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Table of Contents   
ORIGINAL RESEARCH  
Year : 2013  |  Volume : 24  |  Issue : 2  |  Page : 164-167
Evaluation of total antioxidant capacity of saliva and serum in caries-free and caries-active adults: An in-vivo study


1 Department of Conservative Dentistry and Endodontics, A.B.S.M.I.D.S, Mangalore, Karnataka, India
2 Department of Oral and Maxillofacial Surgery, A.B.S.M.I.D.S, Mangalore, Karnataka, India
3 Department of Biochemistry, Central Research Laboratory, Nitte University, Mangalore, Karnataka, India

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Date of Submission25-May-2012
Date of Decision08-Nov-2012
Date of Acceptance14-Dec-2012
Date of Web Publication20-Aug-2013
 

   Abstract 

Context: Oxidative stress is defined as an imbalance between the production of reactive oxygen species and antioxidant defense systems which are widely recognized as an important feature of many diseases including dental caries, suggesting that total antioxidant status may be of importance in determining the progression of caries.
Aim: The aim of this study was to estimate the total antioxidant capacity (TAC) in saliva and serum of caries-free and caries-active adults and to correlate the TAC level with DMFT index (D = decayed, M = missing, F = filled, T = teeth).
Settings and Design: The present study included 100 healthy adults who were divided into four groups: Control, group I, group II, and group III, with a DMFT index 0, <3, <10, and >10, respectively.
Materials and Methods: Saliva and serum samples were collected from all the four groups. The TAC of saliva and serum was estimated by phosphomolybdenum method.
Statistical Analysis Used: One-way analysis of variance (ANOVA) was used to compare the TAC of saliva and serum. Only differences with "P" < 0.05 were considered statistically significant.
Results: The mean TAC level of both saliva and serum increased significantly (P < 0.001) with the DMFT index.
Conclusion: Total antioxidant capacity of saliva has a linear relation with caries, i.e., as the severity of caries increases, the TAC level also increases.

Keywords: Dental caries, saliva, serum, total antioxidant capacity

How to cite this article:
Hegde MN, Hegde ND, Ashok A, Shetty S. Evaluation of total antioxidant capacity of saliva and serum in caries-free and caries-active adults: An in-vivo study. Indian J Dent Res 2013;24:164-7

How to cite this URL:
Hegde MN, Hegde ND, Ashok A, Shetty S. Evaluation of total antioxidant capacity of saliva and serum in caries-free and caries-active adults: An in-vivo study. Indian J Dent Res [serial online] 2013 [cited 2019 Apr 24];24:164-7. Available from: http://www.ijdr.in/text.asp?2013/24/2/164/116670
Despite great achievements in oral health of populations globally, problems still remain in many communities all over the world, particularly among underprivileged groups in the developing countries. Dental caries have historically been considered the most important global oral health burdens. [1] It affects all people regardless of their sex, socioeconomic strata, race, and age. Dental caries is still a major oral health problem in most industrialized countries, affecting 60-90% of school children and a vast majority of adults. [1],[2] The etiology and pathogenesis of dental caries are known to be multifactorial. It is profoundly affected by other factors like oral hygiene and saliva. Changes in salivary components are in accordance with caries formation, and they may be used for recognizing risk in patients and to maintain prevention. [2],[3],[4],[5]

It has been long recognized that saliva serves as a mirror of the body's health as it contains proteins, hormones, antibodies, and other molecules that are frequently measured in standard blood tests to monitor health and disease. However, unlike whole blood, saliva is easy to collect, its collection is not painful to the patient, and it is less infectious for the health care provider. Healthy individuals produce about a liter and a quarter of saliva per day. [6],[7] Nearly all analytes that are in blood are also present in saliva. Although saliva has not been used as much up to this point as a sampling medium, it does have a strong potential, so it can be subjected to the same kinds of tests that are done currently in blood. [7]

Consequently, more intense salivary research can be observed in recent decades, which leads to a higher amount of scientific data presented by numerous researchers of this far-reaching field. Recently it has been claimed that the imbalance in levels of free radicals, reactive oxygen species, and antioxidants in saliva plays an important role in the onset and development of dental caries. [8]

