JARID1B represses the osteogenic potential of human periodontal ligament mesenchymal cells.

BACKGROUND: Here, we evaluated whether the histone lysine demethylase 5B (JARID1B), is involved in osteogenic phenotype commitment of periodontal ligament cells (PDLCs), by considering their heterogeneity for osteoblast differentiation. MATERIALS AND METHODS: Epigenetic, transcriptional, and protein levels of a gene set, involved in the osteogenesis, were investigated by performing genome-wide DNA (hydroxy)methylation, mRNA expression, and western blotting analysis at basal (without osteogenic induction), and at the 3rd and 10th days of osteogenic stimulus, in vitro, using PDLCs with low (l) and high (h) osteogenic potential as biological models. RESULTS: h-PDLCs showed reduced levels of JARID1B, compared to l-PDLCs, with significant inversely proportional correlations between RUNX2 and RUNX2/p57. Epigenetically, a significant reduction in the global H3K4me3 content was observed only in h-PDLCs. Immunoblotting data reveal a significant reduction in the global H3K4me3 content, at 3 days of induction only in h-PDLCs, while an increase in the global H3K4me3 content was observed at 10 days for both PDLCs. Additionally, positive correlations were found between global H3K4me3 levels and JARID1B gene expression. CONCLUSIONS: Altogether, our results show the crucial role of JARID1B in repressing PDLCs osteogenic phenotype and this claims to pre-clinical protocols proposing JARID1B as a potential therapeutic target.

Protease-activated receptor type 2 activation downregulates osteogenesis in periodontal ligament stem cells.

Protease-activated receptor-2 (PAR2) is associated with the pathogenesis of many chronic diseases with inflammatory characteristics, including periodontitis. This study aimed to evaluate how the activation of PAR2 can affect the osteogenic activity of human periodontal ligament stem cells (PDLSCs) in vitro. PDLSCs collected from three subjects were treated in osteogenic medium for 2, 7, 14, and 21 days with trypsin (0.1 U/mL), PAR2 specific agonist peptide (SLIGRL-NH2) (100 nM), and PAR2 antagonist peptide (FSLLRY-NH2) (100 nM). Gene (RT-qPCR) expression and protein expression (ELISA) of osteogenic factors, bone metabolism, and inflammatory cytokines, cell proliferation, alkaline phosphatase (ALP) activity, alizarin red S staining, and supernatant concentration were assessed. Statistical analysis of the results with a significance level of 5% was performed. Activation of PAR2 led to decreases in cell proliferation and calcium deposition (p < 0.05), calcium concentration (p < 0.05), and ALP activity (p < 0.05). Additionally, PAR2 activation increased gene and protein expression of receptor activator of nuclear factor kappa-Β ligand (RANKL) (p < 0.05) and significantly decreased the gene and protein expression of osteoprotegerin (p <0. 05). Considering the findings, the present study demonstrated PAR2 activation was able to decrease cell proliferation, decreased osteogenic activity of PDLSCs, and upregulated conditions for bone resorption. PAR2 may be considered a promising target in periodontal regenerative procedures.

Protease-activated receptor type 2 activation downregulates osteogenesis in periodontal ligament stem cells

Abstract Protease-activated receptor-2 (PAR2) is associated with the pathogenesis of many chronic diseases with inflammatory characteristics, including periodontitis. This study aimed to evaluate how the activation of PAR2 can affect the osteogenic activity of human periodontal ligament stem cells (PDLSCs) in vitro. PDLSCs collected from three subjects were treated in osteogenic medium for 2, 7, 14, and 21 days with trypsin (0.1 U/mL), PAR2 specific agonist peptide (SLIGRL-NH2) (100 nM), and PAR2 antagonist peptide (FSLLRY-NH2) (100 nM). Gene (RT-qPCR) expression and protein expression (ELISA) of osteogenic factors, bone metabolism, and inflammatory cytokines, cell proliferation, alkaline phosphatase (ALP) activity, alizarin red S staining, and supernatant concentration were assessed. Statistical analysis of the results with a significance level of 5% was performed. Activation of PAR2 led to decreases in cell proliferation and calcium deposition (p < 0.05), calcium concentration (p < 0.05), and ALP activity (p < 0.05). Additionally, PAR2 activation increased gene and protein expression of receptor activator of nuclear factor kappa-Β ligand (RANKL) (p < 0.05) and significantly decreased the gene and protein expression of osteoprotegerin (p <0. 05). Considering the findings, the present study demonstrated PAR2 activation was able to decrease cell proliferation, decreased osteogenic activity of PDLSCs, and upregulated conditions for bone resorption. PAR2 may be considered a promising target in periodontal regenerative procedures.

