COMPARISON OF THE EFFECTS OF OMEGA 3 AND PROBIOTICS ON ALVEOLAR BONE LOSS IN RATS: AN IMMUNOHISTOCHEMICAL STUDY

Year 2021, Volume: 31 Issue: 2, 167 - 174, 15.04.2021

Burak Doğan Esra Sinem Kemer Doğan Özlem Özmen

https://doi.org/10.17567/ataunidfd.860173

Abstract

Aim: Omega 3 and probiotics have been demonstrated to have beneficial effects in periodontal disease pathogenesis. Inducible nitric oxide synthase is an important mediator regulating immune response. The objective of this study was to compare the impacts of omega 3 and probiotic supplementation on alveolar bone loss and gingival inducible nitric oxide synthase levels in an experimental periodontitis model.
Material and Methods: Thirty-two rats were separated into 4 equal groups as control, periodontitis, omega 3, and probiotic. Omega 3 or probiotic was supplemented by oral gavage and experimental periodontitis was induced to the rats in periodontitis, omega 3, and probiotic groups by ligature. Alveolar bone loss was measured histometrically. The number of neutrophils was counted and gingival inducible nitric oxide synthase levels were analysed immunohistochemically.
Results: Alveolar bone loss significantly increased in periodontitis induced groups, compared to control group and decreased in omega 3 and probiotic groups, compared to the periodontitis group (p <0.05). Probiotic group had further decrease in alveolar bone loss than the omega 3 group (p <0.05). The number of neutrophils and gingival inducible nitric oxide synthase levels were higher in the periodontitis, omega 3, and probiotic groups than those in the control group, and decreased in omega 3 and probiotic groups, compared to the periodontitis group (p <0.05).
Conclusion: We suggest that omega 3 and probiotic administrations could reduce alveolar bone loss by decreasing gingival inducible nitric oxide synthase. Probiotic support seems to have more beneficial reducing alveolar bone loss, compared to omega 3.
Key words: Fatty Acids, Probiotics, Alveolar Bone Loss, Nitric Oxide Synthase Type II, Periodontitis

OMEGA 3 VE PROBİYOTİKLERİN RATLARDA ALVEOLER KEMİK KAYBINA ETKİLERİNİN KARŞILAŞTIRILMASI: İMMÜNOHİSTOKİMYASAL BİR ÇALIŞMA
Amaç: Omega 3 ve probiyotiklerin periodontal hastalık patogenezinde yararlı etkilerinin olduğu gösterilmiştir. İndüklenebilir nitrik oksit sentaz immün yanıtı düzenleyen önemli bir mediyatördür. Bu çalışmanın amacı, deneysel periodontitis modelinde omega 3 ve probiyotik desteğinin alveoler kemik kaybı ve dişeti indüklenebilir nitrik oksit sentaz seviyeleri üzerine olan etkilerini karşılaştırmaktır.
Gereç ve Yöntem: Otuz iki sıçan kontrol, periodontitis, omega 3 ve probiyotik olarak 4 eşit gruba ayrıldı. Omega 3 ve probiyotik oral gavajla uygulandı ve periodontitis, omega 3 ve probiyotik gruplarındaki sıçanlara ligatür kullanılarak deneysel periodontitis oluşturuldu. Alveoler kemik kaybı histometrik olarak ölçüldü. Nötrofil sayısı hesaplandı ve dişeti indüklenebilir nitrik oksit sentaz seviyeleri immünohistokimyasal olarak değerlendirildi.
Bulgular: Alveoler kemik kaybı periodontitis indüklenen gruplarda kontrol grubuna göre anlamlı derecede arttı ve periodontitis grubuna kıyasla omega 3 ve probiyotik gruplarında azaldı (p <0.05). Alveoler kemik kaybı, probiyotik grubunda omega 3 grubuna göre daha fazla azaldı (p <0.05). Nötrofil sayıları ve dişeti indüklenebilir nitrik oksit sentaz seviyeleri kontrol grubuna göre periodontitis, omega 3 ve probiyotik gruplarında daha yüksekti ve periodontitis grubuyla kıyaslandığında omega 3 ve probiyotik gruplarında azaldı (p <0.05).
Sonuç: Omega 3 ve probiyotik desteğinin dişeti indüklenebilir nitrik oksit sentaz seviyelerini azaltarak alveoler kemik kaybını düşürebildiğini öne sürmekteyiz. Alveoler kemik kaybını azaltmada probiyotik desteği, omega 3 ile karşılaştırıldığında daha yararlı görünmektedir.
Anahtar Kelimeler: Yağ Asidleri, Probiyotikler, Alveoler Kemik Kaybı, Nitrik Oksit Sentaz Tip II, Periodontit

