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1.
Int J Nanomedicine ; 15: 9241-9253, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33262586

RESUMO

Purpose: Reducing toxicity, immunogenicity, and costs of small interfering RNAs (siRNA) carrier materials are key goals for RNA interference (RNAi) technology transition from bench to bed. Recently, calcium ions (Ca2+) have garnered attention as a novel, alternative material for delivering siRNA to cells. However, the tolerance for Ca2+ concentration varies in different cell types, which has limited its applications in vivo. Bovine serum albumin (BSA) can bind to Ca2+ through chelation. Moreover, BSA is a favorable coating material for nanoparticles owing to its excellent biocompatibility. Therefore, we hypothesized that coating Ca2+-siRNA with BSA helps buffer Ca2+ toxicity in vivo. Methods: BSA-Ca2+-siRNA nanoparticles were prepared, and the size, shape, encapsulation, and release efficiency were characterized using atomic force microscopy, scanning electronic microcopy, and gel electrophoresis. Binding nanoparticles were evaluated using attenuated total reflection-Fourier-transform infrared spectroscopy. The cellular uptake, intracellular release, cytotoxicity, and gene knockdown of nanoparticles were evaluated in periodontal ligament stem cells (PDLSCs) using laser-scanning confocal microscope, flow cytometry, and real-time quantitative polymerase chain reaction. Results: BSA and Ca2+-siRNA could form a stable nano-scale complex (~140 nm in diameter). The nanocomplexes could maintain siRNA release for more than 1 week in neutral phosphate-buffered saline (PBS) and could induce accelerated degradation in acidic PBS (pH 5.0). The nanoparticles were taken up by the cells, primarily through macropinocytosis, and were then released intracellularly through the acidification of endosomes/lysosomes. Importantly, the BSA-Ca2+ carrier had high transfection efficiency and biocompatibility both in vitro and in vivo. To demonstrate the therapeutic potential of our BSA coating-optimized Ca2+-siRNA technology, we showed that BSA-Ca2+-siWWP1 complexes strongly enhanced the osteogenic differentiation of inflammatory PDLSCs. Conclusion: BSA-Ca2+ could potentially be used for siRNA delivery, which is not only highly efficient and cost-effective but also biocompatible to host tissues owing to the BSA coating.


Assuntos
Cálcio/metabolismo , Técnicas de Transferência de Genes , Nanopartículas/química , Periodontite/terapia , RNA Interferente Pequeno/metabolismo , Soroalbumina Bovina/química , Adulto , Animais , Morte Celular , Diferenciação Celular , Endocitose , Humanos , Camundongos Endogâmicos BALB C , Nanopartículas/ultraestrutura , Osteogênese , Ligamento Periodontal/patologia , Periodontite/patologia , Espectroscopia de Infravermelho com Transformada de Fourier , Células-Tronco/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Adulto Jovem
2.
PLoS Pathog ; 16(10): e1008881, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-33002094

RESUMO

Epidemiological studies reveal significant associations between periodontitis and oral cancer. However, knowledge about the contribution of periodontal pathogens to oral cancer and potential regulatory mechanisms involved is limited. Previously, we showed that nisin, a bacteriocin and commonly used food preservative, reduced oral cancer tumorigenesis and extended the life expectancy in tumor-bearing mice. In addition, nisin has antimicrobial effects on key periodontal pathogens. Thus, the purpose of this study was to test the hypothesis that key periodontal pathogens (Porphyromonas gingivalis, Treponema denticola, and Fusobacterium nucleatum) promote oral cancer via specific host-bacterial interactions, and that bacteriocin/nisin therapy may modulate these responses. All three periodontal pathogens enhanced oral squamous cell carcinoma (OSCC) cell migration, invasion, tumorsphere formation, and tumorigenesis in vivo, without significantly affecting cell proliferation or apoptosis. In contrast, oral commensal bacteria did not affect OSCC cell migration. Pathogen-enhanced OSCC cell migration was mediated via integrin alpha V and FAK activation, since stably blocking alpha V or FAK expression abrogated these effects. Nisin inhibited these pathogen-mediated processes. Further, Treponema denticola induced TLR2 and 4 and MyD88 expression. Stable suppression of MyD88 significantly inhibited Treponema denticola-induced FAK activation and abrogated pathogen-induced migration. Together, these data demonstrate that periodontal pathogens contribute to a highly aggressive cancer phenotype via crosstalk between TLR/MyD88 and integrin/FAK signaling. Nisin can modulate these pathogen-mediated effects, and thus has therapeutic potential as an antimicrobial and anti-tumorigenic agent.


