Antineoplastic therapy is an independent risk factor for dental caries in childhood cancer patients: a retrospective cohort study.

PURPOSE: To evaluate the antineoplastic therapy (AT) as a risk factor for dental caries lesions independent of other risk factors such as income, family education, stimulated salivary flow rate, hygiene habits, frequency of sugar intake, and microbiota in childhood cancer (CC) patients. METHODS: 72 individuals were divided into CC patients (n=36) and healthy individuals (control group - CT n=36). Demographic data, hygiene habits, frequency of sugar intake, CC type, and AT were collected. Stimulated salivary flow rate was measured and the presence and concentration of Streptococcus mutans were assessed using a real-time polymerase chain reaction (qPCR) technique. Clinical evaluations included plaque index (PI) and decayed-missing-filled-teeth index (dmft/DMFT). Descriptive statistics, T-test, Mann-Whitney test, chi-square test, Fisher's exact test, and two-way analysis of variance were used for data analysis (p<0.05). RESULTS: At the time of oral evaluation, both groups exhibited similar ages with means of 12.0±3.9 years old for CC and 12.0±4.0 years old for CT patients. All CC patients underwent chemotherapy with nine also undergoing radiotherapy. Significant differences were observed between the groups in terms of color/race, income, family education, and hygiene habits. However, no statistically significant differences were found between groups regarding the frequency of sugar intake, stimulated salivary flow rate, or the concentration of Streptococcus mutans (qPCR technique). For clinical parameters, the DMF (CC:1.80, CT: 0.75), decayed (CC: 0.88, CT: 0.19), missing (CC: 0.25, CT:0), and PI (CC: 30.5%, CT: 22.6%) were higher in the CC group (p<0.05). CONCLUSION: Childhood cancer (CC) patients undergoing antineoplastic therapy (AT) exhibit a higher prevalence of dental caries, regardless of income/education, frequency of sugar intake, stimulated salivary flow rate, and microbiota.

Clinical, immunological and microbiological evaluation of experimental peri-implant mucositis and gingivitis in subjects with Grade C, stage III/IV periodontitis background.

AIM: To compare individuals with a periodontitis background (Grade C, stage III/IV-formerly generalized aggressive periodontitis) (H-GAP) with periodontally healthy subjects (H-Health) in terms of molecular changes (immunological/microbiological) accompanying experimental peri-implant mucositis and gingivitis. MATERIALS AND METHODS: H-GAP and control (H-Health) subjects were recruited, and experimental mucositis/gingivitis was induced around a single screw-retained implant and one contralateral tooth. Participants refrained from oral hygiene for 21 days in the selected areas, followed by professional prophylaxis and hygiene instructions for 21 days. Clinical parameters, immunological markers (multiplex analysis) and microbial data (16S rRNA gene sequencing) were collected at baseline, during induction (7, 14 and 21 days) and following remission (42 days). RESULTS: Clinically, no significant differences were observed between the groups (n = 10/each group) (H-GAP vs. H-Health) (p > .05, Mann-Whitney test) and the type of site (tooth vs. implant) (p > .05, Wilcoxon test) at the time of onset and resolution, or severity of gingival/mucosal inflammation. H-GAP displayed lower concentrations of the cytokines interleukin (IL)-1B, IL-4, IL-17, tumor necrosis factor-α and interferon-γ around implants than H-Health at baseline and during induction of mucositis (p < .05, Mann-Whitney test). In both groups, implants showed significantly higher inflammatory background at baseline and all subsequent visits when compared with teeth (p < .05, Wilcoxon test). Alpha and ß-diversity metrics showed a significant shift in the microbiome composition and abundances of core species during induction and resolution of peri-implant mucositis and gingivitis (p < .05, restricted maximum likelihood method of Shannon and Bray-Curtis indices, respectively). Differences were not significant for these parameters between the H-Health and H-GAP groups when the periodontal and peri-implant microbiomes were compared separately; however, at each time point, the peri-implant microbiome differed significantly from the periodontal microbiome. CONCLUSIONS: Within the limitations of this pilot study (e.g. low power), it can be concluded that different microbial shifts contribute to the onset and progression of inflammatory responses around teeth and implants and that history of periodontal disease experience plays an additional role in modulating the immune response of peri-implant and periodontal tissues to biofilm accumulation.

Microbial colonization in the partially exposed nonabsorbable membrane during alveolar ridge preservation.