Keywords

Periodontitis, Fatty Acids, Alveolar Bone Loss, itric Oxide Synthase Type II,

References

• 1. Kassebaum NJ, Bernabe E, Dahiya M, Bhandari B, Murray CJ, Marcenes W. Global burden of severe periodontitis in 1990-2010: a systematic review and meta-regression. J Dent Res 2014;93:1045-53.
• 2. Kononen E, Gursoy M, Gursoy UK. Periodontitis: A multifaceted disease of tooth-supporting tissues. J Clin Med 2019;8:1135.
• 3. Connor WE. Importance of n-3 fatty acids in health and disease. Am J Clin Nutr 2000;71:171S-5S.
• 4. Chanda W, Joseph TP, Guo XF, Wang WD, Liu M, Vuai MS, Padhiar AA, Zhong MT. Effectiveness of omega-3 polyunsaturated fatty acids against microbial pathogens. J Zhejiang Univ Sci B 2018;19:253-62.
• 5. Hill C, Guarner F, Reid G, Gibson GR, Merenstein DJ, Pot B, Morelli L, Canani RB, Flint HJ, Salminen S, Calder PC, Sanders ME. Expert consensus document. The International Scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nat Rev Gastroenterol Hepatol 2014;11:506-14.
• 6. Arat Maden E, Altun C. Probiyotikler ve ağız sağlığı. Atatürk Üniv Diş Hek Fak Derg 2012;22:334-9.
• 7. Sanders ME, Merenstein DJ, Reid G, Gibson GR, Rastall RA. Probiotics and prebiotics in intestinal health and disease: from biology to the clinic. Nat Rev Gastroenterol Hepatol 2019;16:605-16.
• 8. Castro Dos Santos NC, Andere NM, Araujo CF, de Marco AC, Kantarci A, Van Dyke TE, Santamaria MP. Omega-3 PUFA and aspirin as adjuncts to periodontal debridement in patients with periodontitis and type 2 diabetes mellitus. Randomized clinical trial. J Periodontol 2020.
• 9. Deore GD, Gurav AN, Patil R, Shete AR, Naiktari RS, Inamdar SP. Omega 3 fatty acids as a host modulator in chronic periodontitis patients: a randomised, double-blind, palcebo-controlled, clinical trial. J Periodontal Implant Sci 2014;44:25-32.
• 10. Elgendy EA, Kazem HH. Effect of Omega-3 Fatty Acids on Chronic Periodontitis Patients in Postmenopausal Women: A Randomised Controlled Clinical Study. Oral Health Prev Dent 2018;16:327-32.
• 11. Invernici MM, Salvador SL, Silva PHF, Soares MSM, Casarin R, Palioto DB, Souza SLS, Taba M, Jr., Novaes AB, Jr., Furlaneto FAC, Messora MR. Effects of bifidobacterium probiotic on the treatment of chronic periodontitis: A randomized clinical trial. J Clin Periodontol 2018;45:1198-210.
• 12. Meenakshi SS, Varghese S. Adjunctive effect of probiotic (lactobacillus casei shirota) to scaling and root planing in the management of chronic periodontitis. Drug Invention Today 2018;10:1381-6.
• 13. Kuru BE, Laleman I, Yalnizoglu T, Kuru L, Teughels W. The influence of a bifidobacterium animalis probiotic on gingival health: A randomized controlled clinical trial. J Periodontol 2017;88:1115-23.
• 14. Ince G, Gursoy H, Ipci SD, Cakar G, Emekli-Alturfan E, Yilmaz S. Clinical and Biochemical Evaluation of Lozenges Containing Lactobacillus reuteri as an Adjunct to Non-Surgical Periodontal Therapy in Chronic Periodontitis. J Periodontol 2015;86:746-54.
• 15. Tekce M, Ince G, Gursoy H, Dirikan Ipci S, Cakar G, Kadir T, Yilmaz S. Clinical and microbiological effects of probiotic lozenges in the treatment of chronic periodontitis: a 1-year follow-up study. J Clin Periodontol 2015;42:363-72.
• 16. Kesavalu L, Vasudevan B, Raghu B, Browning E, Dawson D, Novak JM, Correll MC, Steffen MJ, Bhattacharya A, Fernandes G, Ebersole JL. Omega-3 fatty acid effect on alveolar bone loss in rats. J Dent Res 2006;85:648-52.
• 17. Bendyk A, Marino V, Zilm PS, Howe P, Bartold PM. Effect of dietary omega-3 polyunsaturated fatty acids on experimental periodontitis in the mouse. J Periodontal Res 2009;44:211-6.