Assuntos
Infecções por Bacteroidaceae/tratamento farmacológico , Carcinoma de Células Escamosas/tratamento farmacológico , Neoplasias Bucais/tratamento farmacológico , Periodontite/tratamento farmacológico , Porphyromonas gingivalis/efeitos dos fármacos , Probióticos/farmacologia , Animais , Apoptose , Infecções por Bacteroidaceae/metabolismo , Infecções por Bacteroidaceae/microbiologia , Infecções por Bacteroidaceae/patologia , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/microbiologia , Carcinoma de Células Escamosas/patologia , Movimento Celular , Proliferação de Células , Quinase 1 de Adesão Focal/genética , Quinase 1 de Adesão Focal/metabolismo , Humanos , Integrinas/genética , Integrinas/metabolismo , Camundongos , Camundongos Nus , Neoplasias Bucais/metabolismo , Neoplasias Bucais/microbiologia , Neoplasias Bucais/patologia , Fator 88 de Diferenciação Mieloide/genética , Fator 88 de Diferenciação Mieloide/metabolismo , Periodontite/metabolismo , Periodontite/microbiologia , Periodontite/patologia , Porphyromonas gingivalis/patogenicidade , Receptor 2 Toll-Like/genética , Receptor 2 Toll-Like/metabolismo , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de Xenoenxerto
3.
PLoS One ; 15(8): e0232731, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32817640

RESUMO

This study evaluated the effects of the chronic consumption of different concentrations of alcohol on the experimental periodontitis (EP). 160 rats were divided into 4 groups: (EP-NT) rats with EP and no alcohol exposure; (EP-A14) rats with EP exposed to 14% alcohol; (EP-A25) rats with EP exposed to 25% alcohol; (EP-A36) rats with EP exposed to 36% alcohol. The animals from the EP-A14, EP-A25 and EP-A36 groups were subjected to different concentrations of alcohol 30 days before EP induction. The histological characteristics, percentage of bone in the furcation (PBF) and bone metabolism in the furcation region were evaluated. The PBF and tartrate-resistant acid phosphatase (TRAP) data were subjected to statistical analysis. The EP-A14, EP-A25 and EP-A36 groups had lower PBFs compared with the EP-NT group. A more severe inflammatory process and a greater number of TRAP+ cells were also observed. In the EP-A14, EP-A25 and EP-A36 groups, the inflammatory process became more severe as the ingested alcoholic concentration increased. An increase in RANKL immunolabeling and a significantly higher number of TRAP+ cells were also observed. We conclude that chronic alcohol consumption increases the severity of experimental periodontitis in a dose-dependent manner by increasing the magnitude of local inflammatory responses and stimulating alveolar bone resorption.


Assuntos
Perda do Osso Alveolar/patologia , Etanol/efeitos adversos , Periodontite/patologia , Consumo de Bebidas Alcoólicas/efeitos adversos , Consumo de Bebidas Alcoólicas/fisiopatologia , Animais , Masculino , Ratos , Ratos Wistar , Fosfatase Ácida Resistente a Tartarato/metabolismo
4.
Acta Odontol Latinoam ; 33(1): 50-55, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-32621600

RESUMO

The purpose of this study was to evaluate aortic wall thickness after periodontal disease and/or obesity induction in a Wistar rat model.Sixty male Wistar rats were randomly divided into four groups: control (CT), periodontal disease (PD), obesity (OB), and obesity plus periodontal disease (OB+PD). Groups OB and OB+PD received cafeteria diet for 17 weeks. After they had acquired obesity (week 12), periodontal disease was induced by placing a silk ligature on the maxillary right second molar of groups PD and OB+PD. During the experimental period, body weight and Lee index were assessed. Mean alveolar bone loss (ABL) was evaluated, and aortas were prepared for histometric analysis of the aortic wall by ImageJ software. Body weight and Lee index increased in rats exposed to cafeteria diet. Mean ABL was higher in Groups PD and OB+PD than in control and OB (p<0.05). ABL was 18% higher in Group OB+PD than in Group PD, with statistically significant difference (p<0.001). Aortas were thicker in Groups OB and OB+PD than in control and PD groups, respectively (2.31mm ± 0.28 and 2.33 ± 0.29 vs. 2.18 ± 0.26 and 2.14 ± 0.27). Group OB differed significantly from the control group (p=0.036), and OB+PD and OB differed significantly from PD (p=0.004 and p= 0.001, respectively). Obesity alters aortic wall thickness in Wistar rats. However, the presence of periodontal disease did not affect the aortic wall thickness under the conditions of the present study.


Assuntos
Perda do Osso Alveolar/etiologia , Aterosclerose , Ligadura/efeitos adversos , Obesidade/complicações , Periodontite/complicações , Perda do Osso Alveolar/patologia , Animais , Modelos Animais de Doenças , Masculino , Periodontite/patologia , Ratos , Ratos Wistar
5.
PLoS One ; 15(7): e0236161, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32730269

RESUMO

BACKGROUND: Periodontitis is a multifactorial inflammatory disease of tooth supporting tissues caused by oral biofilms, influenced by environmental and genetic factors, among others. Ethanol consumption has been considered a factor that enhances alveolar bone loss, especially in high doses. The present study aims to investigate the changes promoted by ethanol binge drinking per se or associated with ligature-induced periodontal breakdown on alveolar bone loss. MATERIALS AND METHODS: Thirty-two Wistar rats were randomly allocated into four groups: control (C), ethanol (3g/kg/day; 3 days On-4 days Off protocol by gavage for 28 days, EtOH), experimental periodontitis (EP) and experimental periodontitis plus ethanol administration (EP+EtOH). On day 14th, periodontitis was induced by ligatures that were placed around the lower first molars. On day 28th, the animals were euthanized and mandibles were submitted to stereomicroscopy for exposed root area analysis and micro-computed tomography (micro-CT) for the evaluation of alveolar bone loss and microstructural parameters. RESULTS: The results revealed that ligature-induced alveolar bone loss is aggravated by ethanol binge drinking compared to controls (1.06 ± 0.10 vs 0.77 ± 0.04; p<0.0001). In addition, binge drinking per se altered the alveolar bone quality and density demonstrating a reduction in trabecular thickness, trabecular number parameter and bone density percentual. Periodontal disorder plus ethanol binge drinking group also demonstrated reduction of the quality of bone measured by trabecular thickness. CONCLUSIONS: In conclusion, intense and episodic ethanol intake decreased alveolar bone quality in all microstructural parameters analyzed which may be considered a modifying factor of periodontitis, intensifying the already installed disease.


