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Original Article · Open access

Evaluation of the effects of thymoquinone on fracture healing in experimental bone fractures in rats

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Duygu İlke Yıldırım

Department of Family Medicine, Selçuk University Faculty of Medicine, Konya, Türkiye

ORCID: 0000-0002-3893-5173

Ahmet Yıldırım

Department of Orthopedics and Traumatology, Konya Health Application and Research Center, University of Health Sciences, Konya, Türkiye

ORCID: 0000-0002-3953-091X

Mehmet Sedat Durmaz

Department of Radiology, Selçuk University Faculty of Medicine, Konya, Türkiye

ORCID: 0000-0002-1340-2477

Fikret Akyürek

Department of Biochemistry, Selçuk University Faculty of Medicine, Konya, Türkiye

ORCID: 0000-0002-8091-7737

Nejat Ünlükal

Department of Histology and Embryology, Selçuk University Faculty of Medicine, Konya, Türkiye

ORCID: 0000-0002-8107-4882

Seda Şimşek

Department of Histology and Embryology, Selçuk University Faculty of Medicine, Konya, Türkiye

ORCID: 0000-0003-4237-5831

Serhat Ayan

Department of Microbiology, Faculty of Veterinary Medicine, Selçuk University, Konya, Türkiye

ORCID: 0000-0001-6234-0703

Abstract

Objectives: This study aims to investigate whether thymoquinone (TQ) accelerates fracture healing and modulates inflammatory and regenerative markers in a rat model.

Materials and methods: Forty-five male Wistar-Albino rats were randomly divided into seven groups and administered intraperitoneal doses of TQ. A femur fracture was surgically induced, and healing was assessed through radiological, histological, and biochemical analyses. The study included seven groups: a sham-operated control (Group 1), untreated fracture controls at two and four weeks (Groups 2 and 5), and fracture groups treated with TQ at 2.5 or 5 mg/kg/day for two weeks (Groups 3 and 4) or four weeks (Groups 6 and 7). Fracture healing was evaluated at two time points: at the end of two weeks (Groups 1-4) and at the end of four weeks (Groups 5-7). Intraperitoneal administration was chosen to ensure accurate and consistent dosing, as oral administration could lead to variability in bioavailability and potential loss during ingestion. Radiological assessments included measuring total callus diameter, radiopacity ratios, and callus surface area using high-resolution tomography. Histological analysis involved hematoxylin-eosin and immunohistochemical staining. Biochemical analyses measured complete blood count, sedimentation rate, C-reactive protein (CRP), total protein, albumin, calcium, alkaline phosphatase (ALP) and phosphorus.

Results: Significant differences in callus diameter were observed between the high dose TQ group (Group 7) and the untreated control (Group 5), while callus area showed significant differences among Groups 5, 6 and 7, and callus surface differed significantly between Groups 3 and 6. In Group 6, total protein and calcium levels increased, while albumin and ALP were lower in Group 7. Histological findings demonstrated that TQ treatment reduced dense fibrous tissue, with initial formation of spongy structures followed by trabecular bone development, indicating enhanced bone formation. Immunohistochemical analysis showed increased interleukin (IL)-10 and proliferating cell nuclear antigen (PCNA) levels, supporting improved fracture healing in high-dose groups.

Conclusion: Thymoquinone enhanced fracture healing, particularly in the fourth-week groups, improving callus formation, biochemical markers, and histological outcomes.

Declarations

Author Contributions

Idea/concept: D.İ.Y., A.Y.; Design, literature review, critical review, references and fundings, materials: D.İ.Y., A.Y., M.S.D., F.A., N.Ü., S.S., S.A.; Control/ supervision, writing the article, data collection and/or processing: D.İ.Y., A.Y., M.S.D., F.A., N.Ü., S.S.; Analysis and/or interpretation: D.İ.Y., A.Y., M.S.D., N.Ü., S.S.

Conflict of Interest

The authors declared no conflicts of interest with respect to the authorship and/or publication of this article.

Financial Disclosure

The authors received no financial support for the research and/or authorship of this article.

Data Sharing Statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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