Influence of orthodontic loading on biomarkers levels around miniscrews.

OBJECTIVE: The aim of this study was to evaluate the levels of Interleukin-1α (IL-1α), Interleukin-1ß (IL-1ß), Interleukin-1 receptor antagonist (IL-1Ra), Interleukin-10 (IL-10), Interleukin-13 (IL-13), Vascular endothelial growth factor (VEGF), Granulocyte-colony stimulating factor (G-CSF), and Growth related oncogene (GRO) in the peri-miniscrew implant crevicular fluid (MICF) under orthodontic loading. DESIGN: The study sample comprised 14 miniscrews immediately loaded and 17 unloaded ones. A load of 200gF was immediately applied to the miniscrews in the loaded group after the placement surgery. Peri-miniscrew implant crevicular fluid was collected at baseline, at day 7, and at day 21. The levels of the biomarkers were measured using a multiplexed bead immunoassay. Intergroup comparisons were made using Mann-Whitney test. Friedman and Dunn's multiple comparison tests were used to evaluate intragroup differences over time. RESULTS: Although no statistical differences were observed between the groups at any time point for any of the 8 biomarkers evaluated, there was a statistically significant increase (p < 0.02) in the levels of all the biomarkers over time on both groups. CONCLUSIONS: An immediate loading of 200gF does not alter the balance in the inflammatory response in peri-miniscrew tissues.

Microbial Ecosystem Analysis in Root Canal Infections Refractory to Endodontic Treatment.

INTRODUCTION: The purpose of this study was to combine multiple displacement amplification and checkerboard DNA-DNA hybridization to qualitatively and quantitatively evaluate the microbiota present in infections refractory to endodontic treatment. METHODS: The subjects of this study were 40 patients presenting with periapical lesions refractory to endodontic treatment. Samples were taken by scraping or filing root canal walls with a #10 K-type hand file. Sample DNA was amplified by multiple displacement amplification, and the levels of 107 bacterial taxa were analyzed by checkerboard DNA-DNA hybridization. The taxa were divided into 3 distinct microbial populations depending on their mean proportion in samples (% DNA probe counts ± standard error of the mean) as follows: dominant (≥3.0%), subdominant (>1.6%-3.0%), and residual (≤1.6%) populations. The significance of differences was determined using the Mann-Whitney test. RESULTS: The taxa present with the highest mean proportions (constituting the dominant population) were Corynebacterium diphtheriae (8.03 ± 0.98), Porphyromonas gingivalis (5.42 ± 2.09), Streptococcus sobrinus (5.33 ± 0.69), and Stenotrophomonas maltophilia (4.72 ± 1.73). Among the subdominant population were Eubacterium saphenum (3.85 ± 1.06), Helicobacter pylori (3.16 ± 0.62), Dialister pneumosintes (3.12 ± 1.1), Clostridium difficile (2.74 ± 0.41), Enterobacter agglomerans (2.64 ± 0.54), Salmonella enterica (2.51 ± 0.52), Mobiluncus mulieris (2.44 ± 0.6), and Klebsiella oxytoca (2.32 ± 0.66). In the population of bacteria present at the lowest mean proportions (the residual population), Bacteroides ureolyticus (0.04 ± 0.01), Haemophilus influenzae (0.04 ± 0.02), and Prevotella oris (0.01 ± 0.01) were found at the lowest mean proportions. Enterococcus faecalis was detected in the residual population (0.52 ± 0.26). CONCLUSIONS: The microbial climax community in teeth refractory to endodontic treatment not only harbors medically important species but also contains distinct microbial consortia present with different population levels.

Peri-implant crevicular fluid biomarkers as discriminants of peri-implant health and disease.

