RESUMO
Tyrosine-protein phosphatase non-receptor type 2 (PTPN2) is an important protection factor for diabetes and periodontitis, but the underlying mechanism remains elusive. This study aimed to identify the substrate of PTPN2 in mediating beneficial effects of 25-Hydroxyvitamin D3 (25(OH)2D3 ) on diabetic periodontitis. 25(OH)2D3 photo-affinity probe was synthesized with the minimalist linker and its efficacy to inhibit alveolar bone loss, and inflammation was evaluated in diabetic periodontitis mice. The probe was used to pull down the lysates of primary gingival fibroblasts. We identified PTPN2 as a direct target of 25(OH)2D3 , which effectively inhibited inflammation and bone resorption in diabetic periodontitis mice. In addition, we found that colony-stimulating factor 1 receptor (CSF1R) rather than JAK/STAT was the substrate of PTPN2 to regulate bone resorption. PTPN2 direct interacted with CSF1R and dephosphorylated Tyr807 residue. In conclusion, PTPN2 dephosphorylates CSF1R at Y807 site and inhibits alveolar bone resorption in diabetic periodontitis mice. PTPN2 and CSF1R are potential targets for the therapy of diabetic periodontitis or other bone loss-related diseases.
Assuntos
Perda do Osso Alveolar/enzimologia , Calcifediol/uso terapêutico , Diabetes Mellitus Experimental/complicações , Periodontite/enzimologia , Proteína Tirosina Fosfatase não Receptora Tipo 2/metabolismo , Receptores de Fator Estimulador das Colônias de Granulócitos e Macrófagos/metabolismo , Perda do Osso Alveolar/tratamento farmacológico , Perda do Osso Alveolar/microbiologia , Perda do Osso Alveolar/fisiopatologia , Animais , Calcifediol/química , Células Cultivadas , Citocinas/metabolismo , Fibroblastos/enzimologia , Fibroblastos/metabolismo , Fibroblastos/patologia , Gengiva/citologia , Gengiva/enzimologia , Gengiva/metabolismo , Gengiva/patologia , Inflamação/genética , Inflamação/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Periodontite/tratamento farmacológico , Periodontite/metabolismo , Periodontite/microbiologia , Porphyromonas gingivalis , Proteína Tirosina Fosfatase não Receptora Tipo 2/genética , RNA Interferente Pequeno , Tirosina/metabolismoRESUMO
To evaluate the degree of periodontal tissue destruction, aspartate aminotransferase (AST) levels in the gingival crevicular fluid (GCF) are utilized as a predictor of periodontal therapy. We have previously shown that the usefulness of AST activities [periodontal tissue monitor (PTM) values] using a PTM-kit to evaluate the effects of initial periodontal therapy and periodontal regeneration therapy by enamel matrix derivative (EMD). This prospective, longitudinal study was conducted using 38 healthy and 80 periodontitis sites with probing depth (PD) of 5-10 mm for guided tissue regeneration (GTR) and EMD from 36 patients. GCF samples were used to evaluate PTM values at base line (BL) and after 6 months of surgeries (re-evaluation: RE), and periodontal examinations were performed concurrently. PTM values at BL were statistically improved at RE, accompanied by the improvement of periodontal parameters in both groups. PTM values and PD, and the clinical attachment level (CAL) showed high correlations. PD, CAL and bleeding on probing (BOP) were highly correlated with PTM values in both groups, whereas only PD showed a significant correlation with PTM values at RE in the GTR group. Change in the amounts of PD, CAL and BOP between BL and RE in both groups showed no correlation with PTM values. In the negative PTM value sites at BL in EMD group, the mean PD was significantly reduced at RE compared with positive PTM sites at BL. PTM values are able to be utilized as the biochemical predictor of prognosis after periodontal regeneration therapy.
Assuntos
Perda do Osso Alveolar/enzimologia , Perda do Osso Alveolar/cirurgia , Aspartato Aminotransferases/análise , Líquido do Sulco Gengival/química , Regeneração Tecidual Guiada Periodontal , Perda da Inserção Periodontal/enzimologia , Perda da Inserção Periodontal/cirurgia , Adulto , Idoso , Feminino , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Índice Periodontal , Prognóstico , Estudos ProspectivosRESUMO
AIM: Leukotrienes (LTs) are pro-inflammatory lipid mediators formed by the enzyme 5-lipoxygenase (5-LO). The involvement of 5-LO metabolites in periodontal disease (PD) is not well defined. This study aimed to assess the role of 5-LO in experimental PD induced by Aggregatibacter actinomycetemcomitans (Aa). MATERIAL AND METHODS: In vivo experiments were carried out using SV129 wild-type (WT) and 5-LO-deficient (5lo-/- ) mice inoculated with Aa. Osteoclasts were stimulated in vitro with AaLPS in the presence or not of selective inhibitors of the 5-LO pathway, or LTB4 or platelet-activating factor (PAF), as PAF has already been shown to increase osteoclast activity. RESULTS: In 5lo-/- mice, there were no loss of alveolar bone and less TRAP-positive osteoclasts in periodontal tissues, after Aa inoculation, despite local production of TNF-α and IL-6. The differentiation and activity of osteoclasts stimulated with AaLPS were diminished in the presence of BLT1 antagonist or 5-LO inhibitor, but not in the presence of cysteinyl leukotriene receptor antagonist. The osteoclast differentiation induced by PAF was impaired by the BLT1 antagonism. CONCLUSION: In conclusion, LTB4 but not CysLTs is important for Aa-induced alveolar bone loss. Overall, LTB4 affects osteoclast differentiation and activity and is a key intermediate of PAF-induced osteoclastogenesis.