• 18. Kesavalu L, Bakthavatchalu V, Rahman M, Su J, Raghu B, Dawson D, Fernandes G, Ebersole J. Omega‐3 fatty acid regulates inflammatory cytokine/mediator messenger RNA expression in Porphyromonas gingivalis‐induced experimental periodontal disease. Oral microbiology and immunology 2007;22:232-9.
• 19. Oliveira LF, Salvador SL, Silva PH, Furlaneto FA, Figueiredo L, Casarin R, Ervolino E, Palioto DB, Souza SL, Taba M, Jr., Novaes AB, Jr., Messora MR. Benefits of Bifidobacterium animalis subsp. lactis Probiotic in Experimental Periodontitis. J Periodontol 2017;88:197-208.
• 20. Foureaux Rde C, Messora MR, de Oliveira LF, Napimoga MH, Pereira AN, Ferreira MS, Pereira LJ. Effects of probiotic therapy on metabolic and inflammatory parameters of rats with ligature-induced periodontitis associated with restraint stress. J Periodontol 2014;85:975-83.
• 21. Garcia VG, Knoll LR, Longo M, Novaes VC, Assem NZ, Ervolino E, de Toledo BE, Theodoro LH. Effect of the probiotic Saccharomyces cerevisiae on ligature-induced periodontitis in rats. J Periodontal Res 2016;51:26-37.
• 22. Keskiner I, Saygun I, Bal V, Serdar M, Kantarci A. Dietary supplementation with low-dose omega-3 fatty acids reduces salivary tumor necrosis factor-alpha levels in patients with chronic periodontitis: a randomized controlled clinical study. J Periodontal Res 2017;52:695-703.
• 23. Martinez GL, Koury JC, Martins MA, Nogueira F, Fischer RG, Gustafsson A, Figueredo CM. Serum level changes of long chain-polyunsaturated fatty acids in patients undergoing periodontal therapy combined with one year of omega-3 supplementation: a pilot randomized clinical trial. J Periodontal Implant Sci 2014;44:169-77.
• 24. Montero E, Iniesta M, Rodrigo M, Marin MJ, Figuero E, Herrera D, Sanz M. Clinical and microbiological effects of the adjunctive use of probiotics in the treatment of gingivitis: A randomized controlled clinical trial. J Clin Periodontol 2017;44:708-16.
• 25. Pelekos G, Ho SN, Acharya A, Leung WK, McGrath C. A double-blind, paralleled-arm, placebo-controlled and randomized clinical trial of the effectiveness of probiotics as an adjunct in periodontal care. J Clin Periodontol 2019;46:1217-27.
• 26. Vardar-Sengul S, Buduneli E, Turkoglu O, Buduneli N, Atilla G, Wahlgren J, Sorsa T, Baylas H. The effects of selective COX-2 inhibitor/celecoxib and omega-3 fatty acid on matrix metalloproteinases, TIMP-1, and laminin-5gamma2-chain immunolocalization in experimental periodontitis. J Periodontol 2008;79:1934-41.
• 27. Cinelli MA, Do HT, Miley GP, Silverman RB. Inducible nitric oxide synthase: Regulation, structure, and inhibition. Med Res Rev 2020;40:158-89.
• 28. Kim YS, Pi SH, Lee YM, Lee SI, Kim EC. The anti-inflammatory role of heme oxygenase-1 in lipopolysaccharide and cytokine-stimulated inducible nitric oxide synthase and nitric oxide production in human periodontal ligament cells. J Periodontol 2009;80:2045-55.
• 29. Hirose M, Ishihara K, Saito A, Nakagawa T, Yamada S, Okuda K. Expression of cytokines and inducible nitric oxide synthase in inflamed gingival tissue. J Periodontol 2001;72:590-7.
• 30. Aldridge C, Razzak A, Babcock TA, Helton WS, Espat NJ. Lipopolysaccharide-stimulated RAW 264.7 macrophage inducible nitric oxide synthase and nitric oxide production is decreased by an omega-3 fatty acid lipid emulsion. J Surg Res 2008;149:296-302.
• 31. Yang M, Li L, Soh Y, Heo SM. Effects of omega-3 fatty acids and aspirin on Porphyromonas gingivalis-induced periodontitis in rats. J Periodontol 2019;90:1307-19.
