Comparative molecular analysis of bacterial species associated with periodontal disease.

Periodontal disease is an inflammatory disorder affecting the supporting teeth structures, including gingiva, periodontal ligament and alveolar bone, causing loss of connective tissue, reabsorption of alveolar bone and formation of periodontal pockets. The aim of this study is to find a correlation between bacterial growth and periodontal disease. Fifty-seven patients aged between 21 and 65 years, median age 46 years, were enrolled. According to gingival pocket depth, ranging from 3 to 7 mm, patients were divided into two groups: the first (30 patients, 53%) with deep pockets ³ 5 mm and the second (27 patients, 47%) less than 5 mm. The samples taken were processed for microbiological analysis by absolute quantitative real-time Taq-Man technique. Patients affected by periodontal disease were 32 (56%) and patients with gingival bleeding were 35 (61%). This data showed that the presence, the type and the bacterial load in gingival pockets were strongly correlated with gingival depth, periodontal disease and gingival bleeding. Quantitative microbiological analysis is a key point to improve patient compliance, allowing to choose the specific antibiotic treatment. avoiding antibiotic resistance and ensuring the successful outcome of therapy for periodontal disease.

Complementary clinical effects of red complex bacteria on generalized periodontitis in a caucasian population.

OBJECTIVES: To relate five periodontopathogenic bacteria, including the red complex, to the severity, extent, and inflammation of the periodontal lesion in Caucasian patients with generalized aggressive and chronic periodontitis and to explore whether tobacco use is associated with a specific bacterial profile. MATERIALS AND METHODS: A cross-sectional and analytic study was conducted in patients with aggressive and chronic periodontitis. Data were gathered on socio-demographic and periodontal variables, and RH-PCR was used to determine subgingival bacterial profile. Linear and logistic regression analyses were performed. RESULTS: The study included 60 patients with aggressive and 123 with chronic periodontitis. Total red complex bacteria count was higher in aggressive periodontitis, mainly due to T. denticola (P = 0.015). In both periodontitis types, models showed an association between T. forsythia count and probing depth (B = 0.157, P = 0.030) and between T. denticola count and higher bleeding scores (B = 2.371, P = 0.027). Smoking did not affect the red complex bacteria count in either disease. CONCLUSIONS: The prevalence of red complex bacteria was similar between aggressive and chronic periodontitis, but their count was higher in the former. In both diseases, T. forsythia was associated with greater severity and T. denticola with more severe bleeding. Tobacco smoking was not associated with the presence of red complex bacteria in either disease.

Clinical and antimicrobial efficacy of a controlled-release device containing chlorhexidine in the treatment of chronic periodontitis.

A controlled-release device (CRD) containing chlorhexidine gluconate, such as PerioCol(™)CG (Eucare Pharmaceuticals Pvt. Ltd,, Chennai, India), for subgingival application has little reported data with clinical as well as antimicrobial efficacy. This study evaluated clinical and subgingival microbial changes on using indigenously developed PerioCol™CG as an adjunct to scaling and root planing (SRP) in the treatment of chronic periodontitis. Forty posterior first molar sites having probing pocket depth ≥ 5 mm were selected and divided into two groups, with 20 sites in each group, in a split-mouth design. Group A (test site) was treated with SRP and PerioCol(™)CG, while group B (control site) was treated with SRP alone. Subgingival microbial samples were collected at baseline and 1 month after the initial SRP, while probing depth (PD), clinical attachment level (CAL) and gingival index (GI) were recorded at baseline, after 1 month and after 3 months. Microbial detection of Porphyromonas gingivalis (P. gingivalis) and Tannerella forsythia (T. forsythia) was done by means of polymerase chain reaction (PCR). A significant improvement was observed in all clinical measures in sites treated with PerioCol(™)CG as compared to the control sites during the study period. Also, there was a statistically significant reduction in the proportion of occurrence of P. gingivalis and T. forsythia after intervention in test sites as compared to control sites. Our data suggest that SRP combined with subgingival administration of PerioCol™CG has a significantly better and prolonged effect compared to SRP alone on the PD, clinical attachment loss and elimination of periodontopathogens.

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.

Response of chronic gingivitis to hygiene therapy and experimental gingivitis. Clinical, microbiological and metabonomic changes.

