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SIGNIFICANCE: FT-IR is an important and emerging tool, providing information related to the biochemical composition of biofluids. It is important to demonstrate that there is an efficacy in separating healthy and diseased groups, helping to establish FT-IR uses as fast screening tool. AIM: Via saliva diagnosis evaluate the accuracy of FT-IR associate with machine learning model for classification among healthy (control group), diabetic (D) and periodontitis (P) patients and the association of both diseases (DP). APPROACH: Eighty patients diagnosed with diabetes and periodontitis through conventional methods were recruited and allocated in one of the four groups. Saliva samples were collected from participants of each group (n = 20) and were processed using Bruker Alpha II spectrometer in a FT-IR spectral fingerprint region between 600 and-1800 cm-1, followed by data preprocessing and analysis using machine learning tools. RESULTS: Various FTI-R peaks were detectable and attributed to specific vibrational modes, which were classified based on confusion matrices showed in paired groups. The highest true positive rates (TPR) appeared between groups C vs D (93.5 % ± 2.7 %), groups C vs. DP (89.2 % ± 4.1 %), and groups D and P (90.4 % ± 3.2 %). However, P vs DP presented higher TPR for DP (84.1 % ±3.1 %) while D vs. DP the highest rate for DP was 81.7 % ± 4.3 %. Analyzing all groups together, the TPR decreased. CONCLUSION: The system used is portable and robust and can be widely used in clinical environments and hospitals as a new diagnostic technique. Studies in our groups are being conducted to solidify and expand data analysis methods with friendly language for healthcare professionals. It was possible to classify healthy patients in a range of 78-93 % of accuracy. Range over 80 % of accuracy between periodontitis and diabetes were observed. A general classification model with lower TPR instead of a pairwise classification would only have advantages in scenarios where no prior patient information is available regarding diabetes and periodontitis status.
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Periodontite , Saliva , Humanos , Periodontite/diagnóstico , Feminino , Masculino , Saliva/química , Pessoa de Meia-Idade , Adulto , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Diabetes Mellitus/diagnóstico , Aprendizado de Máquina , Estudos de Casos e ControlesRESUMO
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.
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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.
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Osteogênese , Ligamento Periodontal , Humanos , Diferenciação Celular , Receptor PAR-2/metabolismo , Cálcio , Células-Tronco , Proliferação de Células , Células CultivadasRESUMO
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.
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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.
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Sistema de Sinalização das MAP Quinases , Receptor PAR-1 , Diferenciação Celular , Células Cultivadas , Expressão Gênica , Humanos , Osteogênese , Ligamento Periodontal , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas , Receptor PAR-1/genética , Receptor PAR-1/metabolismoRESUMO
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.
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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.
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Perda de Dente , Seguimentos , Humanos , Cooperação do Paciente , Bolsa Periodontal , Estudos Prospectivos , Estudos Retrospectivos , Perda de Dente/etiologiaRESUMO
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.
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Subunidade alfa 1 de Fator de Ligação ao Core/genética , Células-Tronco Mesenquimais/fisiologia , MicroRNAs/metabolismo , Osteogênese , Ligamento Periodontal/citologia , Processamento Pós-Transcricional do RNA , RNA Longo não Codificante/metabolismo , Células Cultivadas , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Humanos , Carioferinas/genética , Carioferinas/metabolismo , MicroRNAs/genética , Ligamento Periodontal/metabolismo , Fenótipo , RNA Longo não Codificante/genética , Fator de Transcrição Sp7/genética , Fator de Transcrição Sp7/metabolismo , Transcrição Gênica , Transcriptoma , Adulto JovemRESUMO
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.
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Obesidade/microbiologia , Periodontite/microbiologia , Periodontite/terapia , Adulto , Aggregatibacter actinomycetemcomitans/isolamento & purificação , Análise de Variância , Antropometria , Índice de Placa Dentária , Feminino , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Obesidade/fisiopatologia , Índice Periodontal , Porphyromonas gingivalis/isolamento & purificação , Estudos Prospectivos , Fatores de Risco , Estatísticas não Paramétricas , Tannerella forsythia/isolamento & purificação , Fatores de Tempo , Resultado do Tratamento , Treponema denticola/isolamento & purificaçãoRESUMO
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