AIM: This study evaluated the impact of the partial exposition of the nonabsorbable membrane (dPTFE) on microbial colonization during bone healing. MATERIALS AND METHODS: Patients indicated for tooth extraction were randomized to dPTFE group (n = 22) - tooth extraction and alveolar ridge preservation (ARP) using an intentionally exposed dPTFE membrane and USH group (n = 22) - tooth extraction and unassisted socket healing. Biofilm samples were collected at the barrier in the dPTFE and on the natural healing site in the USH after 3 and 28 days. Samples from the inner surface of the dPTFE barrier were also collected (n = 13). The microbiome was evaluated using the Illumina MiSeq system. RESULTS: Beta diversity was different from 3 to 28 days in both groups, and at 28 days, different microbial communities were identified between therapies. The dPTFE was characterized by a higher prevalence and abundance of gram-negative and anaerobic species than USH. Furthermore, the inner surface of the dPTFE membrane was colonized by a different community than the one observed on the outer surface. CONCLUSION: Intentionally exposed dPTFE membrane modulates microbial colonization in the ARP site, creating a more homogeneous and anaerobic community on the inner and outer surfaces of the membrane. CLINICAL RELEVANCE: DPTFE promoted faster biofilm colonization and enrichment of gram-negative and anaerobes close to the regenerated site in the membrane's inner and outer surfaces. dPTFE membrane can be used exposed to the oral site, but approaches for biofilm control should still be considered. The study was retrospectively registered at Clinicaltrials.gov (NCT04329351).

Experimental Inoculation of Aggregatibacter actinomycetemcomitans and Streptococcus gordonii and Its Impact on Alveolar Bone Loss and Oral and Gut Microbiomes.

Oral bacteria are implicated not only in oral diseases but also in gut dysbiosis and inflammatory conditions throughout the body. The periodontal pathogen Aggregatibacter actinomycetemcomitans (Aa) often occurs in complex oral biofilms with Streptococcus gordonii (Sg), and this interaction might influence the pathogenic potential of this pathogen. This study aims to assess the impact of oral inoculation with Aa, Sg, and their association (Aa+Sg) on alveolar bone loss, oral microbiome, and their potential effects on intestinal health in a murine model. Sg and/or Aa were orally administered to C57Bl/6 mice, three times per week, for 4 weeks. Aa was also injected into the gingiva three times during the initial experimental week. After 30 days, alveolar bone loss, expression of genes related to inflammation and mucosal permeability in the intestine, serum LPS levels, and the composition of oral and intestinal microbiomes were determined. Alveolar bone resorption was detected in Aa, Sg, and Aa+Sg groups, although Aa bone levels did not differ from that of the SHAM-inoculated group. Il-1ß expression was upregulated in the Aa group relative to the other infected groups, while Il-6 expression was downregulated in infected groups. Aa or Sg downregulated the expression of tight junction genes Cldn 1, Cldn 2, Ocdn, and Zo-1 whereas infection with Aa+Sg led to their upregulation, except for Cldn 1. Aa was detected in the oral biofilm of the Aa+Sg group but not in the gut. Infections altered oral and gut microbiomes. The oral biofilm of the Aa group showed increased abundance of Gammaproteobacteria, Enterobacterales, and Alloprevotella, while Sg administration enhanced the abundance of Alloprevotella and Rothia. The gut microbiome of infected groups showed reduced abundance of Erysipelotrichaceae. Infection with Aa or Sg disrupts both oral and gut microbiomes, impacting oral and gut homeostasis. While the combination of Aa with Sg promotes Aa survival in the oral cavity, it mitigates the adverse effects of Aa in the gut, suggesting a beneficial role of Sg associations in gut health.

Parents with periodontitis drive the early acquisition of dysbiotic microbiomes in their offspring.

AIM: To evaluate the microbial colonization in different dentition phases on individuals from 0 to 18 years of age belonging to families with a history of periodontitis compared to descendants of periodontally healthy parents. MATERIALS AND METHODS: The offspring of subjects with periodontitis ('Perio' group) and the offspring of periodontally healthy subjects ('Healthy' group), matched for gender and age, were included in this cross-sectional study and divided according to the dentition phase: pre-dentate, primary, mixed and permanent. The patients were clinically assessed, and their saliva was collected. DNA was extracted, and V1-V3 and V4-V5 regions of the 16S rRNA gene were sequenced. RESULTS: Fifty children of parents with periodontitis and 50 from healthy parents were included in the study and divided according to the dentition phase: pre-dentate (n = 5/group), primary dentition (n = 15/group), mixed dentition (n = 15/group) and permanent dentition (n = 15/group) in each group. The microbiome composition was different between dentitions for both groups. Children of the Perio group presented a microbial diversity different from that of the Healthy group in mixed and permanent dentitions. The more intense shift in the community occurred between primary and mixed dentition in the Perio group, while the transition between mixed and permanent dentition was the period with greater changes in the microbiome for the Healthy group. Furthermore, a pathogen-rich environment-higher prevalence and abundance of periodontitis-associated species such as Prevotella spp., Selenomonas spp., Leptotrichia spp., Filifactor alocis, Prevotella intermedia, Treponema denticola and Tannerella forsythia- was observed in the Perio group. CONCLUSIONS: The parents' periodontal status significantly affects the microbiome composition of their offspring from an early age. The mixed dentition was the phase associated with establishing a dysbiotic and pathogen-rich microbiome in descendants of parents with periodontitis.

