RESUMO
OBJECTIVE: The present study aimed to compare levels of matrix metalloproteinase-9 (MMP-9) and myeloperoxidase (MPO) in gingival crevicular fluid (GCF) from subjects with controlled and noncontrolled Type 2 Diabetes Mellitus (T2D), with and without stage 2 grade B periodontitis (POD2B) versus healthy (H) subjects. METHODS: The levels of both enzymes, from 80 GCF samples collected with PerioPaper strips, were analyzed by a Multiplex/Luminex assay. Five groups were formed, all current patients at the Institutional Dentistry Service, and distributed as follows: two groups of diabetics (one controlled and one poorly controlled); two groups with the previous conditions and diagnosed with POD2B; and one H group. RESULTS: The highest concentration of MMP-9 corresponded to the H group, while the lowest corresponded to the T2D controlled group. Regarding MPO levels, the highest levels were associated with the T2D controlled with POD2B group and the lowest with the T2D controlled group. CONCLUSIONS: No apparent relationship between the elevation of MMP-9 and MPO levels was observed among subjects with T2D, with and without POD2B, compared to H subjects.
Assuntos
Diabetes Mellitus Tipo 2/enzimologia , Líquido do Sulco Gengival/enzimologia , Metaloproteinase 9 da Matriz/metabolismo , Periodontite/enzimologia , Peroxidase/metabolismo , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
OBJECTIVE: In this study, we aimed to histologically and immunologically evaluate the effect of diode laser treatment when applied adjunctive to scaling and root planing (SRP) in an experimental periodontitis model. MATERIALS AND METHODS: We used Wistar-Albino rats (n=60) with average weight of 230 g. Experimental periodontitis was induced by ligature at the right and left first mandibular molar teeth in all rats. After 11 days, the ligature was removed and rats were divided into two groups. The control group (n=30) received only SRP treatment, while the laser group (n=30) received a diode laser (GaAlAs, 810 nm, 1 W, 10 J, 20 s) treatment adjunctive to SRP. Ten rats in each group were sacrificed after 7, 15, and 30 days. Histopathological examination was performed in the left mandible of rats. Myeloperoxidase (MPO) was evaluated by western blot in the gingival specimens from the right mandible. RESULTS: MPO levels in the laser group were statistically significantly lower compared with the control group (p≤0.05). There was no statistically significance at any time between MPO levels in the control group (p>0.05). MPO levels in the laser group at the 7th day were statistically significantly higher compared to the 15th (p≤0.05) and the 30th day (p≤0.05). Inflammatory cell infiltration decreased over time in both groups and was statistically significantly lower in the laser group than in the control group at all times (p≤0.01). CONCLUSIONS: Within the limits of this study, we suggest that diode laser application is an adjunctive treatment because it reduced inflammation and MPO when applied in addition to SRP. On the other hand, more studies are needed for the assessment of the effects of diode laser application to periodontal tissues.
Assuntos
Raspagem Dentária/métodos , Lasers Semicondutores/uso terapêutico , Terapia com Luz de Baixa Intensidade/métodos , Periodontite/patologia , Periodontite/terapia , Peroxidase/análise , Animais , Western Blotting , Terapia Combinada , Modelos Animais de Doenças , Ligadura , Periodontite/enzimologia , Distribuição Aleatória , Ratos Wistar , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do TratamentoRESUMO
Owing to its molecular stability in body fluids, soluble urokinase-type plasminogen activator receptor (suPAR) is used as a biomarker for the level of systemic inflammation. This study compares the suPAR levels in serum with those in the saliva of adolescents and evaluates their association with the periodontal conditions. Adolescents identified as screen positive (n = 87) or screen negative (n = 73) for periodontitis had saliva and serum samples taken, along with subgingival plaque samples. The concentrations of suPAR were determined in saliva and serum, and 18 microbial species and the immunoglobulin response to them was evaluated. Factor analyses were used to reduce the number of variables within each of the domains of clinical, microbiological, and immunological findings. The median salivary suPAR concentration was 13.18 [(interquartile range (IQR): 6.20-23.36] µg l-1 and was not associated with the serum suPAR levels (median 2.05; IQR: 1.62-2.46 µg l-1 ). Linear regression analysis showed that the log10 (salivary suPAR concentration) was statistically significantly positively associated with the clinical phenotype 'Periodontitis Extent' (ß = 0.28; 95% CI: 0.16-0.39) along with 'Putative periodontopathogens' (ß = 0.65; 95% CI: 0.51-0.79). The study represents the first determination of salivary suPAR concentration in a larger well-defined adolescent population. Our results suggest the potential for clinical use of suPAR in saliva as an inflammatory risk indicator/biomarker of periodontitis.