Antioxidants are found in all biological species and they protect against the potentially harmful effects of processes or reactions that cause excessive oxidations. Therefore, biological antioxidants form an important part of our diet, and together with intracellular antioxidants and antioxidant enzyme systems, may prevent various pathological diseases. [9],[10] Antioxidants interact with free radicals and terminate the chain reaction. However, since free radicals/reactive oxygen species and antioxidant systems appear to act in concert rather than alone, investigations of individual antioxidant activity may be misleading and the measurement of any individual antioxidant may be less representative of the whole antioxidant status. Research is now being directed toward assays that evaluate the so-called "total antioxidant capacity" (TAC) of biological fluids. [11],[12],[13] Hence, evaluation of TAC in saliva can pave way in understanding the risk of individuals to dental caries. As for the literature available, very little has been discussed about dental caries and antioxidants in adults, and no data regarding saliva and serum total antioxidants and dental caries in adults are available. The aims of the present study were the following:

  • To estimate the total antioxidant levels in saliva and serum of healthy and diseased individuals
  • To correlate total antioxidant levels in saliva and serum with the prevalence of dental caries

   Materials and Methods Top


This study was conducted in the Central Research Laboratory of the university after getting approval from the institutional ethical committee.

Subjects

A total of 100 healthy adult patients belonging to South Canara region, coming to the OPD of Department of Conservative Dentistry and Endodontics, in the age group of 25-50 years were included in the study based on the following inclusion and exclusion criteria:

Inclusion criteria

  • Patients free from systemic or local disease which affects salivary secretions.
  • The caries status was assessed according to the WHO criteria.
Exclusion criteria

  • Patients with periodontal disease, hypertension, diabetes, and those on radiotherapy and chemotherapy.
  • Patients with systemic disease of the vital organs.
  • Those with history of long-term medications.
Procedure

One hundred healthy adults were selected for the study. Detailed case history of the patient was taken. A case history format was filled, with an informed consent which was duly signed by each patient (ethical committee protocol number: Cert. No. ABSM/EC/72/2009). The patients were grouped as Control, group I, group II, and group III, based on the DMFT index 0, <3, <10, and >10, respectively.

Calculation of DMFT

The smooth and occlusal surfaces of teeth were cleaned with soft bristle brush, dried, and examined. The DMFT score was calculated.

Grouping

The subjects were divided into three groups based on the DMFT score [Table 1].
Table 1: Selection of subjects

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Collection of saliva for salivary analysis

Resting saliva: Resting saliva was collected by allowing the patients to drool saliva into a collection cup.

Stimulated saliva: Stimulated saliva was collected by instructing the patients to chew on a piece of wax (to stimulate saliva). After 30 sec, they expectorated into spittoons. Chewing was continued for a further 5 min and the saliva was collected into the collection cup at regular intervals.

Sample collection for estimation of TAC

Five milliliters of saliva was collected from the patient, centrifuged, and the supernatant obtained was stored at 4°C for subsequent analysis. Also, 5 ml venous blood was collected from the patient, centrifuged, and the serum obtained was stored at 4°C for subsequent analysis.

TAC of saliva and serum by phosphomolybdenum method

This quantitative assay is based on the conversion of molybdenum (Mo VI) by reducing agents like antioxidants to molybdenum (Mo V), which further reacts with phosphate under acidic pH resulting in the formation of a green-colored complex, the intensity of which can be read spectrophotometrically at 695 nm. [14]

Statistical analysis

One-way analysis of variance (ANOVA) was used to correlate between total antioxidant level and dental caries between the control and the three study groups. A "P " value of 0.05 or less was considered significant. Results are presented as mean ± standard deviation (X ± SD).


   Results Top


The mean total antioxidant level in saliva was 0.34 ± 0.13 μM/L in the control group. It was found that total antioxidant levels in the study groups increased with caries prevalence. The mean total antioxidant levels in groups I, II, and III were 0.44 ± 0.13, 0.56 ± 0.26, and 0.59 ± 0.18 μM/L, respectively, and the P value was statistically significant (P < 0.05) [Table 2], [Figure 1].
Figure 1: Mean total antioxidant capacity of saliva in control and study groups

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Table 2: Comparison of total antioxidant level in saliva and serum

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Similar results were obtained in the total antioxidant levels of serum. The total antioxidant levels in the study groups I, II, and III were 0.85 ± 0.21, 1.01 ± 0.20, and 1.03 ± 0.23 μM/L, respectively, and that of control group was 0.76 ± 0.12 μM/L, and it was statistically significant (P < 0.05) [Table 2], [Figure 2].
Figure 2: Mean total antioxidant capacity of serum in control and study groups