Assuntos
Perda do Osso Alveolar/etiologia , Bebedeira/complicações , Densidade Óssea/efeitos dos fármacos , Estimulantes do Sistema Nervoso Central/toxicidade , Etanol/toxicidade , Periodontite/complicações , Animais , Masculino , Periodontite/induzido quimicamente , Periodontite/patologia , Ratos , Ratos Wistar
6.
Braz Oral Res ; 34: e038, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32374812

RESUMO

The possible role of B-cell growth and differentiation-related cytokines on the pathogenesis of diabetes-related periodontitis has not been addressed so far. The aim of this study was to evaluate the effects of diabetes mellitus (DM) on the gene expression of proliferation-inducing ligand (APRIL) and B-lymphocyte stimulator (BLyS), two major cytokines associated to survival, differentiation and maturation of B cells in biopsies from gingival tissue with periodontitis. Gingival biopsies were obtained from subjects with periodontitis (n = 17), with periodontitis and DM (n = 19) as well as from periodontally and systemically healthy controls (n = 10). Gene expressions for APRIL, BLyS, RANKL, OPG, TRAP and DC-STAMP were evaluated using qPCR. The expressions APRIL, BLyS, RANKL, OPG, TRAP and DC-STAMP were all higher in both periodontitis groups when compared to the control group (p < 0.05). Furthermore, the expressions of BLyS, TRAP and RANKL were significantly higher in the subjects with periodontitis and DM when compared to those with periodontitis alone (p < 0.05). The mRNA levels of BLyS correlated positively with RANKL in the subjects with periodontitis and DM (p < 0.05). BLyS is overexpressed in periodontitis tissues of subjects with type 2 DM, suggesting a possible role of this cytokine on the pathogenesis DM-related periodontitis.


Assuntos
Fator Ativador de Células B/análise , Diabetes Mellitus Tipo 2/complicações , Periodontite/imunologia , Periodontite/patologia , Adulto , Idoso , Biomarcadores/análise , Biópsia , Estudos de Casos e Controles , Citocinas/análise , Citocinas/fisiologia , Diabetes Mellitus Tipo 2/imunologia , Feminino , Expressão Gênica , Gengiva/imunologia , Gengiva/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Osteogênese/imunologia , RNA Mensageiro/análise , Reação em Cadeia da Polimerase em Tempo Real , Valores de Referência , Estatísticas não Paramétricas , Membro 13 da Superfamília de Ligantes de Fatores de Necrose Tumoral/análise
7.
Sci Rep ; 10(1): 7823, 2020 05 08.
Artigo em Inglês | MEDLINE | ID: mdl-32385413

RESUMO

This study investigates the role of NLRP3 inflammasome and its main effector Caspase-1 in inflammation and alveolar bone resorption associated with periodontitis. Heat-killed Aggregatibacter actinomycetemcomitans (Aa) was injected 3x/week (4 weeks) into gingival tissues of wild-type (WT), Nlrp3-KO and Caspase1-KO mice. Bone resorption was measured by µCT and osteoclast number was determined by tartrate-resistant acid phosphatase (TRAP) staining. Inflammation was assessed histologically (H/E staining and immunofluorescence of CD45 and Ly6G). In vitro studies determined the influence of Nlrp3 and Caspase-1 in Rankl-induced osteoclast differentiation and activity and on LPS-induced expression of inflammation-associated genes. Bone resorption was significantly reduced in Casp1-KO but not in Nlrp3-KO mice. Casp1-KO mice had increased in osteoclast numbers, whereas the inflammatory infiltrate or on gene expression were similar to those of WT and Nlrp3-KO mice. Strikingly, osteoclasts differentiated from Nlrp3-deficient macrophages had increased resorbing activity in vitro. LPS-induced expression of Il-10, Il-12 and Tnf-α was significantly reduced in Nlrp3- and Casp1-deficient macrophages. As an inceptive study, these results suggest that Nlrp3 inflammasome does not play a significant role in inflammation and bone resorption in vivo and that Caspase-1 has a pro-resorptive role in experimental periodontal disease.