Assuntos
Aggregatibacter actinomycetemcomitans/patogenicidade , Perda do Osso Alveolar/enzimologia , Perda do Osso Alveolar/microbiologia , Araquidonato 5-Lipoxigenase/farmacologia , Animais , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Hidroxiureia/análogos & derivados , Hidroxiureia/farmacologia , Interleucina-6/metabolismo , Camundongos , Osteoclastos/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real , Fator de Necrose Tumoral alfa/metabolismoRESUMO
AIM: The aim of the present study was to estimate the gingival crevicular fluid (GCF) levels of matrix metalloproteinases (MMP)-3 and -13 in periodontally-healthy controls and chronic periodontitis (CP) patients, and also to investigate the effect of phase 1 periodontal therapy on MMP-3 and -13 levels in CP patients. METHODS: Fifty-five systemically-healthy patients were divided into two groups: group 1 (healthy) and group 2 (CP). The recording of clinical parameters and GCF sampling was done at baseline for both groups and again at 6 weeks post-therapy for group 2. The MMP level was determined by ELISA. RESULTS: A significant increase in the mean MMP-3 and -13 was found between healthy and CP patients. There was a statistically-significant reduction of GCF MMP-3 and -13 concentration after periodontal therapy in the CP group. A positive correlation was found between clinical parameters and GCF MMP-3 and -13 levels. CONCLUSIONS: A lower concentration of GCF MMP-3 and -13 was found in healthy patients, and a higher concentration was noted for CP patients, which was reduced after periodontal therapy. This indicates the important role played by these MMP in periodontal destruction. Thus, MMP-3 and -13 could be used as inflammatory biomarkers in diagnosing periodontal disease severity.
Assuntos
Periodontite Crônica/terapia , Líquido do Sulco Gengival/enzimologia , Metaloproteinase 13 da Matriz/análise , Metaloproteinase 3 da Matriz/análise , Adulto , Perda do Osso Alveolar/enzimologia , Perda do Osso Alveolar/terapia , Biomarcadores/análise , Periodontite Crônica/enzimologia , Índice de Placa Dentária , Feminino , Seguimentos , Humanos , Mediadores da Inflamação/análise , Masculino , Pessoa de Meia-Idade , Perda da Inserção Periodontal/enzimologia , Perda da Inserção Periodontal/terapia , Índice Periodontal , Bolsa Periodontal/enzimologia , Bolsa Periodontal/terapia , Adulto JovemRESUMO
BACKGROUND: The present study aims to examine the inhibitory effect of cyclosporin-A (CsA) on periodontal breakdown and to further explore the correlations of CsA-induced attenuation of periodontal bone loss with the expressions of gelatinases (i.e., matrix metalloproteinase [MMP]-2 and MMP-9) and extracellular matrix metalloproteinase inducer (EMMPRIN). METHODS: Forty Sprague-Dawley rats were randomly divided into four groups: 1) control; 2) CsA; 3) ligature (Lig); and 4) ligature plus CsA (Lig + CsA). The CsA group received 10 mg â Kg(-1) â d(-1) CsA for 8 days. The Lig group received silk ligature on selected molars. The Lig + CsA group received silk ligature and CsA treatment. The inhibitory effects of CsA on the ligature-induced periodontal breakdown was examined with microcomputed tomography (micro-CT) and histometric analyses to analyze the amount of attachment loss, crestal bone loss, connective tissue attachment, and the surface area with inflammatory cell infiltration. The effects of CsA on ligature-induced expressions of gelatinases and EMMPRIN in gingival tissues were examined with Western blotting and zymography, respectively. RESULTS: By micro-CT and histology, the Lig + CsA group had significantly more periodontal breakdown than the control and CsA groups but less periodontal breakdown than the Lig group. Consistent results were found for the expressions of gelatinases and EMMPRIN among the groups demonstrating that the Lig + CsA group had significantly less gingival protein expression of gelatinases and EMMPRIN than the Lig group. CONCLUSIONS: CsA inhibited the expressions of gelatinase MMPs and EMMPRIN and partially prevented the periodontal breakdown in ligature-induced experimental periodontitis. The CsA-induced attenuation of periodontal bone loss was strongly correlated positively with the expressions of MMP-2, MMP-9, and EMMPRIN in gingiva.
Assuntos
Basigina/efeitos dos fármacos , Ciclosporina/farmacologia , Inibidores Enzimáticos/farmacologia , Metaloproteinase 2 da Matriz/efeitos dos fármacos , Metaloproteinase 9 da Matriz/efeitos dos fármacos , Periodontite/enzimologia , Perda do Osso Alveolar/enzimologia , Perda do Osso Alveolar/prevenção & controle , Animais , Tecido Conjuntivo/efeitos dos fármacos , Gengiva/efeitos dos fármacos , Gengiva/enzimologia , Gengivite/enzimologia , Gengivite/prevenção & controle , Masculino , Inibidores de Metaloproteinases de Matriz/farmacologia , Perda da Inserção Periodontal/enzimologia , Perda da Inserção Periodontal/prevenção & controle , Periodontite/prevenção & controle , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Microtomografia por Raio-X/métodosRESUMO
BACKGROUND AND OBJECTIVE: It has been demonstrated that periodontitis induces a systemic inflammation, which may impair endothelial function. Cyclooxygenase-2 (COX-2) is an important enzyme in the inflammatory process and is responsible for prostacyclin production. We hypothesised that in periodontitis, an increase in vascular COX-2 expression may occur, which in turn may have a role in vascular homeostasis. Thus, we evaluated the vascular effects of COX-2 inhibition in an experimental rat model of periodontitis. MATERIAL AND METHODS: Experimental periodontitis was induced in rats by placing a cotton ligature around the cervix of both sides of the mandibular first molars and maxillary second molars. Sham-operated rats had the ligature removed immediately after the procedure. Mesenteric vessels were obtained for the study of COX-2 expression, and blood samples were collected for nitric oxide quantification. In another set of experiments, animals received etoricoxib (10 mg/kg/d, v.o.) or vehicle, and alveolar bone loss and cardiovascular parameters were evaluated. RESULTS: We observed an increase in COX-2 expression in mesenteric vessels harvested from animals with periodontitis, which was accompanied by a reduction in nitric oxide content. Etoricoxib treatment impaired the endothelium-dependent reduction in blood pressure in rats with periodontitis. CONCLUSION: Periodontitis increases vascular COX-2 expression, which is important in the maintenance of vascular homeostasis in this model. Despite the limitations of an animal study, these findings may have important implications regarding the safety of using selective COX-2 inhibitors in patients with periodontitis.
