Comparison of the effect of postoperative care agents on human gingival fibroblasts: a preliminary study

Abstract

Purpose The aim of this study is to compare effects of postoperative care agents; chlorhexidine, octenidine dihydrochloride and hyaluronic acid on human gingival fibroblasts’ viability, proliferation, apoptosis and migration. Material and Methods After cell culturing; chlorhexidine, octenidine dihydrochloride and hyaluronic acid solutions were applied on cells and nothing was applied for control group. The cells were monitored to investigate cytotoxicity; the percentage of apoptotic, living and dead cells at the time of 24, 48, and 72 hours (h). A scratch wound assay was performed to detect cell migration and cells were monitored at baseline, at 24 and 48h. Results At 24h, chlorhexidine showed statistically lower percentage of total apoptotic cells’ than octenidine dihydrochloride (p=0.049), hyaluronic acid (p=0.049) and control (p=0.049). At 48h, hyaluronic acid showed statistically lower percentage than chlorhexidine (p=0.049), and control (p=0.049). All agents were found to have statistically and significantly more cytotoxic than control. However, there was no difference between experimental groups for proliferation rate. Octenidine dihydrochloride showed statistically negative effects on cell migration than chlorhexidine and hyaluronic acid at 24h. Chlorhexidine and hyaluronic acid maintained migration ability of cells than octenidine dihydrochloride at 48h. Conclusion All agents have similar effects on cell behavior such as viability, apoptosis and cell proliferation. However, octenidine dihydrochloride showed statistically negative effects on migration ability than chlorhexidine and hyaluronic acid.

Keywords

• Chlorhexidine,hyaluronic acid,octenidine dihydrochloride,cell viability,cell migration

References

1. 1. Cabral CT, Fernandes MH. In vitro comparison of chlorhexidine and povidone-iodine on the long- term proliferation and functional activity of human alveolar bone cells. Clin Oral Investig 2007;11(2):155-64. [CrossRef]
2. 2. Gocmen G, Gonul O, Oktay NS, Yarat A, Goker K. The antioxidant and anti-inflammatory efficiency of hyaluronic acid after third molar extraction. J Craniomaxillofac Surg 2015;43(7):1033-7. [CrossRef]
3. 3. Schmidt J, Zyba V, Jung K, Rinke S, Haak R, Mausberg RF, Ziebolz D. Cytotoxic effects of octenidine mouth rinse on human fibroblasts and epithelial cells - an in vitro study. Drug Chem Toxicol 2016;39(3):322-30. [CrossRef]
4. 4. Löe H, Schiott CR. The effect of mouthrinses and topical application of chlorhexidine on the development of dental plaque and gingivitis in man. J Periodontal Res 1970;5(2):79-83. [CrossRef]
5. 5. Li YC, Kuan YH, Lee SS, Huang FM, Chang YC. Cytotoxicity and genotoxicity of chlorhexidine on macrophages in vitro. Environ Toxicol 2014;29(4):452-8. [CrossRef]
6. 6. Hidalgo E, Dominguez C. Mechanisms underlying chlorhexidineinduced cytotoxicity. Toxicol In Vitro 2001;15(4-5):271-6. [CrossRef]
7. 7. Koray M, Ofluoglu D, Onal Ea, Ozgul M, Ersev H, Yaltirik M, Tanyeri H. Efficacy of hyaluronic acid spray on swelling, pain, and trismus after surgical extraction of impacted mandibular third molars. Int J Oral Maxillofac Surg 2014;43(11):1399-403. [CrossRef]
8. 8. Al-Bayaty1 FH, Taıyeb-Ali TB, Abdulla MA, Mahmud ZB. Antibacterial effects of Oradex, Gengigel and Salviathymol-n mouthwash on dental biofilm bacteria. Afr J Microbiol Res 2011;5(6): 636-642.