AIM: The objective of this cross-sectional study was to examine the potential of peri-implant crevicular fluid (PICF) analytes to discriminate between peri-implant health and disease using a multi-biomarker approach. METHODS: We collected PICF samples from the mesio-buccal site of every implant (n = 145) from 52 subjects with peri-implantitis and measured the levels of 20 biomarkers using Luminex. We grouped implants and subjects based on the clinical characteristic of the sampled sites and implants into: healthy sites from healthy implants (HH), diseased sites from diseased implants (DD) and healthy sites from diseased implants (HD). The significance of the differences between the HH and DD groups was determined using general linear models controlling for false discovery rate. We used logistic regression to determine the best multi-biomarker models that could distinguish HH from DD subjects and HH from HD subjects. RESULTS: There were statistically significant differences between HH and DD groups for 12/20 biomarkers. Logistic regression resulted in a 6-biomarker model (Flt-3L, GM-CSF, IL-10, sCD40L, IL-17 and TNFα) that discriminated HH from DD subjects (AUC = 0.93) and a 3-biomarker model (IL-17, IL-1ra and vascular endothelial growth factor) that distinguished HH from DD subjects (AUC = 0.90). CONCLUSION: PICF biomarkers might help discriminate peri-implant health from disease.

Matrix metalloproteinases -1, -2, -3, -7, -8, -12, and -13 in gingival crevicular fluid during orthodontic tooth movement: a longitudinal randomized split-mouth study.

This randomized split-mouth study aimed to examine the levels of matrix metalloproteinases (MMPs) -1, -2, -3, -7, -8, -12, and -13 in the gingival crevicular fluid (GCF) at different time points during orthodontic tooth movement. A total of 16 healthy orthodontic subjects (7 females, 9 males; mean age, 17.7 years) who needed their first upper premolars extracted were enrolled. One randomly chosen maxillary canine was subjected to a distalizing force and was considered to be the test side. The contralateral canine, which was not subjected to any force but was included in the orthodontic appliance, was used as a control side. GCF sampling was performed at both the mesial (tension) and distal (pressure) test and control sites at baseline, immediately before applying the orthodontic appliance, and after 1 and 24 hours and 7, 14, and 21 days. A multiplexed bead immunoassay was used to analyse the GCF samples. The mean levels of the MMP-1, -2, -3, -7, -8, -12, and -13 were not significantly different between the test and control groups in each time showed. The comparisons between the tension and pressure sites were also not significantly different at each individual time. A few variations focused on MMP-1 and -3, but the expression of MMP-8 was higher than that of the other MMPs. MMPs are released in sufficient quantities such that tooth movement occurs but with no significant increase in GCF levels.

Effects of calcium hydroxide on cytokine expression in endodontic infections.

INTRODUCTION: The use of calcium hydroxide is an effective step in killing bacteria that remain after cleaning and shaping procedures. It also induces hard-tissue formation and is effective for stopping inflammatory exudates. METHODS: The aim of this study was to assay and to compare the influence of calcium hydroxide on periapical interstitial fluid from human root canals. The mRNA expression levels of the cytokines interferon (IFN)-γ, tumor necrosis factor-α, interleukin (IL)-1ß, IL-17A, and IL-10 as well as the chemokine MCP-1 were assayed by real-time polymerase chain reaction immediately after root canal cleaning and 15 days later. RESULTS: Levels of IL-1ß, IFN-γ, IL-10, and the chemokine CCL2/MCP-1 were increased in teeth without endodontic dressings. With calcium hydroxide interappointment dressings, no statistically significant changes were observed in cytokine mRNA expression. However, when comparing teeth that received the medication with those that did not, expression levels of IL-1ß, IFN-γ, and IL-10 were statistically lower in those teeth that received calcium hydroxide. CONCLUSIONS: Analyses of cytokines and the chemokine CCL-2/MCP-1 demonstrated the benefits of calcium hydroxide as a root canal dressing because it impedes the increase of all mediators during the experimental time.

T-lymphocyte and cytokine expression in human inflammatory periapical lesions.

INTRODUCTION: Lymphocytes, among many cells, express different sets of cytokines, chemokines, and receptors, which are considered important mediators of periapical immune response to infection. METHODS: The aim of this study was to evaluate the mRNA expression of CD4(+)CD28(+) and CD8(+) T genes and the gene expression of interferon-γ, tumor necrosis factor-α, interleukin (IL)-1ß, IL-17A, IL-10, CCL2/MCP-1, CCL4, CCL5, CXCR4, CCR5, and receptor activator for nuclear factor kappa B ligand (RANKL) in periapical interstitial fluid from human root canal infections. The samples were collected immediately after root canal cleaning and 7 days later (restrained root canal bacterial load) to characterize those gene expressions. RESULTS: Real-time polymerase chain reaction demonstrated significantly higher levels of CD4(+)CD28(+) and CD8(+) T-cell markers in the former root canal condition and an increase of IL-10 and CXCR4, followed by a decrease of proinflammatory cytokines such as RANKL, interferon-γ, IL-1ß, and CCL5. CONCLUSIONS: Analyses of T-lymphocyte and cytokine expression in periapical area were able to show that distinct root canal conditions might play regulatory roles in controlling local immune/inflammatory processes.

