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).

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.

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.

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.

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.

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.

Triclosan toothpaste as an adjunct therapy to plaque control in children from periodontitis families: a crossover clinical trial.

OBJECTIVES: Studies have demonstrated that children from aggressive periodontitis (AgP) parents presented precocious alterations in their periodontal condition, and the use of chemical agents in association to plaque control could be useful to control these alterations. This study aimed to evaluate the effect of Triclosan toothpaste to modulate the clinical and subgingival condition in children from AgP parents. METHODS: Fifteen children from AgP parents and 15 from periodontally healthy parents were included in this crossover placebo study. Children were randomly allocated into triclosan or placebo therapy, using selected toothpaste for 45 days. After 15 days of wash-out, groups were crossed, changing the used toothpaste. Clinical examination and saliva, crevicular gingival fluid (GCF), and subgingival biofilm collection were performed at baseline and 45 days of each phase. GCF cytokines' levels were analyzed by Luminex/MAGpix platform and subgingival and salivary periodontal pathogens' levels by qPCR. RESULTS: At baseline, AgP group presented higher plaque index (PI), gingival index (GI), and bleeding on probing (BoP), higher Aggregatibacter actinomycetemcomitans (Aa) abundance in saliva and subgingival biofilm, and lower levels of INF-É£, IL-4, and IL-17 in GCF. Placebo therapy only reduced PI in both groups. Triclosan toothpaste reduced PI and GI in both groups. Triclosan promoted reduction of BoP and probing depth (PD), Aa salivary, and IL-1ß levels in AgP group. In health group, triclosan reduced INF-É£ and IL-4 concentration. CONCLUSION: Triclosan toothpaste demonstrated to be more effective than placebo toothpaste to control the periodontal condition in children from AgP parents, by reducing the BoP, PD, salivary Aa, and IL-1ß. CLINICAL RELEVANCE: Triclosan toothpaste can improve oral conditions in higher-risk population for AgP. TRIAL REGISTRATION: This study was registered at ClinicalTrials.gov with the identifier NCT03642353.

Does enamel matrix derivative application improve clinical outcomes after semilunar flap surgery? A randomized clinical trial.

OBJECTIVES: To evaluate the treatment of gingival recessions by semilunar coronally positioned flap plus enamel matrix derivative (SCPF + EMD). MATERIALS AND METHODS: Thirty patients with class I localized gingival recession were included. They were randomly allocated in two groups: SCPF + EMD and SCPF. Recession height (RH), recession width (RW), width of keratinized tissue (WKT), thickness of keratinized tissue (TKT), probing depth (PD), and clinical attachment level (CAL) were measured at baseline, 6 and 12 months post-surgery. Patient/professional evaluation of esthetics and root sensitivity was performed. RESULTS: After 12 months, mean root coverage was 1.98 ± 0.33 mm for SCPF + EMD (90.86 ± 14.69%) and 1.85 ± 0.41 mm (79.76 ± 17.44%) for SCPF (p > 0.05). The esthetic evaluation by the patient showed preference for SCPF + EMD. According to the professional evaluation (QCE), the use of EMD decreases the appearance of postoperative scar tissue line. There was a significant reduction in root hypersensitivity with no further complaints by the patients. CONCLUSIONS: The addition of EMD provides significantly better esthetics to SCPF, according to patient and professional assessments. SCPF + EMD is effective but not superior to SCPF for root coverage, after 12 months. CLINICAL RELEVANCE: Previous clinical trials showed that the combination of EMD with coronally advanced flaps may enhance the outcome of root coverage. There is a lack of studies testing the combination of EMD with SCPF. The combination SCPF + EMD provides better esthetics when compared to the SCPF and is effective, but not superior, to SCPF for root coverage, after 12 months. TRIAL REGISTRATION: NCT02459704.

Homeopathic medicine Cantharis modulates uropathogenic E. coli (UPEC)-induced cystitis in susceptible mice.

