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Table of Contents   
ORIGINAL RESEARCH  
Year : 2014  |  Volume : 25  |  Issue : 5  |  Page : 613-616
Comparative evaluation of serum superoxide dismutase and glutathione levels in periodontally diseased patients: An interventional study


1 Departments of Periodontics, A B Shetty Institute of Dental Sciences, Mangalore, Karnataka, India
2 Department of Oral and Maxillofacial Surgery, A B Shetty Institute of Dental Sciences, Mangalore, Karnataka, India
3 Department of Biochemistry, K.S. Hegde Academy of Medical Sciences, Derlakatte, Mangalore, Karnataka, India

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Date of Submission09-Jan-2014
Date of Decision10-Mar-2014
Date of Acceptance16-Sep-2014
Date of Web Publication16-Dec-2014
 

   Abstract 

Background: Periodontal disease is an immune-inflammatory disease characterized by connective tissue breakdown, loss of attachment, and alveolar bone resorption. Under normal physiological conditions, a dynamic equilibrium is maintained between the reactive oxygen species (ROS) and antioxidant defense capacity. Oxidative stress occurs when this equilibrium shifts in favor of ROS. Oxidative stress is thought to play a causative role in the pathogenesis of periodontal diseases. Aim: The present study was designed to estimate and compare the superoxide dismutase (SOD) and glutathione (GSH) levels in the serum of periodontitis, gingivitis, and healthy individuals before and after nonsurgical periodontal therapy. Materials and Methods: The present study was conducted in the Department of Periodontics, A. B. Shetty Memorial Institute of Dental Sciences, Deralakatte, Mangalore. The study was designed as a single blinded interventional study comprising 75 subjects, inclusive of both sexes and divided into three groups of 25 patients each. Patients were categorized into chronic periodontitis, gingivitis, and healthy. The severity of inflammation was assessed using gingival index and pocket probing depth. Biochemical analysis was done to estimate the SOD and GSH levels before and after nonsurgical periodontal therapy. Results obtained were then statistically analyzed using ANOVA test and paired t-test. Results: The results showed a higher level of serum SOD and GSH in the healthy group compared to the other groups. The difference was found to be statistically significant (P < 0.0001). The post-treatment levels of SOD were statistically higher than the pre-treatment levels in periodontitis and gingivitis group.

Keywords: Antioxidant defense, glutathione, interventional study, periodontitis, superoxide dismutase

How to cite this article:
Biju T, Shabeer M M, Amitha R, Rajendra B P, Suchetha K. Comparative evaluation of serum superoxide dismutase and glutathione levels in periodontally diseased patients: An interventional study. Indian J Dent Res 2014;25:613-6

How to cite this URL:
Biju T, Shabeer M M, Amitha R, Rajendra B P, Suchetha K. Comparative evaluation of serum superoxide dismutase and glutathione levels in periodontally diseased patients: An interventional study. Indian J Dent Res [serial online] 2014 [cited 2019 Nov 22];25:613-6. Available from: http://www.ijdr.in/text.asp?2014/25/5/613/147105
Periodontitis is an immunoinflammatory disease initiated by the plaque biofilm that results in loss of periodontal attachment to the root surface and adjacent alveolar bone, which ultimately lead to tooth loss. Plaque biofilm is the main etiological factor for periodontal diseases. Neutrophils, which are considered as the first line of defense, produce free radicals. These respond to the plaque biofilm by formation of several reactive oxygen species (ROS) and lipid peroxidation products that destroy the microorganisms. This could also lead to periodontal tissue destruction caused by an inappropriate host response to the plaque biofilm. [1] The ROS are required in physiological quantities by the human body, but it has been well established that over-production of ROS occurs at sites of chronic inflammation. The human body contains an array of antioxidant defense mechanisms including both nonenzymatic and enzymatic antioxidants to counter this excessive production of harmful ROS as soon as they are formed and to prevent their deleterious effects. The enzymatic antioxidants include superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSHPx) while the nonenzymatic antioxidants include Vitamins E and C, and reduced glutathione (GSH). [1] Thus, the present study was designed to estimate and compare the SOD and GSH levels in the serum of periodontitis, gingivitis, and healthy individuals before and after nonsurgical periodontal therapy.