Assuntos
Perda do Osso Alveolar/genética , Caspase 1/genética , Inflamação/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Periodontite/genética , Aggregatibacter actinomycetemcomitans , Perda do Osso Alveolar/microbiologia , Perda do Osso Alveolar/patologia , Animais , Diferenciação Celular/efeitos dos fármacos , Modelos Animais de Doenças , Gengiva/crescimento & desenvolvimento , Gengiva/microbiologia , Humanos , Inflamação/microbiologia , Inflamação/patologia , Interleucina-10/genética , Interleucina-12/genética , Camundongos , Camundongos Knockout , Osteoclastos/microbiologia , Osteoclastos/patologia , Periodontite/microbiologia , Periodontite/patologia , Ligante RANK/genética , Fator de Necrose Tumoral alfa/genética
8.
BMC Oral Health ; 20(1): 95, 2020 04 03.
Artigo em Inglês | MEDLINE | ID: mdl-32245460

RESUMO

BACKGROUND: The decision to initiate dialysis treatment via haemodialysis (HD) or peritoneal dialysis (PD) often involves the consideration of complex factors and remains a matter of debate. The purpose of this study was to quantify the inflammatory burden that periodontitis causes in dialysis patients and to examine whether patients on PD and HD differ in terms of the periodontal inflamed surface area (PISA), which can be helpful for selecting the most appropriate dialysis modality. METHODS: A cross-sectional study was performed on 58 consecutive patients on HD and 31 consecutive patients on PD. PISA was calculated using measurements of the clinical attachment level, recession and bleeding on probing. We performed the primary analysis using multivariable robust regression. RESULTS: Patients on PD had a 746 mm2 (93%) lower mean PISA than patients on HD after adjustment for 20 possible confounders, including the duration of dialysis. The type of dialysis was independently correlated with the PISA (semipartial correlation: - 0.50; p = 0.017; false discovery rate < 5%). After adjusting for confounding factors, the correlation between the duration and type of dialysis was not significant (F (2,44) = 0.01; p = 0.994; η2 = 0.00). Differences in the PISA between patients who had undergone dialysis for less than a year, 2-3 years or ≥ 3 years were not significantly different in either of the two dialysis groups. CONCLUSIONS: PISA levels in Croatian patients on dialysis indicate a high need for periodontal treatment. PD is associated with a smaller PISA independent of many sociodemographic, lifestyle, laboratory and clinical factors. The duration of dialysis does not influence PISA levels. TRIAL REGISTRATION: ISRCTN17887630. A clinical study to investigate gum infection in patients undergoing kidney dialysis.


Assuntos
Falência Renal Crônica/terapia , Periodontite/complicações , Periodontite/patologia , Diálise Peritoneal/efeitos adversos , Diálise Renal/efeitos adversos , Adolescente , Adulto , Idoso , Estudos Transversais , Feminino , Humanos , Falência Renal Crônica/complicações , Masculino , Pessoa de Meia-Idade , Periodontite/sangue
9.
Braz Oral Res ; 34: e015, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32130362

RESUMO

We sought to compare the characteristics and clinical significance of neutrophil extracellular traps in gingival samples from patients with periodontitis and those with gingivitis. The clinical indexes of gingival samples from patients with periodontitis and gingivitis were measured; the expression of TNF-alpha and IL-8 was measured by real-time fluorescent quantitative PCR; and the expression of TLR-8 and MMP-9 was measured by western blotting assays. Chemotaxis, phagocytosis and phagocytic activity of neutrophils were measured. Compared with the healthy group, the expression of TNF-α and IL-8 in the periodontitis group and the gingivitis group increased significantly (p < 0.05), and TNF-α in the gingivitis group was significantly lower than that in the healthy group (p < 0.05). The expression of IL-8 in the periodontitis group was significantly higher than that in the periodontitis group (p < 0.05). Furthermore, the expression of TLR-8 and MMP-9 in the periodontitis group was different from that in the gingivitis group and the healthy group, and the expression of TLR-8 and MMP-9 in the gingivitis group was significantly different from that in the healthy group (p < 0.05). In addition, the neutrophil mobility index in healthy people was 3.02 ± 0.53, that in the periodontitis group was 2.21 ± 0.13, and that in the gingivitis group was 2.31 ± 0.12. In conclusion, the chemotaxis of neutrophils in gingival samples of patients with periodontitis and gingivitis was decreased, the phagocytotic ability and activity of neutrophils were reduced, and the release of the extracellular trap-releasing inducible factors TNF-alpha and IL-8 also declined.


Assuntos
Armadilhas Extracelulares , Gengivite/patologia , Neutrófilos/patologia , Periodontite/patologia , Actinas/análise , Adulto , Western Blotting , Estudos de Casos e Controles , Eletroforese em Gel de Ágar , Feminino , Humanos , Interleucina-8/análise , Masculino , Metaloproteinase 9 da Matriz/análise , Pessoa de Meia-Idade , Índice Periodontal , RNA/análise , Reação em Cadeia da Polimerase em Tempo Real , Valores de Referência , Receptor 8 Toll-Like/análise , Fator de Necrose Tumoral alfa/análise , Adulto Jovem
10.
PLoS One ; 15(3): e0230334, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32168352

RESUMO

AIM: The aim of the present study was to assess the feasibility and diagnostic contribution of protein profiling using MALDI-TOF mass spectrometry applied to saliva, gingival crevicular fluid (GCF) and dental plaque from periodontitis and healthy subjects. We hypothesized that rapid routine and blinded MALDI-TOF analysis could accurately classify these three types of samples according to periodontal state. MATERIALS AND METHODS: Unstimulated saliva, GCF and dental plaque, collected from periodontitis subjects and healthy controls, were analyzed by MALDI-TOF MS. Based on the differentially expressed peaks between the two groups, diagnostic decision trees were built for each sample. RESULTS: Among 141 patients (67 periodontitis and 74 healthy controls), the decision trees diagnosed periodontitis with a sensitivity = 70.3% (± 0.211) and a specificity = 77.8% (± 0.165) for saliva, a sensitivity = 79.6% (± 0.188) and a specificity = 75.7% (± 0.195) for GCF, and a sensitivity = 72.1% (± 0.202) and a specificity = 72.2% (± 0.195) for dental plaque. The sensitivity and specificity of the tests were improved to 100% (CI 95% = [0.91;1]) and 100% (CI 95% = [0.92;1]), respectively, when two samples were tested. CONCLUSION: We developed, for the first time, diagnostic tests based on protein profiles of saliva, GCF and dental plaque between periodontitis patients and healthy subjects. When at least 2 of these samples were tested, the best results were obtained.


