with history of P. aeruginosa colonization and 15 non-CF subjects. Presence of P. aeruginosa was evaluated by using conventional culture methods and molecular methods. RESULTS: Levels of IL-1ß, HNE, and HBD1 in CF patients were significantly higher than non-CF subjects. However, IL-10 level was significantly lower in CF patients. Increased pro-inflammatory (IL-1ß) and decreased anti-inflammatory (IL-10) cytokine levels were observed in GCF samples from CF patients, irrespective of their periodontal status. P. aeruginosa were detected in four samples of 18 CF patients, and all were negative in non-CF group. CONCLUSIONS: As a result of this study, CF coexists increasing pro-inflammatory and decreasing anti-inflammatory response locally. Due to increasing pro-inflammation, CF patients should be followed-up more often than non-CF children.

Influence of restorative margins position on one-stage laser-microgrooved implants-supported single screwed crowns: A clinical, biochemical, and microbiological analysis.

AIM: To clinically, biochemically, and microbiologically evaluate the influence of crown margins position on one-stage laser-microgrooved implants. MATERIALS AND METHODS: Twenty-one-stage titanium implants with a laser-microgrooved collar surface, supporting screwed, single crown restorations, were placed in 20 partially edentulous patients and evaluated. Clinical parameters included modified plaque index, modified gingival index, peri-implant probing pocket depth, bleeding on probing, and distance between implant shoulder and mucosal margin. The parameters were recorded at baseline (crowns delivery) and at every 6-month recall visit, until the end of the 3 years follow-up period. At the same time intervals, radiographic marginal bone levels were assessed at the mesial and distal aspect of the implant sites. For biochemical analysis, the volume of the peri-implant sulcus fluid, and its levels of interleukin-1beta (IL-1ß), interleukin-6 (IL-6), and of tumor necrosis factor-α, were utilized to evaluate the peri-implant health conditions at the end of the 3-year follow-up period. At the same time, microbiological analysis, including the concentration of five putative periodontal pathogens (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Treponema denticola, and Tannerella forsythensis), were assessed. The crown margins positions were classified into four groups (A = intracrevicular position >2 mm, B = intracrevicular position ≤2 mm/<1 mm, C = intracrevicular position ≤1 mm/<0 mm, and D = extracrevicular position), and the biochemical, and microbiological parameters were evaluated at 3 years. RESULTS: No statistical differences of clinical and biochemical parameters were found between the four groups. In group A, compared to groups B, C, and D, a statistically significant higher concentration of putative periodontal pathogens was found. CONCLUSIONS: Results showed that the intracrevicular deeper position of the restoration margin does not influence the clinical and biochemical peri-implant parameters. The deeper position of the crown margin is associated with a greater amount of putative periodontal pathogenic microflora colonization.

An Analytical Method for Assessing Optimal Storage Conditions of Gingival Crevicular Fluid and Disclosing a Peptide Biomarker Signature of Gingivitis by MALDI-TOF MS.

PURPOSE: Gingival crevicular fluid (GCF) is an important diagnostic source of biomarkers for both periodontitis and gingivitis. However, GCF peptide signature may change depending on factors such as handling and storage. Here we propose a standardized methodology for GCF analysis by MALDI-TOF/TOF-MS in order to distinguish a characteristic peptide signature of gingivitis. EXPERIMENTAL DESIGN: The best storage/handling conditions which may ensure the stability of the endogenous peptidome in GCF is determined and then MALDI-TOF MS comparative analysis is performed. Reproducible GCF MALDI-TOF signatures between two groups of gingivitis (n = 10) and healthy (n = 10) subjects are compared. RESULTS: A pattern of five peptides resulted differentially expressed between gingivitis and healthy groups. Interestingly, among these biomarkers the C-terminal fragment of alpha-1-antitrypsin (AAT) namely C-36 peptide and two different PTMs of the full-length S100A9 protein are found. CONCLUSIONS AND CLINICAL RELEVANCE: The method described provides a rapid comparative analysis of GCF signatures between periodontally healthy and gingivitis subjects. A pattern based on the expression of endogenous peptides and their PTMs is identified in GCF as putative biomarkers of gingivitis. These findings improve the knowledge of the inflammatory, immune, and structural substrates which might have a key role in the pathogenesis of gingivitis.