PURPOSE: To compare the clinical, microbiological and metabonomic profiles of subjects with high and low levels of chronic gingival bleeding during a controlled oral hygiene regimen intervention including sequential phases of rigorous therapeutic oral hygiene followed by experimental gingivitis (EG). METHODS: Two cohorts of qualified study subjects with differences in gingival bleeding on probing levels at their baseline clinical examination were entered into the study. These two cohorts were followed through three separate study phases including a 1-week baseline phase, a 2-week phase of rigorous oral hygiene including dental prophylaxis, and a 3-week EG phase of no oral hygiene to encourage relapse of gingivitis. The 58 subjects were assessed during each phase of the study for clinical presentation of gingivitis and concurrently had plaque sampled for real-time polymerase chain reaction (RTPCR) microbiological characterization and salivary lavage samples for 'systems biology' metabonomics assessment by 1H-NMR. RESULTS: Subjects presenting with different levels of gingival bleeding on probing when they entered the study responded differently to rigorous oral hygiene and EG. Specifically, the high bleeding cohort responded sluggishly to rigorous oral hygiene and exhibited markedly greater relapse to gingivitis during EG. RTPCR analysis showed changes in bacterial populations that were associated with study phases, particularly the increases in putative periodontal pathogens during EG. However, the microbiological profiles of high- and low-susceptibility gingival bleeding patients were largely similar. Metabonomic analysis likewise revealed significant changes in metabolite composition during study phases associated with differences in plaque toxicity, especially the short chain carboxylic acids propionate and n-butyrate, which tracked clinical changes in gingivitis severity. Systems analysis of metabonomic changes suggested differences between cohorts, although analysis to date has not elucidated whether these differences are causative (population predictive) or simply diagnostic of clinical status within populations.

Clinical and microbiological efficacy of systemic roxithromycin as an adjunct to non-surgical periodontal therapy in treatment of chronic periodontitis. A randomized, double-blinded, placebo-controlled clinical trial.

PURPOSE: The objective of this randomized clinical trial was to evaluate the clinical and microbiological effects of systemic administration of roxithromycin (RXM) as an adjunct to non-surgical periodontal therapy (NSPT) in the treatment of individuals with moderate to severe chronic periodontitis (CP). METHODS: 70 individuals (38 males and 32 females, aged 25 to 60 years) with moderate to severe CP were randomly allocated into two groups. 35 individuals were allocated to full mouth SRP+RXM while 35 individuals were allocated to SRP+ Placebo group. The clinical parameters evaluated were probing depth (PD), clinical attachment level (CAL), gingival index (GI), plaque index (PI) and % bleeding on probing sites (%BOP) at baseline (B/L), 1-, 3- and 6-month intervals while microbiologic parameters included percentage of sites positive for periodontopathic bacteria A. actinomycetemcomitans, P. gingivalis and T. forsythia at B/L, 3 and 6 months using polymerase chain reaction. RESULTs: Both groups showed improved clinical and microbiologic parameters over 6 months. RXM group showed a statistically significant reduction in mean PD and CAL gain as compared to the placebo group (P < 0.0001). There was reduction in percentage of sites positive for periodontopathic bacteria over the duration of the study in both groups and a statistically significant reduction in the number of sites positive for A. actinomycetemcomitans in RXM group (P < 0.001).

Effects of doxycycline on clinical, microbiological and immunological parameters in well-controlled diabetes type-2 patients with periodontal disease: a randomized, controlled clinical trial.

AIM: To evaluate the clinical, microbiological and immunological effects of systemic doxycycline as an adjunct to scaling and root planing (SRP) in chronic periodontitis patients with well-controlled type 2 diabetes. MATERIALS AND METHODS: Sixty-six patients compliant to oral hygiene (Hygiene Index <20%) allocated to either a test (systemic doxycycline for 21 days) or a control (placebo) group participated in the present randomized controlled trial (RCT). Clinical assessments were recorded at baseline, 3 and 6 months after therapy and included clinical attachment level (CAL), set as the primary outcome of the study, probing pocket depth (PPD), recession (RE) and bleeding on probing (BOP). At the same time points, counts of 15 subgingival species were evaluated by "checkerboard" DNA-DNA hybridization, gingival crevicular fluid samples were analysed for matrix metalloproteinase-8 (MMP-8) by ELISA and HbA1c levels were determined. Comparisons between and within groups were performed by non-parametric tests (Mann-Whitney, Wilcoxon signed-ranks and z-test for proportions with Bonferroni corrections) at the 0.05 level. RESULTS: No major differences were noticed in clinical and microbiological parameters of periodontal disease or levels of MMP-8 between the two groups. CONCLUSIONS: Adjunctive systemic doxycycline does not seem to significantly enhance the effects of SRP in well-controlled diabetes type 2 patients.