Assuntos
Periodontite/enzimologia , Receptores de Ativador de Plasminogênio Tipo Uroquinase/metabolismo , Adolescente , Chile , Placa Dentária/enzimologia , Placa Dentária/microbiologia , Análise Fatorial , Feminino , Humanos , Imunoglobulina G/sangue , Masculino , Periodontite/microbiologia , Saliva/enzimologia , Adulto JovemRESUMO
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
Immune-inflammatory processes are trigged in chronic periodontitis (CP), where matrix metalloproteinases (MMPs) are released and involved in the degradation of the extracellular matrix components that can be detected in the gingival crevicular fluid (GCF). The purpose of the study was to determine the levels of MMP-3 and MMP-8 in GCF, before and after nonsurgical periodontal treatment (NSPT), to evaluate disease activity and therapy response. Eleven patients with PC and eleven healthy controls were selected. Clinical measurements to evaluate gingival index (GI), plaque index (PI), probing depth (PD) and clinical attachment loss (CAL) were made in all the teeth of each individual and in six sites per tooth. GCF samples were taken from one tooth per quadrant, with a pocket depth > or =4 mm and a clinical attachment loss > or =5 mm, and the levels of MMP-3 and MMP-8 measured using an ELISA test. Statistically significant differences in clinical parameters were observed (p < 0.05) between patients with CP and control groups before the periodontal treatment, with significant decrease in all indexes after the NSPT. The initial concentrations of MMP-3 and MMP-8 were significantly higher than those obtained after the NSPT and in the control group, without observing a correlation between the clinical parameters and the levels of MMPs. Increased levels of MMP-3 and MMP-8 in the GCF of patients with PC declined significantly after NSPT, and the difference between the levels in healthy individuals and patients, suggests the important participation of these MMPs in tissue destruction in PC disease..
Assuntos
Líquido do Sulco Gengival/enzimologia , Metaloproteinase 3 da Matriz/análise , Metaloproteinase 8 da Matriz/análise , Periodontite/enzimologia , Adulto , Biomarcadores , Doença Crônica , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Índice Periodontal , Periodontite/terapiaRESUMO
En la periodontitis crónica (PC) se desencadenan procesos inmunoinflamatorios, donde se liberan metaloproteinasas de la matriz (MMPs), enzimas involucradas en la degradación de la matriz extracelular, las cuales pueden ser detectadas en el fluido gingival crevicular (FGC). El propósito del estudio fue determinar los niveles de MMP-3 y MMP-8 en FGC, antes y después del tratamiento periodontal no quirúrgico (TPNQ) para evaluar actividad de la enfermedad y respuesta terapéutica. Once pacientes con periodontitis crónica y 11 controles sanos fueron seleccionados. Se evaluaron los parámetros clínicos: índice gingival, índice de placa, profundidad al sondaje y pérdida de inserción; en todos los dientes de cada individuo y en seis sitios por diente. Muestras de FGC fueron tomadas de un diente por cuadrante, con profundidad de saco ≥ 4mm y pérdida de inserción ≥ 5 mm, los niveles de MMP-3 y MMP-8 fueron determinados por ELISA. Diferencias estadísticamente significativas fueron observadas entre los parámetros clínicos del grupo control y los pacientes con PC antes del tratamiento, registrándose posterior al TPNQ disminución significativa de todos los índices. Las concentraciones iniciales de MMP-3 y 8 en el grupo con PC fueron significativamente mayores a las obtenidas luego del TPNQ y en el grupo control, sin observar correlación entre parámetros clínicos y niveles de MMPs. La disminución significativa de los valores de MMP-3 y 8 en FGC de los pacientes con PC, posterior al TPNQ, indican la participación importante de estas enzimas en la degradación del tejido, y la efectividad del tratamiento periodontal para su control.
Immune-inflammatory processes are trigged in chronic periodontitis (CP), where matrix metalloproteinases (MMPs) are released and involved in the degradation of the extracellular matrix components that can be detected in the gingival crevicular fluid (GCF). The purpose of the study was to determine the levels of MMP-3 and MMP-8 in GCF, before and after nonsurgical periodontal treatment (NSPT), to evaluate disease activity and therapy response. Eleven patients with PC and eleven healthy controls were selected. Clinical measurements to evaluate gingival index (GI), plaque index (PI), probing depth (PD) and clinical attachment loss (CAL) were made in all the teeth of each individual and in six sites per tooth. GCF samples were taken from one tooth per quadrant, with a pocket depth ≥ 4 mm and a clinical attachment loss ≥ 5 mm, and the levels of MMP-3 and MMP-8 measured using an ELISA test. Statistically significant differences in clinical parameters were observed (p < 0.05) between patients with CP and control groups before the periodontal treatment, with significant decrease in all indexes after the NSPT. The initial concentrations of MMP-3 and MMP-8 were significantly higher than those obtained after the NSPT and in the control group, without observing a correlation between the clinical parameters and the levels of MMPs. Increased levels of MMP-3 and MMP-8 in the GCF of patients with PC declined significantly after NSPT, and the difference between the levels in healthy individuals and patients, suggests the important participation of these MMPs in tissue destruction in PC disease.