Cytokines and MMPs levels in gingival crevicular fluid from patients with chronic periodontitis before and after non-surgical periodontal therapy

The study of host response in periodontal disease may provide a mechanism to monitor disease progression. the purpose of the present research was to determine the levels of IL-1alfa, IL-1beta, TNF-alfa, IL-6, IL-6sR, IL-8, IL-10, MMP-3 and MMP-8 in gingival crevicular fluid (GCF), before and after non-surgical periodontal treatment (NSPT) in order to evaluate therapy response. methodology: eleven patients diagnosed with chronic periodontitis and eleven healthy subjects were selected for this study. clinical measurements, including probing depth (PD) and clinical attachment loss (CAL) were carried out in patients diagnosed with chronic periodontitis and periodontal healthy controls. the clinical indexes evaluated were: gingival index (GI) and plaque index (PI). samples of GCF were taken from one tooth per quadrant before and 45 days after NSPT. the levels of inflammatory mediators were measured by ELISA. results: the values of all clinical parameters decressed significsntly after treatment. the concentration levels of all cytokines and MMP-3 and MMP-8 in the GCF sample were higher in patients diagnosed with chronic periodontitis compared to the healthy group. all inflammatory mediators decreased after therapy, but did not reach control values; IL-6, Il-6sR, IL-10 and TNF-alfa, attained the highest reduction (70 percent -54 percent); the vales of MMP3, IL-1alfa, IL-1beta and IL-8 were reduced between 50 percent ­ 34 percent; and MMP-8 showed the lowest decrease (28 percent). conclusion: all clinical parameters and cytokines levels decreased after NSPT. the mediators TNF-alfa IL-6, IL-6sR, and IL-10 showed the largest variation between before and after NSPT and could thus be used to evaluate therapy response.

Intraoral versus extraoral cementation of implant-supported single crowns: Clinical, biomarker, and microbiological comparisons.

OBJECTIVES: Implant supported single metal-ceramic crowns cemented either extraorally or intraorally were comparatively evaluated by clinical, radiologic, biomarker, and microbiological parameters. MATERIALS AND METHODS: Twelve patients with bilateral single tooth gap in the maxillary posterior region received two locking-taper implants; 4.5 mm width, 8 mm length. Selection of intraoral (IOC) or extraoral cementation (EOC) using screwless titanium abutments was done randomly. Peri-implant crevicular fluid (PICF), gingival crevicular fluid (GCF) samples were collected from the implants, adjacent teeth, and bleeding on probing, soft tissue thickness, keratinized tissue width were recorded before starting the prosthetic procedures (baseline) and 3, 6 months after implant loading. Crestal bone loss was measured on radiographs taken immediately and 6 months after cementation. Cytokine levels, amounts of bacteria were determined in PICF/GCF samples. Data were tested by appropriate statistical analyses. RESULTS: Clinical findings were similar in the crowns cemented extraorally or intraorally at all times (P < .05). PICF and GCF data were similar. At 3 month, interleukin-17E and osteoprotegerin levels were lower in the intraorally cemented crowns. CONCLUSION: Extraorally and intraorally cemented crowns exhibited similar crestal bone loss after loading. Higher amount of osteoprotegerin at 3 month at the EOC than the IOC sites might bode well for good osseointegration.

Association of Distinct Fine Specificities of Anti-Citrullinated Peptide Antibodies With Elevated Immune Responses to Prevotella intermedia in a Subgroup of Patients With Rheumatoid Arthritis and Periodontitis.

