J Periodontal Implant Sci.  2013 Jun;43(3):121-129. 10.5051/jpis.2013.43.3.121.

Locally delivered antioxidant gel as an adjunct to nonsurgical therapy improves measures of oxidative stress and periodontal disease

• Chandra RV
• Srinivas G
• Reddy AA
• Reddy BH
• Reddy C
• Nagarajan
• Naveen A

• KMID: 1798485
• DOI: http://doi.org/10.5051/jpis.2013.43.3.121

Abstract

PURPOSE
The present study has two aims; firstly, it attempts to verify the presence of oxidative stress by estimating the reactive oxygen species (ROS) levels in periodontal pockets > or =5 mm as compared to controls. The second aim is to evaluate the effect of lycopene as a locally delivered antioxidant gel on periodontal health and on the gingival crevicular fluid (GCF) levels of 8-hydroxydeoxyguanosine (8-OHdG), a marker of oxidative injury.
METHODS
Thirty-one subjects participated in this study. In the pretreatment phase, the ROS levels in pockets > or =5 mm were measured by flow cytometry. Three sites in each subject were randomly assigned into each of the following experimental groups: sham group, only scaling and root planing (SRP) was done; placebo group, local delivery of placebo gel after SRP; and lycopene group, local delivery of lycopene gel after SRP. Clinical parameters included recording site-specific measures of GCF 8-OHdG, plaque, gingivitis, probing depth, and clinical attachment level.
RESULTS
The gel, when delivered to the sites with oxidative stress, was effective in increasing clinical attachment and in reducing gingival inflammation, probing depth, and 8-OHdG levels as compared to the placebo and sham sites.
CONCLUSIONS
From this trial conducted over a period of 6 months, it was found that locally delivered lycopene seems to be effective in reducing the measures of oxidative stress and periodontal disease.

Keyword

Antioxidants; Oxidative stress; Periodontitis

MeSH Terms

Antioxidants
Carotenoids
Deoxyguanosine
Flow Cytometry
Gingival Crevicular Fluid
Gingivitis
Inflammation
Oxidative Stress
Periodontal Diseases
Periodontal Pocket
Periodontitis
Reactive Oxygen Species
Root Planing
Salicylamides
Antioxidants
Carotenoids
Deoxyguanosine
Reactive Oxygen Species
Salicylamides

Figure

• 

  Figure 1 Bar graph representing the percentage of cells showing increased reactive oxygen species (ROS) production compared to controls. Values are reported as fluorescence intensity. Duplicate samples were run to confirm the assay's repeatability, and the mean was taken. DCF FI: dichlorofluorescein fluorescence intensity. a)Highly significant as compared to the control group.

• 

  Figure 2 Effects of lycopene, placebo, and sham delivery on the 8-hydroxydeoxyguanosine (8-OHdG) levels at different time periods. a)Significant intragroup reduction as compared to the baseline levels. b)Significant intragroup reduction as compared to the levels at 1 week. c)Intergroup comparison at the first week shows a highly significant reduction in 8-OHdG in the lycopene-treated sites.

Reference

1. Kiyoshima T, Enoki N, Kobayashi I, Sakai T, Nagata K, Wada H, et al. Oxidative stress caused by a low concentration of hydrogen peroxide induces senescence-like changes in mouse gingival fibroblasts. Int J Mol Med. 2012. 30:1007–1012.

2. Battino M, Bullon P, Wilson M, Newman H. Oxidative injury and inflammatory periodontal diseases: the challenge of anti-oxidants to free radicals and reactive oxygen species. Crit Rev Oral Biol Med. 1999. 10:458–476.

3. 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.

4. Masi S, Salpea KD, Li K, Parkar M, Nibali L, Donos N, et al. Oxidative stress, chronic inflammation, and telomere length in patients with periodontitis. Free Radic Biol Med. 2011. 50:730–735.