OBJECTIVE: This is a random blinded placebo controlled murine experimental model to study the effects of Cantharis 6 CH, a homeopathic medicine, on E coli-induced cystitis. METHODS: 24 adult susceptible female BALB/c mice were inoculated with E coli - UPEC O4:K-:H5 by a transurethral catheter. Cantharis 6cH or vehicle (placebo) was offered to mice by free access into the drinking water (1:100), during 24 h after infection. Spleen, bladder and kidneys were processed for quantitative histopathology after immunohistochemistry, using anti-CD3, CD79, MIF, NK and VEGF antibodies; the cytokines present in the bladder washing fluid were measured using a LUMINEX-Magpix KIT. Mann-Whitney and Fisher exact test were used as statistical analysis. RESULTS: Cantharis 6 CH increased IL12p40, IFN-γ and decreased IL10 concentrations in the bladder fluid (p⩽0.05); in the bladder mucosa, it increased the ratio between B and T lymphocytes (31%) and between B lymphocytes and MIF+ macrophages (57%, p⩽0.05). In the pelvis, instead, it decreased the B/T cells ratio (41%, p⩽0.05) and increased the M1/M2 macrophage ratio (42%, p⩽0.05). No differences were seen in the kidney and spleen analysis. CONCLUSION: The inverted balance of inflammatory cells and cytokines in bladder and pelvis mucosa shows specific local immune modulation induced by Cantharis 6cH.

Effect of Resveratrol on Critical-Sized Calvarial Defects of Diabetic Rats: Histometric and Gene Expression Analysis.

PURPOSE: This study investigated the influence of resveratrol (RESV) on the repair of bone critical defects in calvaria of animals with induced diabetes mellitus (DM). MATERIAL AND METHODS: One hundred rats were divided into 5 groups: induced DM + RESV administration (DM + RESV; n = 20); induced DM plus placebo solution administration (DM + PLAC; n = 20); induced DM plus insulin therapy (DM + INS; n = 20); induced DM plus administration of RES and INS (DM + RESV + INS; n = 20); and nondiabetic controls (NDM; n = 20). DM was induced by intraperitoneal injection of streptozotocin 50 mg/kg 3 days before the surgical procedures. Two critical calvarial defects were created in each animal at the start of the study (day 0). Treatments were administered from day 0 to day 30 of the experiment, when animals were euthanized. One defect was processed for histometric analysis to measure closure of the bone defect. The tissue of the other defect was analyzed for quantification of bone morphogenetic protein-2 (BMP-2), osteopontin, osteoprotegerin, receptor activator of nuclear factor-κB ligand, runt-related transcription factor-2, osterix (Osx), ß-catenin, lipoprotein receptor-related protein-5, and dikkop-1 mRNA by quantitative polymerase chain reaction. RESULTS: Histometric results showed that the DM + RESV, DM + RESV + INS, and NDM groups exhibited greater closure of the bone defects compared with the PLAC- or INS-treated groups (P < .05). Diabetic animals treated with RESV plus INS showed higher levels of BMP-2 and Osx; Osx also was positively increased in animals treated with INS alone (P < .05). CONCLUSIONS: The use of RESV, regardless of the presence of INS, positively influenced bone repair in animals with induced DM. Further, the combination of INS plus RESV was necessary for the modulation of BMP-2 gene expression.

Short-term microbiological effects of photodynamic therapy in non-surgical periodontal treatment of residual pockets: A split-mouth RCT.

