Nicotine reduces cell viability and induces oxidative stress in human gingival fibroblasts

Main Article Content

Sabrina Azmi
Restu Syamsul Hadi
Indra Kusuma
Yulia Suciati
Wening Sari


Nicotine, as the main component of cigarettes, is known to interfere with the proliferation of human gingival fibroblasts (HGFs) and can trigger oxidative stress. This study aimed to analyze the impact of nicotine on viability, expression of the antioxidant Nrf2, levels of the product of oxidative stress malondialdehyde (MDA), and the migration capacity of HGFs.

An experimental laboratory study used fibroblasts isolated from healthy human gingiva. The cells were grouped into the non-treatment control group (NTC), the solvent control (SC), and the treatment groups, exposed to nicotine at various concentrations for twenty-four hours. Cell viability was assesed using the cell counting kit-8 (CCK-8), Nrf2 expression was examined using ELISA, MDA level was measured using an MDA kit, and migration capacity was assessed using a scratch assay. Statistical analysis used one-way Anova or Kruskal-Wallis test. A p-value of <0.05 was expressed statistically significant.

The Cell viability was substantially reduced in the nicotine group compared to the untreated group, accompanied by changes in cell morphology. In contrast, Nrf2 expression increased significantly (p=0.010) in the 5 mM nicotine group compared with the control group. The MDA levels were not significantly distinct across groups (p=0.056). Cell migration was delayed significantly in the 5 mM nicotine group at 72 hours after scratching compared to the control group.

Nicotine decreased HGFs viability and increased Nrf2 expression significantly in a dose-dependent manner. Nicotine at 5 mM concentration did not alter MDA levels but delayed cell migration.

Article Details

How to Cite
Azmi, S. ., Hadi, R. S. . . ., Kusuma, I., Suciati, Y. ., & Sari, W. (2024). Nicotine reduces cell viability and induces oxidative stress in human gingival fibroblasts. Universa Medicina, 43(1), 20–30.
Original Articles


Dai X, Gil GF, Reitsma MB, et al. Health effects associated with smoking: a burden of proof study. Nat Med 2022;28:2045–55. doi: 10.1038.s41591-022-01978-x.

Li Y, Hecht SS. Carcinogenic components of tobacco and tobacco smoke: a 2022 update. Food Chem Toxicol 2022;165:113179. doi: 10.1016/ j.fct.2022.113179.

World Health Organization. Tobacco. Geneva: World Health Orgaization; 2023.

Survei Sosial Ekonomi Nasional. [Percentages of smokers in population aged ≥15 years by province (Percentage), 2021-2023.] Jakarta: Badan Pusat Statistik. 2024.Indonesian.

Esfahrood ZR, Zamanian A, Torshabi M, Abrishami M. The effect of nicotine and cotinine on human gingival fibroblasts attachment to root surfaces. J Basic Clin Physiol Pharmacol 2015;26: 517–22. doi: 10.1515/jbcpp-2014-0120.

Trybek G, Preuss O, Aniko-Wlodarczyk M, et al. The effect of n‪icotine on oral health. Balt J Health Phys Act 2018;10:7–13. doi: 10.29359/bjhpa. 10.2.01.‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬‬

Katti N, Asif K, Mohanty D, Shatapathy N. Determination of efficacy of root planing in removal of nicotine from periodontally involved teeth of smokers. Chronicles Young Scientists 2012;3:160. doi: 10.4103/2229-5186.98691.

Madi M, Smith S, Alshehri S, Zakaria O, Almas K. Influence of smoking on periodontal and implant therapy: a narrative review. Int J Environ Res Public Health 2023;20:5368. doi: 10.3390/ ijerph20075368.

Chen CS, Lee SS, Yu HC, Huang FM, Chang YC. Effects of nicotine on cell growth, migration, and production of inflammatory cytokines and reactive oxygen species by cementoblasts. J Dent Sci 2015;10:154–60. doi: 10.1016/j.jds.2014.04. 002.

