Evaluation of Gingival Crevicular Interleukin-1β Levels in Patients Undergoing Clear Aligners vs. Fixed Orthodontic Therapy

Areeba  Khan; Rabiya  Saif; Muhammad Mohsin  Kamal; Muhammad Iftikhar  Ahsen; Ghazala  Hassan; Eesha  Muneeb; Ejaz  Ahmed

doi:10.36283/ziun-pjmd14-4/067

Authors

Areeba Khan Superior University ,Lahore, Pakistan.
Rabiya Saif de'Montmorency College of Dentistry, Lahore, Pakistan.
Muhammad Mohsin Kamal Fatima Memorial Dental Hospital, Lahore, Pakistan.
Muhammad Iftikhar Ahsen Rahbar College of Dentistry, Lahore, Pakistan.
Ghazala Hassan Fatima Memorial College of Medicine and Dentistry, Lahore, Pakistan.
Eesha Muneeb Rahbar College of Dentistry , Lahore, Pakistan.
Ejaz Ahmed International Max Planck Research Institute, Berlin, Germany.

DOI:

https://doi.org/10.36283/ziun-pjmd14-4/067

Keywords:

Orthodontic Appliances, Gingival Crevicular Fluid, Interleukin-1beta, Periodontal Index

Abstract

Background: Tooth movements induced by orthodontic force cause an inflammatory response in the periodontium that is mediated by cytokines, including Interleukin-1beta (IL-1β). Fixed appliances produce different biomechanical forces, which have the potential to modulate GCF cytokines compared to clear aligners. The study aimed to determine the levels of gingival crevicular fluid (GCF) IL-1β in patients receiving clear aligner treatment compared to those receiving fixed appliances.

Methods: This cross-sectional analytical research was conducted at a tertiary care dental hospital between From February to June 2025. 100 (50 clear aligner patients and 50 fixed appliance patients), 18-30 years old participants were recruited. Samples of GCF were obtained as a baseline and 1 month after treatment. IL-1β levels were measured by Enzyme-Linked Immunosorbent Assay (ELISA). The significance level was set as p < 0.05.

Results: No significant difference was found between baseline IL-1 levels (54.7 ± 9.3 vs. 56.1 ± 10.1 pg/L, CI: 2.5-5.3, p = 0.47). At 1 month, IL-1β levels were significantly elevated in the non-MAR group (92.4 ± 12.6 vs. 71.8 ± 10.9 pg/L, CI: 16.3-24.9, p < 0.001). Increases within the group were noted in both groups (clear aligners: 17.1 ± 6.2; fixed appliances: 36.3 ± 7.8, both p < 0.001). The difference between the groups persisted after controlling for confounders (CI: 15.7-23.0, p < 0.001).

Conclusion: Both appliance systems increased GCF IL-1β levels, but fixed appliances produced a significantly larger inflammatory response than clear aligners, even after adjustment for potential confounders.

Author Biographies

Areeba Khan, Superior University ,Lahore, Pakistan.

Department of Dental,
Rabiya Saif, de'Montmorency College of Dentistry, Lahore, Pakistan.

Department of Oral Pathology,
Muhammad Mohsin Kamal, Fatima Memorial Dental Hospital, Lahore, Pakistan.

Department of Community and Preventive Dentistry,
Muhammad Iftikhar Ahsen, Rahbar College of Dentistry, Lahore, Pakistan.

Department of Periodontology,
Ghazala Hassan, Fatima Memorial College of Medicine and Dentistry, Lahore, Pakistan.

Department of Oral Biology,
Eesha Muneeb, Rahbar College of Dentistry , Lahore, Pakistan.

Department of Orthodontics,
Ejaz Ahmed, International Max Planck Research Institute, Berlin, Germany.

School of Pathology,

References

1. Sadek KM, El Moshy S, Radwan IA, Rady D, Abbass MMS, El-Rashidy AA, et al. Molecular Basis beyond Interrelated Bone Resorption/Regeneration in Periodontal Diseases: A Concise Review. Int J Mol Sci. 2023 Feb 27;24(5):4599. doi: 10.3390/ijms24054599.

