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Table of Contents   
ORIGINAL ARTICLE  
Year : 2022  |  Volume : 33  |  Issue : 1  |  Page : 41-45
Evaluation of the effect of nonsurgical periodontal therapy on malondialdehyde and 8-hydroxy deoxyguanosine levels in chronic periodontitis


1 Department of Periodontology, Faculty of Dental Sciences, Institute of Medical Sciences, BHU, Uttar Pradesh, India
2 Department of Conservative Dentistry and Endodontics, Faculty of Dental Sciences, Institute of Medical Sciences, BHU, Uttar Pradesh, India
3 Department of Biochemistry, Faculty of Dental Sciences, Institute of Medical Sciences, BHU, Uttar Pradesh, India
4 Department of Community Medicine, Division of Biostatistics, Faculty of Dental Sciences, Institute of Medical Sciences, BHU, Uttar Pradesh, India
5 Oral Pathology and Microbiology, Faculty of Dental Sciences, Institute of Medical Sciences, BHU, Uttar Pradesh, India

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Date of Submission15-Sep-2021
Date of Decision08-Jan-2022
Date of Acceptance19-Apr-2022
Date of Web Publication09-Aug-2022
 

   Abstract 


Background: Reactive oxygen species released on stimulation by periodontal pathogens cause oxidation of biomolecules and play significant role in periodontal disease pathogenesis. Aim: This study aimed to evaluate the levels of oxidative by-products malondialdehyde (MDA) and 8-hydroxy deoxyguanosine (8-OHdG) as biomarkers in chronic periodontitis patients compared to healthy as well as before and after nonsurgical periodontal therapy. The correlation between biomarkers and clinical attachment level was also evaluated. Settings and Design: A total of 112 subjects were included in this study. The subjects were divided into two groups (Group I included 56 healthy subjects and Group II constituted 56 chronic periodontitis patients) on the basis of clinical periodontal parameters. Materials and Methods: Group I subjects received no treatment and were evaluated once only for clinical and oxidative stress biomarker parameters. Nonsurgical periodontal therapy was carried out in Group II patients and they were evaluated at baseline and 3 months after therapy. Results: Both salivary and serum levels of MDA and 8-OHdG were found to be increased in chronic periodontitis patients as compared to healthy subjects. After nonsurgical periodontal therapy, the levels of MDA and 8-OHdG significantly reduced. Linear correlation between clinical attachment level and oxidative stress parameters was found to be positive and highly significant. Conclusion: It can be concluded that periodontal therapy is effective in improving the oxidative stress condition.

Keywords: Biomarkers, DNA oxidation, lipid peroxidation, oxidative stress, periodontitis

How to cite this article:
Gautam A, Mittal N, Mishra SP, Singh TB, Chandra A. Evaluation of the effect of nonsurgical periodontal therapy on malondialdehyde and 8-hydroxy deoxyguanosine levels in chronic periodontitis. Indian J Dent Res 2022;33:41-5

How to cite this URL:
Gautam A, Mittal N, Mishra SP, Singh TB, Chandra A. Evaluation of the effect of nonsurgical periodontal therapy on malondialdehyde and 8-hydroxy deoxyguanosine levels in chronic periodontitis. Indian J Dent Res [serial online] 2022 [cited 2022 Oct 4];33:41-5. Available from: https://www.ijdr.in/text.asp?2022/33/1/41/353546



   Introduction Top


Periodontitis is an inflammatory disease primarily initiated by bacterial infection and subsequently modified by anomalous host response that leads to loss of periodontal attachment, alveolar bone, and ultimately teeth. These anomalous host responses may be in various forms, one of them is oxidative stress. Reactive oxygen species (ROS) are produced in physiological quantities thus body contains antioxidants that remove ROS as they formed. Antioxidants prevent the harmful effects of ROS and repair damage caused by them, maintaining a dynamic equilibrium, but if it shifts in favor of ROS either by increase in ROS activity or production or by diminished antioxidant defence, it leads to oxidative stress.[1]