OBJECTIVE: In addition to the long-established link with smoking, periodontitis (PD) is a risk factor for rheumatoid arthritis (RA). This study was undertaken to elucidate the mechanism by which PD could induce antibodies to citrullinated peptides (ACPAs), by examining the antibody response to a novel citrullinated peptide of cytokeratin 13 (CK-13) identified in gingival crevicular fluid (GCF), and comparing the response to 4 other citrullinated peptides in patients with RA who were well-characterized for PD and smoking. METHODS: The citrullinomes of GCF and periodontal tissue from patients with PD were mapped by mass spectrometry. ACPAs of CK13 (cCK13), tenascin-C (cTNC5), vimentin (cVIM), α-enolase (CEP-1), and fibrinogen ß (cFIBß) were examined by enzyme-linked immunosorbent assay in patients with RA (n = 287) and patients with osteoarthritis (n = 330), and cross-reactivity was assessed by inhibition assays. RESULTS: A novel citrullinated peptide cCK13-1 (444 TSNASGR-Cit-TSDV-Cit-RP458 ) identified in GCF exhibited elevated antibody responses in RA patients (24%). Anti-cCK13-1 antibody levels correlated with anti-cTNC5 antibody levels, and absorption experiments confirmed this was not due to cross-reactivity. Only anti-cCK13-1 and anti-cTNC5 were associated with antibodies to the periodontal pathogen Prevotella intermedia (P = 0.05 and P = 0.001, respectively), but not with antibodies to Porphyromonas gingivalis arginine gingipains. Levels of antibodies to CEP-1, cFIBß, and cVIM correlated with each other, and with smoking and shared epitope risk factors in RA. CONCLUSION: This study identifies 2 groups of ACPA fine specificities associated with different RA risk factors. One is predominantly linked to smoking and shared epitope, and the other links anti-cTNC5 and cCK13-1 to infection with the periodontal pathogen P intermedia.

The correlation between periodontal health status and suspectibility to infections associated with craniomaxillofacial osteosynthesis plates.

AIM: The aim of the present study was to demonstrate the possible relation between periodontal health status and infections associated with osteosynthesis materials (OMs) used in the oral and maxillofacial reconstruction. MATERIAL AND METHODS: The study group consisted of 32 individuals which were subdivided into two groups regarding their PSI scores. After the removal of the osteosynthesis plates, microbial colonization was assessed via microbiological cultivation, fluorescence microscopy and scanning electron microscopy. In addition, samples obtained from gingival crevicular fluids were investigated by fluorescence microscopy. RESULTS: A total of 118 osteosynthesis plates were examined. 8.5% (n = 10) of the plates were associated at least one of the clinical signs of infection. There was a positive correlation between periodontal disease and clinical signs of infection (p = 0.022). Patients with infection signs also had a higher number of smoking history (pack years, p = 0.010). Intraorally placed osteosynthesis plates showed wide range of bacterial colonizations compared to extraorally inserted osteosynthesis materials (p = 0.004). CONCLUSION: Patients with poor periodontal health might be potential candidates for OM related infections. Early removal of OMs in patients with poor periodontal health status and/or heavy smokers would have clinical benefits. In addition, preferation of extraoral access to the fracture line might decrease the possibility of plate related infections.

Comparison of clinical parameters, microbiological effects and calprotectin counts in gingival crevicular fluid between Er: YAG laser and conventional periodontal therapies: A split-mouth, single-blinded, randomized controlled trial.