Recovery of putative periodontal pathogens from curette sampling at different depths of periodontal lesions: an in vivo cross-sectional clinical study.

OBJECTIVE: The aim of this study was to evaluate the effect of the depth of curette sample collection from periodontal lesions on the recovery of putative periodontal pathogens using real-time polymerase chain reaction (PCR). METHODS: Twenty-two periodontal pockets 6 to 8 mm deep with bleeding on probing at a single-rooted tooth were sampled, yielding 66 separate samples. Curette samples were obtained at three different levels of the periodontal lesion (orifice, shallow--2 mm into the pocket; or base of lesion), and processed using PCR to identify 10 periodontal pathogens. The chi-square procedure was used to determine whether probe depth affected the distribution of bacterial counts observed. A repeated measures analysis of variance tested the hypotheses related to level of probe and microorganism on mean rank of bacterial counts. RESULTS: The effect of probe level on mean bacterial counts depends on the type of microorganism. Likewise, the effect of microorganism type on mean bacterial counts significantly depends on probe level, where sampling from 2 mm into the periodontal pocket was found to yield significantly higher results than sampling from the orifice. Overall mean counts of pathogenic microorganisms were found to differ significantly across the three probe depths. The microorganisms differed in their observed levels over all three probe levels. Further analysis found several significant differences that characterize the nature of the interaction between probe level and microorganism type. CONCLUSION: There is significant difference in the amount of putative periodontal pathogens at varying depths of the pocket when sampled with a periodontal curette.

Clinical and microbiological effects of levofloxacin in the treatment of Aggregatibacter actinomycetemcomitans-associated periodontitis: a randomized placebo-controlled clinical trial.

AIM: To evaluate the clinical and microbiological effects of systemic levofloxacin (LFX) in subjects with Aggregatibacter actinomycetemcomitans-associated chronic periodontitis (AA-ACP). MATERIALS AND METHODS: Subjects with severe periodontitis with subgingival detection of A. actinomycetemcomitans were randomly divided into two treatment groups; a test group (n = 35) that received scaling and root planing (SRP) and LFX (500 mg o.d.) and a control group (n = 34) that received SRP and placebo (o.d.) for 10 days. Plaque index (PI), gingival index (GI), percent of sites with bleeding on probing (% BoP), probing depth (PD) and clinical attachment level (CAL) were recorded and subgingival plaque samples were cultivated for detection of A. actinomycetemcomitans at baseline to 6 months at various intervals. RESULTS: Subjects receiving LFX showed the greatest improvements in mean PD and CAL. The difference in the reduction of PD and CAL in the two groups was significant at 1, 3 and 6 months for PD and 3 and 6 months for CAL (p < 0.05). The inter-group difference in PI, GI and % BoP was not significant at any interval. Detectable levels of A. actinomycetemcomitans were significantly less in the test group 3 and 6 months post-therapy. CONCLUSION: Systemic LFX as an adjunct to SRP improves clinical outcomes and suppresses A. actinomycetemcomitans below detectable levels.

Effect of 2 elastomeric ligatures on microbial flora and periodontal status in orthodontic patients.

INTRODUCTION: The aim of this study was to compare the effects of a nonconventional elastomeric ligature (Slide; Leone, Florence, Italy) with those of a conventional elastomeric ligature (Ormco, Orange, Calif) on microbial flora and periodontal status in orthodontic patients. METHODS: A total of 13 orthodontic patients scheduled for fixed orthodontic treatment were selected for this study. The use of Slide and conventional elastomeric ligatures in fixed orthodontic appliances was tested. Microbial and periodontal records were obtained before bonding and 1 and 5 weeks after bonding. For the statistical analysis and calculations, SPSS software (version 15.0; SPSS, Chicago, Ill) was used. In the statistical decisions, P <0.05 values were accepted as significantly different. RESULTS: No significant differences between Slide and conventional elastomeric ligatures were evident at 1 week or 5 weeks after bonding, with regard to gingival index, plaque index, gingival bleeding index, or pocket depth scores (P >0.05). Similarly, aerobic and anaerobic bacteria counts did not differ significantly on the surface or on the elastics (P >0.05). CONCLUSIONS: Although the Slide ligatures cover the total surface of the bracket, they do not cause significantly more plaque accumulation or periodontal problems than do the conventional elastomeric ligatures.