9. 9. Eick S, Goltz S, Nietzsche S, Jentsch H, Pfister W. Efficacy of chlorhexidine digluconate- containing formulations and other mouthrinses against periodontopathogenic microorganisms. Quintessence Int 2011;42(8):687-700.
10. 10. Van Meerloo J, Kaspers GJ, Cloos J. Cell sensitivity assays: the MTT assay. Methods Mol Biol 2011;731:237–45. [CrossRef]
11. 11. Stockert Jc, Blázquez-Castro A, Ca-ete M, Horobin Rw, Villanueva A. MTT assay for cell viability: Intracellular localization of the formazan product is in lipid droplets. Acta Histochem 2012;114(8):785-96. [CrossRef]
12. 12. Pietkiewicz S, Schmidt JH, Lavrik IN. Quantification of apoptosis and necroptosis at the single cell level by a combination of Imaging Flow Cytometry with classical Annexin V/propidium iodide staining. J Immunol Methods 2015;423:99-103. [CrossRef]
13. 13. Liang CC, Park AY, Guan JL. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc 2007;2(2):329-33. [CrossRef]
14. 14. Felice F, Zambito Y, Belardinelli E, Fabiano A, Santoni T, Di Stefano R. Effect of different chitosan derivatives on in vitro scratch wound assay: a comparative study. Int J Biol Macromol 2015;76:236-41. [CrossRef]
15. 15. Bucalo B, Eaglstein WH, Falanga V. Inhibition of cell proliferation by chronic wound fluid. Wound Repair Regen 1993;1(3):181-6. [CrossRef]
16. 16. Thomas GW, Rael LT, Bar-Or R, Shimonkevitz R, Mains CW, Slone DS, Craun ML, Bar-Or D. Mechanisms of delayed wound healing by commonly used antiseptics. J Trauma 2009;66(1):82-90; discussion 90-1. [CrossRef]
17. 17. Seymour RA, Heasman PA, Macgregor IDM Anti-plaque and anti-calculus agents. In: Seymour RA, Heasman PA (eds) Drugs, diseases and the periodontium. Oxford Medical Publications, Hong Kong, 1992, pp 153–179.
18. 18. Cousido MC, Tomás Carmona I, García-Caballero L, Limeres J, Alvarez M, Diz P. In vivo substantivity of 0.12% and 0.2% chlorhexidine mouthrinses on salivary bacteria. Clin Oral Investig 2010;14(4):397-402. [CrossRef]
19. 19. Park JB, Lee G, Yun BG, Kim CH, Ko Y. Comparative effects of chlorhexidine and essential oils containing mouth rinse on stem cells cultured on a titanium surface. Mol Med Rep 2014;9(4):1249-53. [CrossRef]
20. 20. Müller G, Kramer A. Biocompatibility index of antiseptic agents by parallel assessment of antimicrobial activity and cellular cytotoxicity. J Antimicrob Chemother 2008;61(6):1281-7. [CrossRef]
21. 21. Foschi D, Castoldi L, Radaelli E, Abelli P, Calderini G, Rastrelli A, Mariscotti C, Marazzi M, Trabucchi E. Hyaluronic acid prevents oxygen free-radical damage to granulation tissue: a study in rats. Int J Tissue React 1990;12(6):333-9.
22. 22. Brown JA. The role of hyaluronic acid in wound healing’s proliferative phase. J Wound Care 2004;13(2):48-51. [CrossRef]
23. 23. Presti D, Scott JE. Hyaluronan-mediated protective effect against cell damage caused by enzymatically produced hydroxyl (OH.) radicals is dependent on hyaluronan molecular mass. Cell Biochem Funct 1994;12(4):281-8. [CrossRef]
24. 24. Mendoza G, Prieto JG, Real R, Pérez M, Merino G, Alvarez AI. Antioxidant profile of hyaluronan: physico-chemical features and its role in pathologies. Mini Rev Med Chem. 2009;9(13):147988. [CrossRef]
25. 25. Ye J, Wu H, Wu Y, Wang C, Zhang H, Shi X, Yang J. High molecular weight hyaluronan decreases oxidative DNA damage induced by EDTA in human corneal epithelial cells. Eye (Lond) 2012;26(7):1012-20. [CrossRef]