Shang J, Liu H, Zheng Y, Zhang Z. Role of oxidative stress in the relationship between periodontitis and systemic diseases. Front Physiol 2023;14:1210449. doi: 10.3389/fphys.2023. 1210449.

Sczepanik FSC, Grossi ML, Casati M, et al. Periodontitis is an inflammatory disease of oxidative stress: we should treat it that way. Periodontol 2000 2020;84:45-68. doi: 10.1111/ prd.12342Vol.84,

Kumari R, Jat P. Mechanisms of cellular senescence: cell cycle arrest and senescence associated secretory phenotype. Front Cell Dev Biol 2021;9:645593. doi: 10.3389/fcell.2021. 645593.

Klran TR, Otlu O, Karabulut AB. Oxidative stress and antioxidants in health and disease. J Lab Med 2023;47:1-11.

Haro Girón S, Monserrat Sanz J, Ortega MA, et al. Prognostic value of malondialdehyde (MDA) in the temporal progression of chronic spinal cord injury. J Pers Med 2023;13:626. doi: 10.3390/ jpm13040626.

Cui X, Gong J, Han H, et al. Relationship between free and total malondialdehyde, a well-established marker of oxidative stress, in various types of human biospecimens. J Thorac Dis 2018;10: 3088–197. doi: 10.21037/jtd.2018.05.92.

Wang Y, Andrukhov O, Rausch-Fan X. Oxidative stress and antioxidant system in periodontitis. Front Physiol 2017;8:910. doi: 10.3389/fphys. 2017.00910.

Ngo V, Duennwald ML. Nrf2 and oxidative stress: a general overview of mechanisms and implications in human disease. Antioxidants (Basel) 2022;11:2345. doi: 10.3390/ antiox11122345.

Chiu AV, Al Saigh M, McCulloch CA, Glogauer M. The role of NrF2 in the regulation of periodontal health and disease. J Dent Res 2017; 96:975–83. doi: 10.1177/0022034517715007.

Ying S, Tan M, Feng G, et al. Low-intensity pulsed ultrasound regulates alveolar bone homeostasis in experimental Periodontitis by diminishing oxidative stress. Theranostics. 2020; 10:9789–807. doi: 10.7150/thno.42508.

Egea G, Jiménez‐Altayó F, Campuzano V. Reactive oxygen species and oxidative stress in the pathogenesis and progression of genetic diseases of the connective tissue. Antioxidants (Basel) 2020;9:1013. doi: 10.3390/antiox 9101013.

Soares ASLS, Scelza MZ, Spoladore J, et al. Comparison of primary human gingival fibroblasts from an older and a young donor on the evaluation of cytotoxicity of denture adhesives. J Appl Oral Sci 2018;26:e20160594. doi: 10.1590/1678-7757-2016-0594.

Pensalfini M, Tepole AB. Mechano-biological and bio-mechanical pathways in cutaneous wound healing. PLoS Comput Biol 2023;19:e1010902. doi: 10.1371/journal.pcbi.1010902.

Kanmaz M, Kanmaz B, Buduneli N. Periodontal treatment outcomes in smokers: a narrative review. Tob Induc Dis 2021;19:77. doi: 10.18332/tid/142106.

Hardin LT, Vang D, Thor D, et al. Cigarette smoking exposure disrupts the regenerative potential of dental pulp stem cells. Tob Induc Dis 2023;21:101. doi: 10.18332/tid/168125.

Dinos ME, Borke JL, Swiec GD, McPherson JC, Goodin JL, Chuang AH. In vitro study of the adverse effect of nicotine and physical strain on human gingival fibroblasts as a model of the healing of wounds commonly found in the military. Mil Med 2015;180:86–91. https:// 00382.