Salivary annexin A1: A candidate biomarker for periodontitis.

AIM: To compare the salivary proteomic profile of periodontitis-affected (PA) parents and their offspring to periodontally healthy (PH) dyads in the pursuit of possible biomarkers for early diagnosis of this disease. MATERIALS AND METHODS: Unstimulated saliva samples collected from 17 pairs of PA or PH individuals and their children were submitted to mass spectrometric analyses followed by proteomic analyses. Primary PA fibroblasts were triggered towards having an inflammatory response, and an immunoenzymatic assay of its supernatant was performed to validate the obtained data. RESULTS: ANXA1, KRT4, GSTP1, HPX, A2M and KRT13 were lower in PA parents and their children, and IGHG1, CSTB, KRT9, SMR3B, IGHG4 and SERPINA1 were higher. ANXA1 presented the highest fold change, 7.1 times less produced in children of PA parents, and was selected as a potential biomarker for periodontitis. The in vitro assay also showed lower ANXA1 production by cells of PA patients. CONCLUSION: Before any clinical sign of periodontal loss, descendants of PA patients have an altered proteomic profile compared to PH individuals, presenting a lower abundance of ANXA1. This protein is suggested as a potential biomarker for periodontitis.

BMP-2 and asporin expression regulate 5-aza-dC-mediated osteoblast/cementoblast differentiation of periodontal dental ligament mesenchymal progenitor cells.

Periodontal dental ligament (PDL) is composed of heterogeneous population of mesenchymal progenitor cells. The mechanisms that regulate the differentiation of these cells towards osteoblast/cementoblast phenotype are not fully understood. Some studies have demonstrated that is possible to change the pattern of cell differentiation via epigenetic mechanisms. The proposal of this study was to investigate whether 5-aza-2'-deoxycytidine (5-aza-dC) treatment would stimulate the osteoblast/cementoblast differentiation of periodontal ligament mesenchymal progenitor cells (PDL-CD105+ enriched cells), characterized as low osteoblast potential, through bone morphogenetic protein-2 (BMP-2) modulation. PDL-CD105+ cells from a single donor were cloned and characterized in two populations as high osteoblast/cementoblast potential (HOP) and low osteoblast/cementoblast potential (LOP) by mineralization in vitro and expression of osteogenic gene markers, such as runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin (OCN), bone morphogenetic protein 2 (BMP-2) and asporin (ASPN). Next, two LOP clones (L1 and L2) were pretreated with 5-aza-dC (10 µM) for 48 h, cultured under osteogenic condition and evaluated for mineralized matrix in vitro, transcription modulation of osteogenic gene markers, methylated and hydroxymethylated DNA levels of BMP-2 and ASPN and intracellular/extracellular expression of BMP-2 protein. LOP clones showed high expression of ASPN transcripts associated with low mRNA levels of BMP-2, RUNX2, ALP, and OCN. 5-aza-dC treatment raised hydroxymethylated DNA levels of BMP-2 and increased the expression of BMP-2 transcripts in both LOP clones. However, BMP-2 protein (intracellular and secreted forms) was detected only in L1 cell clones, in which it was observed an increased expression of osteoblast/cementoblast markers (RUNX2, ALP, OCN) associated with higher mineralization in vitro. In L2 cell clones, 5-aza-dC increased gene expression of ASPN, with no great change in for osteoblast/cementoblast differentiation potential. These data show that 5-aza-dC improves osteoblast/cementoblast differentiation of PDL-CD105+ cells via BMP-2 secretion, and this effect depends on low levels of ASPN expression.

Antineoplastic therapy in childhood cancer patients presents a negative impact in the periodontal tissues: a cohort study.