   Materials and methods Top


The subjects were selected from the Department of Periodontics A. B. Shetty Memorial Institute of Dental Sciences, Deralakatte, Mangalore, India. Ethical clearance was taken from the Institutional Ethical Committee before commencing the study, and written consent was taken from each patient prior to sample collection. A total of 75 patients was divided into three groups. Namely Group A consisting 25 subjects with chronic periodontitis, Group B consisting 25 subjects with chronic gingivitis, and Group C (Control Group) with a total of 25 periodontally healthy subjects. The patient's medical history, dental history, Gingival index (as given by Loe and Silness 1967), probing depth, and loss of attachment were recorded with Williams periodontal probe prior to the study. The subjects were categorized according to the following criteria: All subjects were systemically healthy, with no medical condition that would affect their participation in the study with a minimum complement of 20 teeth. Controls were periodontally healthy. Gingivitis was defined as patients with bleeding on probing in more than 30% of sites and with a mean Loe and Sillness gingival index score ≥1. Periodontitis was defined as the presence of at least 30% of sites with clinical attachment loss ≥5 mm measured with a Williams periodontal probe. The subjects with a history of any antibiotic or anti-inflammatory therapy within 2 months prior to study, pregnant, pre-eclamptic or lactating women, subjects who received periodontal therapy in the past 3 months, smokers and subjects taking vitamin supplements were excluded from the study. All measurements and readings were done before the collection of blood samples. Group B and Group C patients underwent nonsurgical periodontal therapy and were followed-up after 3 weeks for respective blood parameters.


   Results Top


In the present study, the SOD levels in subjects of periodontitis group (P < 0.001) and gingivitis group (P < 0.001) was found to be significantly lower than in the control group [Table 1], Graph 1]. The study also found a significant increase (P ≤ 0.001) in the post-treatment serum levels of SOD in periodontitis groups and gingivitis group (P ≤ 0.001) [Table 2], Graph 2].

The results of this study demonstrated lower levels of GSH in serum of patients with periodontitis and gingivitis as compared to control group. The results were found to be statistically significant (P ≤ 0.001) among the groups [Table 1], Graph 3]. The post-therapy values were found to be significant in the periodontitis group (P = 0.003) and not in the gingivitis group (P = 0.935) [Table 3], Graph 4].










   Discussion Top


Inflammatory and immune reactions to microbial plaque are the predominant features of gingivitis and periodontitis. There are reports demonstrating the ability of perio-pathogens and their products to induce the generation of ROS by polymorphonuclear leukocytes (PMNs). Immune responses to microorganisms will mainly be directed against outer membrane proteins and polysaccharides of bacteria and extracellularly released enzymes and toxins. One of the host defense mechanism involves the production of powerful oxidizing agents which is characterized by a rapid increase in the uptake of oxygen. This phenomenon is known as the respiratory burst and results in the production of reactive oxidants such as superoxide, hydrogen peroxide, and hydroxyl radicals. Chronic exposure to ROS can initiate a broad spectrum of pathologic reactions in the adjacent tissues. [2],[3] Thus, normal tissue destruction is part of the typical inflammatory response. The prevention of lipid peroxidation is an essential process in all the aerobic organisms, as lipid peroxidation products can cause DNA damage. Increased lipid peroxidation and decreased antioxidant protection frequently occurs.

Thus, the present study was aimed to evaluate the significance of SOD and GSH as an antioxidant in serum by estimating their levels in patients with chronic periodontitis, gingivitis, and periodontally healthy individuals. The interventional part of the study was designed to estimate the levels of serum SOD and GSH along with the periodontal parameters following nonsurgical periodontal therapy.