Prevalence and diversity of Synergistetes taxa in periodontal health and disease.

BACKGROUND AND OBJECTIVE: Members of the phylum Synergistetes have previously been identified within periodontitis subgingival plaque and are considered putative periodontopathogens. This study compared the diversity of subginigval Synergistetes in a cohort of subjects with periodontitis (n = 10) vs. periodontitis-free controls (n = 10). MATERIAL AND METHODS: Pooled subgingival plaque samples from all deep periodontal pockets or all sulci were collected from the periodontitis and periodontitis-free subjects, respectively. Bacterial 16S rRNA genes were PCR-amplified from purified subgingival plaque DNA using a Synergistetes 'selective' primer set. PCR products were cloned and sequenced to analyze the prevalence and diversity of Synergistetes operational taxonomic units (OTUs) present in plaque samples of both subject groups. RESULTS: A total of 1030 non-chimeric 16S rRNA clones were obtained, of which 162 corresponded to members of the phylum Synergistetes. A significantly larger number of Synergistetes clones were obtained from periodontitis subgingival plaque than from periodontitis-free controls (25.4% vs. 5.9%, p < 0.001). All Synergistetes clones corresponded to cluster A oral Synergistetes, and fell into 31 OTUs (99% sequence identity cut-off). Twenty-nine Synergistetes OTUs were detected in the periodontitis group while eight were detected in the periodontitis-free group (p < 0.001). Five Synergistetes OTUs; including one OTU corresponding to the recently-characterized species Fretibacterium fastidiosum, were more prevalent in the periodontitis subjects (p < 0.05). CONCLUSION: OTUs belonging to oral Synergistetes cluster A were more readily detectable and were more diverse in subgingival plaque from periodontitis subjects compared with periodontitis-free controls. Specific Synergistetes OTUs appear to be associated with periodontitis.

Evaluation of three selective media for isolation of Aggregatibacter actinomycetemcomitans.

BACKGROUND AND OBJECTIVE: Aggregatibacter actinomycetemcomitans is a pathogen in oral and nonoral infections. Detection and quantification of this pathogen can be performed using selective culture techniques. The aim of this study was to establish the efficacy of two known selective media in their ability to select and support the growth of A. actinomycetemcomitans. MATERIAL AND METHODS: Trypticase soy bacitracin vancomycin (TSBV) medium and brain-heart infusion agar with vancomycin (Dentaid-1), as well as a modified Dentaid-1 medium (in which the brain-heart infusion agar was substituted with brain-heart infusion broth), were compared. Two-hundred and eighteen clinical samples were used to establish the recovery rate, the number of colony-forming units (CFUs) of A. actinomycetemcomitans as well as the total number of CFUs on the three different types of medium. In addition, the numbers of gram-negative aerobic rods and yeasts were determined. RESULTS: Both types of Dentaid-1 medium showed a higher recovery of A. actinomycetemcomitans compared with TSBV. However, these differences did not reach statistical significance. The total number of CFUs of A. actinomycetemcomitans recovered was significantly higher on Dentaid-1 compared with TSBV (p = 0.029). The mean number of gram-negative aerobic rods recovered was statistically higher on both types of Dentaid-1 medium in comparison with TSBV. Low numbers of yeasts were recovered occasionally on all test plates. CONCLUSION: Dentaid-1 is a low-cost effective alternative to TSBV for the isolation and growth of A. actinomycetemcomitans from clinical samples, such as dental plaque, which contain a complex microflora.

Comprehensive microbiological findings in peri-implantitis and periodontitis.

