RESUMO
OBJECTIVE: This study aims to evaluate the clinical and biochemical (oxidative stress and pro-inflammatory mediators) effects of the gaseous ozone use accompanied by scaling and root planning (SRP) in periodontal treatment. MATERIAL AND METHODS: The study population consisted of 40 patients with chronic periodontitis (CP) randomly sorted into two groups of 20. The experimental group received SRP plus 3 watts gaseous ozone in two separate applications five days apart, whereas the control group received SRP plus placebo. Clinical periodontal parameters were assayed and saliva samples were taken before the initial and one month after the second treatment. Periodontal examination assessed plaque index (PI), gingival index (GI), probing depth, and clinical attachment level (CAL). Total antioxidant status (TAS), total oxidant status (TOS), nitric oxide (NO), 8-hydroxy-2'-deoxyguanosine (8-OHdG), myeloperoxidase (MPO), glutathione (GSH), malondialdehyde (MDA), and transforming growth factor-beta (TGF-ß) levels were evaluated from saliva samples. RESULTS: Changes following treatment in PI, GI, probing depth, and CAL scores were similar for both groups (p>0.05). Of note, TGF-ß levels were observed to be higher in the treatment group than in controls (p<0.05). Changes in 8-OHdG, TAS, TOS, NO, MPO, GSH and MDA levels, however, were not significantly different between groups (p>0.05). CONCLUSION: The findings of this study indicate that SRP plus gaseous ozone versus SRP alone does not correlate to a significant improvement in periodontal recovery.
Assuntos
Periodontite Crônica/terapia , Oxidantes Fotoquímicos/uso terapêutico , Ozônio/uso terapêutico , Aplainamento Radicular/métodos , 8-Hidroxi-2'-Desoxiguanosina , Adulto , Antioxidantes/análise , Periodontite Crônica/patologia , Índice de Placa Dentária , Desoxiguanosina/análogos & derivados , Desoxiguanosina/análise , Ensaio de Imunoadsorção Enzimática , Feminino , Glutationa/análise , Humanos , Masculino , Malondialdeído/análise , Pessoa de Meia-Idade , Óxido Nítrico/análise , Oxidantes/antagonistas & inibidores , Índice Periodontal , Peroxidase/análise , Reprodutibilidade dos Testes , Saliva/química , Estatísticas não Paramétricas , Fatores de Tempo , Fator de Crescimento Transformador beta/análise , Resultado do TratamentoRESUMO
Abstract Objective: This study aims to evaluate the clinical and biochemical (oxidative stress and pro-inflammatory mediators) effects of the gaseous ozone use accompanied by scaling and root planning (SRP) in periodontal treatment. Material and Methods: The study population consisted of 40 patients with chronic periodontitis (CP) randomly sorted into two groups of 20. The experimental group received SRP plus 3 watts gaseous ozone in two separate applications five days apart, whereas the control group received SRP plus placebo. Clinical periodontal parameters were assayed and saliva samples were taken before the initial and one month after the second treatment. Periodontal examination assessed plaque index (PI), gingival index (GI), probing depth, and clinical attachment level (CAL). Total antioxidant status (TAS), total oxidant status (TOS), nitric oxide (NO), 8-hydroxy-2'-deoxyguanosine (8-OHdG), myeloperoxidase (MPO), glutathione (GSH), malondialdehyde (MDA), and transforming growth factor-beta (TGF-β) levels were evaluated from saliva samples. Results: Changes following treatment in PI, GI, probing depth, and CAL scores were similar for both groups (p>0.05). Of note, TGF-β levels were observed to be higher in the treatment group than in controls (p<0.05). Changes in 8-OHdG, TAS, TOS, NO, MPO, GSH and MDA levels, however, were not significantly different between groups (p>0.05). Conclusion: The findings of this study indicate that SRP plus gaseous ozone versus SRP alone does not correlate to a significant improvement in periodontal recovery.
