Salivary Superoxide Dismutase Levels Research Articles

Salivary Oxidative Stress Assessment in Tobacco Users with and without Potentially Malignant Disorders and Micronuclei Estimation using Fluorescent Microscopy

Abstract Background: Consumption of tobacco in either smoking or nonsmoking form produces free radicals, contributing to an increased risk of oral cancer as a result of oxidative stress. Biochemical changes in antioxidant enzymes are responsible for tobacco-associated oral carcinogenesis. Due to the production of reactive oxygen species and oxidative stress, the promotion of carcinogenesis occurs in the cells. Oral potentially malignant disorders (OPMDs) is an assembly of benign lesions with morphologically altered clinical or histopathological tissue that has a greater than normal risk of transforming into malignant lesions after diagnosis at a later date. Micronuclei are the goal site for early genotoxic events and are involved in carcinogenic events. Micronuclei in exfoliated cells are useful biomarkers in assessing cytogenetic damage. Early detection will help to devise appropriate treatment plans, thereby improving patient survival rates. Aim and Objectives: To estimate the salivary superoxide dismutase (SOD) levels and the expression of micronuclei in exfoliated buccal cells of tobacco users before the onset of tobacco-induced oral lesions and correlating the same with patients with OPMDs to determine if this can be useful in predicting early malignant potential in subjects with habitual tobacco usage. Materials and Methods: The study included three groups. Group I – Tobacco users with OPMD (n = 50), Group II – Tobacco users without OPMD (n = 50), and Group III – Healthy controls (n = 50). Sandwich enzyme-linked immunosorbent assay was done for saliva samples for quantification of salivary SOD. Exfoliated buccal cells were smeared and stained with acridine orange stain and viewed under a fluorescent microscope for micronuclei estimation. Results: The study showed a significant difference in salivary SOD levels and increased micronuclei expression among tobacco users than healthy controls. Conclusion: Our study results revealed a gradual and steady decrease of SOD levels and increased micronuclei expression from healthy control to tobacco users without lesions and tobacco users with lesion, and this can be used as an effective noninvasive early diagnostic biomarker in assessing malignant progression in tobacco users. Furthermore, the evaluation of micronuclei estimation can serve as a simple, quick, and noninvasive chair-side procedure for regular oral cancer screening in patients with tobacco usage.

Evaluation of Salivary Superoxide Dismutase Levels in Oral Submucous Fibrosis – A Cross Sectional Study

Background: Oral Submucous Fibrosis (OSMF) is a potentially malignant disorder with areca nut being the main etiological agent. Increased risk of malignant potential of OSMF necessitates the early diagnosis of the condition. Superoxide dismutase (SOD) scavenges oxygenated free radicals and can be used as a potential biomarker for oral cancer detection.
 Objectives: The present study was conducted to estimate and compare salivary SOD levels in patients with OSMF and healthy controls and to compare its levels in different clinical and functional stages of OSMF.
 Methodology: The study group (Group 1) included 30 patients of OSMF, and the control group (Group 2) was formed by age and sex-matched healthy individuals. Salivary SOD levels were determined by Nitro Blue Tetrazolium (NBT) chloride method. The levels of salivary SOD were compared between the two groups and the data thus obtained was subjected to statistical analysis using independent sample t-test and one way Analysis of variance (ANOVA) test.
 Results: The mean salivary SOD in the study group was 0.583 IU/mL and 0.864 IU/mL in control group and the difference was found to be statistically significant. Thus, the patients with OSMF had significantly lower levels of salivary SOD compared to the controls.
 Conclusion: The decrease in the salivary SOD levels in patients with OSMF suggests the role of oxidative stress in the pathogenesis of OSMF and initiation and progression of carcinogenesis. Thus, it can be used as a prognostic marker for early detection of carcinoma in patients with OSMF and thereby, improving the quality of life patients.

Dental Pulp Mesenchymal Stem Cells as a Treatment for Periodontal Disease in Older Adults.