AIM: The microbial differences between peri-implantitis and periodontitis in the same subjects were examined using 16S rRNA gene clone library analysis and real-time polymerase chain reaction. MATERIALS AND METHODS: Subgingival plaque samples were taken from the deepest pockets of peri-implantitis and periodontitis sites in six subjects. The prevalence of bacteria was analysed using a 16S rRNA gene clone library and real-time polymerase chain reaction. RESULTS: A total of 333 different taxa were identified from 799 sequenced clones; 231 (69%) were uncultivated phylotypes, of which 75 were novel. The numbers of bacterial taxa identified at the sites of peri-implantitis and periodontitis were 192 and 148 respectively. The microbial composition of peri-implantitis was more diverse when compared with that of periodontitis. Fusobacterium spp. and Streptococcus spp. were predominant in both peri-implantitis and periodontitis, while bacteria such as Parvimonas micra were only detected in peri-implantitis. The prevalence of periodontopathic bacteria was not high, while quantitative evaluation revealed that, in most cases, prevalence was higher at peri-implantitis sites than at periodontitis sites. CONCLUSIONS: The biofilm in peri-implantitis showed a more complex microbial composition when compared with periodontitis. Common periodontopathic bacteria showed low prevalence, and several bacteria were identified as candidate pathogens in peri-implantitis.

Differential expression of mitogen activating protein kinases in periodontitis.

AIM: Following toll-like receptor (TLR) engagement, lipopolysaccharide (LPS) can stimulate the expression of pro-inflammatory cytokines thus activating the innate immune response. The production of inflammatory cytokines results, in part, from the activation of kinase-induced signalling cascades and transcriptional factors. Of the four distinct classes of mitogen-activated protein kinases (MAPK) described in mammals, p38, c-Jun N-terminal activated kinases (JNK1-3) and extracellular activated kinases (ERK1,2) are the best studied. Previous data have established that p38 MAPK signalling is required for inflammation and bone loss in periodontal disease pre-clinical animal models. MATERIALS & METHODS: In this study, we obtained healthy and diseased periodontal tissues along with clinical parameters and microbiological parameters. Excised fixed tissues were immunostained with total and phospho-specific antibodies against p38, JNK and ERK kinases. RESULTS: Intensity scoring from immunostained tissues was correlated with clinical periodontal parameters. Rank correlations with clinical indices were statistically significantly positive (p-value < 0.05) for total p38 (correlations ranging 0.49-0.68), phospho-p38 (range 0.44-0.56), and total ERK (range 0.52-0.59) levels, and correlations with JNK levels also supported association (range 0.42-0.59). Phospho-JNK and phospho-ERK showed no significant positive correlation with clinical parameters of disease. CONCLUSION: These data strongly implicate p38 MAPK as a major MAPK involved in human periodontal inflammation and severity.

Clinical and subclinical effects of power brushing following experimental induction of biofilm overgrowth in subjects representing a spectrum of periodontal disease.

AIM: Investigate short-term effects of power brushing following experimental induction of biofilm overgrowth in periodontal disease states. MATERIALS AND METHODS: Overall, 175 subjects representing each of five biofilm-gingival interface (BGI) periodontal groups were enrolled in a single-blind, randomized study. After stent-induced biofilm overgrowth for 21 days subjects received either a manual or a power toothbrush to use during a 4 weeks resolution phase. At baseline and during induction and resolution, standard clinical parameters were measured. Subclinical parameters included multikine analysis of 13 salivary biomarkers and 16s Human Oral Microbe Identification Microarray (HOMIM) probe analysis of subgingival plaque samples. RESULTS: All groups exhibited significantly greater reductions in bleeding on probing (BOP) (p = 0.002), gingival index (GI) (p = 0.0007), pocket depth (PD) (p = 0.04) and plaque index (p = 0.001) with power brushing compared to manual. When BGI groups were combined to form a shallow PD (PD ≤ 3 mm) and a deep PD group (PD > 4 mm) power brushing reduced BOP and GI in subjects with both pocket depths. Power brushing significantly reduced IL-1ß levels at resolution while changes in bacterial levels showed non-significant trends between both brushing modalities. CONCLUSIONS: Short-term changes in select clinical parameters and subclinical salivary biomarkers may be useful in assessing efficacy of power brushing interventions in a spectrum of periodontal disease states.

Subgingival root brushing in deep human periodontal pockets.