FTIR spectroscopy as a point of care diagnostic tool for diabetes and periodontitis: A saliva analysis approach.

Diabetes mellitus (DM) is a general term for heterogeneous metabolic disorders whose main characteristic is chronic hyperglycemia. Considering that conventional diagnostic methods are currently unable of early DM detection and the number of diabetic patients has been increasing worldwide, there is a clinical need for novel diagnostic approaches. Fourier-transform infrared (FT-IR) spectroscopy technique an alternative non-invasive diagnostic method for real-time evaluation of biofluids such as saliva. This study aims evaluate the feasibility of diagnosing diabetes and periodontitis through saliva samples based on their FT-IR spectra. Our first collection and spectral analysis of samples was a pilot study and comprised a total of 23 patients, 2 healthy, 9 with diabetes and 12 with diabetes and periodontitis. By using weighted KNN as classifier, we have obtained an area under the Receiver Operating Characteristic curve of 0.92 and 0.95 when considering the diabetes or diabetes + periodontitis groups as positive groups, respectively.

Protease-activated receptor type 1 (PAR1) increases CEMP1 gene expression through MAPK/ERK pathway.

PAR1 is a G-coupled protein receptor that regulates several cellular metabolism processes, including differentiation and proliferation of osteogenic and cementogenic related cells and our group previously demonstrated the regenerative potential of PAR1 in human periodontal ligament stem cells (hPDLSCs). In this study, we hypothesized that PAR1 regulates the cementogenic differentiation of hPDLSCs. Our goal was to identify the intracellular signaling pathway underlying PAR1 activation in hPDSLC differentiation. hPDLSCs were isolated using the explant technique. Cells were cultured in an osteogenic medium (OST) (α-MEM, 15% fetal bovine serum, L-glutamine, penicillin, streptomycin, amphotericin B, dexamethasone, and beta-glycerophosphate). The hPDLSCs were treated with a specific activator of PAR1 (PAR1 agonist) and blockers of the MAPK/ERK and PI3K pathways for 2 and 7 days. The gene expression of CEMP1 was assessed by RT-qPCR. The activation of PAR1 by its agonist peptide led to an increase in CEMP1 gene expression when compared with OST control. MAPK/ERK blockage abrogated the upregulation of CEMP1 gene expression induced by PAR1 agonist (p < 0.05). PI3K blockage did not affect the gene expression of CEMP1 at any experimental time (p > 0.05). We concluded that CEMP1 gene expression increased by PAR1 activation is MAPK/ERK-dependent and PI3K independent, suggesting that PAR1 may regulate cementogenetic differentiation of hPDLSCs.

Protease-activated receptor type 1 (PAR1) increases CEMP1 gene expression through MAPK/ERK pathway

Abstract: PAR1 is a G-coupled protein receptor that regulates several cellular metabolism processes, including differentiation and proliferation of osteogenic and cementogenic related cells and our group previously demonstrated the regenerative potential of PAR1 in human periodontal ligament stem cells (hPDLSCs). In this study, we hypothesized that PAR1 regulates the cementogenic differentiation of hPDLSCs. Our goal was to identify the intracellular signaling pathway underlying PAR1 activation in hPDSLC differentiation. hPDLSCs were isolated using the explant technique. Cells were cultured in an osteogenic medium (OST) (α-MEM, 15% fetal bovine serum, L-glutamine, penicillin, streptomycin, amphotericin B, dexamethasone, and beta-glycerophosphate). The hPDLSCs were treated with a specific activator of PAR1 (PAR1 agonist) and blockers of the MAPK/ERK and PI3K pathways for 2 and 7 days. The gene expression of CEMP1 was assessed by RT-qPCR. The activation of PAR1 by its agonist peptide led to an increase in CEMP1 gene expression when compared with OST control. MAPK/ERK blockage abrogated the upregulation of CEMP1 gene expression induced by PAR1 agonist (p < 0.05). PI3K blockage did not affect the gene expression of CEMP1 at any experimental time (p > 0.05). We concluded that CEMP1 gene expression increased by PAR1 activation is MAPK/ERK-dependent and PI3K independent, suggesting that PAR1 may regulate cementogenetic differentiation of hPDLSCs.