Periodontal disease (PD) is one of the main causes of tooth loss and is related to oxidative stress and chronic inflammation. Although different treatments have been proposed in the past, the vast majority do not regenerate lost tissues. In this sense, the use of dental pulp mesenchymal stem cells (DPMSCs) seems to be an alternative for the regeneration of periodontal bone tissue. A quasi-experimental study was conducted in a sample of 22 adults between 55 and 64 years of age with PD, without uncontrolled systemic chronic diseases. Two groups were formed randomly: (i) experimental group (EG) n = 11, with a treatment based on DPMSCs; and a (ii) control group (CG) n = 11, without a treatment of DPMSCs. Every participant underwent clinical and radiological evaluations and measurement of bone mineral density (BMD) by tomography. Saliva samples were taken as well, to determine the total concentration of antioxidants, superoxide dismutase (SOD), lipoperoxides, and interleukins (IL), before and 6 months after treatment. All subjects underwent curettage and periodontal surgery, the EG had a collagen scaffold treated with DPMSCs, while the CG only had the collagen scaffold placed. The EG with DPMSCs showed an increase in the BMD of the alveolar bone with a borderline statistical significance (baseline 638.82 ± 181.7 vs. posttreatment 781.26 ± 162.2 HU, p = 0.09). Regarding oxidative stress and inflammation markers, salivary SOD levels were significantly higher in EG (baseline 1.49 ± 0.96 vs. 2.14 ± 1.12 U/L posttreatment, p < 0.05) meanwhile IL1β levels had a decrease (baseline 1001.91 ± 675.5vs. posttreatment 722.3 ± 349.4 pg/ml, p < 0.05). Our findings suggest that a DPMSCs treatment based on DPMSCs has both an effect on bone regeneration linked to an increased SOD and decreased levels of IL1β in aging subjects with PD.

186 - Dental Pulp Mesenchymal Stem Cells for Treatment of Periodontal Disease and its Relationship with Oxidative Stress Markers

Background Periodontal disease (PD) is a chronic disease of the tooth supporting tissues, characterized by gingival bleeding, periodontal pocket formation, destruction of connective tissue attachment and alveolar bone loss. In this sense, it has been pointed out that oxidative stress (OxS) is involving in the pathogenesis of PD. A therapeutic option to regenerate tissues for loss due to PD is the placement of mesenchymal stem cells derived from tooth pulp (MSC). MSCs generate a protective environment against OxS, in vitro, where respiratory burst is regulated downwards, thereby protecting the surrounding tissues from this damage. Experimental reports in animal models suggest that MSCs have antioxidant potential and, consequently, the ability to reduce OxS. In light of these findings, a decrease in oxidative damage in periodontal tissues can be expected after application of dental pulp MSC. Objective To determine the effect of Dental Pulp MSC on OxS and PD. Methods It was carried out a cases series of 10 patients with PD. Clinical measurements and saliva samples were collected before and after 3 and 6-months of Dental Pulp MSC application. Thiobarbituric acid reactive substances (TBARS), superoxide dismutase (SOD) and total antioxidant capacity (TAC) were spectrophotometrically assayed in saliva. Results Application of Dental Pulp MSC showed a decreased in mean of clinical parameters at 6 months (4.6±0.9 to 3.8±0.8 mm). It was also found a statistically significant decrease in TBARS at 6 months (0.0826±0.018 to 0.0381±0.042 μmol/L). Salivary SOD levels were significantly higher at 3 months (1.500±0.777 to 2.754±1.32 U/l) (p>0.05). Conclusions Our findings suggest that the treatment with DPMSC for periodontal disease has effect antioxidant. Trial registry: ISRCTN 12831118

Dietary supplementation with low-dose omega-3 fatty acids reduces salivary tumor necrosis factor-α levels in patients with chronic periodontitis: a randomized controlled clinical study.