Intracellular SOD is the most prominent antioxidant in mammalian tissue. Several authors have proven the significance of extracellular SOD in plasma and other body fluids. SOD activity in periodontal health and disease in serum and saliva has been evaluated with inconclusive results. Within the periodontal tissues, SOD has been immune-localized predominantly in the periodontal ligament, in association with collagen fibrils and fibroblasts. [4] In the present study, the SOD levels in subjects of periodontitis group (P < 0.001) and gingivitis group (P < 0.001) was found to be significantly lower than the control group [Table 1], Graph 1]. This is in accordance with the study where Ellis et al. [5] saw a decrease in the SOD activity with an increase in probing depth. Decreased SOD activity and increased number of leukocytes were considered to lead to an increase in the ROS level, resulting in tissue breakdown.

The study also found a significant increase (P ≤ 0.001) in the post-treatment serum levels of SOD in periodontitis groups and gingivitis group (P ≤ 0.001) [Table 2], Graph 2]. Our results are in accordance with previous studies which demonstrated a lower antioxidant activity in saliva of chronic periodontitis patients compared to periodontally healthy subjects. [6],[7],[8] Depressed levels of serum antioxidant levels were recorded in chronic periodontitis patients in comparison to periodontally healthy subjects by various authors. [9],[10],[11],[12] The alteration in serum antioxidant capacity can be explained by the fact that chronic periodontal disease is associated with peripheral neutrophils that are hyper-reactive and are responsible for the production of ROS in response to Fc gamma-receptor stimulation. [1]

Glutathione peroxidase is a selenium-containing peroxidase, which catalyze the reduction of a variety of hydroperoxides (ROOH and H 2 O 2 ) using GSH, thereby protecting mammalian cells against oxidative damage. Although GSHPx shares the substrate, H 2 O 2 , with CAT, it alone can react effectively with lipid and other organic hydroperoxides. The GSH redox cycle is a major source of protection against low levels of oxidant stress, whereas CAT becomes more significant in protecting against severe oxidant stress. Hence, level of GSH is expected to be inversely related to periodontal disease severity.

The results of this study demonstrated lower levels of GSH in serum of patients with periodontitis and gingivitis as compared to control group. The results were found to be statistically significant (P ≤ 0.001) among the groups [Table 1], Graph 3]. The post-therapy values were found to be significant in the periodontitis group (P = 0.003) and not in the gingivitis group (P = 0.935) [Table 3], Graph 4]. This is in accordance with earlier studies. [1],[8],[13] This could be explained by a reduction in the bacterial load and the varied degree of destruction among the groups which warrants the difference seen.
Table 1: Comparison of serum SOD (U/mgHb) and GSH (mg/L) levels among the three groups: One - way ANOVA with Tukey post-hoc test


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Table 2: Comparison of the pre and the post - serum SOD (U/mgHb) values in gingivitis and periodontitis using paired t-test


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Table 3: Comparison of the pre and the post - serum GSH values (mg/L) in gingivitis and periodontitis using paired t-test


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Thus, in health there is a delicate balance between ROS and the antioxidant defense mechanism of tissue destruction which if tipped in favor of tissue damage could lead to significant attachment loss. Novel small molecular weight compounds that function as SOD mimetics may offer reliable benefits due to the catalytic properties that could permit enzymatic detoxification. Misaki et al. in an intervention study done on rats demonstrated a curative effect of SOD on Porphyromonas gingivalis-induced inflammation and periodontal wound healing. [14]

However, it is only as data becomes available from other long-term interventional trials done on humans that we will see whether these compounds live up to their potential for treatment of periodontal diseases.


   Conclusion Top


Serum levels of SOD and GSH are assessed to be decreased in the periodontally compromised groups (Group A and B) compared to the control group. The post-treatment serum levels of SOD and GSH showed a significant increase compared to the pre-treatment levels in the periodontitis group. The post-treatment serum levels showed a significant increase in SOD in the gingivitis group. As periodontitis is a multifactorial disease, there are various factors that account for the breakdown of periodontal tissues.

