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.

Association of active oxygen-releasing gel and photodynamic therapy in the treatment of residual periodontal pockets in type 2 diabetic patients: A randomized controlled clinical study.

BACKGROUND: The aim of this study was to evaluate the effect of active oxygen-releasing gel as an adjuvant, with and without antimicrobial photodynamic therapy (aPDT), in the treatment of residual pockets in periodontal patients with type 2 diabetes mellitus (DM2). METHODS: Patients with residual pockets with probing depth (PD) ≥4 mm and bleeding on probing (BOP) were divided into the following groups: SI (n = 17)-subgingival instrumentation in a single session; BM (n = 17)-SI followed by local application of active oxygen-releasing gel inside the periodontal pocket for 3 min; BM + aPDT (n = 17)-SI followed by application of BM for 3 min and pocket irrigation with methylene blue, and 660-nm diode laser irradiation at 100 mW for 50 s. The periodontal clinical parameters, serum levels of glycated hemoglobin, and immunological analysis of crevicular fluid were evaluated. All data were submitted to statistical analysis (α = 5%). RESULTS: A significant reduction in BOP was verified at 90 and 180 days in the BM + aPDT group. The percentage of sites with PD ≥ 4 mm was significantly reduced at 90 days in BM + aPDT and BM, whereas after 180 days only BM showed a significant reduction. In the BM + aPDT group, there was a significant reduction in tumor necrosis factor α levels at 90 days. There were no differences between the treatments. CONCLUSION: The use of adjuvant active oxygen-releasing gel, with or without aPDT, resulted in the same clinical benefits as SI in the treatment of residual pockets in poorly controlled DM2 patients.

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.

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.

Immune response characterization of primary gingival fibroblasts from Grade C periodontitis patients.

BACKGROUND: Grade C, Stage 3-4 Periodontitis (Perio4C) is a rapidly destructive disease caused by an unequilibrated immune response that starts after the primary contact of the periodontopathogens with the gingival tissue. However, it is still unclear how this imbalanced response initiates and what is the role of the connective tissue cells in the progression of this disease. Thus, this study aims to assess the local immune response of Perio4C patients through the exposure of primary gingival fibroblast cells (GFs) with Aggregatibacter actinomycetemcomitans protein extract (AaPE) and the quantification of the inflammatory cytokines interleukin (IL)-4, IL-17, tumor necrosis factor (TNF)-α, IL-1ß, interferon (IFN)-γ, and IL-10 super-family members (IL-10, IL-19, and IL-24) secreted by them. METHODS: Gingival biopsies from nine periodontally health (PH) and eight Perio4C patients were harvested, and the primary culture of GFs was obtained. The cells were exposed to AaPE (5 and 20 µg/ml) and 12-myristate 13-phorbol acetate and ionomycin - calcium salt (PMA). The supernatant was collected after 1.5 and 3 h, and a cytokine panel was evaluated. RESULTS: Clustering analysis indicated dissimilar and stimuli-dependent cytokine production between Perio4C and PH subjects. Perio4C GFs presented lower production of IL-4, TNF-α, IFN-γ, IL-17, IL-10, IL-24, and IL-19, while IL-1ß levels were similar to the PH group, leading to a disruption in the pro-/anti-inflammatory cytokine ratio (p < 0.05). IL-1ß and IL-10 super-family were the most discriminative representants for PH and Perio4C, respectively. CONCLUSION: GFs from individuals with Perio4C tended to hypo-respond to stimulation with AaPE, producing lower concentrations of some pro- and anti-inflammatory molecules, trending to develop a pro-inflammatory extracellular environment.

Anti-Inflammatory Effects of (3S)-Vestitol on Peritoneal Macrophages.