Assuntos
Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Líquido do Sulco Gengival/enzimologia , /análise , /análise , Periodontite/enzimologia , Biomarcadores , Doença Crônica , Ensaio de Imunoadsorção Enzimática , Índice Periodontal , Periodontite/terapiaRESUMO
La periodontitis constituye la infección bacteriana más prevalente a nivel mundial y representa un factor de riesgo para diversas patologías sistémicas. El estado de inflamación y destrucción periodontal se manifiestan a través de la presencia de biomarcadores en el suero y fluidos orales, tales como el fluido gingival crevicular (FGC), saliva y enjuague oral. Enzimas como las metaloproteinasas de matriz (MMP) y mieloperoxidasa, constituyen biomarcadores potenciales para ensayos moleculares complementarios a la clínica de uso en el sillón dental. A continuación se presenta una revisión de la literatura respecto de la aplicación potencial del análisis de metaloproteinasas de matriz extracelular (MMPs) en el diagnóstico complementario de las enfermedades periodontales. Se ha demostrado que los niveles de MMP-9, -13 y particularmente de MMP-8, se asocian con el grado de inflamación periodontal, y pueden diferenciar entre sujetos sanos, con gingivitis, periodontitis y peri-implantitis, mientras que la mejoría de los parámetros clínicos en respuesta al tratamiento periodontal se asocia con la reducción de la activación y niveles de estas enzimas en FGC, como así también en el suero. Se concluye que la determinación, particularmente de MMP-8 en fluidos orales presenta un elevado potencial como complemento de los métodos clínicos tradicionales para identificar a los pacientes con periodontitis o en riesgo de desarrollar la enfermedad, monitorear fases del tratamiento y mejoría de signos periodontales e incluso evaluar el estado de inflamación sistémica.
Periodontal disease is the most common bacterial infection worldwide and it can contribute to enhance the risk for the development of several systemic diseases. The status of periodontal inflammation and destruction can be reflected in biomarker measurement in serum and oral fluids, like gingival crevicular fluid (GCF), saliva and mouth-rinse. Some enzymes, such as matrix metalloproteinases (MMPs) and myeloperoxidase are potential candidates for chair-side point-of-care oral fluid assays. This review is focused on the utility of matrix metalloproteinase (MMP) analysis in oral fluid as a complementary diagnostic method to chronic periodontitis. Levels of MMP-9,-13 and specially of MMP-8, reflect oral inflammatory status and discriminate among healthy, gingivitis, periodontitis and periimplantitis individuals, whereas MMP levels and activation in GCF and serum are in line with the improvement of clinical parameters in response to periodontal treatment. As a conclusion, MMP-8 assessment in GCF could represent a helpful adjunctive method to traditional diagnostics to identify periodontitis or patients at risk to develop the disease, monitor treatment phases, improvement of periodontal signs and even screen the systemic inflammation status.
Assuntos
Humanos , Colagenases/análise , Líquido do Sulco Gengival/enzimologia , Periodontite/diagnóstico , Periodontite/enzimologia , Doenças Cardiovasculares/diagnóstico , Doenças Cardiovasculares/enzimologia , Biomarcadores/análise , /análise , Sistemas Automatizados de Assistência Junto ao Leito , Fatores de TempoRESUMO
OBJECTIVE: In a previous study we observed that parotid glands from rats with experimental periodontitis showed an increase in basal amylase release as a result of an increase in cAMP accumulation induced by PGE(2) production. The aim of this work was to study whether this change in amylase release influences the secretory effect of carbachol. DESIGN: Experimental periodontitis was induced through placing a black thread around the cervix of the two lower first molars. Experiments were done 22 days after ligature induced periodontitis. Amylase release was evaluated in vitro and determined using a colorimetric method which uses starch as substrate. RESULTS: The effect of carbachol was increased in parotid glands from periodontitis rats. The effect of 10(-6)M carbachol was inhibited by 4-DAMP (10(-6)M), U-73122 (5 × 10(-6)M) and trifluoperazine (5 × 10(-6)M) in both groups. No changes were observed in the binding sites and affinity in parotid membranes from rats with experimental periodontitis. The inhibition of the adenylyl cyclase and the cyclooxygenase induced a right shift of the carbachol concentration-response curve in periodontitis group whilst the opposite effect was observed in control group in the presence of db-cAMP and PGE(2). CONCLUSIONS: Parotid glands from rats with experimental periodontitis release more amylase in response to carbachol suggesting an interaction between Ca(2+) and cAMP in the fusion/exocytosis step of secretory vesicles.
