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Submitted: 14 Jun 2025
Revision: 28 Oct 2025
Accepted: 07 Nov 2025
ePublished: 31 Dec 2025
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Avicenna J Med Biochem. 2025;13(2): 89-96.
doi: 10.34172/ajmb.2676
  Abstract View: 4
  PDF Download: 1

Original Article

Western Blotting of Salivary Interleukin-1β and Tumor Necrosis Factor-α for Non-Invasive Diagnosis of Orthodontically Induced Root Resorption

Homa Farhadifard 1 ORCID logo, Parnia Yari 1 ORCID logo, Amir Taherkhani 2,3* ORCID logo, Vahid Siavashi 4 ORCID logo, Parzhin Moradi 1 ORCID logo

1 Department of Orthodontics, School of Dentistry, Hamadan Dental Research Center, Hamadan University of Medical Sciences, Hamadan, Iran
2 Research Center for Molecular Medicine, Institute of Cancer, Avicenna Health Research Institute, Hamadan University of Medical Sciences, Hamadan, Iran
3 Urology and Nephrology Research Center, Avicenna Institute of Clinical Sciences, Hamadan University of Medical Sciences, Hamadan, Iran
4 Azma Teb Gostar Sorena Research Company, Basic Medical Science Research Center, Tehran, Iran
*Corresponding Author: Amir Taherkhani, Email: a.taherkhani@umsha.ac.ir, Email: amir.007.taherkhani@gmail.com

Abstract

Introduction: Orthodontically induced inflammatory root resorption (OIIRR) is an unpredictable side effect of orthodontic treatment, typically identified only after irreversible damage. This study sought to assess whether the salivary concentrations of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) could function as dependable diagnostic indicators for OIIRR. It was hypothesized that OIIRR patients would demonstrate higher salivary TNF-α and IL-1β than controls, and that these cytokines would be accurate diagnostic biomarkers.

Methods: A case-control study was conducted involving 24 orthodontic patients, 12 with radiographically verified OIIRR and 12 controls free of root resorption, comparable in age and gender. Whole saliva samples were obtained, and protein expression of IL-1β and TNF-α was quantified via Western blotting using β-actin as a reference. Statistical methods included independent-samples t-tests, Cohen’s d for effect size estimation, and receiver operating characteristic (ROC) curve analysis.

Results: Salivary IL-1β was significantly higher in cases (1.36±0.33) than in controls (0.68±0.16; t(22)=6.32, P<0.0001; Cohen’s d=2.58). Likewise, TNF-α was notably elevated in cases (1.38±0.29) versus controls (0.80±0.15; t(22)=6.27, P<0.0001; Cohen’s d=2.56). Moreover, ROC analysis revealed outstanding diagnostic performance for both markers, with AUC values of 0.986 for IL-1β and 0.951 for TNF-α. Optimal cutoffs were also 0.875 and 1.1 for IL-1β and TNF-α, respectively.

Conclusion: Our findings indicated that both cytokines represent valuable non-invasive salivary biomarkers for the early diagnosis and monitoring of OIIRR, potentially supporting safer orthodontic care. Nonetheless, larger prospective studies are needed for confirmation.



Please cite this article as follows: Farhadifard H, Yari P, Taherkhani A, Siavashi V, Moradi P. Western blotting of salivary interleukin-1β and tumor necrosis factor-α for non-invasive diagnosis of orthodontically induced root resorption. Avicenna J Med Biochem 2025;13(2): 89-96. doi:10.34172/ajmb.2676
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