Assuntos
Humanos , Masculino , Feminino , Adulto , Oxidantes Fotoquímicos/uso terapêutico , Ozônio/uso terapêutico , Aplainamento Radicular/métodos , Periodontite Crônica/terapia , Saliva/química , Fatores de Tempo , Ensaio de Imunoadsorção Enzimática , Índice Periodontal , Índice de Placa Dentária , Reprodutibilidade dos Testes , Fator de Crescimento Transformador beta/análise , Resultado do Tratamento , Oxidantes/antagonistas & inibidores , Peroxidase/análise , Estatísticas não Paramétricas , Desoxiguanosina/análise , Desoxiguanosina/análogos & derivados , Periodontite Crônica/patologia , Glutationa/análise , Malondialdeído/análise , Pessoa de Meia-Idade , Óxido Nítrico/análise , Antioxidantes/análiseRESUMO
Crack cocaine is a very toxic product derived from cocaine. The aim of this study was to evaluate genetic damage in multiple organs of rats following acute exposure to crack cocaine. A total of 20 Wistar rats were distributed into four groups (n = 5), as follows: 0, 4.5, 9, and 18 mg/kg body weight (b.w.) of crack cocaine administered by intraperitoneal route (i.p.). All animals were killed 24 h after intraperitoneal (i.p.) injection. The results showed that crack cocaine increased the number of micronucleated cells in bone marrow cells exposed to 18 mg/kg crack cocaine (p < 0.05). Peripheral blood and liver cells presented genetic damage as depicted by single cell gel (comet) assay at 9 and 18 mg/kg doses (p < 0.05). Immunohistochemistry data revealed significant increase in 8-hydroxy-20-deoxyguanosine (8-OHdG) immunoexpression in hepatocytes of animals exposed to crack cocaine at 9 and 18 mg/kg (p < 0.05) when compared with negative controls. Taken together, our results demonstrate that crack cocaine is able to induce genomic damage in multiple organs of Wistar rats.
Assuntos
Cocaína Crack/toxicidade , 8-Hidroxi-2'-Desoxiguanosina , Animais , Células da Medula Óssea/efeitos dos fármacos , Desoxiguanosina/análogos & derivados , Desoxiguanosina/análise , Imuno-Histoquímica , Ratos , Ratos WistarRESUMO
BACKGROUND: 8-Hydroxy-2'-deoxyguanosine (8-OHdG) and human neutrophil elastase/α1-proteinase inhibitor (HNE/α1-PI) complex have been regarded as reliable biomarkers of oxidative stress in inflammatory conditions. This study investigates whether the salivary levels of these two analytes may be linked with periodontal health status. METHODS: One hundred ten patients with chronic periodontitis (CP) and 50 healthy controls were selected. Periodontal status was assessed by criteria based on probing depth, clinical attachment level, and extent and severity of periodontal breakdown. 8-OHdG and HNE/α1-PI salivary levels were analyzed by enzyme-linked immunosorbent assay. The association of these analytes with CP was analyzed individually and adjusted for confounding factors using a multivariate binary logistic regression model. RESULTS: Significantly higher levels of both markers were detected in the CP group in comparison to controls. Weak-to-moderate positive significant correlations between salivary biomarkers and clinical parameters were observed. After binary logistic regression analysis, salivary levels of 8-OHdG >17.35 ng/mL and HNE/α1-PI complex >158.28 ng/mL were independently associated with disease status. Interaction effects among candidate prognostic variables were also noted. CONCLUSIONS: Increased salivary levels of 8-OHdG and HNE/α1-PI complex may be strong, independent prognostic indicators of the amount and extent of oxidative stress-induced periodontal breakdown. In addition, unstimulated whole saliva samples might reflect a synergistic biologic interactive effect of HNE/α1-PI associated with the aging and smoking cumulative characteristics of periodontal damage.
Assuntos
Periodontite Crônica/diagnóstico , Desoxiguanosina/análogos & derivados , Elastase de Leucócito/análise , 8-Hidroxi-2'-Desoxiguanosina , Biomarcadores/análise , Estudos de Casos e Controles , Desoxiguanosina/análise , Humanos , Estresse Oxidativo , Prognóstico , Saliva/químicaRESUMO
Burn injuries (BIs) result in both local and systemic responses distant from the site of thermal injury, such as skeletal muscle. The purpose of this study was to investigate the expression of cyclooxygenase-2 (COX-2) and hydroxy-2'-deoxyguanosine (8-OHdG) as a result of inflammation and reactive oxygen species production, respectively. A total of 16 male rats were distributed into two groups: control (C) and submitted to BI. The medial part of gastrocnemius muscle formed the specimens, which were stained with hematoxylin and eosin and were evaluated. COX-2 and 8-OHdG expressions were assessed by immunohistochemistry, and cell profile area and density of muscle fibers (number of fibers per square millimeter) were evaluated by morphometric methods. The results revealed inflammatory infiltrate associated with COX-2 immunoexpression in BI-gastrocnemius muscle. Furthermore, a substantial decrease in the muscle cell profile area of BI group was noticed when compared with the control group, whereas the density of muscle fibers was higher in the BI group. 8-OHdG expression in numerous skeletal muscle nuclei was detected in the BI group. In conclusion, the BI group is able to induce skeletal muscle degeneration as a result of systemic host response closely related to reactive oxygen species production and inflammatory process.
