Investigation of Effects of Local Vitamin C Application on Inflammatory Response and Periodontal Tissue Destruction in Rat Periodontitis Model

Year 2023, Volume: 10 Issue: 2, 371 - 376, 21.08.2023

Zeliha Aytekin , Ayşe Toraman , Kübra Karaçam

https://doi.org/10.15311/selcukdentj.1115391

Abstract

Background: The present study aims to evaluate the effect of local vitamin C application on inflammatory response and periodontal tissue destruction in rats with experimental periodontitis.
Methods: A total of 21 animals, 7 rats in each group, were used in the study: 1- Non-ligation (NL), 2-experimental periodontitis (EP), and 3- local vitamin C (VtC) groups. Experimental periodontitis was induced in EP and VtC group rats. After 11 days, the ligature was removed. 50 mL of saline solution and vitamin C were injected locally into the vestibule sulcus of the EP and VtC group rats, respectively. After the rats were sacrificed, blood and gingival tissue samples were taken. TNF-α levels were analyzed biochemically in serum and 8-OHdG and MMP-8 were analyzed immunohistochemically in gingival tissue. The mandibular specimens were histologically evaluated for bone destruction and attachment loss.
Results: The numeric density of 8-OHdG and MMP-8 immunopositive cells were found to be statistically significantly lower in the Vtc group than in the EP group. Vitamin C administration significantly reduced attachment loss and bone loss in the VtC group as compared to the EP group. However, there was no significant difference in serum TNF-α level in the VtC group compared to the EP group.
Conclusions: Local application of vitamin C may reduce inflammation-related bone destruction by supporting the inflammatory response with the antioxidant activity of vitamin C and contributing to the resolution of inflammation with its potential immunomodulatory effect. Vitamin C may be a therapeutic agent that can be used in the treatment of periodontitis.

Keywords

antioxidants, Experimental periodontitis, oxidative stress, vitamin C

Sıçan Periodontitis Modelinde Lokal C Vitamini Uygulamasının İnflamatuar Yanıt ve Periodontal Doku Yıkımı Üzerine Etkilerinin Araştırılması

Abstract

Amaç: Bu çalışmanın amacı deneysel periodontitisli ratlarda lokal C vitamini uygulamasının inflamatuar yanıt ve periodontal doku yıkımı üzerine olan etkisini değerlendirmektir.
Gereç ve Yöntemler: Çalışmada her grupta 7 sıçan olmak üzere toplam 21 hayvan kullanıldı: 1-Ligatür bağlanmayan (NL) 2-deneysel periodontitis (EP) ve 3-lokal vitamin C (VtC) grupları. EP ve VtC grubu sıçanlarda deneysel periodontitis oluşturuldu. Ligatürler 11 gün sonra ligatür çıkarıldı. EP ve VtC grubu sıçanların vestibül sulkusuna sırasıyla 50 µL salin solüsyonu ve C vitamini lokal olarak enjekte edildi. Ratlar sakrifiye edildikten sonra kan ve dişeti dokusu örnekleri alındı. TNF-α seviyesi serumda biyokimyasal olarak, 8-OHdG ve MMP-8 düzeyleri ise dişeti dokusunda immünohistokimyasal olarak analiz edildi. Mandibula örnekleri kemik yıkımı ve ataşman kaybı açısından histolojik olarak analiz edildi.
Bulgular: 8-OHdG ve MMP-8 immünopozitif hücre yoğunluğunun Vtc grubunda EP grubuna göre istatistiksel olarak anlamlı derecede daha düşük olduğu bulundu. C vitamini uygulaması, EP grubuna kıyasla VtC grubunda ataşman kaybını ve kemik kaybını önemli ölçüde azalttı. Ancak EP grubuna göre VtC grubunda serum TNF-α düzeyinde anlamlı bir fark yoktu.
Sonuç: Lokal C vitamini uygulaması, C vitamininin antioksidan aktivitesi ile inflamatuar yanıtı destekleyerek ve potansiyel immünomodülatör etkisi ile inflamasyonun çözülmesine katkıda bulunarak inflamasyona bağlı kemik yıkımını azaltabilir. C vitamini periodontitis tedavisinde kullanılabilecek terapötik bir ajan olabilir.