BACKGROUND: The erbium-doped yttrium, aluminum, and garnet (Er:YAG) laser is thought to be the most promising laser for periodontal treatment; however, its application is still under consideration. The aim of this study was to compare Er:YAG laser monotherapy with conventional scaling and root planing (SRP) for chronic periodontitis using clinical parameters, the detection rate of periodontal pathogens, and the calprotectin level in gingival crevicular fluid. METHODS: Twenty-seven participants with moderate-to-advanced chronic periodontitis were included. In a split-mouth design, the 2 half-mouths of each participant were randomly assigned to Er:YAG laser or SRP (combination of ultrasonic and manual instruments) treatment. Clinical parameters were recorded at baseline, 6 weeks, and 3 and 6 months after treatment. At the same time points, gingival crevicular fluid was collected to analyze the detection rate of 6 periodontal pathogens by polymerase chain reaction and the levels of calprotectin by enzyme-linked immunosorbent assay. RESULTS: Both treatment groups showed significant reductions in probing depth (PD), bleeding index (BI), and clinical attachment level (CAL) from baseline to 6 months. For sites with 4 mm ≤ PD ≤ 6 mm at baseline, SRP resulted in a greater reduction in PD and CAL than Er:YAG laser treatment, and the difference remained at 6 months post-treatment (P = .01 and P < .01, respectively). For sites with PD ≥7 mm at baseline, the clinical parameters showed similar results between the 2 groups. SRP resulted in a lower detection rate of Porphyromonas gingivalis at 6 months post-treatment. The levels of calprotectin were significantly decreased from baseline to 6 months in both groups, without a significant difference between the groups. CONCLUSION: For mild pockets, conventional SRP may still be the preferred choice. For deep pockets, Er:YAG laser treatment could be an effective alternative. Studies are needed to explore more advanced instruments and new application methods for the Er:YAG laser for periodontal treatment in deep pockets.

Impact of heavy smoking on the clinical, microbiological and immunological parameters of patients with dental implants: a prospective cross-sectional study.

AIM: The aim of the present study was to investigate how heavy smoking influences the clinical, microbiological, and host-response characteristics in peri-implant sulcus fluid of patients with healthy dental implants. METHODS: A total of 29 individuals with 74 dental implants were included in the present study; 20 implants were in heavy smokers and 54 were in non-smokers. The modified gingival index, modified plaque index, and probing pocket depth were evaluated. Periodontopathogenic bacteria Tannerella forsythia, Treponema denticola, and Porphyromonas gingivalis were evaluated, together with the total bacterial load. Peri-implant sulcus fluid samples were analyzed for the quantification of interleukin-8, interleukin-1ß, interleukin-6, interleukin-10, and tumor necrosis factor-α. RESULTS: No significant differences in the clinical parameters evaluated were found between the groups, although smokers had poorer peri-implant parameters. Among the smokers, subgingival microbiota was composed of a greater number of periodontal pathogens; these differences were not statistically significant. Smokers showed a greater expression of interleukin-1ß, interleukin-6, interleukin-10, and tumor necrosis factor-α, but interleukin-8 was slightly higher among non-smokers, but not significantly. CONCLUSIONS: Although smokers presented deeper probing depths, bleeding on probing, and peri-implant microbiota composed of a greater number of periodontal pathogens than in non-smoking patients, these data did not show significant differences. In the present study, and in relation to the samples analyzed, smoking alone did not influence the immunological and microbiological parameters in dental implants with healthy peri-implant tissues. Further studies with larger samples are required to better evaluate the influence of smoking on dental implants.

Salivary IL-1ß and red complex bacteria as predictors of the inflammatory status in sub-peri-implant niches of subjects with peri-implant mucositis.