2. Jiang Q, Li J, Mei L, Du J, Levrini L, Abbate GM, et al. Periodontal health during orthodontic treatment with clear aligners and fixed appliances: A meta-analysis. J Am Dent Assoc. 2018 Aug;149(8):712-720.e12. doi: 10.1016/j.adaj.2018.04.010.

3. Chelărescu S, Șurlin P, Decusară M, Oprică M, Bud E, Teodorescu E, et al. Evaluation of IL1β and IL6 gingival crevicular fluid levels during the early phase of orthodontic tooth movement in adolescents and young adults. Appl. Sci. 2021 Jan 7;11(2):521. doi: 10.3390/APP11020521.

4. Dipalma G, Inchingolo AD, Fiore A, Balestriere L, Nardelli P, Casamassima L, et al. The Differential Impact of Clear Aligners and Fixed Orthodontic Appliances on Periodontal Health: A Systematic Review. Children (Basel). 2025 Jan 26;12(2):138. doi: 10.3390/children12020138.

5. Kredig C, Peuckert E, Schmidtmann I, Drechsler T, Erbe C. Oral health in adolescents: periodontal inflammatory biomarkers during orthodontic clear aligner therapy. Clin Oral Investig. 2025 Mar 5;29(3):168. doi: 10.1007/s00784-025-06212-9.

6. Pascoal S, Gonçalves A, Brandão A, Rocha D, Oliveira S, Monteiro F, et al. Human Interleukin-1β Profile and Self-Reported Pain Monitoring Using Clear Aligners with or without Acceleration Techniques: A Case Report and Investigational Study. Int J Dent. 2022 Aug 31;2022:8252696. doi: 10.1155/2022/8252696.

7. Alnazeh AA, Kamran MA, Aseeri Y, Alrwuili MR, Aljabab MA, Baig EA, et al. Levels of Inflammatory and Bone Metabolic Markers in the Gingival Crevicular Fluid of Individuals Undergoing Fixed Orthodontic Treatment in Comparison to Those Utilizing Invisalign. Medicina (Kaunas). 2023 Dec 1;59(12):2107. doi: 10.3390/medicina59122107.

8. Gonçalves A, Mathelié-Guinlet Q, Ramires F, Monteiro F, Carvalho Ó, Silva FS, et al. Biological alterations associated with the orthodontic treatment with conventional appliances and aligners: A systematic review of clinical and preclinical evidence. Heliyon. 2024 Jun 13;10(12):e32873. doi: 10.1016/j.heliyon.2024.e32873.

9. Inchingolo F, Inchingolo AM, Malcangi G, Ferrante L, Trilli I, Di Noia A, et al. The interaction of cytokines in orthodontics: A systematic review. Appli. Sci. 2024 Jun 13;14(12):5133. doi: 10.3390/APP14125133.

10. Sharma S, Pamnani M. Evaluation of salivary cytokines in patients with fixed orthodontic treatment. Int. J. of Health Sci. 2022(III).:2250-6. doi: 10.53730/ijhs.v6nS3.5996.

11. Bhardwaj R, Agrawal U, Vashist P, Manna S. Determination of sample size for various study designs in medical research: A practical primer. J Family Med Prim Care. 2024 Jul;13(7):2555-2561. doi: 10.4103/jfmpc.jfmpc_1675_23.

12. Kitaura H, Ohori F, Marahleh A, Ma J, Lin A, Fan Z, et al. The Role of Cytokines in Orthodontic Tooth Movement. Int J Mol Sci. 2025 Jul 11;26(14):6688. doi: 10.3390/ijms26146688.

13. Di Spirito F, D'Ambrosio F, Cannatà D, D'Antò V, Giordano F, Martina S. Impact of Clear Aligners versus Fixed Appliances on Periodontal Status of Patients Undergoing Orthodontic Treatment: A Systematic Review of Systematic Reviews. Healthcare (Basel). 2023 May 6;11(9):1340. doi: 10.3390/healthcare11091340.

14. Giannini L, Galbiati G, Tartaglia FC, Grecolini ME, Maspero C, Biagi R. Orthodontic Treatment with Fixed Appliances Versus Aligners: An Experimental Study of Periodontal Aspects. Dent J (Basel). 2025 Feb 4;13(2):70. doi: 10.3390/dj13020070.