OBJECTIVES: To investigate the effect of antineoplastic therapy (AT) in the periodontal tissues of childhood cancer (CC) patients. MATERIALS AND METHODS: Seventy-two individuals were divided into CC (n=36) and healthy individuals (control group-CG, n=36). Demographics, hygiene habits, CC type, and AT were collected. Salivary flow and the presence and concentration of Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Tannerella forsythia, and Fusobacterium nucleatum were analyzed. Clinical evaluation included plaque (PI) and gingival indexes (GI), periodontal probing depth (PPD), and clinical attachment level (CAL). Patients were classified into periodontal health, gingivitis, or periodontitis. Descriptive statistics, T test, Mann-Whitney test, chi-square, Fisher's exact test, and two-way analysis of variance were used (p<0.05). RESULTS: The mean age of the patients was similar (CC 12.0±3.9 years and CG 12.0±4.0 years). In the CC group, all patients underwent chemotherapy and nine radiotherapy. Color/race, income, and family education showed significant differences between groups. There was no difference between groups in salivary flow. Higher levels of Fusobacterium nucleatum were seen in CC (p=0.02). Significant difference between groups was found for PI (CC: 30.5%, CG: 22.6%), GI (CC: 28.8%, CG: 17.3%), PPD (CC: 1.77 mm, CG: 1.61 mm), and CAL (CC: 1.77 mm, CG: 1.57 mm), periodontal health (CC: 3, CG: 7), gingivitis (CC: 16, CG: 24), or periodontitis (CC: 17, CG: 5). CONCLUSION: AT in CC patients presents a negative impact in the periodontal and microbiological parameters. CLINICAL RELEVANCE: Childhood cancer individuals showed worse periodontal parameters and higher levels of Fusobacterium nucleatum in the saliva when compared to healthy individuals.

Effects of oral administration of Bifidobacterium animalis subsp. lactis HN019 on the treatment of plaque-induced generalized gingivitis.

OBJECTIVES: This double-blind, randomized, placebo-controlled clinical trial evaluated the adjuvant effects of Bifidobacterium lactis HN019 on the treatment of plaque-induced generalized gingivitis. MATERIALS AND METHODS: Sixty patients were submitted to professional supragingival scaling and prophylaxis. They were randomly assigned to test (probiotic lozenges containing B. lactis HN019, n = 30) or control (placebo lozenges, n = 30) groups. Lozenges were consumed twice a day for 8 weeks. Bleeding on probing (BoP), Gingival Index (GI), Plaque Index (PI), probing depth (PD), and clinical attachment level (CAL) were evaluated at baseline and after 2 and 8 weeks. Gingival crevicular fluid (GCF) was collected at baseline and at 8 weeks for analysis of the inflammatory mediators IL-1ß, IL-1α, IL-8, MCP-1, and MIP-1ß. Data were statistically analyzed (p < 0.05). RESULTS: After 8 weeks, both groups showed reduction in the percentage of PI, with no significant difference between groups (p = 0.7423). The test group presented a lower percentage of BoP and a higher percentage of sites with GI ≤ 1 when compared with the control group at the end of the study (p < 0.0001). At 8 weeks, the test group had a greater number of patients without generalized gingivitis than the control group (20 and 11 patients, respectively; p < 0.05). The test group presented significantly lower levels of IL-1α, IL-1ß, and MCP-1 in GCF than the control group at the end of the study (p < 0.05). CONCLUSION: The adjunct use of B. lactis HN019 promotes additional clinical and immunological benefits in the treatment of generalized gingivitis. CLINICAL RELEVANCE: B. lactis HN019 can be an efficient and side-effect-free adjunct strategy in the treatment of generalized gingivitis.

Smoking negatively impacts the clinical, microbiological, and immunological treatment response of young adults with Grade C periodontitis.

OBJECTIVE: This study aimed to investigate the influence of smoking on clinical, microbiological and immunological parameters in young adult with stage III-IV Grade C periodontitis after full-mouth ultrasonic debridement (FMUD) associated with Amoxicillin and Metronidazole (AMX + MTZ), comparing smokers (PerioC-Y-Smk) with non-smokers (PerioC-Y-NSmk). MATERIALS AND METHODS: Fifteen PerioC-Y-NSmk and 14 PerioC-Y-Smk patients underwent FMUD associated with AMX + MTZ for 10 days. All parameters were collected at baseline and 3 and 6 months after treatment. Plaque index (PI), bleeding on probing (BoP), probing depth (PD), clinical attachment level (CAL)- the primary variable-, and gingival recession (GR) were clinically assessed. The impact of PI on CAL change at 6-month was verified by a regression analysis. Samples of the subgingival biofilm was collected for detection of levels of Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans), Porphyromonas gingivalis (P.gingivalis), Tannerella forsythia (T. forsythia), and Fusobacterium nucleatum ssp (F. nucleatum), and were analyzed by real-time qPCR; gingival crevicular fluid was collected for detection of levels of interleukin (IL)-1ß, IL-4, IL-6, IL-10, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ, which were analyzed using an enzyme immunoassay. RESULTS: PerioC-Y-Smk had significantly higher PI, BOP, and GR at baseline compared to non-smokers (p < .05). PerioC-Y-Smk presented higher PD, CAL, and GR at 3 and 6 months (p < .05) compared with PerioC-Y-NSmk in the same periods; PI negatively affected CAL gain in PerioC-Y-NSmk at 6-month follow-up (p = .052) and did not impact on clinical response in PerioC-Y-Smk (p = .882). Lower levels of IFN-γ, IL1-ß, and IL-4 were observed at 3 months in the PerioC-Y-NSmk (p < .05) compared with PerioC-Y-Smk. Lower proportions of P. gingivalis were observed in PerioC-Y-NSmk at baseline and at 3 months (p < .05) and lower proportions of F. nucleatum were observed at 6 months, in the PerioC-Y-NSmk (p < .05). CONCLUSIONS: PerioC-Y-Smk presents an unfavorable clinical, microbiological, and immunological response after 3 and 6 months after FMUD associated with AMX + MTZ. CLINICAL RELEVANCE: Smoking worsens periodontal condition of young treated adults presenting stage III/IV Grade C periodontitis.