5. Marchetti E, Monaco A, Procaccini L, Mummolo S, Gatto R, Tete S, et al. Periodontal disease: the influence of metabolic syndrome. Nutr Metab (Lond). 2012. 9:88.

6. Govindaraj P, Khan NA, Gopalakrishna P, Chandra RV, Vanniarajan A, Reddy AA, et al. Mitochondrial dysfunction and genetic heterogeneity in chronic periodontitis. Mitochondrion. 2011. 11:504–512.

7. Sugiyama S, Takahashi SS, Tokutomi FA, Yoshida A, Kobayashi K, Yoshino F, et al. Gingival vascular functions are altered in type 2 diabetes mellitus model and/or periodontitis model. J Clin Biochem Nutr. 2012. 51:108–113.

8. Esen C, Alkan BA, Kırnap M, Akgul O, Isıkoglu S, Erel O. The effects of chronic periodontitis and rheumatoid arthritis on serum and gingival crevicular fluid total antioxidant/oxidant status and oxidative stress index. J Periodontol. 2012. 83:773–779.

9. Kawaguchi K, Matsumoto T, Kumazawa Y. Effects of antioxidant polyphenols on TNF-alpha-related diseases. Curr Top Med Chem. 2011. 11:1767–1779.

10. Soory M. Relevance of nutritional antioxidants in metabolic syndrome, ageing and cancer: potential for therapeutic targeting. Infect Disord Drug Targets. 2009. 9:400–414.

11. Bagaitkar J, Williams LR, Renaud DE, Bemakanakere MR, Martin M, Scott DA, et al. Tobacco-induced alterations to Porphyromonas gingivalis-host interactions. Environ Microbiol. 2009. 11:1242–1253.

12. Kornman KS, Page RC, Tonetti MS. The host response to the microbial challenge in periodontitis: assembling the players. Periodontol 2000. 1997. 14:33–53.

13. Darveau RP, Tanner A, Page RC. The microbial challenge in periodontitis. Periodontol 2000. 1997. 14:12–32.

14. Giannelli M, Chellini F, Margheri M, Tonelli P, Tani A. Effect of chlorhexidine digluconate on different cell types: a molecular and ultrastructural investigation. Toxicol In Vitro. 2008. 22:308–317.

15. D'Aiuto F, Nibali L, Parkar M, Patel K, Suvan J, Donos N. Oxidative stress, systemic inflammation, and severe periodontitis. J Dent Res. 2010. 89:1241–1246.

16. Wood N, Johnson RB. The relationship between tomato intake and congestive heart failure risk in periodontitis subjects. J Clin Periodontol. 2004. 31:574–580.

17. Chandra RV, Prabhuji ML, Roopa DA, Ravirajan S, Kishore HC. Efficacy of lycopene in the treatment of gingivitis: a randomised, placebo-controlled clinical trial. Oral Health Prev Dent. 2007. 5:327–336.

18. Chandra RV, Sandhya YP, Nagarajan S, Reddy BH, Naveen A, Murthy KR. Efficacy of lycopene as a locally delivered gel in the treatment of chronic periodontitis: smokers vs nonsmokers. Quintessence Int. 2012. 43:401–411.

19. Maruyama T, Tomofuji T, Endo Y, Irie K, Azuma T, Ekuni D, et al. Supplementation of green tea catechins in dentifrices suppresses gingival oxidative stress and periodontal inflammation. Arch Oral Biol. 2011. 56:48–53.

20. Fuhrman B, Elis A, Aviram M. Hypocholesterolemic effect of lycopene and beta-carotene is related to suppression of cholesterol synthesis and augmentation of LDL receptor activity in macrophages. Biochem Biophys Res Commun. 1997. 233:658–662.

21. Dalcico R, de Menezes AM, Deocleciano OB, Oria RB, Vale ML, Ribeiro RA, et al. Protective Mechanisms of Simvastatin in Experimental Periodontal Disease. J Periodontol. 2012. 11. 26. [Epub]. http://dx.doi.org/10.1902/jop.2012.120114.