BACKGROUND AND OBJECTIVE: Photodynamic therapy (PDT) has been used as a therapeutic alternative to treat periodontitis, especially in challenging sites that require additional periodontal therapy such as residual pockets. The aim of this split-mouth randomized trial was to evaluate the microbiological and clinical effects of PDT on non-surgical treatment of unresponsive pockets. STUDY DESIGN/MATERIALS AND METHODS: A split-mouth, randomized controlled clinical trial was conducted in 15 patients presenting at least two residual pockets (probing pocket depth [PPD] ≥5 mm with bleeding on probing [BoP]) in single-rooted teeth in supportive periodontal therapy. The selected sites randomly received: (1) SRP + PDT: scaling and root planing combined with photodynamic therapy (methylene blue as a photosensitizer), or (2) SRP: scaling and root planing alone. The concentrations of Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans were evaluated using a Real-time PCR technique at baseline and 3, 7, 14, and 90 days. Clinical parameters were assessed at baseline and 3 months post-therapies. RESULTS: Both treatments promoted clinical improvements, with additional benefits to the SRP + PDT group in PPD reduction and clinical attachment level gain (P < 0.05) after 3 months. Only the SRP + PDT group exhibited a statistically significant reduction in the levels of A. actinomycetemcomitans on the 3rd and 7th days after therapy (P < 0.05), and a lower concentration of this pathogen was detected at 7 days in the SRP + PDT group when compared to the other therapy (P < 0.05). CONCLUSION: The combined therapeutic approach SRP + PDT may reduce A. actinomycetemcomitans levels for a short-term period, associated with additional improvement in clinical parameters in treating residual pockets. Lasers Surg. Med. 48:944-950, 2016. © 2015 Wiley Periodicals, Inc.

Full-mouth ultrasonic debridement associated with povidone iodine rinsing in GAgP treatment: a randomised clinical trial.

OBJECTIVE: This study evaluated the clinical, immunological and microbiological results of full-mouth ultrasonic debridement (FMUD) with 10 % povidone iodine (PVPI) as the cooling liquid in the treatment of generalised aggressive periodontitis (GAgP). MATERIAL AND METHODS: Twenty-eight patients presenting GAgP were randomly assigned to one of the following groups for evaluation: FMUD + SS (n = 14)--single session of FMUD with 0.9 % saline solution as cooling agent and FMUD + PVPI (n = 14)--single session of FMUD with PVPI solution as cooling agent. Probing depth (PD), relative clinical attachment level (RCAL), relative position of gingival margin, plaque index (FMPI) and bleeding score (FMBS), immunological (interleukin-10 and interleukin-1ß concentrations in gingival crevicular fluid) and microbiological (Aa and Pg amounts) parameters were evaluated at baseline, first, third and sixth months after treatment. RESULTS: The two groups presented reduction of FMPI and FMBS and had statistically significant PD reductions, RCAL gains and gingival recession (p < 0.05). Both therapies reduced Pg levels in deep and in moderate pockets (p < 0.05). FMUD + PVPI reduced Aa levels in deep pockets. However, no inter-group differences in clinical, immunological and microbiological parameters were observed (p > 0.05). CONCLUSIONS: It could be concluded that 10 % PVPI used as an irrigant solution in FMUD decreased Aa levels in deep pockets but had no additional benefits when compared with saline solution irrigation in terms of clinical, microbiological and immunological results. CLINICAL RELEVANCE: The FMUD is a valid option for the treatment of GAgP, but the use of 10 % PVPI did not improve the results of the periodontal therapy.

Dentin hypersensitivity reduction using an arginine-based approach after non-surgical periodontal treatment.