BACKGROUND AND OBJECTIVES: Salivary biomarkers may enhance diagnostic sensitivity for peri-implant disease assessment. This study aimed to investigate the association of salivary periodontopathogen count and salivary interleukin-1beta (IL-1ß) level with the peri-implant crevicular fluid IL-1ß response at peri-implant mucositis (PM) sites among subjects with differing periodontal disease susceptibility. MATERIALS AND METHODS: Eighty-seven partially edentulous subjects having at least one implant with peri-implant mucositis were included: 40 with history of chronic periodontitis (P) and 47 with no history of periodontitis (NP). Salivary IL-1ß, peri-implant crevicular fluid (PICF) IL-1ß, and salivary red complex pathogen counts were recorded. Subjects were scored according to a threshold salivary pathogen level of more than 5log (10) counts and assigned a "red complex score." Quartiles of salivary and PICF IL-1ß levels were also scored. Area under receiver operating curve (AUC) was computed to predict the highest PICF IL-1ß score using salivary biomarker as predictors and age-adjusted logistic regression performed for the significant predictors. RESULTS: In the NP group, red complex score (AUC = 0.758 P = 0.010) (odds ratio = 1.377) and salivary IL-1ß (AUC = 0.708 P = 0.038) (odds ratio = 2.506) were significant predictors of highest PICF IL-1ß quartile score. In the P group, no significant associations were noted. CONCLUSIONS: Salivary biomarkers could distinguish the "high" pro-inflammatory responders at PM sites only in subjects without inherent periodontal disease susceptibility. Periodontal susceptibility may impact the immuno-inflammatory response in sub-peri-implant niches of those with peri-implant mucositis.

Chlamydia pneumoniae Clinical Isolate from Gingival Crevicular Fluid: A Potential Atherogenic Strain.

Chlamydia pneumoniae has been associated to atherosclerotic cardiovascular diseases. The aim of our study was to characterize, for the first time, a C. pneumoniae strain isolated from the gingival crevicular fluid of a patient with chronic periodontitis, described as a risk factor for cardiovascular diseases. C. pneumoniae isolate was characterized and compared to the respiratory AR-39 strain by VD4-ompA genotyping and by investigating the intracellular growth in epithelial and macrophage cell lines and its ability to induce macrophage-derived foam cells. Inflammatory cytokine levels were determined in the gingival crevicular fluid sample. C. pneumoniae isolate showed a 99% similarity with the AR-39 strain in the VD4-ompA gene sequence and shared a comparable growth kinetic in epithelial cells and macrophages, as evidenced by the infectious progeny and by the number of chlamydial genomic copies. C. pneumoniae isolate significantly increased the number of foam cells as compared to uninfected and LDL-treated macrophages (45 vs. 6%, P = 0.0065) and to the AR-39 strain (45 vs. 30%, P = 0.0065). Significantly increased levels of interleukin 1-ß (2.1 ± 0.3 pg/µL) and interleukin 6 (0.6 ± 0.08 pg/µL) were found. Our results suggest that C. pneumoniae may harbor inside oral cavity and potentially be atherogenic, even though further studies will be needed to clarify the involvement of C. pneumoniae in chronic periodontitis as a risk factor for cardiovascular diseases.

Periodontal disease and bone pathogenesis: the crosstalk between cytokines and porphyromonas gingivalis

Periodontal disease is the most frequent cause of tooth loss among adults. It is defined as a plaque-induced inflammation of the periodontal tissues that results in a loss of support of the affected teeth. This process is characterized by destruction of the periodontal attachment apparatus, increased bone resorption with loss of crestal alveolar bone, apical migration of the epithelial attachment, and formation of periodontal pockets. Although the presence of periodontal pathogens such as Porphyromonas gingivalis is a prerequisite, the progression of periodontal disease is dependent on the host response to pathogenic bacteria that colonize the tooth surface. Nowadays, a growing body of literature has accumulated to investigate the association between bone diseases, periodontal pathogens and periodontal diseases. The integration of pathogen-associated molecular patterns from microorganisms with their surface receptors in the immune cells, induces the production of several cytokines and chemokines that present either a pro- and/or anti-inflammatory role and the activation of mechanisms of controlling this and the related disease, such as osteoporosis and rheumatoid arthritis. This review focuses on the evidence and significance of bone host cell invasion by Porphyromonas gingivalis in the pathogenesis of bone disorders, as well as the different lines of evidence supporting the role of cytokines in bone diseases.

Alcohol Consumption and Periodontitis: Quantification of Periodontal Pathogens and Cytokines.