Assuntos
Pressão Arterial/fisiologia , Ciclo-Oxigenase 2/fisiologia , Periodontite/enzimologia , Perda do Osso Alveolar/enzimologia , Perda do Osso Alveolar/prevenção & controle , Animais , Anti-Inflamatórios/farmacologia , Pressão Arterial/efeitos dos fármacos , Ciclo-Oxigenase 2/efeitos dos fármacos , Inibidores de Ciclo-Oxigenase 2/farmacologia , Modelos Animais de Doenças , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/enzimologia , Etoricoxib , Frequência Cardíaca/efeitos dos fármacos , Masculino , Artérias Mesentéricas/enzimologia , Óxido Nítrico/sangue , Periodontite/prevenção & controle , Piridinas/farmacologia , Distribuição Aleatória , Ratos Wistar , Sulfonas/farmacologia , Vasoconstritores/farmacologia , Vasodilatadores/farmacologiaRESUMO
BACKGROUND: The aim of the present study is to investigate matrix metalloproteinase (MMP)-8 and tissue inhibitor of MMP-1 (TIMP-1) gene polymorphisms in generalized aggressive periodontitis (GAgP) and to assess the effects of MMP-8 and TIMP-1 genotypes on the outcomes of non-surgical periodontal therapy. METHODS: Genomic DNA was obtained from peripheral blood of 100 patients with GAgP and 167 periodontally healthy controls. MMP-8 +17 C/G, -799 C/T, -381 A/G and TIMP-1 372 T/C, *429 T/G polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism. Patients with GAgP received non-surgical periodontal therapy and were followed for 6 months. Clinical periodontal parameters and gingival crevicular fluid (GCF) samples were collected at baseline and at follow-up visits. GCF biomarkers were analyzed by immunofluorescence assay and enzyme-linked immunosorbent assay. RESULTS: Distribution of the MMP-8 -799 C/T genotypes was significantly different between the GAgP and control groups (P <0.005). TIMP-1 372 T/C and *429 T/G genotypes in males were also significantly different between study groups (P <0.004). GCF MMP-8 levels decreased until 3 months after non-surgical therapy compared with baseline in T and G alleles, as well as G and C allele carriers (P <0.0125), whereas no significant decreased was observed in non-carriers (P >0.0125). CONCLUSION: On the basis of the present findings, it can be suggested that MMP-8 -799 C/T and TIMP-1 372 T/C, *429 T/G gene polymorphisms in males may be associated with the susceptibility to GAgP in the Turkish population.
Assuntos
Periodontite Agressiva/enzimologia , Líquido do Sulco Gengival/enzimologia , Metaloproteinase 8 da Matriz/genética , Polimorfismo de Nucleotídeo Único/genética , Inibidor Tecidual de Metaloproteinase-1/genética , Adenina , Adolescente , Adulto , Periodontite Agressiva/genética , Periodontite Agressiva/terapia , Perda do Osso Alveolar/enzimologia , Perda do Osso Alveolar/terapia , Biomarcadores/análise , Citosina , Feminino , Seguimentos , Frequência do Gene/genética , Predisposição Genética para Doença/genética , Variação Genética/genética , Genótipo , Guanina , Humanos , Masculino , Pessoa de Meia-Idade , Perda da Inserção Periodontal/enzimologia , Perda da Inserção Periodontal/terapia , Fatores Sexuais , Timidina , Resultado do Tratamento , Adulto JovemRESUMO
IL-1ß contributes to connective tissue destruction in part by up-regulating stromelysin-1 (MMP-3), which in fibroblasts is a focal adhesion-dependent process. Protein tyrosine phosphatase-α (PTPα) is enriched in and regulates the formation of focal adhesions, but the role of PTPα in connective tissue destruction is not defined. We first examined destruction of periodontal connective tissues in adult PTPα(+/+) and PTPα(-/-) mice subjected to ligature-induced periodontitis, which increases the levels of multiple cytokines, including IL-1ß. Three weeks after ligation, maxillae were processed for morphometry, micro-computed tomography and histomorphometry. Compared with unligated controls, there was â¼1.5-3 times greater bone loss as well as 3-fold reduction of the thickness of the gingival lamina propria and 20-fold reduction of the amount of collagen fibers in WT than PTPα(-/-) mice. Immunohistochemical staining of periodontal tissue showed elevated expression of MMP-3 at ligated sites. Second, to examine mechanisms by which PTPα may regulate matrix degradation, human MMP arrays were used to screen conditioned media from human gingival fibroblasts treated with vehicle, IL-1ß or TNFα. Although MMP-3 was upregulated by both cytokines, only IL-1ß stimulated ERK activation in human gingival fibroblasts plated on fibronectin. TIRF microscopy and immunoblotting analyses of cells depleted of PTPα activity with the use of various mutated constructs or with siRNA or PTPα(KO) and matched wild type fibroblasts were plated on fibronectin to enable focal adhesion formation and stimulated with IL-1ß. These data showed that the catalytic and adaptor functions of PTPα were required for IL-1ß-induced focal adhesion formation, ERK activation and MMP-3 release. We conclude that inflammation-induced connective tissue degradation involving fibroblasts requires functionally active PTPα and in part is mediated by IL-1ß signaling through focal adhesions.