Assuntos
Amilases/biossíntese , Carbacol/farmacologia , Ativação Enzimática/efeitos dos fármacos , Glândula Parótida/enzimologia , Periodontite/enzimologia , Análise de Variância , Animais , Colorimetria/métodos , AMP Cíclico/fisiologia , Masculino , Glândula Parótida/efeitos dos fármacos , Periodontite/metabolismo , Ratos , Ratos WistarRESUMO
OBJECTIVE: In this study we investigated the activity of the nitric oxide synthase (NOS) in parotid glands from rats with experimental periodontitis and controls. METHODS: Periodontitis was produced by a ligature placed around the cervix of the two lower first molar. Experiments were carried out 22 days after the ligature. RESULTS: Ligation caused an increase in parotid NOS activity. The selective blocker of the inducible isoform of the enzyme partially inhibited its activity in parotid glands from rat with ligature. In controls, the activity was partially inhibited by the antagonists of the selective neural and endothelial isoforms. NOS activity in rats with ligature was cyclic adenosine monophosphate (cAMP)-dependent while in controls it was calcium-dependent. Prostaglandin E2 concentration was increased in parotid gland from rats with ligature. The inhibitor of prostaglandin production, FR 122047, diminished both, prostaglandin production and NOS activity. In rats with ligature unstimulated amylase released is increased. Both, prostaglandin and NOS were involved in the increment of amylase release. CONCLUSION: It can be concluded that in parotid glands from ligated rats, prostaglandin E2 production is increased and, through cAMP accumulation, activates the inducible NOS isoform. The increment of nitric oxide production participates in the increase in basal amylase release.
Assuntos
Óxido Nítrico Sintase/metabolismo , Glândula Parótida/enzimologia , Periodontite/enzimologia , Proteínas e Peptídeos Salivares/metabolismo , Adenina/análogos & derivados , Adenina/farmacologia , Amilases/metabolismo , Animais , Cálcio/farmacologia , Subunidades Catalíticas da Proteína Quinase Dependente de AMP Cíclico/antagonistas & inibidores , Subunidades Catalíticas da Proteína Quinase Dependente de AMP Cíclico/farmacologia , Inibidores de Ciclo-Oxigenase/farmacologia , Dinoprostona/antagonistas & inibidores , Dinoprostona/metabolismo , Modelos Animais de Doenças , Ácido Egtázico/farmacologia , Inibidores Enzimáticos/farmacologia , Guanidinas/farmacologia , Indazóis/farmacologia , Indometacina/farmacologia , Masculino , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase Tipo I/antagonistas & inibidores , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Óxido Nítrico Sintase Tipo III/antagonistas & inibidores , Tamanho do Órgão , Ornitina/análogos & derivados , Ornitina/farmacologia , Glândula Parótida/efeitos dos fármacos , Piperazinas/farmacologia , Prostaglandina-Endoperóxido Sintases/efeitos dos fármacos , Ratos , Ratos Wistar , Proteínas e Peptídeos Salivares/efeitos dos fármacos , Tiazóis/farmacologia , ômega-N-Metilarginina/farmacologiaRESUMO
BACKGROUND: The present study investigated the effects of venlafaxine, an antidepressant drug with immunoregulatory properties on the inflammatory response and bone loss associated with experimental periodontal disease (EPD). MATERIALS AND METHODS: Wistar rats were subjected to a ligature placement around the second upper left molar. The treated groups received orally venlafaxine (10 or 50 mg/kg) one hour before the experimental periodontal disease induction and daily for 10 days. Vehicle-treated experimental periodontal disease and a sham-operated (SO) controls were included. Bone loss was analyzed morphometrically and histopathological analysis was based on cell influx, alveolar bone, and cementum integrity. Lipid peroxidation quantification and immunohistochemistry to TNF-alpha and iNOS were performed. RESULTS: Experimental periodontal disease rats showed an intense bone loss compared to SO ones (SO = 1.61 +/- 1.36; EPD = 4.47 +/- 1.98 mm, p < 0.001) and evidenced increased cellular infiltration and immunoreactivity for TNF-alpha and iNOS. Venlafaxine treatment while at low dose (10 mg/kg) afforded no significant protection against bone loss (3.25 +/- 1.26 mm), a high dose (50 mg/kg) caused significantly enhanced bone loss (6.81 +/- 3.31 mm, p < 0.05). Venlafaxine effectively decreased the lipid peroxidation but showed no significant change in TNF-alpha or iNOS immunoreactivity. CONCLUSION: The increased bone loss associated with high dose venlafaxine may possibly be a result of synaptic inhibition of serotonin uptake.