15. Kumar BD, Singh N, Verma SK, Singh S, Thakur S. A Study to Evaluate Il1 And Il6 Gingival Crevicular Fluid Levels in Adolescents and Young Adults During the Early Phase of Orthodontic Tooth Movement. J Pharm Bioallied Sci. 2022 Jul;14(Suppl 1):S494-S497. doi: 10.4103/jpbs.jpbs_52_22.

16. Kamran MA, Alnazeh AA, Almagbol M, Almoammar S, Alhaizaey AHA, Alshahrani I. Role of six cytokines and bone metabolism biomarkers in gingival crevicular fluid in patients undergoing fixed orthodontic appliance treatment in comparison with aligners: a clinical study. Angle Orthod. 2023 May 1;93(3):335-340. doi: 10.2319/091722-649.1.

17. Luo L, Xu C, Li Y, Hao C, Zheng J, Jin X, et al. The gingival crevicular fluid biomarkers with micropulse vibration device: A pilot study. Heliyon. 2024 May 28;10(12):e31982. doi: 10.1016/j.heliyon.2024.e31982.

18. Baeshen HA. GCF cytokine profile comparison between patients with lingual fixed appliances and aligners. J King Saud Univ Sci. 2022 Aug 1;34(6):102075. doi: 10.1016/J.JKSUS.2022.102075.

19. Liu Q, Wang A, Guo D, Luo H, Fang S, Song Z, et al. Characterization and correlation analysis of oral NET markers and inflammatory factor levels in patients after orthodontic treatment: a pilot study. Front Immunol. 2025 May 6;16:1490637. doi: 10.3389/fimmu.2025.1490637.

20. Chen Y, Mei L, Qian Y, Zhou X, Zhao Z, Zheng W, et al. Integrated bioinformatic analysis of protein landscape in gingival crevicular fluid unveils sequential bioprocess in orthodontic tooth movement. Prog Orthod. 2024 Sep 23;25(1):37. doi: 10.1186/s40510-024-00536-0.

21. Debnath P, Bangi SL, Hussain MF, Rafiq S, Tousifulla S, Abdu M, et al. The Evaluation of Gingival Crevicular Fluid Biomarkers as Predictors of Gingival Enlargement in Patients Undergoing Fixed Orthodontic Treatment: A Prospective Study. Cureus. 2024 Nov 22;16(11):e74281. doi: 10.7759/cureus.74281.

22. Bud A, Lazăr L, Mârțu MA, Dakó T, Suciu M, Vlasiu A, et al. Challenges and Perspectives Regarding the Determination of Gingival Crevicular Fluid Biomarkers During Orthodontic Treatment: A Narrative Review. Medicina (Kaunas). 2024 Dec 4;60(12):2004. doi: 10.3390/medicina60122004.

23. Nemec M, Ringl P, Spettel K, Schneider L, Kriz R, Galazka S, et al. Exploring the impact of orthodontic appliances on the oral microbiome and inflammatory parameters. Prog Orthod. 2025 Apr 7;26(1):13. doi: 10.1186/s40510-025-00560-8. Erratum in: Prog Orthod. 2025 Aug 21;26(1):29. doi: 10.1186/s40510-025-00577-z.

24. Cm AR, Ghonmode S, Powar S, Rajput P, Chaudhary P. Association Between Different Biomarkers and Initial Orthodontic Tooth Movement in Children and Adults: A Systematic Review and Meta-Analysis. Cureus. 2025 Feb 4;17(2):e78483. doi: 10.7759/cureus.78483.

25. Hussein S, Ismail H. Influence of Reminder on Enhancing Compliance in Patients with Fixed Orthodontic Appliance Treatment (a Randomized Controlled Clinical Trial). Patient Prefer Adherence. 2023 Jul 20;17:1759-1769. doi: 10.2147/PPA.S418109.

26. Cuevas-González MV, Cuevas-González JC, Espinosa-Cristóbal LF, Tovar-Carrillo KL, Saucedo-Acuña RA, García-Calderón AG, et al. The Potential of Gingival Crevicular Fluid as a Tool for Molecular Diagnosis: A Systematic Review. Biomed Res Int. 2024 Oct 15;2024:5560866. doi: 10.1155/2024/5560866.