Assuntos
Periodontite/diagnóstico , Proteínas/genética , Saliva/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Adulto , Feminino , Regulação da Expressão Gênica/genética , Humanos , Masculino , Pessoa de Meia-Idade , Periodontite/genética , Periodontite/patologia , Proteínas/isolamento & purificação , Dente/metabolismo , Dente/patologia
11.
Braz Oral Res ; 34: e012, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32049112

RESUMO

Lipoproteins are important bacterial immunostimulating molecules capable of inducing receptor activator of nuclear factor-κB (RANKL) and osteoclast formation in vitro and in vivo . Although these molecules are present in periodontopathogenic bacteria, their role in periodontitis is not known. In this study, we used Pam2CSK4 (PAM2), a synthetic molecule that mimics bacterial lipoprotein, to investigate the effects of lipoproteins on periodontitis in mice. C57BL/6 male mice were randomly divided into three experimental groups: 1) Negative control group: animals received vehicle injection; 2) Positive control group: animals received injection of Escherichia coli lipopolysaccharide (LPS); 3) PAM2 group: animals received PAM2 injection. All the injections were performed bilaterally every other day into the palatal mucosa between first and second molars. After twenty-four days, the animals were euthanized to assess alveolar bone volume (micro-CT), cellular and extracellular composition in the gingiva (stereometric analysis), and osteoclast numbers (TRAP staining). Treatment with either PAM2 or LPS induced gingival inflammation, as demonstrated by increased infiltration of inflammatory cells and enhanced angiogenesis, associated with a smaller number of fibroblasts and decreased extracellular matrix. Importantly, treatment not only with LPS but also with PAM2 resulted in a larger number of TRAP+ multinucleated osteoclasts and significant loss of alveolar bone. Collectively, our data demonstrate that PAM2 can induce gingival inflammation and bone loss in mice, broadening the avenues of investigation into the role of lipoproteins in the pathogenesis of periodontal disease.


Assuntos
Lipopeptídeos/farmacologia , Periodontite/etiologia , Periodontite/patologia , Receptor 2 Toll-Like/antagonistas & inibidores , Perda do Osso Alveolar/etiologia , Perda do Osso Alveolar/patologia , Processo Alveolar/efeitos dos fármacos , Processo Alveolar/patologia , Animais , Modelos Animais de Doenças , Gengiva/efeitos dos fármacos , Gengiva/patologia , Gengivite/etiologia , Gengivite/patologia , Masculino , Camundongos Endogâmicos C57BL , Osteoclastos/efeitos dos fármacos , Osteoclastos/fisiologia , Periodontite/microbiologia , Distribuição Aleatória , Fosfatase Ácida Resistente a Tartarato , Fatores de Tempo , Microtomografia por Raio-X
12.
Med Sci Monit ; 26: e918932, 2020 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-32057034

RESUMO

BACKGROUND Osteogenic differentiation of periodontal ligament stem cells (PDLSCs) is associated with periodontitis. It has been reported that long noncoding RNA X-inactive specific transcript (lncRNA XIST) is upregulated and microRNA-214-3p (miR-214-3p) is downregulated in PDLSCs after osteogenic induction. However, whether XIST is involved in osteogenic differentiation of PDLSCs via miR-214-3p has not been reported. MATERIAL AND METHODS The protein expressions of osteogenic markers alkaline phosphatase (ALP), osteocalcin (OCN), and runt-related transcription factor 2 (RUNX2) were examined by Western blot. The levels of miR-214-3p and XIST were determined by qRT-PCR. The relationship between miR-214-3p and XIST was evaluated by luciferase reporter, RNA immunoprecipitation, and RNA pulldown assays. RESULTS We found that XIST was increased and miR-214-3p was decreased in PDLSCs after osteogenic stimulation. Silencing of XIST decreased the protein expressions of ALP, OCN, and RUNX2, and also decreased ALP activity. Higher miR-214-3p levels also inhibited osteogenic differentiation of PDLSCs. XIST interacted with miR-214-3p and depletion of miR-214-3p mitigated XIST absence-mediated suppression of osteogenic differentiation. CONCLUSIONS XIST participates in osteogenic differentiation of PDLSCs by sponging miR-214-3p.