Assuntos
Perda do Osso Alveolar/complicações , Perda do Osso Alveolar/tratamento farmacológico , Cicloexanóis/uso terapêutico , Periodontite/complicações , Periodontite/tratamento farmacológico , Perda do Osso Alveolar/enzimologia , Animais , Cicloexanóis/farmacologia , Gengiva/efeitos dos fármacos , Gengiva/patologia , Imuno-Histoquímica , Ligadura , Malondialdeído/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Periodontite/enzimologia , Periodontite/patologia , Ratos , Ratos Wistar , Inibidores Seletivos de Recaptação de Serotonina/farmacologia , Inibidores Seletivos de Recaptação de Serotonina/uso terapêutico , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Cloridrato de VenlafaxinaRESUMO
The mitogen-activated protein (MAP) kinase phosphatase (MKP) family plays an important function in regulating the pro-inflammatory cytokines by deactivating MAP kinases. MKP-1 is essential for the dephosphorylation of p38 MAP kinase that regulates expression of IL-6, TNF-alpha, and IL-1 beta. We hypothesized that MKP-1 regulates inflammatory bone loss in experimental periodontitis. Wild-type and Mkp-1(-/-) mice received A. actinomycetemcomitans LPS injection in the palatal region or PBS control 3 times/wk for 30 days. Mice were killed, and maxillae were assessed by microcomputed tomography, histological analysis, and TRAP staining for measurement of bone loss, extent of inflammation, and degree of osteoclastogenesis. Results indicated that, in LPS-injected Mkp-1(-/-) mice, significantly greater bone loss occurred with more inflammatory infiltrate and a significant increase in osteoclastogenesis compared with Mkp-1(-/-) control sites or either wild-type group. Analysis of these data indicates that MKP-1 plays a key role in the regulation of inflammatory bone loss.
Assuntos
Perda do Osso Alveolar/enzimologia , Fosfatase 1 de Especificidade Dupla/fisiologia , Fosfatase Ácida/análise , Aggregatibacter actinomycetemcomitans/fisiologia , Perda do Osso Alveolar/patologia , Perda do Osso Alveolar/prevenção & controle , Animais , Biomarcadores/análise , Contagem de Células , Linhagem Celular , Tomografia Computadorizada de Feixe Cônico , Fosfatase 1 de Especificidade Dupla/imunologia , Imageamento Tridimensional , Imunidade Inata/imunologia , Interleucina-6/análise , Interleucina-6/metabolismo , Isoenzimas/análise , Leucócitos/patologia , Lipopolissacarídeos/farmacologia , Doenças Maxilares/enzimologia , Doenças Maxilares/patologia , Doenças Maxilares/prevenção & controle , Camundongos , Camundongos Knockout , Osteoclastos/patologia , Palato , Periodontite/enzimologia , Periodontite/microbiologia , Fosforilação , Fosfatase Ácida Resistente a Tartarato , Fatores de Tempo , Microtomografia por Raio-X , Proteínas Quinases p38 Ativadas por Mitógeno/análise , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
It is known that subjects with periodontitis show enhanced amylase concentration in saliva. Our purpose was to analyze the release of amylase in parotid glands from rats with experimental periodontitis and controls. We present evidence that periodontitis induces an increase in resting amylase activity and release without changes in isoproterenol-induced amylase secretion. Changes in amylase were reverted by the inhibition of the adenylyl cyclase by SQ 22536, the cyclooxygenase type 1 by FR 122047 and by blocking the vasoactive intestinal peptide (VIP) receptor with VIP 6-28. Parotid glands from rats with periodontitis showed an increase in cAMP levels that was also reverted in the presence of SQ 22536, FR 122047 and VIP 6-28. We concluded that both PGE(2) and VIP are produced in parotid glands from rats with periodontitis and, by activating their own receptors in acinar cells, induce cAMP accumulation leading to an increase in amylase basal secretion.
Assuntos
Amilases/biossíntese , AMP Cíclico/fisiologia , Glândula Parótida/enzimologia , Periodontite/enzimologia , Amilases/metabolismo , Animais , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/fisiologia , Isoproterenol/farmacologia , Masculino , Glândula Parótida/efeitos dos fármacos , Glândula Parótida/patologia , Periodontite/metabolismo , Periodontite/patologia , Ratos , Ratos WistarRESUMO
OBJECTIVE: Evaluate expression of MMP-13 during the course of two models experimentally induced periodontal disease in rats. DESIGN: Expression of MMP-13 at mRNA and protein levels was studied, respectively, by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. Two experimental models were used: LPS injections and ligature placement. 30mug of LPS from Eschericia coli was injected twice a week into the palatal aspect of upper molars. Ligatures were placed at the gingival margin around lower first molars. Controls received injections of PBS vehicle and no ligatures on lower molars. Samples were collected 5, 15 and 30 days after initiation of periodontal disease and processed for extraction of total RNA, total protein, and routinely processed for histology. RESULTS: Both experimental models produced a significant increase on the inflammatory infiltrate that paralleled elevated levels of MMP-13 mRNA and protein at 5 and 15 days. The LPS model was associated with a sustained level of inflammation and increased MMP-13 mRNA throughout the 30 days, whereas the ligature model showed a decrease on the severity of inflammation and MMP-13 mRNA at the 30-day period. Interestingly, MMP-13 protein levels were diametrically contrary to the mRNA levels. CONCLUSION: MMP-13 expression during LPS- and ligature-induced experimental periodontal disease follows the increase on severity of inflammation at the earliest periods. At 30 days, there is a decrease on the severity of inflammation on the ligature model associated with decreased MMP-13 mRNA. There is a lack of transcription-translation coupling of MMP-13 gene in both experimental models.
