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Copyright: ©Author(s) 2026. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution-NonCommercial (CC BY-NC 4.0) license. No commercial re-use. See permissions. Published by Baishideng Publishing Group Inc.
World J Exp Med. Jun 20, 2026; 16(2): 118828
Published online Jun 20, 2026. doi: 10.5493/wjem.118828
Comparison of ozone administration routes on liver, lung, and brain damage after spinal cord ischemia-reperfusion injury
Necmiye Şengel, Zeynep Köksal, Aysegül Küçük, Şaban Cem Sezen, Işın Güneş, Muharrem Atlı, Gülay Kip, Seda Gökgöz Acar, Abdullah Özer, Mustafa Kavutçu, Mustafa Arslan
Necmiye Şengel, Department of Oral and Maxillofacial Surgery, Gazi University Faculty of Dentistry, Ankara 06490, Türkiye
Zeynep Köksal, Department of Anesthesiology and Reanimation, Ankara 29 Mayıs State Hospital, Ankara 06105, Türkiye
Aysegül Küçük, Department of Physiology, Bandırma Onyedi Eylül University, Bandırma 10250, Türkiye
Şaban Cem Sezen, Muharrem Atlı, Department of Histology and Embryology, Kırıkkale University Faculty of Medicine, Kırıkkale 71450, Türkiye
Işın Güneş, Department of Anesthesiology and Reanimation, Erciyes University Faculty of Medicine, Kayseri 38030, Türkiye
Gülay Kip, Mustafa Arslan, Department of Anesthesiology and Reanimation, Gazi University Faculty of Medicine, Ankara 06560, Türkiye
Seda Gökgöz Acar, Mustafa Kavutçu, Department of Medical Biochemistry, Gazi University Faculty of Medicine, Ankara 06560, Türkiye
Abdullah Özer, Department of Cardiovascular Surgery, Gazi University Faculty of Medicine, Ankara 06560, Türkiye
Mustafa Arslan, Life Sciences Application and Research Center, Gazi University, Ankara 06560, Türkiye
Mustafa Arslan, Laboratory Animal Breeding and Experimental Research Center (GÜDAM), Gazi University, Ankara 06560, Türkiye
Co-corresponding authors: Necmiye Şengel and Mustafa Arslan.
Author contributions: Şengel NŞ and Arslan M contributed equally to this manuscript and are co-corresponding authors. Arslan M, Kip G, Şengel N, Köksal Z, and Küçük A designed and coordinated the study and analyzed data; Özer A, Arslan M, and Güneş I performed the experiments and acquired data; Sezen ŞC, Atlı M, Acar SG, and Kavutçu M performed the biochemical and histological experiments and interpreted the data; Köksal Z and Kip G collected samples; Şengel NŞ, Arslan M, Küçük A, and Köksal Z wrote the manuscript; all authors approved the final version of the article.
Institutional animal care and use committee statement: All procedures were performed in accordance with the NIH Guide for the Care and Use of Laboratory Animals and relevant national guidelines for the protection of animals used for scientific purposes. The experimental protocol of this study was reviewed and approved by the Gazi University Animal Care and Use Ethics Committee (Approval Date: July 14, 2025; Protocol Code: No. G.Ü.ET-25.065).
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Data sharing statement: The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
Corresponding author: Necmiye Şengel, Assistant Professor, Department of Oral and Maxillofacial Surgery, Gazi University Faculty of Dentistry, Bişkek Cd. (8.Cd.) 1. Sk. No. 8 06490 Emek, Ankara 06490, Türkiye. necmiyesengel@hotmail.com
Received: January 12, 2026
Revised: March 11, 2026
Accepted: May 11, 2026
Published online: June 20, 2026
Processing time: 152 Days and 9.3 Hours
Abstract
BACKGROUND

Spinal cord ischemia-reperfusion injury (IRI), which can occur as a result of temporary aortic occlusion during resection of thoracoabdominal aneurysms, can be an unpredictable and devastating complication of aortic surgery. There are no specific medications or guidelines for the prevention and treatment of spinal cord IRI (SCIRI). It is now known that IRI not only exacerbates local tissue damage when blood flow is restored but also affects distant organs, such as the liver, lungs, and brain, through mediators released into the systemic circulation. Studies show that ozone pretreatment reduces oxidative damage by increasing antioxidant capacity, promotes anti-inflammatory signaling, improves blood circulation, and ameliorates IRI. Our study is one of the first to examine the effects of ozone on remote organs (liver, lung, and brain) when administered via different routes (intrathecal, intraperitoneal, rectal) in a spinal cord ischemia-reperfusion model.

AIM

To determine whether ozone administered by different routes offers multiorgan protection in SCIRI.

METHODS

Thirty adult Wistar albino rats were randomly divided into five groups (n = 6, each): A control (C group), an IR group, an IR rectal ozone (IRRO) group, an IR intrathecal ozone (IRITO), and an IR intraperitoneal ozone (IRIPO). In the IR groups, the spinal cord IR models (a 30-minute ischemia period was applied to the infrarenal abdominal aorta using an atraumatic vascular clamp, and then a 120-minute reperfusion period was applied by removing the clamp) were applied. An ozone-oxygen mixture of 1 mg/kg (50 μg/mL) was administered by rectal insufflation to the IRRO group, 0.7 mg/kg (50 μg/mL) via the peritoneum to the IRIPO group, and 20 μL (20 μg/mL) intrathecally to the IRITO group 30 minutes before midline laparotomy. At the end of the reperfusion procedure, histopathological and biochemical analyses of liver, lung, and brain tissues were performed.

RESULTS

Liver tissue malondialdehyde (MDA) levels were significantly lower, and catalase (CAT) enzyme activities were significantly higher in the IRRO, IRITO, and IRIPO groups than in the IR group. Histopathologically, we had favorable results from all three ozone applications compared to the IR group. Lung tissue MDA levels were significantly lower, and CAT enzyme activities were significantly higher in the IRITO and IRIPO groups than in the IR group. We had more positive results in the IRRO group than in the IR group, but the difference was not found to be significant. Histopathologically, we obtained significantly more positive results in the IRITO and IRIPO groups compared with the IR group regarding all the criteria we evaluated. Our results in the IRRO group were also positive. Brain tissue MDA levels were significantly lower and CAT enzyme activities significantly higher in the IRITO and IRIPO groups than in the IR group. We had positive results in the IRRO group compared with the IR group. Histopathologically, we obtained significantly more positive results in the IRITO group regarding all the criteria we evaluated.

CONCLUSION

We observed histopathologically that single-dose ozone pretreatment administered intrathecally, intraperitoneally, or rectally had positive effects on liver, lung, and brain tissues compared to the IR group in an SCIRI model in rats due to its antioxidant effect. The best histopathological results were obtained with intrathecal, intraperitoneal, and rectally administered ozone, in that order, in all three tissues.

Keywords: Spinal cord; Ischemia reperfusion; Liver; Lung; Brain; Ozone; Rat

Core Tip: Spinal cord ischemia-reperfusion injury (IRI) is a complex process affecting multiple organs. Current preventive strategies remain insufficient. This study investigated the protective effects of single-dose ozone pretreatment via intrathecal, intraperitoneal, and rectal routes on liver, lung, and brain tissues in a rat spinal cord IRI model. Histopathological results demonstrated that ozone pretreatment through all three routes significantly reduced oxidative damage and improved tissue outcomes compared to the IR group. Medical ozone administration may offer a viable therapeutic option with high efficacy and minimal side effects in preventing IRI-related organ damage.

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