Assuntos
Tecido Conjuntivo/enzimologia , Gengiva/enzimologia , Periodontite/enzimologia , Proteínas Tirosina Fosfatases Classe 4 Semelhantes a Receptores/fisiologia , Perda do Osso Alveolar/enzimologia , Animais , Células Cultivadas , Indução Enzimática , MAP Quinases Reguladas por Sinal Extracelular , Fibroblastos/enzimologia , Gengiva/patologia , Humanos , Interleucina-1beta/metabolismo , Metaloproteinase 3 da Matriz/genética , Metaloproteinase 3 da Matriz/metabolismo , Camundongos , Camundongos Knockout , Células NIH 3T3 , Transdução de SinaisRESUMO
BACKGROUND AND OBJECTIVE: Endopeptidases, such as neutral endopeptidase (NEP), endothelin-converting enzyme-1 (ECE-1) and a disintegrin and metalloprotease 17 (ADAM17), are believed to have various important roles in oral mucosal and epidermal tissue for the regulation of defensive biological responses in the oral cavity, and their expression and activity are influenced by various factors, including oral diseases. However, knowledge concerning these endopeptidases in the oral cavity has been minimal until now. This study focused on three metalloendopeptidases - NEP, ECE-1 and ADAM17 - in the oral buccal mucosal epithelium of patients with periodontal diseases and investigated the relationship between their gene-expression levels and periodontal disease. MATERIAL AND METHODS: The levels of expression of NEP, ECE-1 and ADAM17 mRNAs in tissue samples collected from the oral buccal mucosal epithelium of 61 patients were investigated by relative quantification using real-time RT-PCR analysis. information on oral and systemic health was obtained from the clinical record of each patient. RESULTS: Among the three groups, classified based on the diagnosis of periodontal diseases (healthy/gingivitis, early periodontitis and moderate/advanced periodontitis), the relative expression level of NEP mRNA was significantly increased in the early periodontitis group and in the moderate/advanced periodontitis group compared with that in the healthy/gingivitis group. Moreover, the relative expression levels of ECE1 and ADAM17 mRNAs were significantly increased in the moderate/advanced periodontitis group compared with those in the healthy/gingivitis group. The correlation coefficients between the mean relative expression levels of NEP and ECE1 mRNAs, NEP and ADAM17 mRNAs, and ECE1 and ADAM17 mRNAs were r = 0.758, r = 0.707 and r = 0.934, respectively (p < 0.001). Furthermore, among the oral-related factors, there was a significant correlation between the number of sites with probing pocket depths of more than 4 mm and of more than 6 mm and the relative expression levels of NEP, ECE1 and ADAM17 mRNAs. In stepwise logistic regression models, high relative expression levels of ECE1 and ADAM17 mRNAs were significantly associated with moderate/advanced periodontitis. CONCLUSION: The present study suggests that the severity of periodontal disease may be associated with the expression of metalloendopeptidase genes, including NEP, ECE1 and ADAM17, in the buccal mucosal epithelium.
Assuntos
Metaloendopeptidases/genética , Mucosa Bucal/enzimologia , Periodontite/enzimologia , Proteínas ADAM/genética , Proteína ADAM17 , Idoso , Perda do Osso Alveolar/enzimologia , Perda do Osso Alveolar/genética , Ácido Aspártico Endopeptidases/genética , Periodontite Crônica/enzimologia , Periodontite Crônica/genética , Enzimas Conversoras de Endotelina , Epitélio/enzimologia , Feminino , Regulação Enzimológica da Expressão Gênica/genética , Hemorragia Gengival/enzimologia , Hemorragia Gengival/genética , Gengivite/enzimologia , Gengivite/genética , Humanos , Masculino , Pessoa de Meia-Idade , Neprilisina/genética , Bolsa Periodontal/enzimologia , Bolsa Periodontal/genética , Periodontite/genética , Periodonto/enzimologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Fator de Necrose Tumoral alfa/genética , Adulto JovemRESUMO
BACKGROUND AND OBJECTIVE: Periodontitis is more frequently found in subjects with Down's syndrome. The aim was to investigate whether the relationship between MMPs and TIMPs) in the gingival crevicular fluid of subjects with Down's syndrome is altered compared with controls. MATERIAL AND METHODS: Twenty-one adolescents with Down's syndrome and gingivitis (DS-G), 12 subjects with Down's syndrome and periodontitis (DS-P), 26 controls with gingivitis (HC-G) and eight controls with periodontitis (HC-P) were clinically examined. All patients were between 11 and 20 years of age. Gingival crevicular fluid was collected from each subject and the concentrations of MMPs (2, 3, 8, 9 and 13) and TIMPs (1, 2 and 3) (expressed as pg/µL adjusted for volume of gingival crevicular fluid) were determined using multianalyte kits from R&D Systems. RESULTS: The concentrations of MMP-2, MMP-3, MMP-8, MMP-9 and TIMP-2 in gingival crevicular fluid were significantly higher (p < 0.005) in the DS-G group compared with the HC-G group. The correlation coefficient between MMP-8 and TIMP-2 differed significantly (p = 0.006) between the DS-G group and the HC-G group. On the contrary, the correlation coefficients between MMPs and TIMPs did not differ significantly between the DS-P group and the HC-P group. However, the DS-P group exhibited a significantly lower concentration of TIMP-2 in the gingival crevicular fluid compared with the HC-P group. CONCLUSION: Down's syndrome subjects with gingivitis exhibit higher concentrations of MMPs in gingival crevicular fluid with an altered relationship between MMP-8 and TIMP-2, which might impair the periodontal tissue turnover.