IL10 promoter rs6667202 polymorphism is functional in health but not in grade c periodontitis patients: A pilot study.

BACKGROUND AND OBJECTIVE: Previous studies have demonstrated an association between the IL10 promoter rs6667202 (C > A) single-nucleotide polymorphism (SNP) and grade C, stage 3 or 4 periodontitis (Perio4C) in the Brazilian population, where the altered A allele was detected more frequently in these patients. However, no functional analysis of this variation has yet been performed. Thus, the objective of this preliminary study was to evaluate the functionality of rs6667202 in gingival fibroblasts (GFs) of individuals with Perio4C and with periodontal health (PH) stimulated with Aggregatibacter actinomycetencomitans protein extract (AaPE). METHODS: Patients with PH and Perio4C were segregated according to their genotype (AA, AC, or CC), and a biopsy was performed to establish the culture of the GFs. After GFs exposure to AaPE at 5 µg/ml for 1.5 h, RNA was extracted to analyze IL10 expression by qPCR. Aliquots of the cell's supernatant were subjected to immunoenzymatic analysis (MAGpix) to detect interleukin-10 (IL-10). RESULTS: In PH, the genotypes AA and AC are related to less expression of IL10 (p = 0.027 and p < 0.0001) and less production of IL-10 (p = 0.002 and p = 0.001), when compared to CC. In Perio4C, there was no statistical difference between the genotypes (p > 0.05), although a lower IL-10 expression and release compared with PH CC was seen (p = 0.033 and p < 0.001). CONCLUSION: The rs6667202 SNP is functional in PH, as it decreases the expression and production of IL-10. In Perio4C, other factors may be masking its action by altering the IL-10's response.

Adjuvant use of multispecies probiotic in the treatment of peri-implant mucositis: A randomized controlled trial.

AIM: This randomized placebo-controlled clinical trial evaluated the effects of multispecies probiotic containing Lactobacillus rhamnosus HN001™, Lactobacillus paracasei Lpc-37®, and Bifidobacterium animalis subsp lactis HN019™ as an adjunct to mechanical debridement (MD) on changes in bleeding on probing (BOP) in edentulous patients with peri-implant mucositis (PiM). MATERIALS AND METHODS: Patients were randomly assigned to test (probiotic) or control (placebo) groups. All sites with PiM received MD and topical gel application (probiotic or placebo) at baseline and 12 weeks. After initial MD, patients consumed probiotic or placebo capsules twice a day for 12 weeks. Clinical (modified sulcus bleeding index [mSBI]; modified plaque index [mPI]; probing depth [PD]; and BOP) and immunological parameters were collected at baseline and after 12 and 24 weeks. Data were statistically analysed (p < .05). RESULTS: Thirty-six patients with PiM were recruited. The test group presented higher prevalence (p < .05) of cases of restored peri-implant health at 24 weeks than did the control group (72.2% and 33.3%, respectively). No significant difference was observed between test (n = 18) and control (n = 18) groups for mPI and PD. mSBI %-score 0 was higher in the test group than in the control group at 24 weeks (p < .05). When compared with baseline, both groups presented reduced BOP at 12 and 24 weeks (p < .05). BOP was lower in the test group than in the control group at 12 (mean difference = -14.54%; 95% confidence interval [CI] = -28.87 to 0.22; p = .0163) and 24 (mean difference = -12.56%; 95% CI = -26.51 to 1.37; p = .0090) weeks. At 24 weeks, only the test group presented lower levels of interleukin (IL)-1ß, IL-6, IL-8, and tumour necrosis factor (TNF)-α than those at baseline (p < .05). CONCLUSIONS: The multispecies probiotic (administered locally and systemically) containing L. rhamnosus HN001™, L. paracasei Lpc-37®, and B. lactis HN019™ as an adjunct to repeated MD promotes additional clinical and immunological benefits in the treatment of PiM in edentulous patients (ClinicalTrials.gov NCT04187222).

