This article describes a step-by-step procedure for creating a model of testicular ischemia/reperfusion for the production of a reproducible model of testicular torsion in the study of the pathological mechanisms of ischemia-reperfusion injury in the testis.
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Method Article
Anchorage of Twisted Testis: A Rat Model of Ischemia-Reperfusion Injury of the Testis
In This Article
Summary
Abstract
Ischemia-reperfusion (IR) injuries are associated with several pathologies, including testicular injury. Earlier animal models of testicular IR, which involves twisting the spermatic cord to achieve ischemia, are fraught with many technical difficulties that result in the inability to reproduce the model consistently. Hence, we present a simple method for inducing testicular IR in a rat model that can easily be replicated.
This method involves the separation of the gubernaculum from the testis, creation of a scrotal pouch, twisting of the testis 720° clockwise at its lower pole, and anchorage of the testis to the base of the scrotum. This model effectively constricts testicular vessels and induces testicular ischemia. After 1 h of ischemia, the anchoring stitch is removed and the testis untwisted. The cord and its accompanying vessels are inspected to ensure patency. After IR, the testis is collected and sectioned for histopathological evaluation.
Testicular histology was examined for focal hemorrhagic lesions, vascular congestion, widened interstitial space, and infiltration of inflammatory cells. Furthermore, markers of oxidative stress (malondialdehyde, reduced glutathione, catalase, and superoxide dismutase), inflammation (MPO, TNF-α, IL-1β), and apoptosis (caspase 3 expression) were assayed. This method provides a simpler and easily reproducible model to study the pathophysiology of testicular IR injury. More so, this model opens a window for exploring potential therapeutic agents in the management of testicular IR injury.
Introduction
Torsion of the testis (TT) is an acute urological emergency in which the spermatic cord, along with its accompanying vessels, twists, resulting in ischemia of the testis. It is characterized by severe abdominal pain, nausea, and vomiting1. The ischemia, if unresolved by timely detorsion, results in necrosis and loss of the testis concerned2. However, timely surgical intervention by untwisting and restoration of blood flow to the testis results in ischemia-reperfusion injury (IRI), which results in histological and biochemical disruption of the testis and its functions3,
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Protocol
The study was approved by the Ethics Review Committee of the Faculty of Basic Medical Sciences, Ladoke Akintola University of Technology, Ogbomoso, Oyo State, Nigeria (ERCFBMSLAUTECH:033/05/2024). Twenty male albino Wistar rats, aged 10-12 weeks, and weighing 180 ± 10 g were used for this study. They were acclimated for 2 weeks and then, randomly allotted into either the sham-operated group (SO) or the torsion/detorsion group (T/D) (n = 10 rats per group). The SO rats underwent a similar surgery, just like the T/D rats, but the testes were not twisted or anchored to induce T/D.
1. Preoperative preparation and anesthesia<....
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Results
XO activity, MDA level, and activities of SOD, catalase, and GPx in the testis were employed as markers of oxidative stress. Torsion/detorsion (T/D) markedly increased XO activity and MDA levels and reduced testicular activities of SOD, catalase, and GPx in T/D rats compared with the sham-operated rats (Figure 1), depicting the role of oxidative stress in T/D-induced injury.
Moreover, TNF-α, a media.......
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Discussion
This paper and the accompanying video give a detailed step-by-step description of the surgical induction of TT for the study of testicular IRI using the anchorage approach. The method is particularly useful in that it obviates the prolonged exteriorization of the testis occasioned by clamping techniques7 and at the same time, more closely models the clinical presentation of TT in humans10,12.
When ischaemia has .......
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Disclosures
The authors have no conflicts of interest to declare.
Acknowledgements
We hereby acknowledge the assistance of Mr Ojeniran, head of the audio-visual unit of the College of Health Sciences, who helped in the filming and editing of the procedure.
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Materials
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| 1 cc syringes | Not Applicable | Not Applicable | for anaethetic |
| 2-0 chromic suture | Not Applicable | Not Applicable | For surgery |
| Bard Parker knife handle | Not Applicable | Not Applicable | For surgery |
| Curved Mosquito artery forceps | Not Applicable | Not Applicable | For surgery |
| Dissecting scissors | Not Applicable | Not Applicable | For surgery |
| GraphPad Prism (version 8.0.2) | |||
| Ketamine injection | Swiss Parenterals LTD Gujarat India | 2321154 | Anesthetic |
| Paracetamol injection | Shanxi Zhongbao Pharmaceuticals, China | 220637 | Analgesic |
| Povidone | JAWA International limited Nigeria | 135S | Antiseptic wash |
| Procaine penicillin powder | Anhui Chengshi Pharmaceutical Co Ltd China | 201906 | For surgery |
| Surgical blade | Not Applicable | Not Applicable | For surgery |
| Surgical gloves | Zhenjiang huayang latex products China | 210325 | |
| Stitch scissors | Not Applicable | Not Applicable | For surgery |
| Toothed dissecting forceps | Not Applicable | Not Applicable | For surgery |
| Xylazine injection | Bioveta Czech Republic | 396228A | Anesthetic |
| Light Microscope | Novel Microscope, China | Not applicable | For histopathological evaluation |
| Digital Camera | Hayear, China | Not applicable | For histopathological evaluation |
| TNF-α kit | Elabscience, USA | E-EL-H0109 | For assay |
| Thermo Fischer kit (anti-mouse caspase 3 monoclonal for caspase 3 expression ) | Thermo Fischer Scientific Inc., USA | Not applicable | For Immunohistochemistry (caspase 3) |
References
- Obi, A. O., Okeke, C. J., Ugwuidu, E. I. Acute testicular torsion: A critical analysis of presentation, management, and outcome in southeast Nigeria. Niger J Clin Pract. 23 (11), 1536-1541 (2020).
- Yang, C., Song, B., Tan, J., Liu, X., Wei, G. H.
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