Assuntos
Síndrome de Down/metabolismo , Líquido do Sulco Gengival/química , Metaloproteinase 8 da Matriz/análise , Inibidor Tecidual de Metaloproteinase-2/análise , Adolescente , Perda do Osso Alveolar/enzimologia , Perda do Osso Alveolar/metabolismo , Criança , Estudos Transversais , Síndrome de Down/enzimologia , Feminino , Líquido do Sulco Gengival/enzimologia , Hemorragia Gengival/enzimologia , Hemorragia Gengival/metabolismo , Gengivite/enzimologia , Gengivite/metabolismo , Humanos , Masculino , Metaloproteinase 13 da Matriz/análise , Metaloproteinase 2 da Matriz/análise , Metaloproteinase 3 da Matriz/análise , Metaloproteinase 9 da Matriz/análise , Higiene Bucal , Bolsa Periodontal/enzimologia , Bolsa Periodontal/metabolismo , Periodontite/enzimologia , Periodontite/metabolismo , Inibidor Tecidual de Metaloproteinase-1/análise , Inibidor Tecidual de Metaloproteinase-3/análise , Adulto JovemRESUMO
BACKGROUND AND OBJECTIVE: Matrix metalloproteinases (MMPs) play important roles in extracellular matrix degradation and may be regulated by extracellular matrix metalloproteinase inducer (EMMPRIN). The aim of this study was to investigate the temporal expression and localization of MMP-8 and MMP-13 during the development of ligature-induced periodontitis in rats, and to analyze the correlations of EMMPRIN with MMP-8 and MMP-13 in periodontitis. MATERIAL AND METHODS: Periodontitis was simulated in rats by ligaturing the cervix of the lower first molars, as described in our previous method. The rats were killed 0, 3, 5, 7, 11, 15 and 21 d after ligation. Micro-computed tomography examinations were performed to detect alveolar bone loss. Semiquantitative western blotting was used to assess the temporal changes in the levels of MMP-8, MMP-13 and EMMPRIN proteins in gingival tissue. Immunohistochemistry was applied to detect the expression and locations of MMP-8 and MMP-13 in gingival tissue and alveolar bone. RESULTS: Alveolar bone loss showed an exponential increase from days 3 to 11, followed by a slower rate of loss at subsequent study time points. MMP-8 showed a rapid increase of expression from baseline to a peak on day 3, a gradual decrease from days 5 to 7 and then stabilized thereafter. MMP-8 was predominantly located in neutrophil-like cells. Statistically, the expression of MMP-8 was not correlated with the expression of EMMPRIN. The expression of MMP-13 and of EMMRPIN increased from days 3 to 7, and showed a moderate decrease thereafter. The immunoreactivity of MMP-13 was mainly detected in monocytes/macrophages, on the alveolar bone surface, in osteoclasts and in gingival epithelial cells. Statistically, MMP-13 had a strong, positive correlation with EMMPRIN (r = 0.855, p < 0.01). CONCLUSION: The levels of expression of MMP-8 and MMP-13 are temporally varied at different periods during the development of experimental periodontitis. The level of expression of EMMPRIN is closely associated with the expression of MMP-13, but not with the expression of MMP-8. In addition, MMP-13 might be involved in alveolar bone destruction, as well as in physiological bone remodeling.
Assuntos
Basigina/análise , Metaloproteinase 13 da Matriz/análise , Metaloproteinase 8 da Matriz/análise , Periodontite/enzimologia , Perda do Osso Alveolar/enzimologia , Processo Alveolar/enzimologia , Processo Alveolar/patologia , Animais , Modelos Animais de Doenças , Células Epiteliais/enzimologia , Matriz Extracelular/metabolismo , Fibroblastos/enzimologia , Gengiva/enzimologia , Gengiva/patologia , Macrófagos/enzimologia , Masculino , Monócitos/enzimologia , Neutrófilos/enzimologia , Osteoclastos/enzimologia , Periodontite/imunologia , Distribuição Aleatória , Ratos , Ratos Wistar , Fatores de Tempo , Microtomografia por Raio-XRESUMO
OBJECTIVE: The study aims to evaluate the effect of alendronate (ALD) on bone-specific alkaline phosphatase (BALP) serum levels on periodontal bone loss in Wistar rats. DESIGN: Periodontitis was induced by ligature around the upper second molar in 36 male Wistar rats (± 200 g). Groups of six animals received 0.9% saline (SAL) or ALD (0.01; 0.05; 0.25 mgkg(-1), s.c.), over 11 days; then they were sacrificed and their maxillae were removed to be defleshed and stained for macroscopic or histopathological analysis. Blood samples were collected for BALP, transaminases and total alkaline phosphatase (TALP) serum dosage, and haematologic study. Rats were weighed daily. RESULTS: Periodontitis induction caused reduction of BALP, intense alveolar bone loss (ABL), cementum and periodontal ligament destructions and intense leucocyte infiltration seen microscopically. Systemically, periodontitis induced leucocytosis, weight loss and TALP reduction. ALD (0.25 mgkg(-1)) prevented BALP reduction (19.17 ± 1.36 Ul(-1)) when compared to SAL (13.6 ± 1.5), as well as prevented ABL, by 57.2%, when compared to SAL (4.80+0.18 mm(2)), which was corroborated by histological findings (ALD 0.25 mgkg(-1)=1.5 (1-2) and SAL=3 (2-3)) (p<0.05). ALD did not alter transaminases but reduced TALP levels (p<0.05). ALD 0.25 mgkg(-1) reduced 6th-h neutrophilia (2.50 ± 0.22cell × 10(3)mm(-3)) and 7th- (12.29 ± 0.66) and 11th-day lymphomonocytosis (15.74 ± 0.52) when compared to SAL (5.20 ± 0.28; 18.24 ± 1.05; and 23.21 ± 1.48, respectively). ALD did not alter the weight loss. CONCLUSION: ALD prevented BALP reduction and ABL and reduced inflammatory infiltrate, without causing systemic alterations.