ROS play a fundamental role in physiologic processes, however, also play a central role in progression of inflammatory diseases.[2] These are generated during normal cellular metabolism as by-products, but overproduction of ROS occurs at sites of inflammation. ROS have been implicated in tissue damage, either directly by protein damage, lipid peroxidation, DNA damage, and oxidation of important enzymes or by stimulation of pro-inflammatory cytokine release.[3] Various studies have been conducted to assess the impact of oxidative stress in periodontal disease pathogenesis.[4],[5],[6],[7],[8] However, the results of studies are not consistent, as some demonstrate positive correlation[9],[10],[11]but others have failed to prove.[5],[6],[12] Given the importance of oxidative stress in pathogenesis and contradictory results, this study aimed to evaluate salivary and serum level of lipid (malondialdehyde – MDA) and DNA by-products (8-hydroxy deoxyguanosine – 8-OHdG) before and after nonsurgical periodontal therapy.


   Materials and Methods Top


This study was carried out on 112 participants, including 56 healthy subjects and 56 patients, with chronic periodontitis who were recruited for the study following the inclusion and exclusion criteria. The study design was approved by institutional ethical committee. A written informed consent was obtained from each study participant after explaining the procedure. The ethical committee approval was obtained on 25/09/2019 (Dean/2019/EC/1503).

The participants included in the study were having age group between 30 and 55 years, minimum of 20 teeth and not having any systemic diseases. The participants were excluded if they underwent any periodontal therapy within the past 6 months or received any medication, such as antibiotics/antioxidant multivitamins (vitamin E and vitamin C) in the past 6 months.

The participants selected for this study were categorized into two groups on the basis of clinical parameters, such as plaque index (PI), probing pocket depth (PPD), gingival index (GI), and clinical attachment level (CAL). Group I consisted of 56 periodontally healthy subjects having no gingival inflammation and with good oral hygiene and Group II consisted of 56 chronic generalized periodontitis patients having 30% or more teeth with ≥5 mm probing depth.

Saliva and blood serum were obtained from study participants of both the groups. Biochemical analyses of oxidative stress biomarkers (MDA and 8-OHdG) were carried out once in Group I and twice (at baseline and 3 months after nonsurgical periodontal therapy) in Group II participants.

Saliva collection

Whole unstimulated saliva was collected after the routine check-up of the subjects, between 9.00 a.m. and 10.00 a.m. following an overnight fasting. The subjects were instructed to rinse their mouth thoroughly with water and be seated straight in a chair and asked to pool saliva in the bottom of the mouth over a period of 5 min and drained out into a collection tube. The collected saliva samples were immediately centrifuged at 3000 rpm for 10 min to eliminate cell debris and then supernatant was stored at –80°C until the biochemical analysis.

Blood collection

Under aseptic conditions, 5 ml of venous blood was collected from ante-cubital fossa in plain vaccutainer. The collected blood was allowed to clot for 1 hour at room temperature and serum was separated then centrifuged at 3000 rpm for 10 min and the supernatant was stored at –80°C until the biochemical analysis.

Periodontal therapy

The subjects of Group I received no treatment and evaluated once only for clinical and oxidative stress biomarkers. Whereas the subjects of Group II received nonsurgical periodontal therapy, including full mouth scaling, root planning (SRP), and oral hygiene instructions. The oral hygiene instruction included brushing twice daily with correct brushing technique. Patients were not given any antioxidants or vitamin supplements and no modification were added to the diet of the study subjects. The patients were followed up for 3 months post-treatment and evaluated for clinical and oxidative stress biomarkers at baseline and 3 months after therapy.

Statistical analysis

The collected data were subjected to statistical analysis through SPSS. The statistical methods used were independent 't' test to compare between two groups and paired 't' test to compare the effect of nonsurgical periodontal therapy, before and after treatment. The correlations between CAL and biomarkers levels were evaluated using the Pearson's correlation coefficient. P value < 0.05 was considered as statistically significant.