The isoflavone (3S)-vestitol, obtained from red propolis, has exhibited anti-inflammatory, antimicrobial, and anti-caries activity; however, few manuscripts deal with its anti-inflammatory mechanisms in macrophages. The objective is to elucidate the anti-inflammatory mechanisms of (3S)-vestitol on those cells. Peritoneal macrophages of C57BL6 mice, stimulated with lipopolysaccharide, were treated with 0.37 to 0.59 µM of (3S)-vestitol for 48 h. Then, nitric oxide (NO) quantities, macrophages viability, the release of 20 cytokines and the transcription of several genes related to cytokine production and inflammatory response were evaluated. The Tukey-Kramer variance analysis test statistically analyzed the data. (3S)-vestitol 0.55 µM (V55) lowered NO release by 60% without altering cell viability and diminished IL-1ß, IL-1α, G-CSF, IL-10 and GM-CSF levels. V55 reduced expression of Icam-1, Wnt5a and Mmp7 (associated to inflammation and tissue destruction in periodontitis) and Scd1, Scd2, Egf1 (correlated to atherosclerosis). V55 increased expression of Socs3 and Dab2 genes (inhibitors of cytokine signaling and NF-κB pathway), Apoe (associated to atherosclerosis control), Igf1 (encoder a protein with analogous effects to insulin) and Fgf10 (fibroblasts growth factor). (3S)-vestitol anti-inflammatory mechanisms involve cytokines and NF-κB pathway inhibition. Moreover, (3S)-vestitol may be a candidate for future in vivo investigations about the treatment/prevention of persistent inflammatory diseases such as atherosclerosis and periodontitis.

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).

Open flap debridement compared to repeated applications of photodynamic therapy in the treatment of residual pockets: A randomized clinical trial.

BACKGROUND: The aim of the present study was to compare repeated applications of antimicrobial photodynamic therapy (aPDT) to open flap debridement (OFD) in the treatment of residual periodontal pockets in non-furcation sites. METHODS: Forty-six subjects with a diagnosis of Stage III or IV Grade C periodontitis, that had been previously treated, participated in the study.  Residual pockets were divided between two groups: (1) aPDT group: received ultrasonic periodontal debridement followed by immediate application of aPDT, and repeated on1st, 2nd, 7th, and 14th days; and (2) OFD group: treated by modified papilla preservation technique, where granulation tissue and visible calculus were removed with hand curettes and an ultrasonic device. Clinical, immunological, and microbiological parameters were evaluated before and after treatment. RESULTS: Both treatments were effective reducing clinical parameters of disease. OFD resulted in a greater mean probing pocket depths (PPD) reduction in deep pockets (p = 0.001). However, aPDT resulted in a lower occurrence of gingival recession (GR), dentin hypersensitivity (DH) and analgesic intake. Reduction in Porphyromonas gingivalis was observed in both groups. Only the OFD group had a significant reduction in Aggregatibacter actinomycetemcomitans. aPDT group had greater increase in interleukin 10 (IL-10) levels and a greater reduction of interleukin 1 beta (IL-1ß) at 14 days when compared to the OFD group (p < 0.05). CONCLUSION: OFD was superior in reducing PPD in deep pockets compared to the aPDT. However, OFD resulted in greater GR.  Both treatments lowered P. gingivalis levels but only OFD reduced levels of A. actinomycemtemcomitans.

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.

The use of probiotics can reduce the severity of experimental periodontitis in rats with metabolic syndrome: An immunoenzymatic and microtomographic study.