Assuntos
MicroRNAs/metabolismo , Osteogênese/genética , Periodontite/genética , RNA Longo não Codificante/metabolismo , Células-Tronco/patologia , Adolescente , Diferenciação Celular/genética , Células Cultivadas , Regulação para Baixo , Técnicas de Silenciamento de Genes , Humanos , MicroRNAs/agonistas , MicroRNAs/antagonistas & inibidores , Osteoblastos/patologia , Ligamento Periodontal/citologia , Ligamento Periodontal/patologia , Periodontite/patologia , Cultura Primária de Células , RNA Longo não Codificante/genética , RNA Interferente Pequeno/metabolismo , Regulação para Cima
13.
Arch Oral Biol ; 112: 104679, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32062102

RESUMO

OBJECTIVE: The overall objective of this study was to investigate the effects of hinokitiol on periodontal bone loss in a murine model of experimental periodontitis and evaluate the anti-inflammatory activity of hinokitiol in vitro. DESIGN: Periodontitis was induced by tying a silk ligature around the maxillary second molar of mice for 8 days. Hinokitiol was injected once a day for 7 days into the palatal gingiva of the ligated molar. Periodontal bone loss was then assessed morphometrically in the maxillary second molar, and the number of tartrate-resistant acid phosphatase positive multinucleated giant cells around the molar was quantified. The bacterial load of the silk ligature was calculated by counting the number of colony-forming units, while the transcription levels of proinflammatory cytokine-related genes in the palatal gingiva were evaluated by real-time qPCR. The activity of hinokitiol against LPS-induced transcription of proinflammatory genes in RAW 264.7 macrophages was also examined. RESULTS: Local treatment with hinokitiol significantly inhibited the alveolar bone loss and osteoclast differentiation induced by tooth ligation. In addition, hinokitiol treatment decreased the oral bacterial load of the silk ligature and downregulated the mRNA levels of inflammatory cytokine-related genes, both in vitro and in vivo. CONCLUSION: The results indicated that hinokitiol exhibits antibacterial and anti-inflammatory activity and exerts a protective effect against periodontitis.


Assuntos
Perda do Osso Alveolar/prevenção & controle , Monoterpenos/uso terapêutico , Periodontite/tratamento farmacológico , Tropolona/análogos & derivados , Animais , Antibacterianos/uso terapêutico , Anti-Inflamatórios/uso terapêutico , Carga Bacteriana , Citocinas/metabolismo , Modelos Animais de Doenças , Ligadura , Camundongos , Periodontite/patologia , Tropolona/uso terapêutico
14.
Biochim Biophys Acta Mol Basis Dis ; 1866(6): 165731, 2020 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-32088316

RESUMO

Outer membrane vesicles (OMVs) are nanosized particles derived from the outer membrane of gram-negative bacteria. Oral bacterium Porphyromonas gingivalis (Pg) is known to be a major pathogen of periodontitis that contributes to the progression of periodontal disease by releasing OMVs. The effect of Pg OMVs on systemic diseases is still unknown. To verify whether Pg OMVs affect the progress of diabetes mellitus, we analyzed the cargo proteins of vesicles and evaluated their effect on hepatic glucose metabolism. Here, we show that Pg OMVs were equipped with Pg-derived proteases gingipains and translocated to the liver in mice. In these mice, the hepatic glycogen synthesis in response to insulin was decreased, and thus high blood glucose levels were maintained. Pg OMVs also attenuated the insulin-induced Akt/glycogen synthase kinase-3 ß (GSK-3ß) signaling in a gingipain-dependent fashion in hepatic HepG2 cells. These results suggest that the delivery of gingipains mediated by Pg OMV elicits changes in glucose metabolisms in the liver and contributes to the progression of diabetes mellitus.


Assuntos
Membrana Externa Bacteriana/metabolismo , Cisteína Endopeptidases Gingipaínas/genética , Periodontite/genética , Porphyromonas gingivalis/genética , Animais , Membrana Externa Bacteriana/patologia , Modelos Animais de Doenças , Cisteína Endopeptidases Gingipaínas/metabolismo , Glicogênio Sintase Quinase 3 beta/genética , Humanos , Resistência à Insulina/genética , Fígado/metabolismo , Fígado/microbiologia , Camundongos , Periodontite/microbiologia , Periodontite/patologia , Porphyromonas gingivalis/metabolismo , Porphyromonas gingivalis/patogenicidade , Proteínas Proto-Oncogênicas c-akt/genética , Transdução de Sinais/genética
15.
PLoS One ; 15(1): e0228206, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31978120

RESUMO

Recent experimental studies indicated that a periodontitis-causing bacterium might be a causal factor for Alzheimer's disease (AD). We applied a two-sample Mendelian randomization (MR) approach to examine the potential causal relationship between chronic periodontitis and AD bidirectionally in the population of European ancestry. We used publicly available data of genome-wide association studies (GWAS) on periodontitis and AD. Five single-nucleotide polymorphisms (SNPs) were used as instrumental variables for periodontitis. For the MR analysis of periodontitis on risk of AD, the causal odds ratio (OR) and 95% confidence interval (CI) were derived from the GWAS of periodontitis (4,924 cases vs. 7,301 controls) and from the GWAS of AD (21,982 cases vs. 41,944 controls). Seven non-overlapping SNPs from another latest GWAS of periodontitis was used to validate the above association. Twenty SNPs were used as instrumental variables for AD. For the MR analysis of liability to AD on risk of periodontitis, the causal OR was derived from the GWAS of AD including 30,344 cases and 52,427 controls and from the GWAS of periodontitis consisted of 12,289 cases and 22,326 controls. We employed multiple methods of MR. Using the five SNPs as instruments of periodontitis, there was suggestive evidence of genetically predicted periodontitis being associated with a higher risk of AD (OR 1.10, 95% CI 1.02 to 1.19, P = 0.02). However, this association was not verified using the seven independent SNPs (OR 0.97, 95% CI 0.87 to 1.08, P = 0.59). There was no association of genetically predicted AD with the risk of periodontitis (OR 1.00, 95% CI 0.96 to 1.04, P = 0.85). In summary, we did not find convincing evidence to support periodontitis being a causal factor for the development of AD. There was also limited evidence to suggest genetic liability to AD being associated with the risk of periodontitis.