BACKGROUND: There are few studies on periodontal status related to microbiologic and immunologic profiles among individuals not or occasionally using alcohol and those with alcohol dependence. The aim of this study is to determine the effect of alcohol consumption on the levels of subgingival periodontal pathogens and proinflammatory cytokines (interleukin [IL]-1ß and tumor necrosis factor [TNF]-α) in the gingival fluid among individuals with and without periodontitis. METHODS: This observational analytic study includes 88 volunteers allocated in four groups (n = 22): individuals with alcohol dependence and periodontitis (ADP), individuals with alcohol dependence and without periodontitis (ADNP), individuals not or occasionally using alcohol with periodontitis (NAP), and individuals not or occasionally using alcohol without periodontitis (NANP). Levels of Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Eikenella corrodens, and Fusobacterium nucleatum were determined by real-time polymerase chain reaction on the basis of the subgingival biofilm, and IL-1ß and TNF-α were quantified by enzyme-linked immunosorbent assay in gingival fluid samples. RESULTS: Individuals with alcohol dependence showed worse periodontal status and higher levels of P. intermedia, E. corrodens, F. nucleatum, and IL-1ß than non-users. No significant correlations between TNF-α and bacterial levels were observed. However, in the ADP group, higher levels of E. corrodens were correlated with higher levels of IL-1ß. CONCLUSION: A negative influence of alcohol consumption was observed on clinical and microbiologic periodontal parameters, as well as a slight influence on immunologic parameters, signaling the need for additional studies.

Cytokine expression in peri-implant crevicular fluid in relation to bacterial presence.

AIM: The aim was to assess clinical inflammatory parameters, cytokine levels and bacterial counts in samples from implant crevicular fluid in cases with untreated peri-implantitis. MATERIAL AND METHODS: Several bacterial species known to up-regulate pro-inflammatory cytokines have been associated with peri-implantitis. The Luminex magnet bead technology was used to study cytokines in crevicular fluid. The checkerboard DNA-DNA hybridization method was used to study bacterial counts in samples from 41 implants (41 individuals). RESULTS: Profuse bleeding and suppuration was found in 25/41 (61.0%) of the implants. The reliability of duplicate cytokine processing was high. In the presence of profuse bleeding, higher pg/ml levels of IL-1ß (p = 0.02), IL-8 (p = 0.04), TNF-α (p = 0.03) and VEGF (p = 0.004) were found. Higher concentrations of IL-1ß were found in the presence of suppuration, and if Escherichia coli (p = 0.001) or Staphylococcus epidermidis (p = 0.05) could be detected. CONCLUSION: Profuse bleeding and/or suppuration in untreated peri-implantitis can be associated with higher concentrations of IL-1ß, IL-8, TNF-α and VEGF in peri-implant crevicular fluid. A higher concentration of IL-1ß in peri-implant crevicular fluid was found in samples that were positive for E. coli or S. epidermidis.

Periodontal status and pathogenic bacteria after gastric bypass: a cohort study.

AIM: The aim this study was to evaluate the influence of gastric bypass surgery (GBS) on periodontal disease and quantify the periodontopathogenic bacteria in patients undergoing this surgery. MATERIAL AND METHODS: This prospective study was composed of 50 patients who underwent bariatric surgery and the data collection was performed in three periods pre-operative, 6 (6M) and 12 months (12 M) postoperative. The oral clinical examination to assess periodontal disease; gingival fluid sample collection for quantification of the periodontopathogenic bacteria Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, and Prevotella intermedia using q-PCR; body mass index (BMI) and for collection of the individual's health-related data from medical files. RESULTS: There was a significant reduction in serum C-reactive protein (CRP) and glucose levels after surgery. The mean probing pocket depth (PPD) and clinical attachment level (CAL) increased significantly in the postoperative period of 6 months (p = 0.001). In the same period, the amount of P. gingivalis increased (p = 0.028) and the other bacteria decreased slightly (p > 0.050). In the presence of P. gingivalis, T. forsythia, T. denticola and P. intermedia, a poor periodontal condition was observed. CONCLUSION: The periodontal disease increased in severity and P. gingivalis increased after GBS. A systemic inflammation resolution due to bariatric surgery in obese subjects does not seem to affect the course of periodontal disease.