Assuntos
Alendronato/farmacologia , Fosfatase Alcalina/sangue , Perda do Osso Alveolar/etiologia , Processo Alveolar/patologia , Conservadores da Densidade Óssea/farmacologia , Doenças Maxilares/etiologia , Periodontite/complicações , Fosfatase Alcalina/efeitos dos fármacos , Perda do Osso Alveolar/tratamento farmacológico , Perda do Osso Alveolar/enzimologia , Processo Alveolar/fisiopatologia , Análise de Variância , Animais , Masculino , Doenças Maxilares/tratamento farmacológico , Doenças Maxilares/enzimologia , Osteoclastos/citologia , Osteoclastos/efeitos dos fármacos , Ratos , Ratos Wistar , Cloreto de SódioRESUMO
PURPOSE: Current clinical procedures to control or regenerate bone loss due to peri-implantitis are not predictable neither accomplish complete resolution. Therefore, early detection of the onset and the active periods of bone loss are crucial for prevention of extensive peri-implant bone resorption. This study aimed to determine a possible association between the presence of collagenases (MMP-1, MMP-8 and MMP-13) in peri-implant sulcular fluid (PISF) and active periods of bone loss by annually adjusted vertical bone loss (AVBL) measurements. METHODS: Intended sample consisted of 76 consecutive patients who received oral implant treatment at the Fixed Prosthodontic Clinic of Okayama University Hospital from 1990 to 2000. Twelve subjects were lost to follow-up or refused to participate. Consequently, the actual sample consisted of 64 patients who were followed-up for at least one year. Those patients with AVBL>0.6mm were included in the severe peri-implantitis group, and randomly selected, age-, gender- and implantation site-matched healthy patients (AVBL<0.3mm) comprised the control group. PISF samples were collected from both groups and further analyzed by western blot for detection of collagenases. RESULTS: Four patients presented severe peri-implantitis. MMP-8 was the only collagenase detected in peri-implant sites with ongoing bone loss. PISF samples from control group showed no positive reactions to any collagenase. CONCLUSION: This study showed MMP-8 as a possible marker for progressive bone loss in peri-implantitis.
Assuntos
Perda do Osso Alveolar/enzimologia , Implantes Dentários , Metaloproteinase 8 da Matriz/análise , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
The tissue destruction that characterizes periodontitis is driven by the host response to bacterial pathogens. Inhibition of glycogen synthase kinase 3ß (GSK3ß) in innate cells leads to suppression of Toll-like receptor (TLR)-initiated proinflammatory cytokines under nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65 transcriptional control and promotion of cyclic adenosine monophosphate response element-binding (CREB)-dependent gene activation. Therefore, we hypothesized that the cell permeable GSK3-specific inhibitor, SB216763, would protect against alveolar bone loss induced by the key periodontal pathogen, Porphyromonas gingivalis (P. gingivalis), in a murine model. B6129SF2/J mice either were infected orally with P. gingivalis ATCC 33277; or treated with SB216763 and infected with P. gingivalis; sham infected; or exposed to vehicle only (dimethyl sulfoxide [DMSO]); or to GSK3 inhibitor only (SB216763). Alveolar bone loss and local (neutrophil infiltration and interleukin [IL]-17) and systemic (tumor necrosis factor [TNF], IL-6, Il-1ß and IL-12/IL-23 p40) inflammatory indices also were monitored. SB216763 unequivocally abrogated mean P. gingivalis-induced bone resorption, measured at 14 predetermined points on the molars of defleshed maxillae as the distance from the cementoenamel junction to the alveolar bone crest (p < 0.05). The systemic cytokine response, the local neutrophil infiltration and the IL-17 expression were suppressed (p < 0.001). These data confirm the relevance of prior in vitro phenomena and establish GSK3 as a novel, efficacious therapeutic preventing periodontal disease progression in a susceptible host. These findings also may have relevance to other chronic inflammatory diseases and the systemic sequelae associated with periodontal infections.
Assuntos
Perda do Osso Alveolar/enzimologia , Perda do Osso Alveolar/microbiologia , Quinase 3 da Glicogênio Sintase/antagonistas & inibidores , Porphyromonas gingivalis/fisiologia , Perda do Osso Alveolar/patologia , Perda do Osso Alveolar/prevenção & controle , Animais , Quinase 3 da Glicogênio Sintase/metabolismo , Glicogênio Sintase Quinase 3 beta , Indóis/farmacologia , Indóis/uso terapêutico , Inflamação/complicações , Inflamação/patologia , Interleucina-17/metabolismo , Maleimidas/farmacologia , Maleimidas/uso terapêutico , Maxila/efeitos dos fármacos , Maxila/patologia , Camundongos , Infiltração de Neutrófilos/efeitos dos fármacos , Porphyromonas gingivalis/efeitos dos fármacos , Receptores Toll-Like/metabolismoRESUMO
INTRODUCTION: Mitogen-activated protein kinase (MAPK) phosphatase-1 (MKP-1) has been shown to be a key negative regulator of the MAPK pathways of the innate immune system. The impact of MKP-1 in an endodontic model has yet to be studied. Thus, the purpose of this study was to determine the role of MKP-1 in a bacterial-driven model of pathologic endodontic bone loss. METHODS: Pulps were exposed in both lower first molars of 10-week-old mkp-1(+/+) and mkp-1(-/-) mice and left open to the oral environment for either 3 or 8 weeks. At death, mandibles were harvested and scanned by micro-computed tomography (µCT) to determine periapical bone loss. Histopathologic scoring was then performed on the samples to determine the amount of inflammatory infiltrate within the periapical microenvironment. RESULTS: Significant bone loss and inflammatory infiltrate were found in all experimental groups when compared with control. No statistical difference was found between mkp-1(+/+) and mkp-1(-/-) at either time point with respect to bone loss or inflammatory infiltrate. At 8 weeks, male mkp-1(-/-) mice were found to have significantly more bone loss and inflammatory infiltrate when compared with female mkp-1(-/-) mice. There was also a significant correlation between an increase in bone loss and increase in inflammatory infiltrate. CONCLUSIONS: A sexual dimorphism exists in the periapical inflammatory process, where male mkp-1(-/-) mice have more inflammation than female mkp-1(-/-) mice. The increase in inflammatory infiltrate correlates to more bone loss in the male mice.