The Effects of Patient Compliance in Supportive Periodontal Therapy on Tooth Loss: A systematic Review and Meta-analysis.

BACKGROUND: The present review aimed to assess the impact of being a complier to supportive periodontal therapy (SPT), when compared to not being a complier, on tooth loss in patients with periodontitis. METHODS: Prospective and retrospective observational studies were included. MEDLINE, EMBASE, and LILACS databases were searched up to May 2019. The odds-ratio (OR) and standard error (SE) values of the studied groups (compliant or non-compliant) were converted to logOR, and the results of individual studies were grouped using a random effects model. RESULTS: From a total of 1815 articles initially searched, 13 retrospective studies and one prospective study comparing tooth loss of complier and non-complier individuals in SPT were included. Meta-analysis of eight studies showed that non-compliers in SPT have an increased risk of tooth loss when compared with compliers. Overall meta-analysis demonstrated that non-compliant patients in SPT have a 26% increased risk of tooth loss when compared with compliant patients (OR = 1.26; 95% CI = 1.06 to 1.51, Heterogeneity: I2 = 0%, p = 0.008). CONCLUSIONS: Patients with periodontitis who do not comply in SPT have a higher risk of tooth loss than compliant patients. Oral health professionals should implement measures to obtain optimal adherence by patients in SPT.

Non-coding RNAs repressive role in post-transcriptional processing of RUNX2 during the acquisition of the osteogenic phenotype of periodontal ligament mesenchymal stem cells.

Mesenchymal stem cells are candidates for therapeutic strategies in periodontal repair due to their osteogenic potential. In this study, we identified epigenetic markers during osteogenic differentiation, taking advantage of the individual pattern of mesenchymal cells of the periodontal ligament with high (h-PDLCs) and low (l-PDLCs) osteogenic capacity. We found that the involvement of non-coding RNAs in the regulation of the RUNX2 gene is strongly associated with high osteogenic potential. Moreover, we evaluated miRs and genes that encode enzymes to process miRs and their biogenesis. Our data show the high expression of the XPO5 gene, and miRs 7 and 22 observed in the l-PDLCs might be involved in acquiring osteogenic potential, suppressing RUNX2 gene expression. Further, an inversely proportional correlation between lncRNAs (HOTAIR and HOTTIP) and RUNX2 gene expression was observed in both l- and h-PDLCs, and it was also related to the distinct osteogenic phenotypes. Thus, our results indicate the low expression of XPO5 in h-PDLC might be the limiting point for blocking the miRs biogenesis, allowing the high gene expression of RUNX2. In accordance, the low expression of miRs, HOTAIR, and HOTTIP could be a prerequisite for increased osteogenic potential in h-PDLCs. These results will help us to better understand the underlying mechanisms of osteogenesis, considering the heterogeneity in the osteogenic potential of PDLCs that might be related to a distinct transcriptional profile of lncRNAs and the biogenesis machinery.

Clinical and microbiological evaluation of non-surgical periodontal therapy in obese and non-obese individuals with periodontitis: a 9-month prospective longitudinal study.

Objective Obesity is a chronic disease that negatively affects an individual's general and oral health. The present study aimed to compare the clinical and microbiological effects of non-surgical periodontal therapy with the full mouth disinfection (FMD) protocol on obese and non-obese individuals at 9 months post-therapy. Methodology This clinical study was first submitted and approved by the Ethics Committee. Fifty-five obese patients and 39 non-obese patients with periodontitis were evaluated. The full-mouth periodontal clinical parameters, clinical attachment level (CAL), probing depth (PD), gingival index (GI), and plaque index (PI), were monitored at baseline, 3, 6, and 9 months after periodontal treatment with full mouth disinfection (FMD) protocol. The mean count of Tannerella forsythia , Porphyromonas gingivalis , Treponema Denticola , and Aggregatibacter actinomycetemcomitans was determined by quantitative polymerase chain reaction on subgingival biofilm samples. Demographic data were assessed by Chi-square test. For clinical and microbiological parameters, two-factor repeated-measures ANOVA was used. Results In both groups, periodontal therapy using the one-stage full-mouth disinfection protocol significantly improved CAL, PD, GI, and PI (p<0.05). Obese and non-obese patients equally responded to non-surgical periodontal therapy (p>0.05). Microbial count found no major differences (p>0.05) between obese and non-obese individuals who had undergone non-surgical periodontal therapy. Conclusions Obesity did not affect the clinical and microbiological outcomes of non-surgical periodontal therapy.