Clinical, microbiological, and immunological aspects of healthy versus peri-implantitis tissue in full arch reconstruction patients: a prospective cross-sectional study.

BACKGROUND: Due to the world-wide increase in treatments involving implant placement, the incidence of peri-implant disease is increasing. Late implant failure is the result of the inability to maintain osseointegration, whose most important cause is peri-implantitis. The aim of this study was to analyze the clinical, microbiological, and immunological aspects in the peri-implant sulcus fluid (PISF) of patients with healthy dental implants and patients with peri-implantitis. METHODS: PISF samples were obtained from 24 peri-implantitis sites and 54 healthy peri-implant sites in this prospective cross-sectional study. The clinical parameters recorded were: modified gingival index (mGI), modified plaque index (mPI) and probing pocket depth (PPD). The periodontopathogenic bacteria Tannerella forsythia, Treponema denticola and Porphyromonas gingivalis were evaluated, together with the total bacterial load (TBL). PISF samples were analyzed for the quantification of Interleukin (IL)-8, IL-1ß, IL-6, IL-10 and Tumor Necrosis Factor (TNF)-α using flow cytometry (FACS). RESULTS: The mGI and PPD scores in the peri-implantitis group were significantly higher than the healthy group (p < 0.001). A total of 61.5% of the patients with peri-implantitis had both arches rehabilitated, compared with 22.7% of patients with healthy peri-implant tissues; there was no implant with peri-implantitis in cases that received mandibular treatment exclusively (p < 0.05). Concentrations of Porphyromonas gingivalis (p < 0.01), association with bacteria Porphyromonas gingivalis and Treponema denticola (p < 0.05), as well as the TBL (p < 0.05) are significantly higher in the peri-implantitis group. IL-1ß (p < 0.01), IL-6 (p < 0.01), IL-10 (p < 0.05) and TNF-α (p < 0.01) are significantly higher at the sites with peri-implantitis compared to healthy peri-implant tissue, while IL-8 did not increase significantly. CONCLUSION: The results of the present study involving a limited patient sample suggest that the peri-implant microbiota and which dental arch was rehabilitated involved could contribute to bone loss in peri-implantitis. A significant relationship is observed between the concentration of cytokines (interleukins 1ß, 6 and 10 and TNF-α) and the inflammatory response in peri-implantitis tissue.

Smoking decreases structural and functional resilience in the subgingival ecosystem.

AIMS: Dysbiotic microbial communities underlie the aetiology of several oral diseases, especially in smokers. The ability of an ecosystem to rebound from the dysbiotic state and re-establish a health-compatible community, a characteristic known as resilience, plays an important role in susceptibility to future disease. The present investigation was undertaken to examine the effects of smoking on colonization dynamics and resilience in marginal and subgingival biofilms. MATERIALS AND METHODS: Marginal and subgingival plaque and gingival crevicular fluid samples were collected from 25 current and 25 never smokers with pre-existing gingivitis at baseline, following resolution, after 1, 2 4, 7, 14 and 21 days of undisturbed plaque formation and following resolution. 16S cloning and sequencing was used for bacterial identification and multiplexed bead-based flow cytometry was used to quantify the levels of 27 immune mediators. RESULTS: Smokers demonstrated an early pathogenic colonization that led to sustained pathogen enrichment with periodontal and respiratory pathogens, eliciting a florid immune response. Smokers also demonstrated greater abundance of pathogenic species, poor compositional correlation between marginal and subgingival ecosystems, and significantly greater pro-inflammatory responses following resolution of the second episode of disease. CONCLUSIONS: The ability of the subgingival microbiome to "reset" itself following episodes of disease is decreased in smokers, thereby lowering the resilience of the ecosystem and decreasing its resistance to future disease.