Effects of periodontal therapy on GCF cytokines in generalized aggressive periodontitis subjects.

AIM: To examine changes in levels of gingival crevicular fluid (GCF) cytokines, after periodontal therapy of generalized aggressive periodontitis (GAgP). MATERIALS AND METHODS: Twenty-five periodontally healthy and 24 GAgP subjects had periodontal clinical parameters measured and gingival crevicular fluid (GCF) samples collected from up to 14 sites/subject. GCF samples were analysed using multiplex bead immunoassay for: GM-CSF, IFN-γ, IL-10, IL-1ß, IL-2, IL-6 and TNF-α. Aggressive periodontitis subjects were randomly assigned to either scaling and root planing (SRP) alone or SRP plus systemic amoxicillin (500 mg) and metronidazole (400 mg) 3 times a day for 14 days. Clinical parameters and GCF cytokines were re-measured 6 months after treatment. Differences over time were analysed using the Wilcoxon test and between groups using the Mann-Whitney test. RESULTS: Significant reductions in GCF GM-CSF, IL-1ß and the ratio IL-1ß/IL-10 and increases in GCF IL-6 were detected after therapy. The mean change in GCF cytokines did not differ significantly between groups. CONCLUSIONS: Periodontal therapy improved GCF cytokine profiles by lowering IL-1ß and increasing IL-10 levels. The reduction in GCF GM-CSF after therapy implicates this cytokine in the pathogenesis of GAgP. There was no difference between therapies in changes of GCF cytokines.

Matrix metalloproteinases and chemokines in the gingival crevicular fluid during orthodontic tooth movement.

Matrix metalloproteinases (MMPs) and monocyte chemoattractants are key modulators of the biological mechanisms triggered in the periodontium by mechanical forces. The gingival crevicular fluid (GCF) provides a non-invasive method to assess longitudinally the release of inflammatory mediators during orthodontic tooth movement. The goal of this study was to examine the GCF levels of MMP-3, MMP-9, and MMP-13 and of the chemokines macrophage inflammatory protein (MIP)-1ß, monocyte chemoattractant protein (MCP)-1, and regulated on activation normal T cells expressed and secreted (RANTES) at different time points during orthodontic tooth movement. Fourteen subjects (three males and 11 females, 18.8 ± 4.8 years of age; range from 12 to 28 years) had their maxillary canines retracted. Thirty-second GCF samples were collected from the tension and pressure sides 7 days prior to the activation of the orthodontic appliance, on the day of activation, and after 1 and 24 hours, and 14, 21, and 80 days of constant force application. The volume of GCF was measured and samples analysed using a multiplexed bead immunoassay for the content of the six target molecules. Differences in the mean GFC volumes and mean level for each analyte over time were assessed using the Friedman test, and differences between the tension and pressure sides at each time point with the Mann-Whitney test. The mean levels of the three MMPs changed significantly over time but only at the compression side (P < 0.05, Friedman test). The GCF levels of the three chemokines were not affected by the application of mechanical stress. The levels of MMPs in GCF at the pressure side are modulated by the application of orthodontic force.

Alteração no volume do fluido gengival durante a retração de caninos superiores / Change in the gingival fluid volume during maxillary canine retraction

INTRODUÇÃO: na análise da teoria da pressão/tensão do movimento dentário, a aplicação de uma força ortodôntica causa um deslocamento gradual dos fluidos do ligamento periodontal, acompanhado pela distorção das células e da matriz extracelular. OBJETIVOS: avaliar a quantidade de volume do fluido gengival (FG) nas faces mesiais e distais dos caninos superiores, de 14 pacientes (3 homens e 11 mulheres) submetidos a movimentação ortodôntica. MÉTODOS: o fluido foi coletado com tira de papel absorvente padrão (Periopaper®) e seu volume foi determinado por meio da utilização do medidor Periotron®, em sete tempos distintos (dia -7, dia 0, 1 hora, 24 horas, 14 dias, 21 dias e 80 dias). O teste Friedman foi usado para comparar os dados obtidos (p < 0,01 e p < 0,05). RESULTADOS: os resultados mostraram que houve uma alteração significativa do volume de FG ao longo do tempo, tanto no lado de pressão (p < 0,001) quanto no lado de tensão (p < 0,01). No lado de pressão, o volume de FG foi significativamente menor nos tempos 0 (p < 0,01) e 24h (p < 0,001), comparados com o tempo 80d.