The familial trend of the local inflammatory response in periodontal disease.

OBJECTIVES: The imbalanced host response in front of a dysbiotic biofilm is one of the major aspects of severe periodontitis, which also presents a strong familial aggregation related to the susceptibility factors transmission within family members. This study hypothesized that aggressive periodontitis (GAgP) patients and their descendants could present a similar trend of a local inflammatory response that is different from healthy controls. METHODS: Fifteen GAgP subjects and their children and fifteen healthy subjects and their children were clinically assessed, and the concentration of interferon (IFN)-γ, interleukin (IL)-10, IL-17, IL-1ß, IL-4, IL-6, IL-8, and tumor necrosis factor (TNF)-α was evaluated in the gingival fluid using the multiplexed bead immunoassay. RESULTS: Children from the GAgP group presented lower IL-10 and IFN-γ subgingival concentration than Health children, despite no difference in the clinical parameters. GAgP parents showed a lower IFN-γ, IL-10, and IL-6 than healthy subjects. IL-10/IL-1ß and IFN-γ/IL-4 ratios were reduced in GAgP dyads, suggesting a familial trend in the subgingival cytokine's profile. The cytokines correlated to the clinical data and were predictors of probing depth increase. CONCLUSION: GAgP parents and their children presented a similar cytokine profile and an imbalance in the subgingival response characterized by decreased IFN-γ/IL-4 and IL10/IL-1ß ratios.

Self-ligating brackets exhibit accumulation of high levels of periodontopathogens in gingival crevicular fluid.

Different types of brackets seem to influence the disruption of the oral microbial environment. Therefore, the aim of this study was to evaluate the influence of self-ligating brackets on the gingival crevicular fluid levels of the putative periodontal pathogens Aggregatibacter actinomycetemcomitans sorotype a (Aaa), Tannerella forsythia, Fusobacterium nucleatum, and Porphyromonas gingivalis. Sixty samples of crevicular fluid of twenty patients (11 boys and 9 girls) were analysed at baseline (T0) and after 30 (T1) and 60 (T2) days of bonding of the self-ligating (In-Ovation®R, Dentsply, GAC or SmartClip™, 3 M Unitek, Monrovia, CA, USA) and of one conventional bracket (Gemini™, 3 M Unitek, Monrovia, CA, USA) used with elastomeric ligatures. Total DNA from samples was extracted using CTAB-DNA precipitation method and Real-time PCR was performed to analyse bacterial level. Non-parametric Friedman and Wilcoxon tests were used for data analysis (p value of < 0.05). F. nucleatum presented a different level among the different brackets at T1 (p = 0.025), the highest level in the Gemini™ bracket when compared to the SmartClip™ bracket (p = 0.043). P. ginigvalis levels increased in the In-Ovation®R (p = 0.028) at T1. The subgingival levels of bacterial species associated with periodontal disease P. ginigvalis increased in the self-ligating brackets In-Ovation®R.Clinical Relevance: Some kinds of brackets could provide more retentive sites than others, and it seems to modulate the subgingival microbiota, since, in this study, we could observe the increase of the species associated with periodontal disease. Preventive protocols should be adopted in the use of self-ligating brackets.

Salivary IL-4: A Bleeding Predictor on Probing in Descendants of Severe Periodontitis Patients.

OBJECTIVE: Periodontitis in younger patients can cause severe periodontal destruction, and cases are usually more numerous in members of the same family due to the sharing of susceptibility factors. Thus, the use of a familial study design could improve our understanding of initial alterations in periodontal tissue. This observational study aimed to evaluate the salivary inflammatory pattern in descendants of periodontitis patients and identify any correlation with the clinical periodontal condition. STUDY DESIGN: Fifteen children of Generalized Aggressive Periodontitis (GAgP) patients and 15 children with periodontally healthy parents were evaluated for their plaque index (PI), gingival index (GI), bleeding on probing (BoP), and probing depth (PD). The concentrations of interferon (IFN)-γ, interleukin (IL)-10, IL-17, IL-1ß, IL-4, and tumor necrosis factor (TNF)-α were measured in unstimulated saliva using the Luminex MAGPix platform. RESULTS: Children from the GAgP group presented higher probing depth (PD) and bleeding on probing (BoP) (p<0.05) and lower release of IL-4 in saliva (p<0.05) than the periodontally healthy group. The cytokines IL-10, IFN-γ, IL-17, and IL-4 were negatively correlated with the gingival index, while IL-4 was negatively correlated with BoP. A regression analysis revealed that salivary IL-4 and plaque were predictors of BoP. CONCLUSIONS: Children of GAgP parents presented lower salivary IL-4 and higher BoP and PD than children from periodontally healthy families. Additionally, salivary IL-4 was a predictor of bleeding on probing in the children, suggesting that the lower presence of this anti-inflammatory cytokine is related to higher clinical inflammation.

