Simple and Effective Procedure for Hemostasis in Mouse Arteries

Summary

This study aims to develop an effective method for achieving hemostasis in mouse arteries.

Abstract

During arterial puncture or cannulation in mice, permanent ligation of the artery is often required due to difficulty achieving hemostasis after needle withdrawal. This leads to permanent ischemic injury in the supplied tissue region. Such localized ischemia frequently induces secondary complications, compromising experimental outcomes in animal models. The present method aims to seal the puncture site post-procedure while maintaining normal blood flow, thereby preventing postoperative ischemia. This method involves harvesting an adipose tissue fragment from the experimental animal itself, performing arterial puncture using standard techniques, and then wrapping the puncture site with the fat pad to achieve hemostasis while maintaining blood flow. In this study, bilateral common carotid artery puncture was performed on mice, followed by hemostasis using a fat-wrapping technique. Laser speckle contrast imaging(LSCI) revealed no differences in cerebral blood flow changes between the sham and operated groups of mice, and no significant changes were observed in preoperative versus postoperative cerebral blood flow measurements in the operated group. Histological assessment performed at 7, 14, and 28 days after surgery revealed that: (1) preserved brain architecture (H&E staining), (2) intact neuronal morphology without degenerative changes (Nissl staining), and (3) absence of pathological alterations in brain sections. This arterial hemostasis technique capitalizes on the pliable nature of adipose tissue and physiological coagulation mechanisms. The procedure involves: (1) wrapping autologous fat around the puncture site, (2) securing it with sutures to ensure optimal contact with the arterial adventitia, and (3) allowing escaping blood to permeate the fat-artery interface where it undergoes coagulation. This triple-action mechanism achieves reliable hemostasis while preserving arterial patency and preventing ischemic complications post-operatively. The method demonstrates notable procedural simplicity, clinical efficacy, and excellent translational potential.

Introduction

In animal models, creating arterial injury is an extensively utilized experimental approach for modeling various pathological conditions, including cerebral ischemia-reperfusion models, carotid (CCA) or aortic (AO) atherosclerosis models, and spinal cord ischemia models.

Taking the mice cerebral ischemia-reperfusion (MCAO) model as an example, the current standard modeling approach^1,2,3 involves: isolating the external carotid artery (ECA), ligating its distal end, clamping the proximal ECA with an arterial clip while placing a stay suture, creatin....

Protocol

All experimental procedures were conducted in strict compliance with the Guidelines for the Care and Use of Laboratory Animals (Protocol Approval No. SNLL25040501; Animal Use License No. SYXK(Liao)2021 - 0010) established by the Institutional Animal Care and Use Committee (IACUC) of Shenyang Agricultural University. The reagents and the equipment used are listed in the Table of Materials.

1. Experimental preparation

1. Animal details and documentation: Document all animal research data following ARRIVE guidelines (Animal Research: reporting of In Vivo Experiments). Use male Balb/C mice aged....

Results

Analysis of the pre- and post-operative laser speckle contrast imaging data and relative perfusion measurements in both the sham and operated groups revealed no significant changes, demonstrating that this arterial hemostasis technique successfully sealed the common carotid artery puncture site without compromising cerebral blood flow (Figure 1).

Mice in the operated group were continuously monitored for 28 days post-operatively, with no mortality observed. Their .......

Discussion

In MCAO models, preserving the ECA better recapitulates clinical pathology by preventing facial ischemic damage associated with ECA ligation and transection. While the conventional approach of filament insertion via CCA puncture offers technical simplicity, it necessitates bilateral ligation of the puncture site, resulting in cerebral perfusion that depends on Circle of Willis collateral circulation, a suboptimal representation of true stroke pathophysiology. The fat pad hemostasis technique presented in this st.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
4-0 with thread sewing needle	Shanghai Pudong Jinhuan Medical Supplies Co., Ltd
Balb/C mouse	Beijing Huafukang Biotechnology Co., Ltd		6-8w,male
DHG Series Heating and Drying Oven	Shanghai Yiheng Technology Co., Ltd	DHG-9123A
Hematoxylin and Eosin (H&E) High Definition Staining Kit	Servicebio	G1076-500ML
Isoflurane	Shenzhen Ruiwode Life Technology Co., Ltd	R510-22
Laser speckle analyzer	Shenzhen Ruiwode Life Technology Co., Ltd	RFLSI ZW
Nissl dye solution	Servicebio	G1036-100ML
Organizational embedding machine	Leica	HistoCore Arcadia H
Slide scanning imaging system	Shenzhen Shengqiang Technology Co., Ltd	SQS-40R
Small animal anesthesia machine	Shenzhen Ruiwode Life Technology Co., Ltd	R500
Tissue dehydrator	Leica	HistoCore pearl
Ultrapure Water System	Thermo	50132370
Ultra-Thin Semiautomatic Microtome	Leica	HistoCore MULTICUT