BACKGROUND: This study evaluated the effects of Bifidobacterium animalis subsp. lactis HN019 (B. lactis HN019) in the development of periodontitis (PE), associated or not with metabolic syndrome, (MS) in rats. METHODS: Ninety-six rats were grouped according to a food protocol: high-fat diet for induction of MS or standard diet for the control groups (C). They were subdivided into groups with (+) and without (-) PE, receiving (*) or not (**) probiotic (PROB): C-**, CP-*, PE+**, PEP+*, MS- MSP-*, MSPE+**, and MSPEP+*. PROB administration started on the eighth week of the study and PE was induced on the 14th week by placing ligature on the animals' lower first molars. Euthanasia occurred in the 16th week. Biomolecular analyzes, immunoenzymatic assays, and microtomographic analyses were performed. The data obtained were analyzed statistically (P < 0.05). RESULTS: The PEP and MSPEP groups showed lower levels of alveolar bone loss when compared with the PE and MSPE groups, respectively (P < 0.05). The immunoenzymatic analysis showed higher levels of interleukin (IL)-1ß and a higher receptor activator of NF-kappaB ligand (RANKL)/osteoprotegerin (OPG) ratio in the MSPE group when compared with the MSPEP group (P < 0.05). The PEP group showed lower levels of tumor necrosis factor (TNF)-α and IL-6 when compared with the PE group. The use of PROB attenuated dyslipidemia parameters in animals with MS, with or without PE. CONCLUSION: B. lactis HN019 reduced more significantly the severity of PE in rats with MS, modulating both systemic metabolic and immunoinflammatory parameters in periodontal tissues.

Predicted functional and taxonomic analysis of subgingival biofilm of Grade C periodontitis in young patients under maintenance therapy.

BACKGROUND: In Grade C periodontitis in young patients (PerioC-Y), the functional roles of the subgingival community after years of periodontal treatment are still underexplored. This study evaluated the taxonomic and predicted functional content of the subgingival microbiome of PerioC-Y patients under supportive periodontal therapy (SPT). METHODS: Clinical and microbiological data from subgingival biofilm were assessed from 10 PerioC-Y patients at two time points: at baseline and after 5.7 ± 1.3 years of SPT. This was compared with 15 patients without a history of periodontitis. The V1-V3 and V4-V5 regions of the 16S rRNA were sequenced using the Illumina Miseq. Microbial composition was evaluated by the core microbiome, and alpha- and beta-diversity. The microbiome functional content was predicted using Picrust2, and the gene differential abundance was analyzed with DESeq2. RESULTS: Clinical improvements were seen in PerioC-Y-SPT. Differences in ß-diversity between PerioC-Y and health were observed (health x PerioC-Y-baseline, P = 0.02; health x PerioC-Y-SPT, P = 0.05). Moreover, although ß-diversity did not statistically change between baseline and SPT in PerioC-Y, the microbial correlation evidenced increased Streptococcus and decreased Treponema network contributions during SPT. Based on predicted functional data, treatment induced a reduction in genes related to flagellar protein and signal transduction in PerioC-Y. However, compared with healthy individuals, some genes remained more highly abundant in PerioC-Y-SPT, such as quorum sensing and efflux pump transporters. CONCLUSION: Despite clinical improvements and a shift in taxonomic composition, the PerioC-Y patients' periodontal treatment was not enough to reach a similar microbiome to patients without disease experience. Some functional content in this biofilm remained altered in PerioC-Y regardless of disease control.

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.

Stanniocalcin 2 contributes to aggressiveness and is a prognostic marker for oral squamous cell carcinoma.

Stanniocalcin 2 (STC2), a glycoprotein that regulates calcium and phosphate homeostasis during mineral metabolism, appears to display multiple roles in tumorigenesis and cancer progression. This study aimed to access the prognostic value of STC2 in oral squamous cell carcinoma (OSCC) and its implications in oral tumorigenesis. STC2 expression was examined in 2 independent cohorts of OSCC tissues by immunohistochemistry. A loss-of-function strategy using shRNA targeting STC2 was employed to investigate STC2 in vitro effects on proliferation, apoptosis, migration, invasion, epithelial-mesenchymal transition (EMT) and possible activation of signaling pathways. Moreover, STC2 effects were assessed in vivo in a xenograft mouse cancer model. High expression of STC2 was significantly associated with poor disease-specific survival (HR: 2.67, 95% CI: 1.37-5.21, p = 0.001) and high rate of recurrence with a hazard ratio of 2.80 (95% CI: 1.07-5.71, p = 0.03). In vitro downregulation of STC2 expression in OSCC cells attenuated proliferation, migration and invasiveness while increased apoptotic rates. In addition, the STC2 downregulation controlled EMT phenotype of OSCC cells, with regulation on E-cadherin, vimentin, Snail1, Twist and Zeb2. The reactivation of STC2 was observed in the STC2 knockdown cells in the in vivo xenograft model, and no influence on tumor growth was observed. Modulation of STC2 expression levels did not alter consistently the phosphorylation status of CREB, ERK, JNK, p38, p70 S6K, STAT3, STAT5A/B and AKT. Our findings suggest that STC2 overexpression is an independent marker of OSCC outcome and may contribute to tumor progression via regulation of proliferation, survival and invasiveness of OSCC cells.