OBJECTIVE: The short-term clinical and microbiological effects of patient-applied subgingival root brushing were assessed on untreated deep human periodontal pockets. METHODS: Assessments of plaque, bleeding on probing, probing depth, total cultivable subgingival counts, and cultivable counts and proportions of six putative periodontal pathogens were carried out at baseline and after 14 days on two contralateral > or = 6 mm bleeding interproximal posterior sites in each of 11 adults with untreated chronic periodontitis. One of the sites was randomly assigned to daily patient-applied subgingival root brushing for 14 days, and the other to remain with the patient's pre-existing tooth brushing and flossing regimen. No other periodontal therapy was performed during the 14 test days. RESULTS: Significant reductions in plaque, bleeding on probing, probing depth, total subgingival counts, and levels of putative periodontal pathogens were found after 14 days of subgingival root brushing. Subgingival root brushing nearly eliminated bleeding on probing at test sites, reduced probing depths by a mean of 1.8 mm, and reduced cultivable subgingival proportions of six evaluated putative periodontal pathogens from a cumulative total of 14.1% to 0.8%. In comparison, no significant clinical or microbiological changes were detected after 14 days where the patient's pre-existing oral hygiene regimen remained unaltered. CONCLUSIONS: Subgingival root brushing over 14 days, in properly trained patients, induced favorable clinical and microbiological changes in deep periodontal pockets > or = 6 mm even in the absence of professional subgingival debridement.

Clinical and microbiological results following nonsurgical periodontal therapy with or without local administration of piperacillin/tazobactam.

OBJECTIVES: We assessed if adjunct administration of piperacillin/tazobactam added clinical and microbiological treatment benefits. MATERIALS AND METHODS: Thirty-six subjects (mean age 52.1 years (SD ± 10.3)) (NS by group) with chronic periodontitis were randomly enrolled receiving subgingival debridement and the local administration of piperacillin/tazobactam (test group) or debridement alone (control group). Bleeding on probing (BOP), probing pocket depth (PPD), and microbiological counts of 74 species were studied by checkerboard DNA-DNA hybridization up to month 6 after treatment. RESULTS: Mean PPD changes between baseline and month 6 in the test and control groups were 1.5 and 1.8 mm, respectively (NS between groups). BOP in both groups decreased from about 80 to 40 %. At 4 and 12 weeks, lower counts of the following bacteria were found in the test group (site level): Fusobacterium species, Parvimonas micra, Pseudomonas aeruginosa, Staphylococcus aureus, Tannerella forsythia, Treponema denticola, and a composite load of nine pathogens (p < 0.001). At week 26, subjects receiving local antibiotics had a lower prevalence at tested sites for Fusobacterium nucleatum sp. polymorphum, Fusobacterium periodonticum, P. micra, and T. denticola. CONCLUSIONS: At 26 weeks, treatment with or without piperacillin/tazobactam resulted in similar BOP and PPD improvements. At week 26 and at the subject level, the prevalence of 4/74 pathogens was found at lower counts in the group receiving local antibiotics. CLINICAL RELEVANCE: Administration of piperacillin/tazobactam reduces the prevalence of Fusobacterium, P. micra, and T. denticola to a greater extent than debridement alone but with no short-term differences in PPD or BOP.

Gingival fluid cytokine expression and subgingival bacterial counts during pregnancy and postpartum: a case series.

OBJECTIVES: The aim of this study was to assess gingival fluid (GCF) cytokine messenger RNA (mRNA) levels, subgingival bacteria, and clinical periodontal conditions during a normal pregnancy to postpartum. MATERIALS AND METHODS: Subgingival bacterial samples were analyzed with the checkerboard DNA-DNA hybridization method. GCF samples were assessed with real-time PCR including five proinflammatory cytokines and secretory leukocyte protease inhibitor. RESULTS: Nineteen pregnant women with a mean age of 32 years (S.D. ± 4 years, range 26-42) participated in the study. Full-mouth bleeding scores (BOP) decreased from an average of 41.2% (S.D. ± 18.6%) at the 12th week of pregnancy to 26.6% (S.D. ± 14.4%) at the 4-6 weeks postpartum (p < 0.001). Between week 12 and 4-6 weeks postpartum, the mean probing pocket depth changed from 2.4 mm (S.D. ± 0.4) to 2.3 mm (S.D. ± 0.3) (p = 0.34). Higher counts of Eubacterium saburreum, Parvimonas micra, Selenomonas noxia, and Staphylococcus aureus were found at week 12 of pregnancy than at the 4-6 weeks postpartum examinations (p < 0.001). During and after pregnancy, statistically significant correlations between BOP scores and bacterial counts were observed. BOP scores and GCF levels of selected cytokines were not related to each other and no differences in GCF levels of the cytokines were observed between samples from the 12th week of pregnancy to 4-6 weeks postpartum. Decreasing postpartum counts of Porphyromonas endodontalis and Pseudomonas aeruginosa were associated with decreasing levels of Il-8 and Il-1ß. CONCLUSIONS: BOP decreased after pregnancy without any active periodontal therapy. Associations between bacterial counts and cytokine levels varied greatly in pregnant women with gingivitis and a normal pregnancy outcome. Postpartum associations between GCF cytokines and bacterial counts were more consistent. CLINICAL RELEVANCE: Combined assessments of gingival fluid cytokines and subgingival bacteria may provide important information on host response.