   Results Top


Analysis of clinical periodontal parameters

Comparison of clinical periodontal parameters of Group I and Group II before and 3 months post-treatment are shown in [Table 1]. All the periodontal parameters were significantly improved after nonsurgical periodontal therapy.
Table 1: Comparison of periodontal clinical parameters

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Analysis of oxidative stress parameters

[Table 2] shows the values of saliva and serum level of MDA and 8-OHdG of Group I and Group II before and 3 months after nonsurgical periodontal therapy. It can be observed that the level of oxidative stress biomarkers in both saliva and serum was significantly high in chronic periodontitis patients as compared to healthy subjects. Following nonsurgical periodontal therapy, the levels of MDA and 8-OHdG significantly reduced in both the fluids.
Table 2: Comparison of oxidative stress parameters

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Correlation

The correlation was assessed between CAL and biomarkers of oxidative stress in chronic periodontitis patients before therapy and shown in [Table 3].
Table 3: Clinical attachment level (CAL) and biomarkers of oxidative stress correlation

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The linear relationships between CAL and salivary MDA and 8-OHdG having 'r' values of 0.813 and 0.770, respectively, which showed positive and highly significant correlation (P < 0.001). The linear relationship between the CAL and serum MDA and 8-OHdG having 'r' value of 0.714 and 0.761, respectively, also showed positive and highly significant correlation (P < 0.001).


   Discussion Top


Host response to periodontal pathogens and their products includes the generation of ROS by polymorphonuclear leukocytes. It has been widely accepted that ROS and oxidative stress play an important role in pathogenesis of periodontal disease.[5] It has been a great challenge in clinical periodontics to find biomarker of periodontitis with high specificity for early diagnosis and better treatment outcome assessment. For this reason, various biomarkers of oxidative stress have been studied.[6],[7],[8],[9],[13] These biomarkers can be assessed either by directly measuring ROS and antioxidants or by measuring by-products of oxidative damage to biomolecules, such as lipids, proteins, and DNA.[4],[6],[8] Measurement of ROS can provide direct assessment; however, it is much difficult as ROS have extremely short half-life, they are highly reactive and unstable and their effects are additive.[5] Measurement of antioxidants can be done by measuring the enzymatic antioxidants, such as superoxide dismutase (SOD), catalase, and glutathione peroxidase, and non-enzymatic antioxidants, such as vitamin C, vitamin E, uric acid, reduced glutathione, etc.[4],[7],[13]

Majority of clinical studies assessed oxidative stress by measuring by-products of oxidative tissue destruction, which can provide direct assessment.[5],[7],[8] Polyunsaturated fatty acids in lipid membrane are prime target for lipid peroxidation and form MDA. The 8-OHdG is principal and most commonly evaluated product of oxidative DNA damage.

Various studies have demonstrated alteration in oxidative stress biomarkers in body fluids, such as SOD,[5],[11] myeloperoxidase, oxidized glutathione,[13] reduced glutathione,[4] catalase,[11] glutathione peroxidise, TBARS,[13] MDA,[5],[14] protein carbonyl,[6] 8-OHdG,[9],[12],[15] total oxidant level,[5] and total antioxidant capacity.[16]

Although there are several studies on salivary oxidative stress marker level[9],[16] and serum biomarkers of oxidative stress[7] separately, determining both the saliva and serum biomarkers of oxidative stress is now a current research approach.[5] Additional findings on oxidative stress level in both saliva and serum of chronic periodontitis patients could give new dimensions to systemic influence of periodontitis. Thus, keeping the above-mentioned facts in mind, this study focused on the relationship of chronic periodontitis with local (saliva) and systemic (serum) biomarkers and also the effect of nonsurgical therapy on these biomarkers, MDA – a lipid peroxidation product and 8-OHdG – a DNA oxidation product.

The clinical periodontal parameters were used to assess the effect of the nonsurgical periodontal therapy in patients of Group II. Several studies have concluded that the clinical parameters are altered in chronic periodontitis when compared to healthy controls.[14] In agreement to these studies, it was observed in this study that there was a significant increase in PI, PPD, GI, and CAL in Group II patients as compared to Group I subjects. The nonsurgical periodontal therapy showed improvement in clinical parameters 3 months post-therapy as compared to baseline values. Similar results in improvement in clinical parameters after therapy have been reported by Wei D et al. 2010 and others.[5],[7]

In this study, the level of MDA in saliva was significantly high in Group II (2.659 ± 0.409 nmol/ml) when compared with healthy group (1.318 ± 0.306 nmol/ml). This result was found in accordance with the findings of the previous studies.[16],[17] Also significantly increased level of serum MDA has been observed in this study. (Aziz AS et al. also showed high serum MDA levels in chronic periodontitis.[7]) However, some authors demonstrated no significant difference in serum MDA level.[5],[14]

It has been observed in this study that the serum and salivary level of MDA was significantly decreased after nonsurgical periodontal therapy that is consistent with the findings of Aziz et al.[7] and in contrast to the findings of Wei D et al.[5] who showed nonsignificant reduction after periodontal treatment.