Assuntos
Perda do Osso Alveolar/enzimologia , Fosfatase 1 de Especificidade Dupla/fisiologia , Periodontite Periapical/enzimologia , Caracteres Sexuais , Perda do Osso Alveolar/microbiologia , Animais , Exposição da Polpa Dentária/microbiologia , Fosfatase 1 de Especificidade Dupla/genética , Feminino , Processamento de Imagem Assistida por Computador , Imunidade Inata/imunologia , Inflamação , Sistema de Sinalização das MAP Quinases/imunologia , Masculino , Doenças Mandibulares/enzimologia , Doenças Mandibulares/microbiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Dente Molar/microbiologia , Neutrófilos/patologia , Periodontite Periapical/microbiologia , Fatores de Tempo , Microtomografia por Raio-XRESUMO
The aim of this study was to assess the presence of aspartate aminotransferase (AST) in peri-implant crevicular fluid, with or without clinical signs of mucositis, to determine its predictive diagnostic value, sensitivity, and specificity. The AST levels were determined (at a threshold of 1200 µIU/mL) for 60 clinically successful implants in 25 patients with or without peri-implant mucositis. Samples were taken prior (AST1) to peri-implant probing with a manual constant-pressure probe (0.2 N) and 15 minutes after probing (AST2). Clinical assessments included radiographic determination of preexisting bone loss, probing, and the evaluation of mucositis, plaque, and bleeding upon probing. Analysis was performed at both the level of the implant and the patient as a unit. We detected a significant difference between AST1 and AST2 at both levels. A significant difference was observed at AST1 between implants that bled upon probing and those that did not. However, when we considered the patient as a unit, there were no significant differences. The plaque index was not significant at either level. AST1 had high specificity and positive predictive diagnostic value (80%) for bleeding upon probing. Probing induces a greater release of AST from inflamed tissues compared with healthy tissues in situ but not at the systemic level. At the implant level, the implant position could be responsible for this difference. Aspartate aminotransferase was a reliable predictor of patients with mucositis.
Assuntos
Aspartato Aminotransferases/análise , Implantes Dentários , Líquido do Sulco Gengival/enzimologia , Estomatite/enzimologia , Adolescente , Adulto , Idoso , Perda do Osso Alveolar/enzimologia , Índice de Placa Dentária , Feminino , Hemorragia Gengival/enzimologia , Humanos , Masculino , Pessoa de Meia-Idade , Osseointegração/fisiologia , Índice Periodontal , Bolsa Periodontal/enzimologia , Valor Preditivo dos Testes , Radiografia Interproximal , Sensibilidade e Especificidade , Adulto JovemRESUMO
Molecular mechanisms responsible for periodontal disease (PD) and its worsening in type 1 Diabetes Mellitus (DM1) remain unknown. Cytokine profile and expression levels of collagenases, Mmp14, and tissue inhibitors were determined, as were the numbers of neutrophils and macrophages in combined streptozotocin-induced DM1 and ligature-induced PD models. Increased IL-23 (80-fold) and Mmp8 expression (25-fold) was found in DM1. Ligature resulted in an IL-1ß/IL-6 profile, increased expression of Mmp8, Mmp13, and Mmp14 (but not Mmp1), and transient expression of Timp1 and Reck in non-diabetics. PD in DM1 involved IL-1ß (but not IL-6) and IL-23/IL-17, reduced IL-6 and IL-10, sustained Mmp8 and Mmp14, increased Mmp13 and reduced Reck expression in association with 20-fold higher counts of neutrophils and macrophages. IL-23 and Mmp8 expression are hallmarks of DM1. In association with the IL-1/IL-6 (Th1) response in PD, one found a secondary IL-17 (Th17) pathway in non-diabetic rats. Low IL-6/TNF-α suggest that the Th1 response was compromised in DM1, while IL-17 indicates a prevalence of the Th17 pathway, resulting in high neutrophil recruitment. Mmp8, Mmp13, and Mmp14 expression seems important in the tissue destruction during PD in DM1. PD-associated IL-1/IL-6 (Th1), IL-10, and Reck expression are associated with the acute-to-chronic inflammation transition, which is lost in DM1. In conclusion, IL-23/IL-17 are associated with the PD progression in DM1.
Assuntos
Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Tipo 1/complicações , Gengiva/enzimologia , Gengiva/imunologia , Mediadores da Inflamação/metabolismo , Interleucina-17/metabolismo , Interleucina-23/metabolismo , Metaloproteinase 14 da Matriz/metabolismo , Metaloproteinase 8 da Matriz/metabolismo , Doenças Periodontais/complicações , Perda do Osso Alveolar/enzimologia , Perda do Osso Alveolar/etiologia , Perda do Osso Alveolar/imunologia , Animais , Diabetes Mellitus Experimental/enzimologia , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/imunologia , Diabetes Mellitus Tipo 1/enzimologia , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/imunologia , Progressão da Doença , Proteínas Ligadas por GPI/metabolismo , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Ligadura , Macrófagos/imunologia , Masculino , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinase 14 da Matriz/genética , Metaloproteinase 8 da Matriz/genética , Dente Molar/cirurgia , Infiltração de Neutrófilos , Neutrófilos/imunologia , Doenças Periodontais/enzimologia , Doenças Periodontais/genética , Doenças Periodontais/imunologia , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Células Th17/imunologia , Fatores de Tempo , Proteínas Supressoras de Tumor/metabolismo , Regulação para CimaRESUMO
BACKGROUND: Cathepsin-K is an enzyme involved in bone metabolism which may make this feature important for both natural teeth and dental implants. The aims of the present study are to comparatively analyze the gingival crevicular fluid (GCF)/peri-implant sulcus fluid (PISF) cathepsin-K levels of natural teeth and dental implants, and to assess the potential relationship between this biochemical parameter and alveolar bone loss around natural teeth and dental implants. METHODS: Probing depth, bleeding on probing, gingival index, and plaque index clinical parameters were assessed, and GCF/PISF samples were obtained from natural teeth/dental implants presenting with either clinical health, gingivitis/peri-implant mucositis, or chronic periodontitis/peri-implantitis. Cathepsin-K activity levels of 42 GCF samples and 54 PISF samples were determined, and marginal bone loss (MBL) measures were calculated from digitalized standardized intraoral periapical radiographs obtained from natural teeth and dental implants by using cemento-enamel junction and the actual distance between two consecutive threads of the dental implant as reference points for natural teeth and dental implants, respectively. RESULTS: Comparing the natural teeth group with dental implant group with regard to MBL measure, cathepsin-K activity, and GCF/PISF volume revealed no significant differences. In both natural teeth and dental implant groups, despite higher MBL measures, cathepsin-K activity, and GCF/PISF volumes with the presence of inflammation, it was the presence of alveolar bone loss that lead to significantly higher values for these parameters. CONCLUSION: We suggest cathepsin-K as a biochemical parameter for monitoring periodontal/peri-implant alveolar bone loss.