Enamel matrix derivative effects on palatal mucosa wound healing: Randomized clinical trial.

BACKGROUND: This study was conducted to evaluate the clinical, immunologic, and patient-centered outcomes of enamel matrix protein derivative (EMD) on excisional wounds in palatal mucosa. MATERIALS: Forty-four patients in need of ridge preservation were randomly allocated into two groups: control group (n = 22): open palatal wound after free gingival graft (FGG) harvest and EMD group (n = 22): open palatal wound after FGG harvest that received 0.3 ml of EMD. Clinical and patient-centered parameters were analyzed for 3 months post-treatment. Wound fluid levels of inflammatory markers were assessed 3 and 7 days postoperatively. RESULTS: No significant inter-group difference was observed in remaining wound area and re-epithelialization. EMD and control groups achieved wound closure and re-epithelialization 30 days postoperatively (p < .001), without inter-group differences. Similarly, number of analgesics and Oral Health Impact Profile scores did not present significant inter-group differences (p > .05). EMD appeared to selectively modulate wound fluid levels of monocyte chemoattractant protein-1, macrophage inflammatory protein-1α, matrix metallopeptidase 9, and tissue inhibitor of metalloproteinases-2. CONCLUSION: Within the limits of the present study, it can be concluded that EMD application to excisional palatal wounds using the investigated protocol does not provide clinical healing benefits, despite an apparent modulation of selected inflammatory markers.

Low bone mineral density is associated with severe periodontitis at the end of the second decade of life: A population-based study.

AIM: To evaluate the association between low bone mineral density (BMD) and severe periodontitis at the end of the second decade of life. MATERIALS AND METHODS: This population-based study analysed 2032 youngers (18-19 years old) of the RPS cohort. BMD of lumbar spine (BMD-LS) and of the whole body (BMD-WB) were assessed by dual x-ray emission densitometry. Low BMD-LS (Z-score ≤ -2) and low BMD-WB (Z-score ≤ -1.5) were correlated with severe periodontitis. The extent of periodontal disease was also evaluated as the following outcomes: proportions of teeth affected by clinical attachment loss ≥5 mm and probing depth ≥5 mm. Multivariate models by sex, education, family income, risk of alcohol dependence, smoking, plaque, bleeding index, and body mass index were estimated through logistic regression (binary outcomes) and Poisson regression (continuous outcomes). RESULTS: The prevalence of severe periodontitis was 10.97%. Low BMD-LS (odds ratio [OR] = 2.08, confidence interval [CI] = 1.12-3.85, p = .01) and low BMD-WB (OR = 1.34, CI = 1.001-1.81, p = .04) were associated with severe periodontitis in the final multivariate models. Low BMD-LS and BMD-WB were also associated with a greater extent of periodontitis (p < .05). CONCLUSIONS: Low BMD was found to be associated with the severity and extent of periodontitis in adolescents. Adolescents at peak bone mass age presenting low BMD are more likely to be affected by severe periodontitis.

Subgingival endotoxin and lipoteichoic acid modulate cytokine production in diabetic subjects: A Case-control Study.

BACKGROUND: Periodontal disease and diabetes mellitus (DM) are highly prevalent and interrelated diseases, resulting in altered host response microbiota. Thus, this study aimed to evaluate the impact of DM on local levels of lipopolysaccharide (LPS) and lipoteichoic acid (LTA) and their relationship with cytokines and matrix metalloproteinases' (MMPs) profile. METHODS: This case-control study included diabetic (n = 15) and non-diabetic (n = 15) subjects presenting Stage 3-4, Grade C, Periodontitis. Gingival crevicular fluid (GCF) was collected, and LPS and LTA levels were analyzed by enzyme-linked immunosorbent assay (ELISA), while IFN-γ, IL-10, IL-17, IL-1ß, IL-4, MMP-2, and MMP-9 were measured by LUMINEX/MAGpix. Mann-Whitney and Spearman's correlation tests were used to compared and to correlate variables (p < 0.05). RESULTS: Higher levels of LTA, LPS, IL-10, IL-1ß, and MMP-2 (p < 0.05) and lower levels of IL-17 were found in the DM group (p < 0.05). Non-diabetic subjects presented higher LPS, IFN-γ, IL-17, and MMP-2 levels and lower IL-10 concentration (p < 0.05). No significant correlation was seen between LPS and cytokine profile in non-diabetic. Local levels of LTA were positively correlated with IL-17 and MMP-2 and negatively with IL-10. CONCLUSION: LTA and LPS drove the inflammatory profile through the modulation of cytokines and MMPs in a different manner in DM and non-diabetic subjects.