Connective tissue graft plus resin-modified glass ionomer restoration for the treatment of gingival recession associated with non-carious cervical lesions: microbiological and immunological results.

OBJECTIVES: It was previously reported the clinical results of placing subgingival resin-modified glass ionomer restoration for treatment of gingival recession associated with non-carious cervical lesions. The aim of this study was to evaluate the influence of this treatment on the subgingival biofilm and gingival crevicular fluid (GCF) inflammatory markers. MATERIALS AND METHODS: Thirty-four patients presenting the combined defect were selected. The defects were treated with either connective tissue graft plus modified glass ionomer restoration (CTG+R) or with connective tissue graft only (CTG). Evaluation included bleeding on probing and probing depth, 5 different bacteria targets in the subgingival plaque assessed at baseline, 45, and 180 days post treatments, and 9 inflammatory mediators were also assessed in the GCF. RESULTS: The levels of each target bacterium were similar during the entire period of evaluation (p > 0.05), both within and between groups. The highest levels among the studied species were observed for the bacterium associated with periodontal health. Additionally, the levels of all cyto/chemokines analyzed were not statistically different between groups (p > 0.05). CONCLUSION: Within the limits of the present study, it can be concluded that the presence of subgingival restoration may not interfere with the subgingival microflora and with GCF inflammatory markers analyzed. CLINICAL RELEVANCE: This approach usually leads to the placement of a subgingival restoration. There is a lack of information about the microbiological and immunological effects of this procedure. The results suggest that this combined approach may be considered as a treatment option for the lesion included in this study.

A randomized clinical trial on the clinical and microbiological efficacy of a xanthan gel with chlorhexidine for subgingival use.

BACKGROUND: The main indication of the adjunctive use of local antimicrobials lies around situations in which the outcome of non-surgical mechanical treatment results in a limited number of residual pockets. The purpose of this investigation was to evaluate the clinical and microbiological effects of the subgingival application of a xanthan-based 1.5% chlorhexidine (CHX) gel (Xan-CHX), adjunctive to scaling and root planing (SRP) in localized periodontitis. METHODS: Periodontitis patients with four to ten residual (after conventional SRP) or relapsing (during supportive periodontal treatment) pockets were recruited and randomized to receive SRP plus the subgingival application of (Xan-CHX) or SRP plus a placebo gel. Supragingival plaque, bleeding on probing (BOP), probing pocket depth (PPD), and clinical attachment level were evaluated with a computerized probe at baseline, and after 1, 3, and 6 months. Subgingival samples were also collected for the microbiological analysis. Statistical analysis used ANOVA and chi-square tests. RESULTS: Overall, the clinical results were better in the test group, with significant changes in BOP (between baseline and 3 months) and with a significant increase in the proportion of shallow pockets (1-3 mm) at 6 months. These results did not result in significant intergroup differences. The microbiological impact was limited in both treatment groups. CONCLUSION: The adjunctive use of Xan-CHX may improve, although to a limited extent, the clinical outcomes (BOP and PPD), in chronic periodontitis patients with "residual" or "relapsing" pockets, but no significant differences were detected between groups. No side effects, neither clinical nor microbiological, were detected after the use of the test product. CLINICAL RELEVANCE: Adjunctive use of slow-released chlorhexidine might be considered in the management of periodontal disease and gingival inflammation to reduce the need for periodontal surgery.