Root Coverage Procedures in Noncarious Cervical Lesions With and Without Restoration: A Systematic Review and Meta-Analysis.

The aim of this systematic review was to assess in patients with gingival recessions and noncarious cervical lesions (NCCLs) whether restoration of NCCLs may influence the percentage of root coverage following surgical root coverage procedures compared to surgical root coverage procedures without subsequent restoration. Four studies (randomized controlled trials) assessing the effects of NCCL restoration in combination with surgical root coverage procedures were included. Meta-analyses showed no significant differences in overall root coverage, CAL gain, and KTW change between test and control groups. In teeth with NCCLs and gingival recessions, restoration of NCCLs does not affect the clinical outcomes of surgical root coverage.

Clinical and microbiological evaluation of non-surgical periodontal therapy in obese and non-obese individuals with periodontitis: a 9-month prospective longitudinal study

Abstract Objective Obesity is a chronic disease that negatively affects an individual's general and oral health. The present study aimed to compare the clinical and microbiological effects of non-surgical periodontal therapy with the full mouth disinfection (FMD) protocol on obese and non-obese individuals at 9 months post-therapy. Methodology This clinical study was first submitted and approved by the Ethics Committee. Fifty-five obese patients and 39 non-obese patients with periodontitis were evaluated. The full-mouth periodontal clinical parameters, clinical attachment level (CAL), probing depth (PD), gingival index (GI), and plaque index (PI), were monitored at baseline, 3, 6, and 9 months after periodontal treatment with full mouth disinfection (FMD) protocol. The mean count of Tannerella forsythia , Porphyromonas gingivalis , Treponema Denticola , and Aggregatibacter actinomycetemcomitans was determined by quantitative polymerase chain reaction on subgingival biofilm samples. Demographic data were assessed by Chi-square test. For clinical and microbiological parameters, two-factor repeated-measures ANOVA was used. Results In both groups, periodontal therapy using the one-stage full-mouth disinfection protocol significantly improved CAL, PD, GI, and PI (p<0.05). Obese and non-obese patients equally responded to non-surgical periodontal therapy (p>0.05). Microbial count found no major differences (p>0.05) between obese and non-obese individuals who had undergone non-surgical periodontal therapy. Conclusions Obesity did not affect the clinical and microbiological outcomes of non-surgical periodontal therapy.

Effect of magnification on root coverage surgery: a systematic review

Root coverage surgery can be performed in patients with gingival recession to cover the exposed root aiming to control hypersensitivity and promotes better aesthetic. Optical magnification has been proposed as a refinement in this surgical technique to increase root coverage. This approach may lead to enhanced soft tissue stability, less post-operative discomfort, better predictability and esthetic appearance. Aim: This systematic review aimed to evaluate the effectiveness of magnification on root coverage surgery when compared to procedures performed without magnification. Methods: Randomized controlled trials with a follow-up of at least 6 months that compared surgeries for root coverage performed under optic magnification versus conventional (macro) root coverage surgery were screened. The primary outcome was mean root coverage (mm) (MRC) and secondary outcomes were percentage of root coverage (PRC) and complete root coverage (CRC). Results: Of 569 papers relevant to this review, seven were included. Meta-analysis showed that the use of magnification may favor greater PRC (7.38%, 95% CI 3.66-11.09). Conclusion: Magnification can increase PRC in root coverage surgeries. More randomized trials with the use of magnification may be necessary to verify if this benefit is clinically relevant, in order to justify the use of this device

Effect of smoking cessation on tooth loss: a systematic review with meta-analysis.