Recent studies have demonstrated the beneficial effects of omega-3 polyunsaturated fatty acids (PUFAs) on physiological processes and on a variety of chronic inflammatory diseases, including periodontal diseases. In this study, we evaluated the impact of omega-3 PUFAs in conjunction with scaling and root planing (SRP) on salivary markers in patients with chronic periodontitis. Thirty systemically healthy subjects with chronic periodontitis were enrolled and randomly allocated into two groups. The control group (n = 15) was treated with SRP + placebo whereas the test group was treated with SRP and dietary supplementation of low-dose omega-3 PUFAs (6.25 mg eicosapentaenoic acid and 19.19 mg docosahexaenoic acid). Clinical parameters were taken at baseline, 1, 3 and 6 mo following therapy. Saliva samples were obtained at the same time intervals and analyzed for tumor necrosis factor-α (TNF-α) and superoxide dismutase (SOD). Both groups showed significant changes in clinical parameters in response to treatment compared to baseline with no significant difference between groups. Salivary TNF-α levels showed a statistically significant decrease in the test group at 6 mo compared to the control group. Salivary SOD levels increased significantly at 3 and 6 mo in the test group and at 6 mo in placebo groups compared to baseline with no statistically significant differences between the groups. The results demonstrated that dietary supplementation with low-dose omega-3 PUFAs improves salivary TNF-α without any significant impact on clinical parameters in patients with chronic periodontitis, suggesting that the systemic benefits of dietary omega-3 PUFAs may not be translated to periodontal health. (ClinicalTrials.gov ID NCT02719587).

Status of Serum and Salivary Levels of Superoxide Dismutase in Type 2 Diabetes Mellitus with Oral Manifestations: A Case Control Study.

Type 2 Diabetes Mellitus is a very well known metabolic disorder that has reached epidemic proportions worldwide. Evidence suggests that oxidative stress increases in Diabetes Mellitus because of the excessive production of reactive oxygen species and an impaired antioxidant defence mechanism. This study estimated, compared and correlated the serum and salivary Superoxide dismutase levels in healthy subjects and subjects with Type 2 Diabetes Mellitus having oral manifestations. In this study, serum and salivary Superoxide dismutase levels were estimated in 45 healthy subjects and 45 patients with Type 2 Diabetes Mellitus with oral manifestations. The mean serum and salivary Superoxide dismutase levels were significantly decreased in subjects with Type 2 Diabetes Mellitus with oral manifestations compared to the healthy subjects. Positive correlation in both healthy and diabetics was observed between serum and salivary samples. This study highlights that the abnormally high levels of oxidative stress in diabetics coupled with simultaneous decline in antioxidant defence mechanism results in complications in Diabetes mellitus. Thus exploring saliva for antioxidant markers that accurately reflect the redox status of the body is worthwhile.

The influence of cigarette smoking on blood and salivary super oxide dismutase enzyme levels among smokers and nonsmokers—A cross sectional study

To determine the influence of smoking on blood and salivary superoxide dismutase enzyme levels among smokers, and to demonstrate the significant alterations in the levels of superoxide dismutase in association with patient age, periodontal disease status, smoking duration, and smoking frequency. This study also aimed to evaluate the use of saliva as a biological fluid for disease diagnosis.Ninety males aged 25–56 years were selected and included 30 smokers, 30 nonsmokers with chronic periodontitis, and 30 healthy controls. Clinical parameters such as the gingival index, pocket depth, and clinical attachment loss were recorded. Blood and saliva samples were collected and superoxide dismutase enzyme levels were analyzed using spectrophotometric assay.Superoxide dismutase enzyme levels in the blood and saliva were significantly higher in smokers than in nonsmokers and the controls (p < 0.05). A significant correlation existed between superoxide dismutase levels and clinical parameters. There was also a significant positive correlation between blood and salivary superoxide dismutase levels among the three groups.Systemic and local antioxidant status is affected by periodontal disease and by the impact of smoking. The increased blood and salivary superoxide dismutase enzyme levels in smokers may be an adaptive defense mechanism to counteract the increased reactive oxygen species production induced by smoking. This study emphasizes the importance of saliva as an easy noninvasive tool in diagnosing patients who are more prone to precancerous lesions and conditions, and its importance in patient education and motivation programs for smoking cessation.

Impact of iron deficiency anemia on chronic periodontitis and superoxide dismutase activity: a cross-sectional study.