PURPOSE: To determine the efficacy of two oral hygiene regimens in the reduction of dentin hypersensitivity (DH) on subjects undergoing non-surgical periodontal treatment (NST), over a period of 8 weeks. METHODS: 60 subjects that were randomly assigned to: Test group - NST followed in-office application of an arginine-based professional paste and toothbrushing with arginine-based toothpaste at home (n= 30) and Control group - NST followed in-office application of a fluoride-free prophylaxis paste and toothbrushing with a toothpaste based on sodium monofluorophosphate 0.76%, at home (n= 30). Air blast sensitivity assessments were made using the Schiff scale. The sensitivity parameters were measured at baseline, 1, 4 and 8 weeks. RESULTS: After 1 week, DH reduction was statistically significant for the test group (63.6%) compared to baseline, while no significant reduction was observed for the Control group (4.8%). After 4-8 weeks, the reductions were 81.6%/86.3% for the test group and 9.5%/14.2% for the Control group. When comparing the two groups, the test group showed a superior DH reduction in all evaluation periods (P< 0.05). Within the limits of the present study, it was concluded that the test oral hygiene regimen can effectively reduce dentin hypersensitivity during the most critical period after non-surgical periodontal treatment (up to 8 weeks). CLINICAL SIGNIFICANCE: The arginine-based approach provided significantly greater dentin hypersensitivity (DH) relief after non-surgical periodontal treatment (NST) when compared to the control. The combination of the in-office paste application with the daily used toothpaste may be a useful tool to reduce DH, an unpleasant and common condition that affects a large number of subjects, particularly during the initial weeks following NST.

Smoking Modulates Gene Expression of Type I Collagen, Bone Sialoprotein, and Osteocalcin in Human Alveolar Bone.

PURPOSE: Previous animal studies have shown the negative impact of smoking on bone-to-implant contact, and in humans, a decrease in bone density and implant survival over time. However, the effect of smoking on the human alveolar bone regarding the expression of bone-related markers is unknown. Therefore, the aim of this study was to evaluate the influence of smoking on the gene expression of molecules of bone metabolism in alveolar bone tissue from sites designed to receive dental implants. MATERIALS AND METHODS: Biopsy specimens of alveolar bone were collected from smokers (n = 19) and nonsmokers (n = 19) from areas planned to receive dental implants. Gene expression of tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-ß, osteoprotegerin (OPG), type I collagen (COL-I), bone sialoprotein (BSP), and osteocalcin (OCN) was quantified by quantitative real-time polymerase chain reaction using glyceraldehyde-3-phosphate dehydrogenase as a reference gene. The results were assessed using multiple regression analysis, with a significance level of 5%. RESULTS: Multiple regression analysis indicated that smoking negatively affected mRNA expression of BSP and OCN and positively altered the expression of COL-I (P < .05) despite age, gender, and arch. Moreover, regression analysis did not show a significant correlation between smoking habit and mRNA levels of TNF-α, TGF-ß, and OPG (P > .05). CONCLUSION: These results support the hypothesis that some bone markers in alveolar tissue are modulated by smoking, which could explain the negative impact of smoking on bone healing.

Efficacy of different "in-office" desensitizing treatment methods: An in vitro SEM analysis.

PURPOSE: To evaluate, in vitro, the obliteration of dentin tubules promoted by different desensitizing methods, before and after pH cycling. METHODS: Human dentin blocks of 4x4 mm were randomly divided into: control group (n = 20): no treatment; Group GH (n = 20): surface treatment with a solution containing glutaraldehyde (5.1%), HEMA (36.1%), sodium fluoride (NaF), and deionized water; Group NP (n = 20): surface treatment with a calcium phosphate gel containing nanostructured hydroxyapatite crystals, NaF and NK; and Group ARG (n = 20): surface treatment with a paste containing CaCO3 and 8% arginine. After treatment, 10 samples of each group were evaluated in scanning electron microscopy (SEM) while another 10 were included in a pH cycling procedure for 48 hours, and then analyzed in the SEM. All SEM images were evaluated by three calibrated examiners regarding dentin tubular obliteration. Mann-Whitney and Kruskal-Wallis tests were used for data analysis (P < 0.05). RESULTS: The GH, NP, and ARG groups promoted immediate obliteration higher than the control group (P < 0.05) with NP and ARG groups superior to GH group (P < 0.05). After pH cycling, an increase in tubular obliteration in the GH group was observed, which was similar to the NP and ARG groups (P > 0.05), even though all groups promoted higher obliteration than the control group (P < 0.05). All treatments were effective in a tubular obliteration.