BACKGROUND: Smoking is a major risk factor for periodontitis and tooth loss. Smoking cessation has a positive impact in periodontal treatment. However, so far, no systematic review has evaluated the effect of smoking cessation on tooth loss. Therefore, this review aimed to evaluate if smoking cessation reduces the risk of tooth loss. METHODS: Observational (cross-sectional and longitudinal) studies that investigated the association between smoking cessation and tooth loss were included. MEDLINE, EMBASE and LILACS databases were searched for articles published up to November 2018. Pooled results for subgroups of current and former smokers were compared in meta-analysis. Meta-regression was used to test the influence of smoking status on estimates and explore the heterogeneity. RESULTS: Of 230 potentially relevant publications, 21 studies were included in the qualitative review and 12 in the quantitative analysis. Meta-analysis of cross-sectional studies did not show any differences between former and current smokers in the chance of losing 1 or more teeth (OR = 1.00; 95% CI = 0.80 to 1.24, I2 = 80%), losing more than 8 teeth (OR = 1.02; 95% CI = 0.78 to 1.32, I2 = 0%) or being edentulous (OR = 1.37; 95% CI = 0.94 to 1.99, I2 = 98%). Meta-analysis from longitudinal studies showed that, when compared to never smokers, former smokers presented no increased risk of tooth loss (RR = 1.15; 95% CI = 0.98 to 1.35, I2 = 76%), while current smokers presented an increased risk of tooth loss (RR = 2.60; 95% CI = 2.29 to 2.96, I2 = 61%). Meta-regression showed that, among former smokers, the time of cessation was the variable that better explained heterogeneity (approximately 60%). CONCLUSIONS: Risk for tooth loss in former smokers is comparable to that of never smokers. Moreover, former smokers have a reduced risk of tooth loss, when compared to current smokers.

Efficacy of preprocedural mouthrinses in the reduction of microorganisms in aerosol: A systematic review.

BACKGROUND: The authors of this systematic review aimed to evaluate the efficacy of preprocedural mouthrinses in reducing the number of microorganisms disseminated by means of the aerosol generated via dental procedures when compared with a placebo, water, or no mouthrinse. TYPES OF STUDIES REVIEWED: The authors included only randomized clinical trials. They searched MEDLINE (PubMed), Embase, Google Scholar, and Latin American and Caribbean Health Sciences Literature databases through May 31, 2019. They performed random-effects meta-analysis for reduction of the number of colony-forming units (CFU) in the dental aerosol. RESULTS: Of 770 potentially relevant articles, the authors included 13 randomized clinical trials in which researchers studied the efficacy of chlorhexidine, essential oils, cetylpyridinium chloride, and herbal products. Meta-analysis of 12 studies showed that mouthrinses with chlorhexidine, essential oils, and cetylpyridinium chloride significantly reduced the number of CFU. Overall, the use of a preprocedural mouthrinse resulted in a mean reduction in the number of CFUs of 64.8% (95% confidence interval, 50.4% to 79.3%; I2 = 37%) compared with control. None of the included studies presented a low risk of bias. PRACTICAL IMPLICATIONS: Some dental procedures result in dissemination of microorganisms in the aerosol in the dental office. There is moderate evidence that preprocedural mouthrinses significantly reduce the number of microorganisms in the dental aerosol.

Protease-Activated Receptor Type 1 Activation Enhances Osteogenic Activity in Human Periodontal Ligament Stem Cells.

Protease-activated receptor 1 (PAR1) has been associated to tissue repair and bone healing. The aim of the present study was to evaluate the effect of PAR1 activation on the osteogenic activity of human periodontal ligament stem cells (PDLSCs). PDLSCs were cultured in the presence of PAR1-selective agonist peptide (100 nM), thrombin (0.1 U/mL), or PAR1 antagonist peptide (100 nM). Calcium deposits, calcium concentration (supernatant), alkaline phosphatase activity (ALP), cell proliferation, and gene (qPCR) and protein expression (ELISA assay) of osteogenic factors were assessed at 2, 7, and 14 days. PAR1 activation led to increased calcium deposits (p < 0.05), calcium concentration (p < 0.05), ALP activity (p < 0.05), and cell proliferation (p < 0.05). Further, PAR1 activation may increase gene and protein expression of Runx2 (p < 0.05) and OPG (p < 0.05). In conclusion, PAR1 activation increases osteogenic activity of PDLSCs, providing a possible new strategy for periodontal regenerative therapies.