Both chronic periodontitis (CP) and iron deficiency anemia (IDA) induce oxidative stress in the body and cause an imbalance between reactive oxygen species and antioxidants, such as superoxide dismutase (SOD). This study explored the SOD enzyme activity of saliva and serum in CP patients with and without IDA and analyzed the impact of IDA on CP. A total of 82 patients were divided into four groups: control group (CG, 22), periodontally healthy IDA patients (IDA-PH, 20), CP patients (CP, 20), and IDA patients with CP (IDA-CP, 20). After clinical measurements and samplings, serum and salivary SOD levels were determined using an SOD assay kit. IDA-CP patients exhibited a higher gingival index, bleeding on probing, probing pocket depth, and percentage (%) of sites with a clinical attachment loss (CAL) of ≥6 mm (P<0.008) than CP patients. The mean salivary and serum SOD levels were significantly lower in the IDA-PH, CP, and IDA-CP patients than in the CG group (P<0.008). A significant positive correlation between salivary and serum SOD activity was observed in IDA (P<0.05). Furthermore, serum and salivary SOD levels were significantly and negatively correlated with all periodontal parameters including the percentage of sites with CAL of 4-5 and ≥6 mm (P<0.05) except the significant correlation between salivary SOD activity and mean CAL and the percentage of sites with CAL of 4-5 mm (P>0.05) in these patients. Within the limits of this study, it may be suggested that IDA patients with chronic periodontitis have more periodontal breakdowns than patients with chronic periodontitis. Serum and salivary SOD activity levels were lower in the IDA-PH, CP and IDA-CP groups than in the CG. Iron deficiency anemia influenced the serum SOD activity but did not seem to affect the salivary SOD activity in these patients.

Salivary superoxide dismutase levels in oral leukoplakia and oral squamous cell carcinoma; a clinicopathological study

In the South-East Asian countries oral cancer is one among the commonly occurring malignancies due to tobacco use and quid chewing habits. Early stage diagnosis of the disease is reported to have a better prognosis. Sialochemistry studies involving antioxidants have recently gained importance as biomarkers for oral cancer risk assessment. The aim of this study was to estimate the levels of superoxide dismutase in saliva of patients with oral cancer, oral leukoplakia and healthy controls. Three study groups comprising of 25 oral cancer patients, 25 oral leukoplakia patients and 25 healthy controls were involved in the study. Saliva sample collected from the patients were evaluated superoxide dismutase by nitroblue tetrazolium (NBT) chloride method. The data obtained were analyzed using the one way ANOVA test. There was a significant difference between the levels of salivary superoxide dismutase in between oral cancer, oral leukoplakia groups when compared to healthy controls. The levels of salivary superoxide dismutase were significantly lower in oral cancer when compared to oral leukoplakia. These results suggest that salivary superoxide dismutase level estimation could be used to determine the risk of oral cancer as a non-invasive modality.

Influence of cigarette smoking on blood and salivary super oxide dismutase levels among smokers and non‐smokers

To determine the influence of smoking on the blood and salivary superoxide dismutase levels among smokers, and to demonstrate the significance in alterations in the levels of superoxide dismutase in association with patient age, periodontal disease status, and duration and smoking frequency. Sixty healthy male patients (aged 20-60 years) constituted the target population. Blood and saliva samples were collected, and the periodontal index was measured using a Community Periodontal Index probe. Blood and salivary superoxide dismutase levels were calculated with the reagents prepared, and values were measured in a UV spectrophotometer. There was a significant decrease in mean superoxide dismutase levels with an increase in age and smoking duration and frequency (0.23-0.05 U/mL). There was a range of 0.31-0.06 U/mL variation in the mean superoxide dismutase levels as the periodontal disease condition deteriorated. There was a significant reduction in the superoxide dismutase levels in the blood and saliva of smokers with increased duration and frequency of smoking, and as the periodontal disease condition worsened. Thus, this study emphasizes the importance of saliva as an easy, non-invasive procedure in the diagnosis of patients who are more prone to precancerous lesions and conditions, and its importance in patient education and motivation programs for smoking cessation.