The salivary 8-OHdG level finding in periodontitis group was comparable to the recent studies, which demonstrated significantly higher salivary levels of 8-OHdG in periodontitis when compared with the control group.[9],[16],[18] Ongoz et al.[12] reported nonsignificant increase in 8-OHdG level in saliva. This study result supports the earlier studies,[12] as the serum level of 8-OHdG was significantly increased in periodontitis, and contrast the studies that showed nonsignificant increase[15] or decreased level.[10] SRP resulted in significant decrease in 8-OHdG levels in saliva that seems to confirm the previous studies findings[15],[19] and contrast the findings of other studies.[12] There was decreased serum level of 8-OHdG after therapy that was significant but not reached to level in healthy group.

The linear relationship between the CAL and salivary MDA was found to be positive and highly significant. This supports the findings of the previous studies;[11],[14] however, others reported no correlation.[16] The CAL and serum MDA correlation was positive and significant in this study. Similar results were demonstrated by Fentoglu et al.[8] who showed significant correlation in serum MDA and GI. No correlation between CAL and serum MDA was observed by other studies.[5],[10],[14]

The linear relationship between CAL and salivary 8-OHdG was positive and significant in this study, as demonstrated by some of the studies.[9],[10] but contrast the results of other studies.[12] Regarding serum 8-OHdG and CAL correlation, it was significant in this study. Onder et al.[10] drawn contrast results. Sawamoto et al.[18] reported that salivary 8-OHdG is strongly correlated to periodontal pathogens.

The imbalance between ROS and antioxidants is an important mechanism in periodontal disease pathogenesis. Chronic periodontitis patients showed higher oxidative stress markers both in saliva and in serum in this study and reduced after periodontal therapy. Thus, these markers can be useful for early detection, prevention, and assessment of effect of periodontal treatment. SRP is considered as the initial phase and gold standard of periodontal treatment, as it removes inflammation by removing the causative pathogens from the tooth surfaces. Thus, it may restore the ROS and antioxidant balance and help to control or delay the inflammatory process.


   Conclusion Top


The results of this study support the evidence that oxidative stress can be one of the underlying mechanisms in pathogenesis of chronic periodontitis and also expressed in terms of increased systemic oxidative stress biomarkers that reduce after periodontal therapy. Based on the results, it can be concluded that the periodontal therapy is effective in improving the oxidative stress condition. However, considering the lack of evaluation of correlation of chronic periodontitis severity with oxidative stress biomarkers and selection of panels of biomarkers in this study, future studies can be encouraged, which will provide better insight in to the relationship between oxidative stress and periodontal disease.

Declaration of patient consent

The authors certify that they have obtained all appropriate patient consent forms. In the form the patient(s) has/have given his/her/their consent for his/her/their images and other clinical information to be reported in the journal. The patients understand that their names and initials will not be published and due efforts will be made to conceal their identity, but anonymity cannot be guaranteed.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
   References Top

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2.
Mittal M, Siddiqui MR, Tran K, Reddy SP, Malik AB. Reactive oxygen species in inflammation and tissue injury. Antioxid Redox Sig 2014;20:1126-67.  Back to cited text no. 2
    
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Chapple ILC. Role of free radicals and antioxidants in the pathogenesis of the inflammatory periodontal Diseases. J Clin Pathol Mol Pathol 1996;49:M247-55.  Back to cited text no. 3
    
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Tsai CC. Lipid peroxidation: A possible role in the induction and progression of chronic periodontitis. J Periodontal Res 2005;40:378-84.  Back to cited text no. 4
    