Selective outcome reporting in randomized clinical trials of dental implants.

AIM: Selective outcome reporting (SOR) is a type of bias that occurs when the primary outcome of a trial protocol is changed or omitted in the paper. The purpose of this study was to evaluate the prevalence of SOR in publications of randomized clinical trials (RCT) concerning dental implants. MATERIALS AND METHODS: Two reviewers independently screened protocols registered at ClinicalTrials.gov until February/2019. If the protocol met the eligibility criteria, the reviewers tried to identify the corresponding publication. Data extraction was carried out by the same reviewers. Primary and secondary outcomes were recorded for each trial and compared to outcomes previously described in protocols. RESULTS: A total of 49 protocols were included. SOR was identified in 27 (55.1%) trials. The major discrepancies were as follows: protocol-defined primary outcome omitted in the publication (n = 6, 12.2%), new primary outcome introduced (n = 8, 16.3%), discrepancy in the primary outcome time frame (n = 17, 34.7%) and new secondary outcome introduced (n = 31, 63.3%). SOR was significantly associated with industry funding (p = 0.04) and timing of registration (p = 0.04). CONCLUSIONS: Our results indicate a high rate of SOR in dental implants clinical trials. Use of registry data during the peer-review process may help decreasing SOR.

Comparison of two commercial dentifrices in the reduction of gingivitis and plaque: a pilot study / Comparação de dois dentífricos comerciais na redução da gengivite e da placa bacteriana: um ensaio clínico randomizado controlado

Objectives: Chlorhexidine (CHX) and triclosan are the most used chemical agents in dentistry. However, the combination of these products has never been tested. We hypothesize that the addition of CHX to a triclosan dentifrice formulation may offer additional benefits in the reduction of plaque and gingivitis. Thus, the aim of this study was to compare a commercial dentifrice containing 0.05% chlorhexidine and 0.3% triclosan, with conventional toothpaste containing 0.3% triclosan, in the treatment of gingivitis and plaque reduction. Material and Methods: Thirty volunteers were randomly assigned to receive a dentifrice containing 0.05% CHX and 0.3% triclosan (CHX/ triclosan group) or a dentifrice containing basically 0.3% triclosan (Triclosan group). Subjects received clinical evaluation such as gingival index (GI) and plaque index (PI) at baseline, 30 and 60 days. Results: After 60 days, both treatments led to a significant improvement in GI and PI. There was no significant difference between groups as regards change in GI and PI (p>0.05). Conclusion: The combination of 0.05% CHX with 0.3% triclosan did not offer further benefits to gingival inflammation and plaque control when compared with a dentifrice containing 0.3% triclosan. (AU)

Objetivos: Clorexidina e triclosan são os agentes químicos mais utilizados em odontologia. No entanto, a combinação desses produtos nunca foi testada. Nós levantamos a hipótese de que a adição de clorexidina a um dentifrício contendo triclosan pode oferecer benefícios adicionais na redução de placa e gengivite. Assim, o objetivo deste estudo foi comparar um dentífrico comercial contendo 0,05% de clorhexidina e 0,3% de triclosan, com creme dental convencional contendo 0,3% de triclosan, no tratamento de gengivite e redução da placa. Material e Métodos: trinta voluntários foram distribuídos aleatoriamente para receber um dentifrício contendo 0,05% de clorexidina e 0,3% de triclosan ou um dentifrício contendo basicamente 0,3% de triclosan. Os indivíduos receberam avaliação clínica de índice gengival (IG) e índice de placa (IP) nos tempos 0, 30 e 60 dias. Resultados: após 60 dias, ambos os tratamentos levaram a uma melhora significativa no IG e IP. Não houve diferença significativa entre os grupos no que se refere à mudança no IG e IP (p> 0,05). Conclusão: A combinação de 0,05% de Clorexidina com 0,3% de triclosan não ofereceu benefícios adicionais para a redução de inflamação gengival e o controle da placa quando comparado com um dentifrício contendo 0,3% de triclosan (AU)