Assuntos
Ciclo-Oxigenase 2/análise , Desoxiguanosina/análogos & derivados , Inflamação/metabolismo , Músculo Esquelético/metabolismo , Estresse Oxidativo , 8-Hidroxi-2'-Desoxiguanosina , Animais , Desoxiguanosina/análise , Modelos Animais de Doenças , Imuno-Histoquímica , Ratos , Ratos WistarRESUMO
Aim of this study was to investigate the cytotoxic and genotoxic properties of inorganic and organic mercury compounds, i.e., HgCl(2) and methylmercury (MeHg). In addition, the DNA-protective and antioxidant effects of the flavonoid quercetin (QC) were studied. All experiments were conducted with human-derived liver cells (HepG2), which possess antioxidant and drug-metabolizing enzymes in an inducible form. 8-Hydroxydeoxyguanosine (8-OHdG) and comet formation were monitored as endpoints of DNA damage. The impact of the metal compounds on the redox status was also investigated, since it is assumed that their toxic effects are due to oxidative damage. A number of biochemical parameters related to oxidative stress, namely glutathione, malondialdehyde, protein carbonyl and formation of reactive oxygen species (ROS) were measured after treatment of the cells with the mercury compounds in the presence and absence of quercetin. To elucidate the mechanisms that underlie the effects of QC, three protocols (pre-, simultaneous and post-treatment) were used. Both mercury compounds (range 0.1-5.0µM) caused induction of DNA migration and formation of 8-OHdG. In combination with the flavonoid (range 0.1-5.0µM), DNA-protective effects of QC were observed after pre- and simultaneous treatment but not when the flavonoid was added after treatment with the metal compounds. Exposure to the metal compounds led also to substantial changes of all parameters of the redox status and co-treatment experiments with QC showed that these alterations are reversed by the flavonoid. Taken together, the results of our experiments indicate that these two mercury compounds cause DNA damage and oxidative stress in human-derived liver cells and that the flavonoid reduces these effects. Since the concentrations of the metals and of the flavonoids used in the present work reflect human exposure, our findings can be taken as an indication that QC may protect humans against the adverse effects caused by the metal.
Assuntos
Antimutagênicos/farmacologia , Antioxidantes/farmacologia , Dano ao DNA/efeitos dos fármacos , Cloreto de Mercúrio/toxicidade , Compostos de Metilmercúrio/toxicidade , Oxirredução/efeitos dos fármacos , Quercetina/farmacologia , 8-Hidroxi-2'-Desoxiguanosina , Sobrevivência Celular , Desoxiguanosina/análogos & derivados , Desoxiguanosina/análise , Células Hep G2 , Humanos , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismoRESUMO
Exocyclic DNA adducts produced by exogenous and endogenous compounds are emerging as potential tools to study a variety of human diseases and air pollution exposure. A highly sensitive method involving online reverse-phase high performance liquid chromatography with electrospray tandem mass spectrometry detection in the multiple reaction monitoring mode and employing stable isotope-labeled internal standards was developed for the simultaneous quantification of 1,N(2)-etheno-2'-deoxyguanosine (1,N(2)-epsilondGuo) and 1,N(2)-propano-2'-deoxyguanosine (1,N(2)-propanodGuo) in DNA. This methodology permits direct online quantification of 2'-deoxyguanosine and ca. 500 amol of adducts in 100 microg of hydrolyzed DNA in the same analysis. Using the newly developed technique, accurate determinations of 1,N(2)-etheno-2'-deoxyguanosine and 1,N(2)-propano-2'-deoxyguanosine levels in DNA extracts of human cultured cells (4.01 +/- 0.32 1,N(2)-epsilondGuo/10(8) dGuo and 3.43 +/- 0.33 1,N(2)-propanodGuo/10(8) dGuo) and rat tissue (liver, 2.47 +/- 0.61 1,N(2)-epsilondGuo/10(8) dGuo and 4.61 +/- 0.69 1,N(2)-propanodGuo/10(8) dGuo; brain, 2.96 +/- 1.43 1,N(2)-epsilondGuo/10(8) dGuo and 5.66 +/- 3.70 1,N(2)-propanodGuo/10(8) dGuo; and lung, 0.87 +/- 0.34 1,N(2)-epsilondGuo/10(8) dGuo and 2.25 +/- 1.72 1,N(2)-propanodGuo/10(8) dGuo) were performed. The method described herein can be used to study the biological significance of exocyclic DNA adducts through the quantification of different adducts in humans and experimental animals with pathological conditions and after air pollution exposure.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Adutos de DNA/análise , DNA/química , Desoxiadenosinas/análise , Desoxiguanosina/análogos & derivados , Espectrometria de Massas por Ionização por Electrospray/métodos , Animais , Bovinos , Linhagem Celular , Desoxiguanosina/análise , Humanos , Masculino , Ratos , Ratos Wistar , Espectrometria de Massas em TandemRESUMO