Clinical Investigation of Microbial Profile and Levels of Endotoxins and Lipoteichoic Acid at Different Phases of the Endodontic Treatment in Teeth with Vital Pulp and Associated Periodontal Disease.

INTRODUCTION: The aim of this in vivo study was to investigate the microbial profile as well as the levels of lipopolysaccharide (LPS) and lipoteichoic acid (LTA) at different phases of endodontic treatment in teeth with vital pulp and associated periodontal disease. METHODS: Ten patients were selected for this clinical study. Samples were taken from periodontal pockets (PPs) and root canals (RCs) using sterile paper points before and after chemomechanical preparation and after intracanal medication. For microbiological analysis, nested polymerase chain reaction and checkerboard DNA-DNA hybridization were used. Levels of LPS and LTA were assessed using limulus amebocyte lysate and enzyme-linked immunosorbent assays, respectively. Data were statistically analyzed at a significance level of 5%. RESULTS: Bacterial DNA from 17 of the 17 species investigated was detected in samples of PPs, whereas 6 of the 17 species were not present in the initial samples of RCs using nested polymerase chain reaction. In the initial samples, 38 of 40 probes were detected in PPs, whereas 12 of 40 probes were detected in RCs using checkerboard DNA-DNA hybridization. Overall, endodontic procedures were efficient in modifying the microbiota of PPs and RCs. Levels of LPS and LTA were reduced after the endodontic procedures, although higher concentrations of both had been found in PPs compared with RCs. CONCLUSIONS: The microbiota of PPs and RCs in teeth with vital pulp and associated periodontal disease is polymicrobial, with the presence of gram-negative, gram-positive, facultative, and strict anaerobes. Chemomechanical preparation and calcium hydroxide-based intracanal medication allowed the reduction of infectious content in both sites.

Novel rare frameshift variation in aggressive periodontitis: Exomic and familial-screening analysis.

BACKGROUND: Aggressive periodontitis (AgP), currently periodontitis grade C, presents early onset, rapid progression, and a poorly established genetic association. Thus, this study aimed to identify genetic variants associated with AgP via whole exome sequencing (WES) through a familial screening approach. METHODS: WES was performed in two nuclear families, including a proband and a parent affected by AgP and an unaffected parent and sibling. Common variants among affected individuals, excluding those common to healthy people, from each family, composed the data set associated with AgP. In silico analysis evaluated the impact of each variant on protein structure and protein-protein interactions. Moreover, identified deleterious variants were validated in a populational analysis (n = 96). RESULTS: The missense single nucleotide variations (SNVs) rs142548867 in EEFSEC (c.668C>T), rs574301770 in ZNF136 (c.466C>G), and rs72821893 in KRT25 (c.800G>A) and the frameshift indels rs37146475 in GPRC6A (c.2323-2324insT) and c.1366_1372insGGAGCAG in ELN were identified in AgP and have a predicted functional impact on proteins. In silico analysis indicated that the indel in GPRC6A generates a loss of the C-terminal tail of the Gprca protein. Furthermore, this SNV was significantly associated with AgP in a population-based investigation. CONCLUSION: Novel frameshift variation in GPRC6A (c.2323-2324insT) was identified as a potential genetic alteration associated with AgP occurrence.