Assuntos
Perda do Osso Alveolar/enzimologia , Catepsina K/análise , Implantes Dentários , Líquido do Sulco Gengival/enzimologia , Dente/enzimologia , Adulto , Idoso , Perda do Osso Alveolar/diagnóstico por imagem , Biomarcadores/análise , Periodontite Crônica/enzimologia , Índice de Placa Dentária , Feminino , Hemorragia Gengival/enzimologia , Gengivite/enzimologia , Humanos , Masculino , Pessoa de Meia-Idade , Peri-Implantite/enzimologia , Índice Periodontal , Bolsa Periodontal/enzimologia , Radiografia Interproximal , Estomatite/enzimologia , Adulto JovemRESUMO
BACKGROUND: S-nitrosoglutathione (GSNO) is a nitric oxide donor that may exert antioxidant, anti-inflammatory, and microbicidal actions and is thus a potential drug for the topical treatment of periodontitis. In this study, the effect of intragingival injections of GSNO-containing polyvinylpyrrolidone (PVP) formulations is evaluated in a rat model of periodontitis. METHODS: Periodontal disease was induced by placing a sterilized nylon (000) thread ligature around the cervix of the second left upper molar of the animals, which received intragingival injections of PVP; saline; or PVP/GSNO solutions which corresponded to GSNO doses of 25, 100, and 500 nmol; 1 hour before periodontitis induction, and thereafter, daily for 11 days. RESULTS: PVP/GSNO formulations at doses of 25 and/or 100, but not 500 nmol caused significant inhibition of alveolar bone loss, increase of bone alkaline phosphatase, decrease of myeloperoxidase activity, as well as significant reduction of inflammatory and oxidative stress markers when compared to saline and PVP groups. These effects were also associated with a decrease of matrix metalloproteinases 1 and 8, inducible nitric oxide synthase, and nuclear factor-κB immunostaining in the periodontium. CONCLUSION: Local intragingival injections of GSNO reduces inflammation and bone loss in experimental periodontal disease.
Assuntos
Perda do Osso Alveolar/prevenção & controle , Anti-Inflamatórios/uso terapêutico , Doadores de Óxido Nítrico/uso terapêutico , Periodontite/prevenção & controle , S-Nitrosoglutationa/uso terapêutico , Fosfatase Alcalina/efeitos dos fármacos , Perda do Osso Alveolar/enzimologia , Processo Alveolar/efeitos dos fármacos , Processo Alveolar/enzimologia , Animais , Anti-Inflamatórios/administração & dosagem , Biomarcadores/análise , Reabsorção Óssea/prevenção & controle , Modelos Animais de Doenças , Gengiva , Injeções , Interleucina-1beta/efeitos dos fármacos , Peroxidação de Lipídeos/efeitos dos fármacos , Inibidores de Metaloproteinases de Matriz , NF-kappa B/antagonistas & inibidores , Doadores de Óxido Nítrico/administração & dosagem , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Estresse Oxidativo/efeitos dos fármacos , Peroxidase/antagonistas & inibidores , Excipientes Farmacêuticos , Povidona , Ratos , Ratos Wistar , S-Nitrosoglutationa/administração & dosagem , Cloreto de Sódio , Fator de Necrose Tumoral alfa/efeitos dos fármacosRESUMO
PURPOSE: Cathepsin-K is an enzyme involved in bone metabolism. This feature may make it important both for natural teeth and dental implants. The aims of the present study were to comparatively analyze cathepsin-K levels in gingival crevicular fluid (GCF) and peri-implant sulcus fluid (PISF) and to determine whether GCF and PISF cathepsin-K profiles reflect the clinical periodontal/peri-implant status. MATERIALS AND METHODS: Clinical parameters (probing depth, Gingival Index, Plaque Index, and bleeding on probing) were recorded, and GCF/PISF samples were obtained from natural teeth (group T) and dental implants (group I), which were divided into groups based on health (clinically healthy, gingivitis/peri-implant mucositis, and periodontitis/peri-implantitis). Cathepsin-K activity was determined with a commercially available cathepsin-K activity assay kit (BioVision). RESULTS: Sixty natural teeth and 68 dental implants were examined. Teeth with periodontitis (group T-3) showed significantly higher total cathepsin-K activity (10.39 units) than teeth with gingivitis (group T-2, 1.71 units) and healthy teeth (group T-1, 1.90 units). The difference in cathepsin-K activity between groups T-2 and T-1 was not significant. Implants with peri-implantitis (group I-3) had higher total enzyme activity (10.26 units) than healthy implants (group I-1) (3.44 units). Although the difference between clinical parameters was not significant, group I-3 had higher cathepsin-K levels than group I-2 (4.74 units). When natural teeth (T-1, T-2, T-3) were compared to implants (I-1, I-2, I-3), no significant differences were observed for cathepsin-K levels. CONCLUSION: More cathepsin-K activity was clearly observed with inflammatory periodontal and peri-implant destruction. The highest cathepsin-K levels detected in GCF and PISF samples, obtained from sites with periodontitis and peri-implantitis, suggests the potential involvement of cathespin-K in increased bone metabolism around natural teeth and dental implants.