• 32. Ren J, Chung SH. Anti-inflammatory effect of alpha-linolenic acid and its mode of action through the inhibition of nitric oxide production and inducible nitric oxide synthase gene expression via NF-kappaB and mitogen-activated protein kinase pathways. J Agric Food Chem 2007;55:5073-80.
• 33. Liu Y-W, Ong W-K, Su Y-W, Hsu C-C, Cheng T-H, Tsai Y-C. Anti-inflammatory effects of Lactobacillus brevis K65 on RAW 264.7 cells and in mice with dextran sulphate sodium-induced ulcerative colitis. Beneficial microbes 2016;7:387-96.
• 34. Yu H-S, Lee N-K, Choi A-J, Choe J-S, Bae CH, Paik H-D. Anti-inflammatory potential of probiotic strain Weissella cibaria JW15 isolated from Kimchi through regulation of NF-κB and MAPKs pathways in LPS-induced RAW 264. 7 Cells. J Microbiol Biotechn 2019;29:1022-32.
• 35. Riccia DN, Bizzini F, Perilli MG, Polimeni A, Trinchieri V, Amicosante G, Cifone MG. Anti-inflammatory effects of Lactobacillus brevis (CD2) on periodontal disease. Oral Dis 2007;13:376-85.
• 36. Eratte D, McKnight S, Gengenbach TR, Dowling K, Barrow CJ, Adhikari BP. Co-encapsulation and characterisation of omega-3 fatty acids and probiotic bacteria in whey protein isolate–gum Arabic complex coacervates. Journal of Functional Foods 2015;19:882-92.
• 37. Kirzioglu FY, Tozum Bulut M, Dogan B, Fentoglu O, Ozmen O, Carsancakli SA, Ergun AG, Ozdem M, Orhan H. Anti-inflammatory effect of rosuvastatin decreases alveolar bone loss in experimental periodontitis. J Oral Sci 2017;59:247-55.
• 38. Esposito E, Iacono A, Bianco G, Autore G, Cuzzocrea S, Vajro P, Canani RB, Calignano A, Raso GM, Meli R. Probiotics reduce the inflammatory response induced by a high-fat diet in the liver of young rats. J Nutr 2009;139:905-11.
• 39. Kuo PJ, Fu E, Lin CY, Ku CT, Chiang CY, Fu MM, Fu MW, Tu HP, Chiu HC. Ameliorative effect of hesperidin on ligation-induced periodontitis in rats. J Periodontol 2019;90:271-80.
• 40. Yoshinaga Y, Ukai T, Nakatsu S, Kuramoto A, Nagano F, Yoshinaga M, Montenegro JL, Shiraishi C, Hara Y. Green tea extract inhibits the onset of periodontal destruction in rat experimental periodontitis. J Periodontal Res 2014;49:652-9.
• 41. Antona ME, Ramos C, Stranges A, Monteiro AF, Gonzales Chaves MM, Mandalunis P, Zago V, Friedman SM, Macri EV. Fish oil diet effects on alveolar bone loss, in hypercholesterolemic rats. Arch Oral Biol 2020;109:104553.
• 42. Kobyliak N, Falalyeyeva T, Bodnar P, Beregova T. Probiotics Supplemented with Omega-3 Fatty Acids are More Effective for Hepatic Steatosis Reduction in an Animal Model of Obesity. Probiotics Antimicrob Proteins 2017;9:123-30.
• 43. Herrera BS, Martins-Porto R, Maia-Dantas A, Campi P, Spolidorio LC, Costa SK, Van Dyke TE, Gyurko R, Muscara MN. iNOS-derived nitric oxide stimulates osteoclast activity and alveolar bone loss in ligature-induced periodontitis in rats. J Periodontol 2011;82:1608-15.
• 44. Kirzioglu FY, Ozmen O, Dogan B, Bulut MT, Fentoglu O, Ozdem M. Effects of rosuvastatin on inducible nitric oxide synthase in rats with hyperlipidaemia and periodontitis. J Periodontal Res 2018;53:258-66.
• 45. Paquette DW, Rosenberg A, Lohinai Z, Southan GJ, Williams RC, Offenbacher S, Szabo C. Inhibition of experimental gingivitis in beagle dogs with topical mercaptoalkylguanidines. J Periodontol 2006;77:385-91.
• 46. Doughman SD, Krupanidhi S, Sanjeevi CB. Omega-3 fatty acids for nutrition and medicine: considering microalgae oil as a vegetarian source of EPA and DHA. Curr Diabetes Rev 2007;3:198-203.
• 47. Batista AC, Silva TA, Chun JH, Lara VS. Nitric oxide synthesis and severity of human periodontal disease. Oral Dis 2002;8:254-60.