Assuntos
Metaloproteinase 13 da Matriz/análise , Doenças Periodontais/enzimologia , Animais , Western Blotting , Modelos Animais de Doenças , Epitélio/enzimologia , Epitélio/patologia , Escherichia coli , Regulação Enzimológica da Expressão Gênica/genética , Gengiva/lesões , Gengivite/enzimologia , Gengivite/etiologia , Gengivite/patologia , Ligadura/instrumentação , Lipopolissacarídeos/efeitos adversos , Masculino , Metaloproteinase 13 da Matriz/genética , Dente Molar , Doenças Periodontais/etiologia , Doenças Periodontais/patologia , Periodontite/enzimologia , Periodontite/etiologia , Periodontite/patologia , RNA Mensageiro/análise , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Transcrição Gênica/genéticaRESUMO
RATIONALE: Previous studies have used myeloperoxidase (MPO) as an inflammatory marker to estimate the accumulation of neutrophils in inflamed regions. OBJECTIVE: The aim of this experimental study was to quantify the levels of MPO related to experimental periodontal disease in rats. METHODS: Periodontal disease was induced in a group of rats using placement of a ligature around molar teeth. A group of rats without ligature placement served as a control. Measurements were made on the 3(rd), 7(th), 15(th) and 30(th) day from baseline. Gingival tissues were taken for quantification of MPO levels by ELISA. RESULTS: The rats with induced periodontal disease showed statistically higher MPO levels (p < 0.05) when compared to control rats. A significant increase in the levels of MPO released on days 7 and 30 was observed, with higher levels in the group with induced periodontitis. CONCLUSION: The levels of MPO were found to be higher in rats with induced periodontal disease, confirming the hypothesis that MPO may serve as an inflammatory marker for periodontitis.
Assuntos
Neutrófilos/imunologia , Periodontite/diagnóstico , Peroxidase/análise , Animais , Biomarcadores/análise , Modelos Animais de Doenças , Gengiva/enzimologia , Masculino , Neutrófilos/enzimologia , Periodontite/enzimologia , Peroxidase/imunologia , Ratos , Ratos WistarRESUMO
BACKGROUND AND OBJECTIVE: Periodontal disease is an inflammatory condition of tooth-supporting tissues. Arachidonic acid metabolites have been implicated in development of periodontal disease, especially those derived from the cyclo-oxygenase (COX) pathway. This study investigated the role of inhibitors of cyclo-oxygenases (COX-1 and COX-2) in a model of periodontal disease in rats. MATERIAL AND METHODS: A ligature was placed around the molar of rats. Losses of fiber attachment and of alveolar bone were measured morphometrically in histologically prepared sections. Infiltration of cells into gingival tissue surrounding the ligated tooth was also determined. RESULTS: Systemic and local administration of non-selective and selective COX-2 inhibitors, preventively, resulted in significant reduction of the losses of fiber attachment and alveolar bone, as well as decreased leukocyte numbers in gingival tissue. Preventive selective inhibition of COX-1 was as effective as COX-2 inhibition in reducing local fiber attachment loss and cell migration, but did not prevent alveolar bone loss. CONCLUSION: Our results provide evidence for participation of COX-1 and COX-2 in early stages of periodontal disease in rats. Furthermore, local administration of COX inhibitors reduced the signs of periodontal disease to the same extent as systemic treatment. Therapeutic approaches incorporating locally delivered anti-inflammatory drugs could be of benefit for patients suffering from periodontal disease.