INTRODUCTION: In the analysis of the pressure-tension theory of tooth movement, the application of an orthodontic force causes gradual displacement of fluids of the periodontal ligament, followed by distortion of the cells and extracellular matrix. OBJECTIVES: This study evaluated the gingival fluid volume on the mesial and distal aspects of the maxillary canines of 14 patients (3 males and 11 females) submitted to orthodontic movement. METHODS: The fluid was collected using standard absorbent paper strips (PeriopaperÕ) and the fluid volume was determined using the instrument PeriotronÕ at seven different periods (day -7, day 0, 1 hour, 24 hours, 14 days, 21 days, 80 days). The Friedman test was applied to compare the data achieved (p < 0.01 and p < 0.05). RESULTS: The results revealed a significant change in the gingival fluid volume with time on both the pressure side (p < 0.001) and the tension side (p < 0.01). On the pressure side, the gingival fluid volume was significantly lower at the periods 0 (p < 0.01) and 24hs (p < 0.001) compared to the period 80 days.

Levels of salivary IgA antibodies to Candida spp. in HIV-infected adult patients: a systematic review.

OBJECTIVE: To perform a systematic review of published data with the aim of evaluating the levels of IgA antibodies to Candida albicans in HIV-infected adult patients. METHODS: The search strategy was based on PubMed, Web of Science, Google Scholar, Cochrane and EMBASE databases. Also, the reference lists of included studies were searched. All abstracts found by electronic searches were independently scrutinized by two reviewers. To be eligible for review, the controlled studies had to present the following characteristics: samples of both HIV-infected adults and noninfected adults; appropriate HIV-diagnostic tests for both patient groups (case and control); IgA-diagnostic test applied to a similar population sample. RESULTS: Of 144 studies found, only six met the initial eligibility criteria, but three were excluded after a thorough analysis. To assess the methodological quality of the three remaining studies, they were categorized according the risk of bias. The three selected studies revealed that the levels of C. albicans-specific IgA antibody were higher in HIV-infected individuals compared with the control group. CONCLUSION: Adequate IgA antibody responses to C. albicans appear to be maintained, since the levels of these antibodies were higher in all studies selected. Although the findings of this systematic review are encouraging, the scientific evidence should be interpreted carefully because there are only a few reports in the literature, mostly because of the lack of important methodological details or the varying methodologies employed.

Antimicrobial agents used in the control of periodontal biofilms: effective adjuncts to mechanical plaque control?

The control of biofilm accumulation on teeth has been the cornerstone of periodontal disease prevention for decades. However, the widespread prevalence of gingivitis suggests the inefficiency of self-performed mechanical plaque control in preventing gingival inflammation. This is particularly relevant in light of recent evidence suggesting that long standing gingivitis increases the risk of loss of attachment and that prevention of gingival inflammation might reduce the prevalence of mild to moderate periodontitis. Several antimicrobials have been tested as adjuncts to mechanical plaque control in order to improve the results obtained with oral home care. Recent studies, including meta-analyses, have indicated that home care products containing chemical antimicrobials can provide gingivitis reduction beyond what can be accomplished with brushing and flossing. Particularly, formulations containing chlorhexidine, mouthrinses containing essential oils and triclosan/copolymer dentifrices have well documented clinical antiplaque and antigingivitis effects. In vivo microbiological tests have demonstrated the ability of these antimicrobial agents to penetrate the biofilm mass and to kill bacteria growing within biofilms. In addition, chemical antimicrobials can reach difficult-to-clean areas such as interproximal surfaces and can also impact the growth of biofilms on soft tissue. These agents have a positive track record of safety and their use does not seem to increase the levels of resistant species. Further, no study has been able to establish a correlation between mouthrinses containing alcohol and oral cancer. In summary, the adjunct use of chemical plaque control should be recommended to subjects with well documented difficulties in achieving proper biofilm control using only mechanical means.