Keywords

antioksidanlar, Deneysel periodontitis, oksidatif stres, Vitamin C

References

• 1. Hajishengallis G. Periodontitis: from microbial immune subversion to systemic inflammation. Nat Rev Immunol. Jan 2015;15(1):30-44. doi:10.1038/nri3785
• 2. Chapple I. Reactive oxygen species and antioxidants in inflammatory diseases. J Clin Periodontol. 1997;24(5):287-296.
• 3. Turrens JF. Mitochondrial formation of reactive oxygen species. J Physiol. Oct 15 2003;552(Pt 2):335-44. doi:10.1113/jphysiol.2003.049478
• 4. Chapple IL, Matthews JB. The role of reactive oxygen and antioxidant species in periodontal tissue destruction. Periodontol 2000. 2007;43:160-232. doi:10.1111/j.1600-0757.2006.00178.x
• 5. Chapple IL, Brock GR, Milward MR, Ling N, Matthews JB. Compromised GCF total antioxidant capacity in periodontitis: cause or effect? J Clin Periodontol. Feb 2007;34(2):103-10. doi:10.1111/j.1600-051X.2006.01029.x
• 6. Saglam E, Canakci CF, Sebin SO, et al. Evaluation of oxidative status in patients with chronic periodontitis and polycystic ovary syndrome: A cross-sectional study. J Periodontol. Jan 2018;89(1):76-84. doi:10.1902/jop.2017.170129
• 7. Sezer U, Ciçek Y, Canakçi CF. Increased salivary levels of 8-hydroxydeoxyguanosine may be a marker for disease activity for periodontitis. Dis Markers. 2012;32(3):165-72. doi:10.3233/dma-2011-0876
• 8. Öngöz Dede F, Bozkurt Doğan Ş, Ballı U, Avcı B, Durmuşlar MC. The effect of initial periodontal treatment on plasma, gingival crevicular fluid and salivary levels of 8-hydroxy-deoxyguanosine in obesity. Arch Oral Biol. Feb 2016;62:80-5. doi:10.1016/j.archoralbio.2015.11.014
• 9. Sawamoto Y, Sugano N, Tanaka H, Ito K. Detection of periodontopathic bacteria and an oxidative stress marker in saliva from periodontitis patients. Oral Microbiol Immunol. Aug 2005;20(4):216-20. doi:10.1111/j.1399-302X.2005.00215.x
• 10. Yamasaki N, Tsuboi H, Hirao M, Nampei A, Yoshikawa H, Hashimoto J. High oxygen tension prolongs the survival of osteoclast precursors via macrophage colony-stimulating factor. Bone. Jan 2009;44(1):71-9. doi:10.1016/j.bone.2008.09.015
• 11. Mohammad CA. Efficacy of Curcumin Gel on Zinc, Magnesium, Copper, IL-1β, and TNF-α in Chronic Periodontitis Patients. Biomed Res Int. 2020;2020:8850926. doi:10.1155/2020/8850926
• 12. Kuzmanova D, Jansen ID, Schoenmaker T, et al. Vitamin C in plasma and leucocytes in relation to periodontitis. J Clin Periodontol. Oct 2012;39(10):905-12. doi:10.1111/j.1600-051X.2012.01927.x
• 13. Cathcart RF, 3rd. Vitamin C in the treatment of acquired immune deficiency syndrome (AIDS). Med Hypotheses. Aug 1984;14(4):423-33. doi:10.1016/0306-9877(84)90149-x
• 14. Mason SA, Keske MA, Wadley GD. Effects of Vitamin C Supplementation on Glycemic Control and Cardiovascular Risk Factors in People With Type 2 Diabetes: A GRADE-Assessed Systematic Review and Meta-analysis of Randomized Controlled Trials. Diabetes Care. Feb 2021;44(2):618-630. doi:10.2337/dc20-1893
• 15. Lynch SM, Gaziano JM, Frei B. Ascorbic acid and atherosclerotic cardiovascular disease. Subcell Biochem. 1996;25:331-67. doi:10.1007/978-1-4613-0325-1_17
• 16. Amaliya, Timmerman MF, Abbas F, et al. Java project on periodontal diseases: the relationship between vitamin C and the severity of periodontitis. J Clin Periodontol. Apr 2007;34(4):299-304. doi:10.1111/j.1600-051X.2007.01053.x