Assuntos
Perda do Osso Alveolar/enzimologia , Inibidores de Ciclo-Oxigenase/farmacologia , Perda da Inserção Periodontal/enzimologia , Periodontite/enzimologia , Prostaglandina-Endoperóxido Sintases/metabolismo , Perda do Osso Alveolar/tratamento farmacológico , Animais , Ácido Araquidônico/metabolismo , Celecoxib , Inibidores de Ciclo-Oxigenase/uso terapêutico , Modelos Animais de Doenças , Indometacina/farmacologia , Masculino , Perda da Inserção Periodontal/tratamento farmacológico , Ligamento Periodontal/efeitos dos fármacos , Periodontite/tratamento farmacológico , Pirazóis/farmacologia , Ratos , Ratos Sprague-Dawley , Sulfonamidas/farmacologiaRESUMO
OBJECTIVE: Mucosal surfaces are the primary oral reservoirs of Candida species, but these species can also be found in subgingival biofilm. The present study investigated the genetic diversity and production of exoenzymes of C. albicans and C. dubliniensis isolated from the oral cavity of systemically healthy patients with periodontitis. DESIGN: Fifty-three patients were analysed. Samples were collected from three oral cavity sites (periodontal pocket, gingival sulci and oral mucosa), plated and, after isolation, suspect strains of C. albicans and C. dubliniensis were identified by PCR. The genetic diversity of the isolates was evaluated by RAPD and the activities of the secreted aspartyl proteinases and phospholipases were evaluated by the agar plate method. RESULTS: Twenty-one patients showed positive results for Candida spp. There were no statistically significant differences between genders, or between sites. C. albicans was the most frequently found specie, while C. dubliniensis was isolated from the periodontal pocket of only one patient. Sixteen genotypes were detected among the C. albicans isolates, and one among the C. dubliniensis isolates. The similarity coefficient (S(SM)) values among the C. albicans genotypes ranged from 0.684 to 1.0 with an average of 0.905+/-0.074. All isolates produced high levels of Saps and most of them produced high levels of phospholipases. No relationship was found between the genotypes and the pattern of enzymatic production. There was no association between specific genotypes and their site of isolation. CONCLUSIONS: The results of the present study suggest that genetically homogeneous strains of C. albicans are present in the oral cavity of patients with periodontitis and that these strains are capable of producing high levels of exoenzyme.
Assuntos
Candida/enzimologia , Candidíase Bucal/enzimologia , Boca/microbiologia , Periodontite/microbiologia , Biofilmes , Brasil , Candida/genética , Candidíase Bucal/microbiologia , Feminino , Variação Genética , Genótipo , Humanos , Masculino , Mucosa Bucal/enzimologia , Bolsa Periodontal/enzimologia , Bolsa Periodontal/microbiologia , Periodontite/enzimologiaRESUMO
BACKGROUND AND AIMS: Periodontitis is an infection with an episodic nature of tissue support destruction. The aim of this work was to determine the levels of chemokines, cytokines, matrix metalloproteinase-13, periodontal pathogens and inflammatory cells in periodontal sites characterized by active periodontal connective tissue destruction. MATERIAL AND METHOD: Fifty-six patients with moderate or advanced severity of chronic periodontitis were selected. Periodontitis was characterized by at least six sites with probing depth > or =5 mm, clinical attachment level > or =3 mm and radiographic bone loss. Periodontitis progression was determined by the tolerance method. Receptor activator for nuclear factor kappa B-ligand (RANK-L), monocyte chemoattractant protein-1 (MCP-1), tumour necrosis factor-alpha (TNF-alpha), IL-1beta, MMP-13, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsithia and inflammatory cells levels were determined. Statistical analysis was performed using the Stata 7.0 software. Data were expressed as mean+/-SD and paired samples t-test and chi(2) tests were used. RESULTS: Higher RANK-L, IL-1beta and MMP-13 activity levels were observed in active sites (p<0.05). The proportion of P. gingivalis, A. actinomycetemcomitans, T. forsythia and the number of CD4(+) T were higher in active than in inactive sites (p>0.05). CONCLUSION: The detection of periodontopathic bacteria, host matrix metalloproteinases and cytokines in periodontitis patients with lesions undergoing episodic attachment loss could partially explain the mechanisms associated with the destruction of the supporting tissues of the tooth.