Lallier TE, Moylan JT, Maturin E. Greater sensitivity of oral fibroblasts to smoked versus smokeless tobacco. J Periodontol 2017;88:1356–65.

Nishioka T, Tada H, Ibaragi S, Chen C, Sasano T. Nicotine exposure induces the proliferation of oral cancer cells through the α7 subunit of the nicotinic acetylcholine receptor. Biochem Biophys Res Commun 2019;509:514–20. doi: 10.1016/j.bbrc.2018.12.154.

Park CM, Yoon HS. Strengthened antioxidative potential by Gelidium amansii ethanol extract through the induction of phase II enzymes in human gingival fibroblast cells . Int J Clin Prev Dent 2018;14:157-61. ijcpd.2018.14.3.157.

Xue J, Liao Q, Luo M, et al. Cigarette smoke-induced oxidative stress activates NRF2 to mediate fibronectin disorganization in vascular formation. Open Biol 2022;12:210310. doi: 10.1098/rsob.210310.

Li X, Sun X, Zhang X, et al. Enhanced oxidative damage and Nrf2 downregulation contribute to the aggravation of periodontitis by diabetes mellitus. Oxid Med Cell Longev 2018;2018:1-11. doi: 10.1155/2018/9421019.

Begum SF, Nagajothi G, Latha KS, et al. Possible role of nicotine and cotinine on nitroxidative stress and antioxidant content in saliva of smokeless tobacco consumers. Pract Lab Med 2018;12:e00105. doi: 10.1016/j.plabm.2018. e00105.

Zięba S, Maciejczyk M, Zalewska A. Ethanol- and cigarette smoke-related alternations in oral redox homeostasis. Front Physiol 2022;12: 793028. doi: 10.3389/fphys.2021.793028.

Padhye NM, Padhye AM, Gupta HS. Quantification and comparison of the impact of the smoking status on oral polymorphonuclear leukocyte and malondialdehyde levels in individuals with chronic periodontitis: a double-blinded longitudinal interventional study. Contemp Clin Dent 2019;10:517-24. doi: 10.4103/ccd.ccd_906_18.

Naresh CK, Rao SM, Shetty PR, Ranganath V, Patil AS, Anu AJ. Salivary antioxidant enzymes and lipid peroxidation product malondialdehyde and sialic acid levels among smokers and non-smokers with chronic periodontitis—a clinico-biochemical study. J Family Med Prim Care 2019;8:2960. doi: 10.4103/jfmpc.jfmpc_438_19.

Khademi F, Totonchi H, Mohammadi N, Zare R, Zal F. Nicotine-induced oxidative stress in human primary endometrial cells. Int J Toxicol 2019;38: 202–8. doi: 10.1177/1091581819848081.

Nguyen TT, Huynh NNC, Seubbuk S, Nilmoje T, Wanasuntronwong A, Surarit R. Oxidative stress induced by porphyromonas gingivalis lysate and nicotine in human periodontal ligament fibroblasts. Odontology 2019;107:133–41. doi: 10.1007/s10266-018-0374-1.

SenGupta S, Parent CA, Bear JE. The principles of directed cell migration. Nat Rev Mol Cell Biol 2021;22:529-547. doi: 10.1038/s41580-021-00366-6.

Silva D, Cáceres M, Arancibia R, Martínez C, Martínez J, Smith PC. Effects of cigarette smoke and nicotine on cell viability, migration and myofibroblastic differentiation. J Periodontal Res 2012;47:599–607. doi: 10.1111/j.1600-0765. 2012.01472.x.

Silva H. Tobacco use and periodontal disease—the role of microvascular dysfunction. Biology (Basel) 2021;10:441. doi: 10.3390/biology 10050441.

Missirlis D, Haraszti T, Kessler H, Spatz JP. Fibronectin promotes directional persistence in fibroblast migration through interactions with both its cell-binding and heparin-binding domains. Sci Rep 2017;7:3711. 10.1038/ s41598-017-03701-0.