• 17. Amarasena N, Ogawa H, Yoshihara A, Hanada N, Miyazaki H. Serum vitamin C-periodontal relationship in community-dwelling elderly Japanese. J Clin Periodontol. Jan 2005;32(1):93-7. doi:10.1111/j.1600-051X.2004.00643.x
• 18. Morris MS, Lee Y, Lavin MT, et al. Injectable simvastatin in periodontal defects and alveolar ridges: pilot studies. J Periodontol. Aug 2008;79(8):1465-73. doi:10.1902/jop.2008.070659
• 19. Bradley AD, Zhang Y, Jia Z, et al. Effect of Simvastatin Prodrug on Experimental Periodontitis. J Periodontol. May 2016;87(5):577-82. doi:10.1902/jop.2016.150599
• 20. Oliva F, Maffulli N, Gissi C, et al. Combined ascorbic acid and T(3) produce better healing compared to bone marrow mesenchymal stem cells in an Achilles tendon injury rat model: a proof of concept study. J Orthop Surg Res. Feb 18 2019;14(1):54. doi:10.1186/s13018-019-1098-9
• 21. Souza M, Moraes SAS, de Paula DR, et al. Local treatment with ascorbic acid accelerates recovery of post-sutured Achilles tendon in male Wistar rats. Braz J Med Biol Res. 2019;52(9):e8290. doi:10.1590/1414-431x20198290
• 22. Aytekin Z, Arabacı T, Toraman A, Bayır Y, Albayrak M, Üstün K. Immune modulatory and antioxidant effects of locally administrated vitamin C in experimental periodontitis in rats. Acta Odontol Scand. Aug 2020;78(6):425-432. doi:10.1080/00016357.2020.1734656
• 23. Toraman A, Arabaci T, Aytekin Z, Albayrak M, Bayir Y. Effects of vitamin C local application on ligature-induced periodontitis in diabetic rats. J Appl Oral Sci. 2020;28:e20200444. doi:10.1590/1678-7757-2020-0444
• 24. Unal B, Kara A, Aksak S, Unal D. A stereological assessment method for estimating the surface area of cycloids. Eurasian J Med. Aug 2010;42(2):66-73. doi:10.5152/eajm.2010.20
• 25. Susin C, Rösing CK. Effect of variable moderate chronic stress on ligature-induced periodontal disease in Wistar rats. Acta Odontol Scand. Oct 2003;61(5):273-7. doi:10.1080/00016350310005574
• 26. Grossi SG, Genco RJ. Periodontal disease and diabetes mellitus: a two-way relationship. Ann Periodontol. Jul 1998;3(1):51-61. doi:10.1902/annals.1998.3.1.51
• 27. Ribeiro J, Leão A, Novaes AB. Periodontal infection as a possible severity factor for rheumatoid arthritis. J Clin Periodontol. Apr 2005;32(4):412-6. doi:10.1111/j.1600-051X.2005.00689.x
• 28. Demmer RT, Desvarieux M. Periodontal infections and cardiovascular disease: the heart of the matter. J Am Dent Assoc. Oct 2006;137 Suppl:14S-20S; quiz 38S. doi:10.14219/jada.archive.2006.0402
• 29. Ekuni D, Tomofuji T, Sanbe T, et al. Vitamin C intake attenuates the degree of experimental atherosclerosis induced by periodontitis in the rat by decreasing oxidative stress. Arch Oral Biol. May 2009;54(5):495-502. doi:10.1016/j.archoralbio.2009.02.006
• 30. Tomofuji T, Ekuni D, Irie K, et al. Preventive effects of a cocoa-enriched diet on gingival oxidative stress in experimental periodontitis. J Periodontol. Nov 2009;80(11):1799-808. doi:10.1902/jop.2009.090270
• 31. Kaur G, Kathariya R, Bansal S, Singh A, Shahakar D. Dietary antioxidants and their indispensable role in periodontal health. J Food Drug Anal. 2016/04/01/ 2016;24(2):239-246. doi:https://doi.org/10.1016/j.jfda.2015.11.003
• 32. Chapple IL, Matthews JB. The role of reactive oxygen and antioxidant species in periodontal tissue destruction. Periodontol 2000. 2007;43(1):160-232.