Assuntos
Doença de Alzheimer/patologia , Análise da Randomização Mendeliana/métodos , Periodontite/patologia , Doença de Alzheimer/etiologia , Doença de Alzheimer/genética , Estudos de Casos e Controles , Estudo de Associação Genômica Ampla , Humanos , Razão de Chances , Periodontite/complicações , Periodontite/genética , Polimorfismo de Nucleotídeo Único , Fatores de Risco
16.
PLoS One ; 15(1): e0227905, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31978095

RESUMO

The pathogenesis of periodontitis (PD) involves several molecules of the immune system that interact in a network to eliminate the periodontopathogens, yet, they contribute to periodontal tissue destruction. The different mechanisms that lead to periodontal tissue damage are not clear. Despite this, immune response genes have been related to the development of PD previously, such as those involved in inflammasomes which are multiprotein complexes and cytokines including Interleukin-1. The aim of the study was to evaluate the polymorphisms in NLRP3 inflammasome, cytokine and receptor of cytokines genes in the development of periodontitis. This case-control study was conducted in 186 patients with PD (stage II and III and grade B) and 208 controls (localized gingivitis and periodontally healthy individuals). Genotyping was performed using PCR-RFLP for the SNP rs4612666 in NLRP3 and using PCR-SSP for IL1A, IL1B, IL1R, IL1RN, IL4RA, INFG, TGFB1, TNF, IL2, IL4, IL6, and IL10. Cytokine serum levels were measured using Luminex technology. SNPStats and OpenEpi software were used to perform statistical analysis. The higher frequencies of NLRP3 T/C and IL1B -511 T/T genotypes and IL2 (+166, -330) GT haplotype were observed in patients with PD compared to controls. The SNPs in NLRP3, IL1R +1970, IL6-174, TNF -308, IL2 +166 and -330, TGFB1 +869 and +915, IL4RA +1902, IL4-1098 and -590 were associated to PD in men. In conclusion, polymorphisms in NLRP3, IL1B and IL2 genes were associated to PD susceptibility. Men carrying the NLRP3, IL1R, IL6, TNF, IL2, TGFB1, IL4RA and IL4 polymorphisms had greater susceptibility than women for developing PD.


Assuntos
Interleucina-1beta/genética , Interleucina-2/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Periodontite/genética , Adulto , Citocinas/genética , Feminino , Estudos de Associação Genética , Predisposição Genética para Doença , Genótipo , Haplótipos/genética , Humanos , Inflamassomos/genética , Masculino , Pessoa de Meia-Idade , Periodontite/patologia , Polimorfismo de Fragmento de Restrição/genética , Polimorfismo de Nucleotídeo Único/genética , Caracteres Sexuais , Fator de Crescimento Transformador beta1/genética
17.
Biochem J ; 477(2): 381-405, 2020 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-31899475

RESUMO

As part of the infective process, Porphyromonas gingivalis must acquire heme which is indispensable for life and enables the microorganism to survive and multiply at the infection site. This oral pathogenic bacterium uses a newly discovered novel hmu heme uptake system with a leading role played by the HmuY hemophore-like protein, responsible for acquiring heme and increasing virulence of this periodontopathogen. We demonstrated that Prevotella intermedia produces two HmuY homologs, termed PinO and PinA. Both proteins were produced at higher mRNA and protein levels when the bacterium grew under low-iron/heme conditions. PinO and PinA bound heme, but preferentially under reducing conditions, and in a manner different from that of the P. gingivalis HmuY. The analysis of the three-dimensional structures confirmed differences between apo-PinO and apo-HmuY, mainly in the fold forming the heme-binding pocket. Instead of two histidine residues coordinating heme iron in P. gingivalis HmuY, PinO and PinA could use one methionine residue to fulfill this function, with potential support of additional methionine residue/s. The P. intermedia proteins sequestered heme only from the host albumin-heme complex under reducing conditions. Our findings suggest that HmuY-like family might comprise proteins subjected during evolution to significant diversification, resulting in different heme coordination modes. The newer data presented in this manuscript on HmuY homologs produced by P. intermedia sheds more light on the novel mechanism of heme uptake, could be helpful in discovering their biological function, and in developing novel therapeutic approaches.