Local IL-10 level as a predictive factor in generalized aggressive periodontitis treatment response.

Generalized aggressive periodontitis (GAgP) presents a reduced response to non-surgical therapy. However, it is not clear if the initial clinical, microbiological or immunological characteristics are impacting the worse response to treatment. This study aimed to identify the predictive value of clinical, microbiological and immunological patterns on the clinical response to therapy in GAgP patients. Twenty-four GAgP patients were selected, and gingival crevicular fluid (GCF) and subgingival biofilm were collected. Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythia levels were evaluated by qPCR, and IL-1ß and IL-10 concentration by ELISA. Twelve patients were treated with SRP (scaling and root planning), and twelve with SRP plus 375 mg amoxicillin and 250 mg metronidazole (8/8 hours, 7 days) (SRP + AM). The clinical changes (Probing Pocket Depth [PPD] reduction and Clinical Attachment Level [CAL] gain) 6 months post-treatment were correlated to the initial clinical, inflammatory and microbiological variables using stepwise logistic regression (α = 5%). CAL gain at 6 months was 1.16 ± 0.77 for SRP and 1.74 ± 0.57 mm for SRP + AM (P > .05). PPD reduction was 1.96 ± 0.82 for SRP and 2.45 ± 0.77 mm for SRP + AM (P < .05). In the SRP group, IL-10 showed a predictive value for clinical response. The higher the IL-10 concentration at baseline, the higher the reduction in PPD at 6 months (P = .01, r = .68). However, when antimicrobials were administered, no significant influence was detected (P > .05). It can be concluded that the IL-10 levels in GFC act as a predictor of clinical response to GAgP. Moreover, the intake of antimicrobials appears to overlap the influence of the inflammatory response on clinical response to treatment. Clinical trial registration number: NCT03933501.

Microbiota of periodontal pockets and root canals in induced experimental periodontal disease in dogs.

AIM: To investigate the relationship between the microbiota of periodontal pockets (PP) and root canals (RC) in dogs submitted to experimental periodontal disease (ExPD). METHODS: ExPD was induced by combining cotton and wire ligatures. After 125 days, microbiological samples were collected from PP and RC. Strains isolated from 19 teeth were submitted to DNA extraction, 16S rRNA gene amplification and gene sequencing. Pearson's χ2 - and Fisher's exact tests and McNemar's test were used when appropriate. RESULTS: The number of species in PP was greater than in RC, with prevalence of obligate anaerobes and Gram-negative bacteria. In the PP predominated Fusobacterium necrophorum, Porphyromonas gingivalis, Prevotella loescheii, Campylobacter gracilis and Veillonella parvula. In the RC samples, 9 had microbial growth, with predominance of the following genera: Staphylococcus, Streptococcus and Neisseria. Eight genera were common to both sites in the same tooth. PP presented a greater number of species than the RC. No significant difference was observed in the species found in PP and RC in the same tooth. CONCLUSION: Microbial composition of the RC could be modulated by the presence of periodontal disease, especially in cases of severe periodontal destruction. RC microbiota was less complex and diverse than the PP.

Clinical and molecular analysis in Papillon-Lefèvre syndrome.