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   Discussion Top


Good oral hygiene may contribute to salivary composition as an oxidative stress-decreasing factor in dental diseases. Limited data are available about the effect of oral hygiene on salivary parameters in subjects with caries. Dental caries is a multifactorial disease. [15] Diet, host (saliva and tooth), bacteria, time, and personal factors (oral hygiene) are responsible for the development of dental caries. The first line of defense against dental caries is saliva. The composition and physiology of saliva warrants thorough investigation because it clearly influences oral health. [16]

Cells and biological fluids have an array of protective antioxidant mechanisms, both for preventing the production of free radicals and for repairing oxidative damage. Free radicals are produced during dental decay. The number appears to vary directly with caries activity. Studies have been performed to correlate the total antioxidant level of saliva in children with caries. The authors reported that the TAC increased with increase in caries activity. [10],[12] However, the correlation of total antioxidant levels in saliva and serum in adults with dental caries has not been reported.

In this study, we evaluated the TAC of saliva and serum as it is suggested that free radical/reactive oxygen species and antioxidant system appear to act in concert rather than alone, and measurement of any individual antioxidant may be less representative of the whole antioxidant status. [13]

It was observed in the present study that the total antioxidant level in saliva and serum increased with caries activity. Similar results were obtained in our previous study where we had compared the total antioxidant levels in caries-free and caries-active groups. [17] The increase in TAC could be explained by the fact that the levels of antioxidants could be altered in response to infection or disease. The presence of caries in this study group could be one of the factors for the increased levels of TAC of saliva. An important factor for the increase in TAC of saliva could also be triggered function of salivary peroxidase system which constitutes one of the major salivary antioxidant systems. [13] Salivary peroxidase brings about the control of oral bacteria that form dental plaque, to imbalances in the ecology, and which lead to dental caries. Salivary peroxidase catalyzes the peroxidation of thiocyanate ion (SCN−) to generate more stable oxidation products (OSCN−). These products inturn arrests the growth and metabolism of microorganisms, thereby inhibiting caries or at least slowing down the progress of caries. [18]

Increase in the TAC level can also be attributed to the diet. The sum of endogenous and food-derived antioxidants represents the total antioxidant activity of the system. Uric acid is the major antioxidant present in human saliva, accounting for more than approximately 85% of the TAC of both unstimulated and stimulated saliva, and is mainly derived from the diet. [19] Caries-promoting sugars such as sucrose, glucose, and fructose, present in fruit juices and other sugar-sweetened drinks, are readily metabolized by Streptococcus mutans and lactobacilli to organic acids that demineralize enamel and dentin. One of the striking features of fructose is its ability to produce uric acid. Serum uric acid can increase rapidly after ingestion of fructose, and it has been found in studies in which fructose (or sucrose) diets have been administered that even fasting uric acid levels will increase after several weeks. [20] This might increase the serum uric acid concentration, thereby increasing salivary uric acid concentration, as to a large extent serum composition is reflected in saliva composition, leading to the increase in the TAC of saliva. [21] Thus, the habit of increased sugar intake might be responsible for the increase in uric acid level and TAC, at least in part, being a confounding factor both for the development of caries and increase in uric acid level. Further studies regarding role of uric acid, its contribution in TAC of saliva, and its association with diet are required to better understand this phenomenon. [22]


   Conclusion Top


Dental caries is one of the common diseases in children as well as in adults. Saliva is one of the important factors that influence the development of caries. Oxidative stress which occurs as a result of an imbalance between free radical/reactive oxygen species and antioxidant system has been implicated as one of the important contributory etiologic factors in many of the oral inflammatory pathologies, and dental caries is no exception.

From our results, it can be concluded that the TAC of saliva has a linear relation with caries, i.e., as the severity of caries increases, the TAC level also increases. But in order to extrapolate these findings, studies using larger sample size are needed.

