The effects of hydroxychloroquine-induced oxidative stress on fracture healing in an experimental rat model
Yiğit Önaloğlu1
, Ozan Beytemür2, Elif Yaprak Saraç3, Ozancan Biçer2, Yiğit Güleryüz2, Mehmet Akif Güleç2
1Department of Orthopedics and Traumatology, University of Health Sciences, Başakşehir Çam and Sakura City Hospital, Istanbul, Türkiye
2Department of Orthopedics and Traumatology, University of Health Sciences, Bağcılar Training and Research Hospital, Istanbul, Türkiye
3Department of Molecular Biology and Genetics, Istanbul Technical University, Istanbul, Türkiye
Keywords: Femur, fracture healing, hydroxychloroquine sulfate, oxidative stress, rat.
Abstract
Objectives: The purpose of this study was to investigate whether hydroxychloroquine (HCQ) sulfate causes oxidative stress (OS) and its effect on fracture healing in an experimental rat model.
Materials and methods: In this experimental study, open diaphyseal femur fractures were induced in 24 eight-week-old male rats (mean weight: 225±25 g; range, 200 to 250 g) and then fixed with K-wire. The rats were divided into four groups: HCQ-2, control-2 (C-2), HCQ-4, and control-4 (C-4). During the study period, rats in the HCQ groups received an HCQ solution (160 mg/kg/day), whereas rats in the control groups received saline. The HCQ-2 and C-2 groups were sacrificed on the 14th day, and the HCQ-4 and C-4 groups were sacrificed on the 28th day. After sacrifice, malondialdehyde levels induced by OS were calculated for each rat, and fracture healing was evaluated radiographically, histomorphometrically, histopathologically, and immunohistochemically.
Results: Malondialdehyde levels were higher in the HCQ groups than in the control groups (p<0.05). Hydroxychloroquine caused OS in rats. The ratio of total callus diameter to femur bone diameter was lower in HCQ groups compared to control groups (p<0.05). No differences were observed when comparing radiological and histological healing results between the control and HCQ groups. Alkaline phosphatase levels were lower in the HCQ-4 group than the C-4 group at week four (p<0.05), although osteocalcin and osteopontin levels did not differ between groups (p>0.05). Oxidative stress had no adverse effects on histologic healing outcomes and osteoblast functions. Cathepsin K and tartrate-resistant acid phosphatase-5b levels were higher in the HCQ-4 group than in the C-4 group (p<0.05). While the number and function of osteoclasts increased due to OS in callus tissue, a decrease in the number of chondrocytes was observed.
Conclusion: Hydroxychloroquine-induced OS increases the number and function of osteoclasts and decreases the number of hypertrophic chondrocytes and endochondral ossification but has no significant effect on mid-late osteoblast products and histological fracture healing scores.
Citation: Önaloğlu Y, Beytemür O, Yaprak Saraç E, Biçer O, Güleryüz Y, Güleç MA. The effects of hydroxychloroquineinduced oxidative stress on fracture healing in an experimental rat model. Jt Dis Relat Surg 2024;35(1):146-155. doi: 10.52312/ jdrs.2023.1226.
Approval from the local animal experimentation ethics committee was granted by Bağcılar Training and Research Hospital (date: 10.05.2020; no: 2020/27). The Declaration of Helsinki on the Guide for the Care and Use of Experimental Animals was followed in the conduct of this study.
Idea/Concept/Writing the Article: Y.Ö.; Control/Supervision: O.B.; Data Collection and/ or Processing, literature review: Y.G., O.B.; Analysis and/or interpretation: Y.Ö., E.Y.S.; Critical review: M.A.G.
The authors declared no conflicts of interest with respect to the authorship and/or publication of this article.
The study was carried out with the financial contributions of Bağcılar Training and Research Hospital.
The data that support the findings of this study are available from the corresponding author upon reasonable request.