Highly purified rat-liver nuclei were previously shown to have nuclear ethanol (EtOH) metabolizing system able to bioactivate alcohol to acetaldehyde and 1-hydroxyethyl radicals. These reactive metabolites were able to covalently bind to nuclear proteins and lipids potentially being able to provoke oxidative stress of nuclear components. In this study, the above-mentioned possibility was explored. Sprague Dawley male rats (125-150 g) were fed a standard Lieber and De Carli liquid diet for 28 days. Controls were pair-fed with a diet, in which EtOH was isocalorically replaced with carbohydrate. The presence of a chlorzoxazone hydroxylase activity inducible by the repetitive EtOH drinking further suggested the presence of CYP2E1 in the highly purified nuclei. Nuclei from EtOH-drinking rats evidenced significantly increased susceptibility to a t-butyl hydroperoxide challenge as detected by chemiluminescence emission, increased formation of protein carbonyls, and decreased content of protein sulfhydryls. In contrast, no significant changes in the nuclear lipid hydroperoxides formation or even decreases in the 8-oxo-7,8-dihydro-2-deoxyguanosine were observed. No significant differences were observed in different parameters of the alkaline Comet assay. In immunohistochemical studies performed, no expression of p53 was observed in the livers of the animals under the experimental conditions tested. Since nuclear proteins and lipids are known to play a role in cell growth, differentiation, repair and signaling, their alterations by either oxidative stress, or by covalent binding might be of relevance to liver tumor promotion.
Assuntos
Núcleo Celular/metabolismo , Etanol/administração & dosagem , Fígado/metabolismo , Estresse Oxidativo/efeitos dos fármacos , 8-Hidroxi-2'-Desoxiguanosina , Animais , Clorzoxazona/análogos & derivados , Clorzoxazona/metabolismo , Ensaio Cometa , Citocromo P-450 CYP2E1/metabolismo , Interpretação Estatística de Dados , Desoxiguanosina/análogos & derivados , Desoxiguanosina/análise , Hepatócitos/citologia , Hepatócitos/metabolismo , Imuno-Histoquímica , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/química , Fígado/citologia , Masculino , Microssomos Hepáticos/química , Microssomos Hepáticos/metabolismo , Oxigenases de Função Mista/metabolismo , Carbonilação Proteica/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Compostos de Sulfidrila/metabolismo , Proteína Supressora de Tumor p53 , terc-Butil Hidroperóxido/metabolismoRESUMO
A ruthenium oxide hexacyanoferrate (RuOHCF) modified electrode was developed. Hydrodynamic voltammetry was employed to demonstrate the remarkable electrocatalytic activity toward the oxidation of 2'-deoxyguanosine. The RuOHCF modified electrode was used as amperometric detector for 2'-deoxyguanosine determination in a FIA apparatus. The influence of various experimental conditions was explored for optimum analytical performance, and at these experimental conditions, the method exhibited a linear response range to 2'-deoxyguanosine extending from 3.8 to 252 micromol L(-1) with detection limit of 94 nmol L(-1). Applications in DNA samples were examined, and the results for determination of 2'-deoxyguanosine were in good agreement with those obtained by HPLC analysis. Studies on the kinetics of the in vitro consumption of 2'-deoxyguanosine by acetaldehyde were also performed.
Assuntos
Dano ao DNA , DNA/química , Desoxiguanosina/análise , Ferrocianetos/química , Análise de Injeção de Fluxo/métodos , Potenciometria/métodos , Compostos de Rutênio/química , Catálise , Eletrodos , Análise de Injeção de Fluxo/instrumentação , Potenciometria/instrumentaçãoRESUMO
The present study was undertaken to identify whether inflammation or oxidative stress is the primary abnormality in the kidney in spontaneously hypertensive rats (SHR). Renal inflammation and oxidative stress were evaluated in 2- and 3-week-old prehypertensive SHR and age-matched genetically normotensive control Wistar-Kyoto (WKY) rats. Blood pressure was similar in WKY and SHR rats at 2 and 3 weeks, of age. Renal inflammation (macrophage and nuclear factor-kappaB) was elevated in SHR at 3 weeks, but not at 2 weeks, of age compared with age-matched WKY rats. Renal oxidative stress (nitrotyrosine, 8-hydroxy-2'-deoxyguanosine and p47phox) was also clearly elevated in 3-week-old SHR compared with age-matched WKY rats. Additionally, NADPH oxidase subunit p47phox was found elevated in 2-week-old SHR compared to age-matched WKY rats. Moreover, antioxidant (N-acetyl-L-cysteine and Tempol) treatment reduced renal inflammation in prehypertensive SHR. We therefore conclude that the oxidative stress appears before inflammation as a primary abnormality in the kidney in prehypertensive SHR.