The reduction in the serum levels of SOD and GSH in gingivitis and periodontitis patients has led to an imbalance leading to increased ROS and hence periodontal breakdown. Thus, focus should be shifted to the development of antioxidant-based approaches to periodontal therapy. Further, long-term studies need to be carried out on the efficacy of antioxidant therapies that target the free radicals that lead to periodontal tissue breakdown.


   Acknowledgment Top


Mr. Vishak, Ms.Harshini - Lab technicians.

 
   References Top

1.
Chapple IL, Milward MR, Dietrich T. The prevalence of inflammatory periodontitis is negatively associated with serum antioxidant concentrations. J Nutr 2007;137:657-64.  Back to cited text no. 1
    
2.
Shapira L, Gordon B, Warbington M, Van Dyke TE. Priming effect of Porphyromonas gingivalis lipopolysaccharide on superoxide production by neutrophils from healthy and rapidly progressive periodontitis subjects. J Periodontol 1994;65:129-33.  Back to cited text no. 2
    
3.
Miyasaki KT. The neutrophil: Mechanisms of controlling periodontal bacteria. J Periodontol 1991;62:761-74.  Back to cited text no. 3
    
4.
Jacoby BH, Davis WL. The electron microscopic immunolocalization of a copper-zinc superoxide dismutase in association with collagen fibers of periodontal soft tissues. J Periodontol 1991;62:413-20.  Back to cited text no. 4
    
5.
Ellis SD, Tucci MA, Serio FG, Johnson RB. Factors for progression of periodontal diseases. J Oral Pathol Med 1998;27:101-5.  Back to cited text no. 5
    
6.
Kim SC, Kim OS, Kim OJ, Kim YJ, Chung HJ. Antioxidant profile of whole saliva after scaling and root planing in periodontal disease. J Periodontal Implant Sci 2010;40:164-71.  Back to cited text no. 6
    
7.
Singh N, Chander Narula S, Kumar Sharma R, Tewari S, Kumar Sehgal P. Vitamin E supplementation, superoxide dismutase status, and outcome of scaling and root planing in patients with chronic periodontitis: A randomized clinical trial. J Periodontol 2014;85:242-9.  Back to cited text no. 7
    
8.
Novakovic N, Todorovic T, Rakic M, Milinkovic I, Dozic I, Jankovic S, et al. Salivary antioxidants as periodontal biomarkers in evaluation of tissue status and treatment outcome. J Periodontal Res 2014;49:129-36.  Back to cited text no. 8
    
9.
Babior BM. Phagocytes and oxidative stress. Am J Med 2000;109:33-44.  Back to cited text no. 9
    
10.
Ower PC, Ciantar M, Newman HN, Wilson M, Bulman JS. The effects on chronic periodontitis of a subgingivally-placed redox agent in a slow release device. J Clin Periodontol 1995;22:494-500.  Back to cited text no. 10
    
11.
Chapple IL, Matthews JB. The role of reactive oxygen and antioxidant species in periodontal tissue destruction. Periodontol 2000 2007;43:160-232.  Back to cited text no. 11
    
12.
Czapski G, Aronovitch J, Samuni A, Godinger D, Chevion M. The sensitization of the toxicity of superoxide and vitamin C by copper and iron: A site-specific mechanism. In: Oxyradicals and their Scavenger Systems. Molecular Aspects. Vol. 1. New York: Elsevier; 1983. p. 111-5.  Back to cited text no. 12
    
13.
Chapple IL. Reactive oxygen species and antioxidants in inflammatory diseases. J Clin Periodontol 1997;24:287-96.  Back to cited text no. 13
    
14.
Misaki H, Suzuki M, Yoshie H, Hara K. The effect of superoxide dismutase on the inflammation induced by periodontal pathogenic bacteria and wound healing of gingival incision. Nihon Shishubyo Gakkai Kaishi 1990;32:93-110.  Back to cited text no. 14
    

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Correspondence Address:
M Madani Shabeer
Departments of Periodontics, A B Shetty Institute of Dental Sciences, Mangalore, Karnataka
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0970-9290.147105

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    Tables

  [Table 1], [Table 2], [Table 3]

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