Associations among the use of highly active antiretroviral therapy, oral candidiasis, oral Candida species and salivary immunoglobulin A in HIV-infected children.

OBJECTIVES: The aim was to examine the impact of antiretroviral therapy on the prevalence of oral candidiasis, recovery of oral Candida spp. , and salivary levels of total secretory immunoglobulin A (SIgA) and Candida-specific SIgA in human immunodeficiency virus (HIV)-infected children. STUDY DESIGN: Sixty-six HIV+ and 40 HIV- children were cross-sectionally examined for the presence of oral lesions. Whole stimulated saliva samples were collected for the identification of Candida spp. using culture and measurement of total and specific SIgA using enzyme-linked immunosorbent assay (ELISA). RESULTS: The HIV+ children had a higher prevalence of oral candidiasis (P < .05), higher frequency of detection of Candida spp. (P < .05), and higher levels of total (P < .05) and Candida-specific SIgA (P < .001) than the HIV- children. Among the HIV+ subjects, antiretroviral users had lower viral loads (P < .001) and lower levels of Candida spp. (P < .05) and total SIgA (P < .05) compared with antiretroviral nonusers. CONCLUSIONS: The use of antiretroviral therapy was associated with decreases in the prevalence of oral candidiasis. This diminished exposure to Candida spp. was accompanied by decreases in levels of total and Candida-specific SIgA.

Antimicrobial agents used in the control of periodontal biofilms: effective adjuncts to mechanical plaque control?

The control of biofilm accumulation on teeth has been the cornerstone of periodontal disease prevention for decades. However, the widespread prevalence of gingivitis suggests the inefficiency of self-performed mechanical plaque control in preventing gingival inflammation. This is particularly relevant in light of recent evidence suggesting that long standing gingivitis increases the risk of loss of attachment and that prevention of gingival inflammation might reduce the prevalence of mild to moderate periodontitis. Several antimicrobials have been tested as adjuncts to mechanical plaque control in order to improve the results obtained with oral home care. Recent studies, including meta-analyses, have indicated that home care products containing chemical antimicrobials can provide gingivitis reduction beyond what can be accomplished with brushing and flossing. Particularly, formulations containing chlorhexidine, mouthrinses containing essential oils and triclosan/copolymer dentifrices have well documented clinical antiplaque and antigingivitis effects. In vivo microbiological tests have demonstrated the ability of these antimicrobial agents to penetrate the biofilm mass and to kill bacteria growing within biofilms. In addition, chemical antimicrobials can reach difficult-to-clean areas such as interproximal surfaces and can also impact the growth of biofilms on soft tissue. These agents have a positive track record of safety and their use does not seem to increase the levels of resistant species. Further, no study has been able to establish a correlation between mouthrinses containing alcohol and oral cancer. In summary, the adjunct use of chemical plaque control should be recommended to subjects with well documented difficulties in achieving proper biofilm control using only mechanical means.

Salivary microbiota of HIV-positive children and its correlation with HIV status, oral diseases, and total secretory IgA.

AIM: This study aimed to investigate the prevalence and levels of salivary microorganisms in HIV-positive children, and their correlation to HIV status, oral lesions, and salivary IgA levels. DESIGN: Forty-two HIV-positive and 36 control children were clinically examined, had their saliva collected and processed for the microbiological analysis of 38 bacterial taxa by the checkerboard method, and salivary IgA quantification by ELISA. RESULTS: The majority of the species tested were more prevalent in control children than in the HIV group. Mean concentration of total salivary IgA was similar in both groups. High levels of Veillonella parvula were found in children with cheilitis and herpes. Tannerella forsythia, Eikenella Corrodens, and Propionibacterium acnes were prevalent in children with gingivitis, while Fusobacterium periodonticum, Streptococcus gordonii, and Streptococcus oralis were significantly more frequent in children with no oral lesions. Significant negative correlations between salivary IgA levels and Eubacterium nodatum and oral streptococci were observed (P < 0.05). CONCLUSION: HIV-seropositive children presented significantly lower prevalence and levels of several bacterial species in saliva; HIV-positive children are able to mount a mucosal immune response; HIV-seropositive children under highly active antiretroviral therapy presented low prevalence of oral lesions.