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Wei D, Zhang XL, Wang YZ, Yang CX, Chen G. Lipid peroxidation levels, total oxidant status and superoxide dismutase in serum, saliva and gingival crevicular fluid in chronic periodontitis patients before and after periodontal therapy. Aust Dent J 2010;55:70-8.  Back to cited text no. 5
    
6.
Baltacioglu E, Sukuroglu E. Protein carbonyl levels in serum, saliva and gingival crevicular fluid in patients with chronic and aggressive periodontitis. Saudi Dent J 2019;31:23-30.  Back to cited text no. 6
    
7.
Aziz AS, Kalekar MG, Benjamin T, Suryakar AN, Prakashan MM, Bijle MNA. Effect of nonsurgical periodontal therapy on some oxidative stress markers in patients with chronic periodontitis: A biochemical study. World J Dent 2013;4:17-23.  Back to cited text no. 7
    
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Fentoglu A, Kirzioglu FY, Bulut MT, Doguc DK, Kulac E, Onder C. Evaluation of lipid peroxidation and oxidative DNA damage in patients with periodontitis and hyperlipidemia. J Periodontol 2015;86:682-88.  Back to cited text no. 8
    
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Sezer U, Cicek Y, Canakci CF. Increased salivary levels of 8-hydroxydeoxyguanosine may be a marker for disease activity for periodontitis. Dis Markers 2012;32:165-72.  Back to cited text no. 9
    
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Onder C, Kurgan S, Altingoz SM, Bagis N, Uyanik M, Serdar MA, et al. Impact of non-surgical periodontal therapy on saliva and serum levels of markers of oxidative stress. Clin Oral Investig 2017;21:1961-9.  Back to cited text no. 10
    
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Ongoz Dede F, Otan Ozden F, Avci B. 8-OHdG levels in gingival crevicular fluid and saliva from patients with chronic periodontitis during initial periodontal treatment. J Periodontol 2013;84:821-8.  Back to cited text no. 12
    
13.
Borges I Jr, Moreira EA, Filho DW, de Oliveira TB, da Silva MB, Frode TS. Proinflammatory and oxidative stress markers in patients with periodontal disease. Mediators Inflamm 2007;2007:45794.  Back to cited text no. 13
    
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Akalin FA, Baltacioglu E, Alver A, Karabulut E. Lipid peroxidation levels and total oxidant status in serum, saliva and gingival crevicular fluid in patients with chronic periodontitis. J Clin Periodontol 2007;34:558-65.  Back to cited text no. 14
    
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Hendek MK, Erdemir EO, Kisa U, Ozcan G. Effect of initial periodontal therapy on oxidative stress markers in gingival crevicular fluid, saliva, and serum in smokers and non-smokers with chronic periodontitis. J Periodontol 2015;86:273-82.  Back to cited text no. 15
    
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Miricescu D, Totan A, Calenic B, Mocanu B, Didilescu A, Mohora M, et al. Salivary biomarkers: Relationship between oxidative stress and alveolar bone loss in chronic periodontitis. Acta Odontol Scand 2013;72:42-7.  Back to cited text no. 16
    
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Canakci CF, Cicek Y, Yildirim A, Sezer U, Canakci V. Increased levels of 8-hydroxydeoxyguanosine and malondialdehyde and its relationship with antioxidant enzymes in saliva of periodontitis patients. Eur J Dent 2009;3:100-6.  Back to cited text no. 17
    
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Sawamoto Y, Sugano N, Tanaka H, Ito K. Detection of periodontopathic bacteria and an oxidative stress marker insaliva from periodontitis patients. Oral Microbiol Immunol 2005;20:216-20.  Back to cited text no. 18
    
19.
Kurgan S, Onder C, Altingoz SM, Bagis N, Uyanik M, Serdar MA, et al. High sensitivity detection of salivary 8-hydroxy deoxyguanosine levels in patients with chronic periodontitis. J Periodont Res 2015;50:766-74.  Back to cited text no. 19
    

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Correspondence Address:
Dr. Anju Gautam
Department of Periodontology, Faculty of Dental Sciences, Varanasi, Uttar Pradesh
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ijdr.ijdr_912_21

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