Papillon-Lefèvre syndrome (PLS; MIM#245000) is a rare recessive autosomal disorder characterized by palmar and plantar hyperkeratosis, and aggressively progressing periodontitis leading to premature loss of deciduous and permanent teeth. PLS is caused by loss-of-function mutations in the CTSC gene, which encodes cathepsin C. PLS clinical expressivity is highly variable and no consistent genotype-phenotype correlation has been demonstrated yet. Here we report the clinical and genetic features of five PLS patients presenting a severe periodontal breakdown in primary and permanent dentition, hyperkeratosis over palms and soles, and recurrent sinusitis and/or tonsillitis. Mutation analysis revealed two novel homozygous recessive mutations (c.947T>C and c.1010G>C) and one previous described homozygous recessive mutation (c.901G>A), with parents carrying them in heterozygous, in three families (four patients). The fourth family presented with the CTSC c.628C>T mutation in heterozygous, which was inherited maternally. Patient carrying the CTSC c.628C>T mutation featured classical PLS phenotype, but no PLS clinical characteristics were found in his carrier mother. All mutations were found to affect directly (c.901G>A, c.947T>C, and c.1010G>C) or indirectly (c.628C>T, which induces a premature termination) the heavy chain of the cathepsin C, the region responsible for activation of the lysosomal protease. Together, these findings indicate that both homozygous and heterozygous mutations in the cathepsin C heavy chain domain may lead to classical PLS phenotype, suggesting roles for epistasis or gene-environment interactions on determination of PLS phenotypes.

Two different antibiotic protocols as adjuncts to one-stage full-mouth ultrasonic debridement to treat generalized aggressive periodontitis: A pilot randomized controlled clinical trial.

BACKGROUND: To assess the clinical and microbiological responses of amoxicillin + metronidazole (AMX + MET) versus clarithromycin (CLM) as adjuncts to one-stage full-mouth ultrasonic debridement (FMUD) in the treatment of generalized aggressive periodontitis (GAgP). METHODS: For this parallel, double-masked, pilot randomized clinical trial, 46 patients with GAgP were selected and randomly assigned into two groups: AMX+MET group (n = 23): FMUD associated with AMX (500 mg three times a day) and MET (400 mg three times a day) for 7 days; and CLM group (n = 23): FMUD associated with CLM (500 mg twice a day) for 7 days. Clinical parameters were evaluated at baseline, 3, and 6 months post-treatment. The levels of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, and Fusobacterium nucleatum from subgingival biofilm were determined by quantitative polymerase chain reaction. RESULTS: Both treatments significantly improved all clinical parameters compared with baseline and promoted a significant reduction of A. actinomycetemcomitans and P. gingivalis counts (P > 0.05). CLM succeeded in decreasing T. forsythia at 6 months (P < 0.05), but no antibiotic was able to reduce F. nucleatum. There was no difference between the two protocols regarding the reported adverse effects (P > 0.05). CONCLUSIONS: The results suggest that CLM is not superior than AMX + MET in the treatment of GAgP. However, this antibiotic led to good clinical outcomes and may be a possible alternative to AMX+MET in the treatment of severe periodontitis in young patients. Future studies with larger sample sizes are needed to confirm this statement (NCT02969928).

Pioglitazone abolishes cognition impairments as well as BDNF and neurotensin disturbances in a rat model of autism.

We have shown that exposure of rats to lipopolysaccharide (LPS) during gestation induces autistic-like behaviors in juvenile offspring and pioglitazone post treatment corrects social and communication deficits. The first objective of the present study was to evaluate the cognition of the rats, because this is also a behavioral sphere committed in autism. Second, biomarkers related to pioglitazone pathways and autism were studied to try to understand their mechanisms. We used our rat model of autism and pioglitazone was administered daily to these young offspring. T-maze spontaneous alternations tests, plasma levels of brain-derived neurotrophic factor (BDNF), beta-endorphin, neurotensin, oxytocin, and substance P were all studied. Exposure of rats to LPS during gestation induced cognitive deficits in the young offspring, elevated BDNF levels and decreased neurotensin levels. Daily postnatal pioglitazone treatment abolished cognition impairments as well as BDNF and neurotensin disturbances. Together with our previous studies, we suggest pioglitazone as a candidate for the treatment of autism, because it improved the responses of the three most typical autistic-like behaviors. BDNF and neurotensin also appeared to be related to the autistic-like behaviors and should be considered for therapeutic purposes.