Effects of non-surgical mechanical therapy on the subgingival microbiota of Brazilians with untreated chronic periodontitis: 9-month results.

BACKGROUND: Mechanical periodontal therapy is the most common treatment of periodontal infections. It is directed primarily towards removing biofilm and calculus from the root surfaces, leading to ecological changes in the subgingival environment. Thus, the purpose of this study was to evaluate the effects of scaling and root planing (SRP) on the subgingival microbiota of Brazilian subjects with untreated chronic periodontitis over a 9-month period. METHODS: Twenty-five untreated chronic periodontitis patients (mean age 43 +/- 5 years; 20% smokers; 45% males) were selected from a Brazilian population. At baseline, probing depth (PD), clinical attachment level (CAL), visible supragingival biofilm (SB), bleeding on probing (BOP), and suppuration (SUP) were measured at six sites/tooth. Subgingival plaque samples were obtained from 10 sites with the deepest PD (> or =5 mm) of each subject and tested for the presence of 25 oral species by DNA probes and the checkerboard technique. Patients received full mouth SRP and oral hygiene instructions. Clinical and microbiological assessments were repeated at 3, 6, and 9 months after therapy. During this period, all patients received maintenance therapy, including supragingival prophylaxis and reinforcement in home care procedures. The clinical and microbiological parameters examined were computed for each subject and at each visit. Differences over time were sought using the Friedman test. RESULTS: Significant reductions in mean CAL and PD (P <0.01), percent of sites with SB (P <0.01), BOP and SUP (P <0.05) were observed during the course of the study. In general, microbial changes were more pronounced for the mean counts than for the frequency of the microorganisms, particularly at 3 months post-therapy. Significant reductions in prevalence and levels were observed for certain periodontal pathogens including P. gingivalis (P <0.05; P <0.01), T. forsythensis (P <0.01), C. rectus (P <0.01), and A. actinomycetemcomitans (P <0.01; P <0.05). Nevertheless, the frequency of A. actinomycetemcomitans increased to baseline values at 9 months after therapy. Treponema ssp. and Prevotella spp. showed a modest decrease in prevalence, whereas marked reductions in their levels were observed. In contrast, the frequency and counts of the suspected pathogens P. micros and F. nucleatum increased after treatment. Species considered beneficial including Actinomyces spp., some oral streptococci, and V. parvula increased in prevalence, although these two last species tended to return to baseline levels at 9 months. CONCLUSION: In Brazilians with untreated chronic periodontitis, SRP led to clinical improvement associated with a decrease of certain periodontal pathogens, and an increase of beneficial species for up to 9 months after therapy.

Endodontic therapy associated with calcium hydroxide as an intracanal dressing: microbiologic evaluation by the checkerboard DNA-DNA hybridization technique.

This study evaluated the predominant microbiota of infected necrotic pulps and the effects of calcium hydroxide therapy on these microorganisms by the checkerboard DNA-DNA hybridization technique. Conventional endodontic therapy associated with calcium hydroxide as intracanal dressing was performed in 12 single-rooted teeth with pulp necrosis and periradicular bone lesion. Samples were collected from the canal at baseline and 14 days after therapy, and the presence of 44 bacterial species was determined by the checkerboard method. Significant differences in the microbiota from baseline to post-therapy were sought by the paired-samples t test. The most prevalent species included F. nucleatum ss. vincentii, C. sputigena, C. ochracea, S. constellatus, V. parvula, P. gingivalis, P. melaninogenica, and S. sanguis. Most of the microorganisms were reduced after treatment, particularly A. gerencseriae, A. israelii, A. naeslundii, C. gingivalis, C. ochracea, P. gingivalis, S. noxia, S. sanguis, and S. oralis (p < 0.05). Conversely, A. actinomycetemcomitans, C. sputigena, and E. corrodens increased in numbers after therapy. These results indicate that conventional endodontic therapy with calcium hydroxide results in the reduction of pathogenic species associated with pulp necrosis. However, its use is limited, because it did not eliminate the whole spectrum of microorganisms.