Handcrafted Silicone Coated Filament for Mice Middle Cerebral Artery Occlusion Models

Summary

This protocol describes a straightforward method for creating coated filaments for the middle cerebral artery occlusion (MCAO) model in mice using silicone, nylon sutures, and syringe needles. This method allows for the production of filaments with a consistent diameter and various silicone wrapping lengths tailored to experimental needs.

Abstract

As the global population ages, ischemic stroke has risen to become the second leading cause of disability and mortality worldwide, placing an immense burden on both society and families. Although treatments such as intravenous thrombolysis and endovascular interventions can substantially improve the outcomes for patients with acute ischemic stroke, only a small percentage of individuals benefit from these therapies. To advance our understanding of the disease and to discover more effective treatments, researchers are continuously developing and refining animal models. Among these, the middle cerebral artery occlusion (MCAO) model stands out as the most commonly used model in cerebrovascular disease research. The filament used in this model is crucial for its development. This protocol outlines a method for creating filaments with consistent diameters and varying lengths of silicone coating. The MCAO model produced using this method in C57 mice has demonstrated high success and consistency, offering a valuable tool for tailored investigations into ischemic cerebrovascular diseases.

Introduction

Stroke is one of the most prevalent causes of mortality and disability worldwide. Ischemic and hemorrhagic strokes are the primary types of cerebrovascular event, with ischemic strokes accounting for approximately 87% of cases^1,2,3. Currently, there are two treatment modalities for patients with ischemic stroke: pharmacological therapy with recombinant tissue plasminogen activator (rtPA) and mechanical thrombectomy. However, the narrow therapeutic window and extensive exclusion criteria limit the application of these treatments, benefiting only a minority of patients. This un....

Protocol

All animal procedures adhered to the experimental procedures and standards approved by the Shanxi Provincial People's Hospital Institutional Animal Care and Use Committee (approval number: 2024 Provincial Medical Ethics Committee No. 64). The mice used in this experiment were male C57BL/6 mice, 8-10 weeks old, weighing 24-26 g. Details of the reagents and equipment used are listed in the Table of Materials.

1. Filament preparation

1. Marking the o.......

Discussion

This study demonstrates a simple and cost-effective method for fabricating filament, confirming its feasibility in creating an MCAO model. The length of the filament's silicone coat can be adjusted according to experimental needs, offering additional flexibility. The preparation of a 5 mm filament embolus achieved a 100% success rate without any occurrences of subarachnoid hemorrhage (SAH) in mice. In the group using 10 mm filament emboli, there were instances of SAH, while the rest of the mice showed clear infarctio.......

Materials

Name	Company	Catalog Number	Comments
10 mL Syringe	Haidike Medical Products Co., Ltd.		Instrument for making filaments
2,3,5-Triphenyltetrazolium Chloride (TTC)	Sigma-Aldrich	G3005	Dye for TTC staining
24-well culture plate	Corning	CLS3527	Vessel for TTC staining
26 G syringe needle	Haidike Medical Products Co., Ltd.		Instrument for making filaments
4% paraformaldehyde	Servicebio	G1101	Tissue fixation
6-0 nylon suture	Haidike Medical Products Co., Ltd.		Materials for making filaments
Anesthesia system for isoflurane	Rwd Life Science Co., Ltd.	R610	Anesthetized animal
Bipolar electrocoagulation generator	Yirun Medical Instrument Co., Ltd.	ZG300	Equipment for surgery
Constant temperature water bath	Spring  Instrument Co., Ltd.	HH-M6	TTC staining
Eye ointment	Guangzhou Pharmaceutical	H44023098	Material for surgery
Heat blanket	ZH Biomedical Instrument Co., Ltd.		Maintain body temperatur
Isoflurane	Rwd Life Science Co., Ltd.	R510-22-10	Anesthetized animal
Meloxicam	Boehringer-Ingelheim	J20160020	Analgesia for animal
Microsurgical artery clamp	Shanghai Jinzhong Surgical Instruments Co., Ltd.	W40130	Instrument for surgery
Microsurgical hemostatic clamp forceps	Shanghai Jinzhong Surgical Instruments Co., Ltd.	M-W-0022	Instrument for surgery
Microsurgical instruments set	Rwd Life Science Co., Ltd.	SP0009-R	Equipment for surgery
Mouse thermometer	Hubei Dasjiaer Biotechnology	FT3400	Intraoperative temperature monitoring
Pentobarbital sodium	Sigma-Aldrich	P3761	Euthanized animal
Shaver	Joyu Electrical Appliances	PHC-920	Equipment for surgery
Silicone Sealant	Kafuter	K-704	Materials for making filaments
Stereomicroscope	Rwd Life Science Co., Ltd.	77001S	Equipment for surgery
Suture thread with needle (3-0)	Shanghai Pudong Jinhuan Medical Products Co., Ltd.	F404SUS302	Equipment for surgery