Assuntos
Quimiocinas/análise , Citocinas/análise , Líquido do Sulco Gengival/química , Metaloproteinase 13 da Matriz/análise , Periodontite , Adulto , Quimiocina CCL2/análise , Doença Crônica , Placa Dentária/microbiologia , Métodos Epidemiológicos , Feminino , Gengiva/citologia , Gengiva/cirurgia , Líquido do Sulco Gengival/enzimologia , Humanos , Interleucina-1beta/análise , Masculino , Pessoa de Meia-Idade , Fragmentos de Peptídeos/análise , Periodontite/diagnóstico por imagem , Periodontite/enzimologia , Periodontite/microbiologia , Ligante RANK/análise , Radiografia , Fator de Necrose Tumoral alfa/análiseRESUMO
This present study evaluated the salivary arginase activity (SAA) in patients with chronic periodontitis and the effect of periodontal therapy on the activity of such enzyme. Thirty-six patients (mean age, 45.97 +/- 14.52), 18 chronic periodontitis subjects (test group), and 18 periodontally healthy individuals (control group) participated in the study. Clinical periodontal examinations included measurements of probing pocket depth (PD), clinical attachment level (CAL), plaque (PI), and gingival (GI) indexes. The test group received periodontal therapy according to individual needs. The saliva sample was collected from all study population at baseline (both groups) and 30 days after periodontal therapy (test group). SAA was determined by measuring the L: -ornithine formation from L-arginine and was expressed as mU/ml. The results showed that the mean values of SAA were statistically different between control and test groups. SAA was about 2.5 times higher in test than control groups. Thirty days after periodontal therapy, enzyme levels were 1.56 times lower than before periodontal therapy. We concluded that SAA is increased in chronic periodontitis subjects when compared to periodontally healthy individuals and that periodontal therapy significantly reduced SAA levels. It was suggested that in the near future, SAA may be used as a salivary marker of periodontal status.
Assuntos
Arginase/análise , Periodontite/terapia , Saliva/enzimologia , Doença Crônica , Índice de Placa Dentária , Raspagem Dentária , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Higiene Bucal , Ornitina/análise , Perda da Inserção Periodontal/enzimologia , Perda da Inserção Periodontal/terapia , Índice Periodontal , Bolsa Periodontal/enzimologia , Bolsa Periodontal/terapia , Periodontite/enzimologia , Periodonto/enzimologia , Aplainamento Radicular , Curetagem SubgengivalRESUMO
UNLABELLED: Matrix metalloproteinase (MMP)-13 is a collagenase involved in extracellular matrix degradation either by its direct degradative effects or by processing bioactive substrates. The aim of this study was to determine the levels of MMP-13 and tissue inhibitor of metalloproteinase (TIMP)-1 in gingival crevicular fluid (GCF) and gingival biopsies obtained from active and inactive sites during chronic periodontitis progression. MATERIALS AND METHODS: This was a longitudinal study in which chronic periodontitis patients with moderate to severe disease were included and followed until they developed progression determined by the tolerance method. GCF samples were obtained from periodontitis, active, inactive and healthy sites and additional gingival biopsies were taken from active and inactive sites. MMP-13 and TIMP-1 determinations were carried out by immunodot blots and immunowestern blots. RESULTS: In progressive periodontitis, MMP-13 and TIMP-1 remained unchanged between active and inactive sites, but as the TIMP-1 relative levels increased together with MMP-13 elevation in inactive samples, an inverse correlation was observed in active sites. Besides, MMP-13 was undetectable in healthy controls. CONCLUSION: Chronic periodontitis is characterized by increased MMP-13 expression. During disease progression, active sites tended to decrease TIMP-1 levels in association with MMP-13 elevation.
Assuntos
Metaloproteinase 13 da Matriz/análise , Periodontite/enzimologia , Inibidores de Proteases/análise , Inibidor Tecidual de Metaloproteinase-1/análise , Perda do Osso Alveolar/enzimologia , Biópsia , Western Blotting , Doença Crônica , Progressão da Doença , Feminino , Seguimentos , Gengiva/enzimologia , Líquido do Sulco Gengival/enzimologia , Humanos , Immunoblotting , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Perda da Inserção Periodontal/enzimologia , Bolsa Periodontal/enzimologiaRESUMO
OBJECTIVE: Periodontitis is a well-appreciated example of leukocyte-mediated bone loss and inflammation with pathogenic features similar to those observed in other inflammatory diseases, such as arthritis. Since Tacrolimus, is an immunomodulatory drug used for the treatment of some cases of arthritis, we hypothesized that it may modulate periodontal disease. DESIGN: Using a murine model of ligature-induced periodontal disease, we assessed the effects of daily administrations of Tacrolimus (1mg/kg body weight) on bone loss, enzymatic (myeloperoxidase) analysis, differential white blood cells counts, airpouch exudate and cytokine expression for 5-30 days. RESULTS: Radiographic, enzymatic (myeloperoxidase) and histological analysis revealed that Tacrolimus reduced the severity of periodontitis. More specifically, Tacrolimus suppressed the expression of serum interleukin (IL-1beta), tumour necrosis factor (TNF-alpha), IL-6, airpouch exudate PGE(2) and leukocytosis usually observed after the induction of periodontitis. Tacrolimus treatment in periodontitis-induced rats conferred protection against the inflammation-induced tissue and bone loss associated with periodontitis, through a mechanism involving IL-1beta, TNF-alpha and IL-6. CONCLUSIONS: The effects of Tacrolimus on periodontal disease pathogenesis may provide clues to a novel approach to host modulation therapy in destructive periodontal disease.