Systemic circulating inflammatory burden and periodontitis in adolescents.

OBJECTIVES: To analyze the association between systemic inflammatory burden of cardiovascular disease (CVD) risk and periodontitis in adolescents, including mediating pathways triggered by their common risk factors. MATERIALS AND METHODS: Using a population-based sample study (n = 405) of Brazilian adolescents (17-18 years old), direct and mediation pathways triggered by "Socioeconomic Status," "Adiposity," Smoking, and "Blood Pressure" were modelled for the association between the "Systemic Circulating Inflammatory Burden of CVD Risk" (IL-1ß, IL-6, IL-8, TNF-α) and the "Initial Periodontitis" (bleeding on probing (BoP), probing depth (PD) ≥ 4 mm, clinical attachment loss (CAL) ≥ 4 mm), both as continuous latent variables, using structural equation modeling. Sensitivity analysis was performed for the outcomes "Gingivitis" (visible plaque; BoP); "Moderate Periodontitis" (PD ≥ 5 mm and CAL ≥ 5 mm) and periodontitis (CDC-AAP case definition). RESULTS: Higher "Systemic Circulating Inflammatory Burden of CVD Risk" was directly associated with higher "Initial Periodontitis" (standardized coefficient [SC] = 0.178, P value < 0.001). Lower "Socioeconomic Status" (SC = - 0.022, P value = 0.015) and Smoking (SC = 0.030, P value = 0.021) triggered the "Initial Periodontitis", mediated by "Systemic Circulating Inflammatory Burden of CVD Risk". Sensitivity analysis showed a dose-response relationship between "Systemic Circulating Inflammatory Burden of CVD Risk" and "Moderate Periodontitis" (SC = 0.323, P value = 0.021). CONCLUSIONS: "Systemic Circulating Inflammatory Burden of CVD Risk" appeared as an underlying mechanism of early periodontal breakdown in adolescents, also triggered by social vulnerability and smoking. CLINICAL RELEVANCE: The association between periodontitis and CVD in adulthood seems to establish much earlier in life than had been previously studied, giving impetus to preventive approaches focused on their common risk factors.

Novel 20% doxycycline-loaded PLGA nanospheres as adjunctive therapy in chronic periodontitis in type-2 diabetics: randomized clinical, immune and microbiological trial.

OBJECTIVE: This study evaluated the clinical, microbiological, and immunological results of poly lactic-co-glycolic acid (PLGA) nanospheres containing 20% doxycycline (DOXY) in the treatment of type-2 diabetic patients (DM-2) with chronic periodontitis (CP). MATERIAL AND METHODS: A parallel, double-blind, randomized, placebo-controlled clinical trial was conducted in DM-2 presenting severe and generalized CP. All patients received one-stage full-mouth ultrasonic debridement (FMUD) and they were randomly divided into two groups: PLAC (n = 20)-local application of placebo PLGA nanospheres, and DOXY (n = 20)-local application of doxycycline-loaded nanospheres; both in six non-contiguous sites. Clinical, metabolic (fasting plasma glucose level-FPG and glycated hemoglobin-HbA1c), cytokine pattern (multiplexed bead immunoassay) and microbiological assessments were performed at baseline, and 1, 3, and 6 months after treatment. RESULTS: Both groups showed clinical improvement in all parameters after treatment (p < 0.05). Deep pockets showed improvements in bleeding on probing-BoP (3 and 6 months), PD (at 3 months), and CAL gain (at 1 and 3 months) favoring DOXY (p < 0.05). The percentage of sites presenting PD reduction and CAL gain ≥ 2 mm was higher in DOXY at 3 months (p < 0.05). DOXY group exhibited a significant increase in the levels of anti-inflammatory interleukin (IL)-10 and a reduction in IL-8, IFN-y, IL-6, and IL-17 (p < 0.05), significant reduction in periodontal pathogens (p < 0.05), and a lower mean percentage of HbA1C at 3 months (p < 0.05). CONCLUSION: DOXY nanospheres may be considered a potential adjunct to mechanical debridement in the therapy of periodontitis in DM-2, offering additional benefits in deep pockets, improving the cytokine profile, and reducing periodontal pathogen levels. CLINICAL RELEVANCE: The use of locally applied doxycycline nanospheres may represent an adjunctive therapeutic approach in the treatment of periodontal disease in type-2 diabetic patients, achieving additional benefits in the local modulation of cytokines, microbial reduction, and clinical parameters, especially in deep pockets.