Assuntos
Heme/genética , Hemeproteínas/genética , Periodontite/genética , Prevotella intermedia/genética , Regulação Bacteriana da Expressão Gênica/genética , Heme/química , Hemeproteínas/química , Humanos , Ferro/metabolismo , Periodontite/microbiologia , Periodontite/patologia , Porphyromonas gingivalis/genética , Porphyromonas gingivalis/patogenicidade , Prevotella intermedia/patogenicidade , RNA Mensageiro/genética , Homologia de Sequência de Aminoácidos
18.
AAPS PharmSciTech ; 21(3): 76, 2020 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-31970603

RESUMO

The objectives of the present research work were systematic development of novel in situ gel formulation containing nanoparticles for localised delivery of moxifloxacin against bacterial periodontitis. PLGA nanoparticles were prepared and optimised in a systematic manner. Factor screening was performed with the help of half-factorial design to identify the influential factors, while response surface optimisation of the nanoparticles was conducted using central composite design. The optimum nanoparticle formulation was chosen on the basis of lower particle size, higher drug entrapment and controlled drug release characteristics up to 1 week time period, while the optimum in situ gel was selected on the basis of faster gelling and higher viscosity and gel strength properties for improved retention in the periodontium. In vivo histopathological studies and in vivo gamma scintigraphy studies revealed the extended release, superior efficacy and enhanced retention of nanoparticle-loaded in situ gelling system. Results obtained from in vivo histopathological studies after 1 week treatment with in situ gel formulation containing nanoparticles of moxifloxacin were found to be better than with 3 weeks treatment of marketed gel formulation. Overall, the studies ratify successful development of an effective site-specific drug delivery system with enhanced biopharmaceutical attributes for the periodontitis treatment.


Assuntos
Antibacterianos/uso terapêutico , Moxifloxacina/uso terapêutico , Nanopartículas , Periodontite/tratamento farmacológico , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/uso terapêutico , Animais , Antibacterianos/química , Preparações de Ação Retardada/química , Preparações de Ação Retardada/uso terapêutico , Sistemas de Liberação de Medicamentos/métodos , Feminino , Géis , Moxifloxacina/química , Nanopartículas/química , Tamanho da Partícula , Periodontite/patologia , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Ratos , Ratos Sprague-Dawley , Resultado do Tratamento , Viscosidade
19.
Artigo em Inglês | MEDLINE | ID: mdl-31958293

RESUMO

OBJECTIVE: To determine the efficacy of clindamycin compared with amoxicillin-metronidazole after a 7-day regimen during nonsurgical treatment of periodontitis in patients with type 2 diabetes mellitus. RESEARCH DESIGN AND METHODS: In this double-blind, randomized clinical trial, a total of 42 patients with chronic periodontitis and type 2 diabetes were included. Patients were randomly assigned to treatment with either clindamycin or amoxicillin-metronidazole three times a day during 7 days. Clinical determinations (probing depth, bleeding on probe, and plaque index) were performed to determine the extent and severity of periodontitis before and after the pharmacological treatment. RESULTS: After 7 days of administration of clindamycin or amoxicillin-metronidazole, no differences were observed between the clinical determinations, probing depth (0.44 vs 0.50 mm, p=0.624), plaque index (17.62 vs 15.88%, p=0.910), and bleeding on probing (16.12 vs 22.17%, p=0.163), respectively. There were no adverse events in either group. CONCLUSION: The administration during 7 days of clindamycin or amoxicillin/metronidazole showed the same efficacy for the reduction of probing depth, plaque index, and bleeding on probing in patients with periodontitis and type 2 diabetes.


Assuntos
Amoxicilina/uso terapêutico , Antibacterianos/uso terapêutico , Clindamicina/uso terapêutico , Diabetes Mellitus Tipo 2/complicações , Metronidazol/uso terapêutico , Periodontite/tratamento farmacológico , Adolescente , Adulto , Idoso , Método Duplo-Cego , Quimioterapia Combinada , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Periodontite/etiologia , Periodontite/patologia , Prognóstico , Adulto Jovem
20.
Lasers Med Sci ; 35(4): 991-998, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31955304

RESUMO

This study aimed to histologically and radiographically evaluate the effectiveness of low-intensity laser irradiation of different wavelengths (660 or 808 nm) as an adjunct to scaling and root planing in the treatment of experimental periodontitis in rats. Periodontitis was induced by placing a ligature around the mandibular first molar of the rats. In total, 40 Wistar rats were randomly divided into five groups (n = 8 each): control (CG), periodontal disease (PD), scaling and root planing (SRP), SRP + 660 nm laser (GL660) and SRP + 808 nm laser (GL808). Groups with laser use received radiation at 6 points in the first molar. The animals were euthanized at baseline and at 7 and 14 days after the interventions. Mandibles were surgically removed for histomorphometric and radiographic assessment of periodontal tissues. The GL660 group showed lesser bone loss than the PD group (P < 0.05) and greater alveolar bone margin after 14 days, indicating a better long-term treatment response (P < 0.05). These findings suggest that SRP with the 660 nm laser as an adjunct results in more favorable radiographic and histological responses than the 808 nm laser.


Assuntos
Raspagem Dentária , Ligadura/efeitos adversos , Terapia com Luz de Baixa Intensidade , Periodontite/etiologia , Periodontite/radioterapia , Aplainamento Radicular , Animais , Terapia Combinada , Modelos Animais de Doenças , Masculino , Ligamento Periodontal/diagnóstico por imagem , Ligamento Periodontal/efeitos da radiação , Periodontite/diagnóstico por imagem , Periodontite/patologia , Fotoquimioterapia , Ratos Wistar
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