• 33. Tomofuji T, Ekuni D, Sanbe T, et al. Effects of vitamin C intake on gingival oxidative stress in rat periodontitis. Free Radic Biol Med. Jan 15 2009;46(2):163-8. doi:10.1016/j.freeradbiomed.2008.09.040
• 34. Lihm H, Kim H, Chang H, Yoon M, Lee K, Choi J. Vitamin C modulates lead excretion in rats. Anat Cell Biol. Dec 2013;46(4):239-45. doi:10.5115/acb.2013.46.4.239
• 35. Karabulut-Bulan O, Bolkent S, Yanardag R, Bilgin-Sokmen B. The role of vitamin C, vitamin E, and selenium on cadmium-induced renal toxicity of rats. Drug Chem Toxicol. 2008;31(4):413-26. doi:10.1080/01480540802383200
• 36. Esposito K, Nappo F, Marfella R, et al. Inflammatory cytokine concentrations are acutely increased by hyperglycemia in humans: role of oxidative stress. Circulation. Oct 15 2002;106(16):2067-72. doi:10.1161/01.cir.0000034509.14906.ae
• 37. Rathinasamy K, Ulaganathan A, Ramamurthy S, Ganesan R, Saket P, Alamelu S. Estimation of TNF-α Levels in Saliva and Serum of Patients with Periodontal Health and Chronic Periodontitis: A Case-control Study. J Contemp Dent Pract. Feb 1 2020;21(2):148-151.
• 38. Afacan B, Öztürk V, Paşalı Ç, Bozkurt E, Köse T, Emingil G. Gingival crevicular fluid and salivary HIF-1α, VEGF, and TNF-α levels in periodontal health and disease. J Periodontol. Jul 2019;90(7):788-797. doi:10.1002/jper.18-0412
• 39. Molina N, Morandi AC, Bolin AP, Otton R. Comparative effect of fucoxanthin and vitamin C on oxidative and functional parameters of human lymphocytes. Int Immunopharmacol. Sep 2014;22(1):41-50. doi:10.1016/j.intimp.2014.06.026
• 40. Sorsa T, Tjäderhane L, Konttinen YT, et al. Matrix metalloproteinases: contribution to pathogenesis, diagnosis and treatment of periodontal inflammation. Ann Med. 2006;38(5):306-21. doi:10.1080/07853890600800103
• 41. Hardy DC, Ross JH, Schuyler CA, Leite RS, Slate EH, Huang Y. Matrix metalloproteinase-8 expression in periodontal tissues surgically removed from diabetic and non-diabetic patients with periodontal disease. J Clin Periodontol. Mar 2012;39(3):249-55. doi:10.1111/j.1600-051X.2011.01788.x
• 42. Carr AC, Maggini S. Vitamin C and Immune Function. Nutrients. Nov 3 2017;9(11)doi:10.3390/nu9111211
• 43. Xiao XH, Liao EY, Zhou HD, Dai RC, Yuan LQ, Wu XP. Ascorbic acid inhibits osteoclastogenesis of RAW264.7 cells induced by receptor activated nuclear factor kappaB ligand (RANKL) in vitro. J Endocrinol Invest. Mar 2005;28(3):253-60. doi:10.1007/bf03345382
• 44. Ishikawa S, Iwasaki K, Komaki M, Ishikawa I. Role of ascorbic acid in periodontal ligament cell differentiation. J Periodontol. May 2004;75(5):709-16. doi:10.1902/jop.2004.75.5.709
• 45. Abou Sulaiman AE, Shehadeh RM. Assessment of total antioxidant capacity and the use of vitamin C in the treatment of non-smokers with chronic periodontitis. J Periodontol. Nov 2010;81(11):1547-54. doi:10.1902/jop.2010.100173
• 46. Yussif NM, Abdul Aziz MA, Abdel Rahman AR. Evaluation of the Anti-Inflammatory Effect of Locally Delivered Vitamin C in the Treatment of Persistent Gingival Inflammation: Clinical and Histopathological Study. J Nutr Metab. 2016;2016:2978741. doi:10.1155/2016/2978741
• 47. Greenstein G. Local drug delivery in the treatment of periodontal diseases: assessing the clinical significance of the results. J Periodontol. Apr 2006;77(4):565-78. doi:10.1902/jop.2006.050140