22. Kara A, Akman S, Ozkanlar S, Tozoglu U, Kalkan Y, Canakci CF, et al. Immune modulatory and antioxidant effects of melatonin in experimental periodontitis in rats. Free Radic Biol Med. 2013. 55:21–26.

23. Ku SK, Cho HR, Sung YS, Kang SJ, Lee YJ. Effects of calcium gluconate on experimental periodontitis and alveolar bone loss in rats. Basic Clin Pharmacol Toxicol. 2011. 108:241–250.

24. Di Mascio P, Kaiser S, Sies H. Lycopene as the most efficient biological carotenoid singlet oxygen quencher. Arch Biochem Biophys. 1989. 274:532–538.

25. 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. 3:100–106.

26. Konopka T, Krol K, Kopec W, Gerber H. Total antioxidant status and 8-hydroxy-2'-deoxyguanosine levels in gingival and peripheral blood of periodontitis patients. Arch Immunol Ther Exp (Warsz). 2007. 55:417–422.

27. Turesky S, Gilmore ND, Glickman I. Reduced plaque formation by the chloromethyl analogue of victamine C. J Periodontol. 1970. 41:41–43.

28. Lobene RR, Weatherford T, Ross NM, Lamm RA, Menaker L. A modified gingival index for use in clinical trials. Clin Prev Dent. 1986. 8:3–6.

29. Patel SA, Patel NM, Patel MM. Spectrophotometric methods for the estimation of cephalexin in tablet dosage forms. Indian J Pharma Sci. 2006. 68:278–280.

30. Tuter G, Kurtis B, Serdar M. Interleukin-1beta and thiobarbituric acid reactive substance (TBARS) levels after phase I periodontal therapy in patients with chronic periodontitis. J Periodontol. 2001. 72:883–888.

31. Battino M, Ferreiro MS, Fattorini D, Bullon P. In vitro antioxidant activities of mouthrinses and their components. J Clin Periodontol. 2002. 29:462–467.

32. Neiva RF, Steigenga J, Al-Shammari KF, Wang HL. Effects of specific nutrients on periodontal disease onset, progression and treatment. J Clin Periodontol. 2003. 30:579–589.

33. Watts T. Periodontal treatment and glycemic control in diabetic patients: the problem of a possible Hawthorne effect. J Dent Res. 2006. 85:294.

34. Bansal K, Rawat MK, Jain A, Rajput A, Chaturvedi TP, Singh S. Development of satranidazole mucoadhesive gel for the treatment of periodontitis. AAPS PharmSciTech. 2009. 10:716–723.

35. Halliwell B, Whiteman M. Measuring reactive species and oxidative damage in vivo and in cell culture: how should you do it and what do the results mean? Br J Pharmacol. 2004. 142:231–255.

36. Palozza P, Parrone N, Simone R, Catalano A. Role of lycopene in the control of ROS-mediated cell growth: implications in cancer prevention. Curr Med Chem. 2011. 18:1846–1860.

37. Dede FO, Ozden FO, Avcı B. 8-hydroxy-deoxyguanosine levels in gingival crevicular fluid and saliva in patients with chronic periodontitis after initial periodontal treatment. J Periodontol. 2013. 84:821–828.

38. Jakubowski W, Bartosz G. 2,7-dichlorofluorescin oxidation and reactive oxygen species: what does it measure? Cell Biol Int. 2000. 24:757–760.

39. Wolfe KL, Liu RH. Structure-activity relationships of flavonoids in the cellular antioxidant activity assay. J Agric Food Chem. 2008. 56:8404–8411.

40. Myhre O, Andersen JM, Aarnes H, Fonnum F. Evaluation of the probes 2',7'-dichlorofluorescin diacetate, luminol, and lucigenin as indicators of reactive species formation. Biochem Pharmacol. 2003. 65:1575–1582.