 
   References Top

1.Kathariya R, Pradeep AR. Salivary proteomic biomarkers for oral diseases: A review of literature. Arch Oral Sci Res 2010;1:43-9.  Back to cited text no. 1
    
2.Shafer WG, Hine MK, Levy BM. ED. A Text Book of Oral Pathology. 5 th ed. Philadelphia W. B.: Saunders Company; 1993. p. 567-658.  Back to cited text no. 2
    
3.Lavelle LB. Christopher applied oral physiology, 2 nd ed. United Kingdom: Butterworths and Co (publishers) LTD; 1988. p. 128-41.  Back to cited text no. 3
    
4.Ahmadi Motamayel F, Davoodi P, Dalband M. Hendi SS. Saliva as a Mirror of the Body Health. Dent J Hamadan Univ Med Sci 2010;1:1-15.  Back to cited text no. 4
    
5.Lumikari Lenander M, Loimaranta V. Saliva and dental caries. Adv Dent Res 2000;14:40-474.  Back to cited text no. 5
    
6.Amerongen AV, Bolscher JG, Veerman EC. Salivary proteins: Protective and diagnostic value in cariology. Caries Res 2004;38:247-53.  Back to cited text no. 6
    
7.John T, Mc Devitt. Saliva as the next best diagnostic tool. J Biochem 2006;45:23-5.  Back to cited text no. 7
    
8.Gopinath VK, Arzreanne. Saliva as a diagnostic tool for assessment of dental caries. Arch Orofac Sc 2006;1:57-9.  Back to cited text no. 8
    
9.Battino M, Ferreior MS, Gallardo I, Newman HN, Bullon P. The antioxidant capacity of saliva. J Clin Periodontol 2002;29:189-94.  Back to cited text no. 9
    
10.Tulunoglu O, Demirtas S, Tulunoglu I. Total antioxidant levels of saliva in children related to caries, age and gender. Int J Paed Dent 2006;16:186-19.  Back to cited text no. 10
    
11.Gopinath VK, Arzreanne. Saliva as a diagnostic tool for assessment of dental caries. Arch Orofac Sci 2006;1:57-9.  Back to cited text no. 11
    
12.Uberos J, Alarcon JA, Penalver MA. Influence of the antioxidant content of saliva on dental caries in an at-risk community. Br Dent J 2008;E5:205.  Back to cited text no. 12
    
13.Battino M, Ferreior MS, Gallardo I, Newman HN, Bullon P. The antioxidant capacity of saliva. J Clin Periodontol 2002;29:189-94.  Back to cited text no. 13
    
14.Prieto, Manuel P, Miguel A. Spectrophotometric quantitation of antioxidant capacity through the formation of phosphomolybdenum complex: Specific application to the determination of vitamin E. Anal Biochem 1999;259:337-41.  Back to cited text no. 14
    
15.Tenovuo J. Salivary parameters of relevance for assessing caries activity in individuals and populations. Community Dent Oral Epidemiol 1997;25:82-6.  Back to cited text no. 15
[PUBMED]    
16.John T, Mc Devitt. Saliva as the next best diagnostic tool. J Biochem 2006;45:23-5.  Back to cited text no. 16
    
17.Hegde MN, Kumari S, Hegde N, Moany A. Correlation between total antioxidant level and dental caries in adults-an in vivo study Res J Pharm Biol Chem Sci 2011;2:864-70.   Back to cited text no. 17
    
18.Preethi BP, Pyati A, Dodawad R. Evaluation of flow rate, ph, buffering capacity, calcium, total protein and total antioxidant levels of saliva in caries free and caries active children: An in vivo study. Biomed Res 2010;21:289-29.  Back to cited text no. 18
    
19.Moore S, Calder KAC, Miller NJ, Rice-Evans CA. Antioxidant activity of saliva and periodontal disease. Free Radic Res 1994;21:417-25.  Back to cited text no. 19
    
20.Perheentupa J, Raivio K. Fructose-induced hyperuricaemia. Lancet 1967;2:528-31.  Back to cited text no. 20
[PUBMED]    
21.Reiser S, Powell AS, Scholfield DJ, Panda P, Ellwood KC, Canary JJ. Blood lipids, lipoproteins, apoproteins, and uric acid in men fed diets containing fructose or high-amylose cornstarch. Am J Clin Nutr 1989;49:832-9.  Back to cited text no. 21
[PUBMED]    
22.Kumar D, Pandey KR, Agarval D, Agarval D. An estimation and evaluation of total antioxidant capacity of saliva in children with severe early childhood caries. Int J Paed Dent 2011;21:459-64.  Back to cited text no. 22
    

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Correspondence Address:
Mithra N Hegde
Department of Conservative Dentistry and Endodontics, A.B.S.M.I.D.S, Mangalore, Karnataka
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0970-9290.116670

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