Deep Vein Thrombosis Induced by Stasis in Mice Monitored by High Frequency Ultrasonography

Ryan N. Rys; Mark D. Blostein; Catherine A. Lemarié

doi:10.3791/57392

Method Article

Deep Vein Thrombosis Induced by Stasis in Mice Monitored by High Frequency Ultrasonography

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DOI:

10.3791/57392

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April 13th, 2018

Ryan N. Rys¹ , Mark D. Blostein¹^,² , Catherine A. Lemarié¹^,²

¹Lady Davis Institute for Medical Research, Jewish General Hospital, McGill University, ²Department of Medicine, Jewish General Hospital, McGill University

Summary

The present protocol describes the steps to obtain venous thrombosis using a stasis model. In addition, we are using a non-invasive method to measure thrombus formation and resolution over time.

Abstract

Venous thrombosis is a common condition affecting 1 - 2% of the population, with an annual incidence of 1 in 500. Venous thrombosis can lead to death through pulmonary embolism or results in the post-thrombotic syndrome, characterized by chronic leg pain, swelling, and ulceration, or in chronic pulmonary hypertension resulting in significant chronic respiratory compromise. This is the most common cardiovascular disease after myocardial infarction and ischemic stroke and is a clinical challenge for all medical disciplines, as it can complicate the course of other disorders such as cancer, systemic disease, surgery, and major trauma.

Experimental models are necessary to study these mechanisms. The stasis model induces consistent thrombus size and a quantifiable amount of thrombus. However, it is necessary to systematically ligate side branches of the inferior vena cava to avoid variability in thrombus sizes and any erroneous data interpretation. We have developed a non-invasive technique to measure thrombus size using ultrasonography. Using this technique, we can assess thrombus development and resolution over time in the same animal. This approach limits the number of mice required for quantification of venous thrombosis consistent with the principle of replacement, reduction, and refinement of animals in research. We have demonstrated that thrombus weight and histological analysis of thrombus size correlate with measurement obtained with ultrasonography. Therefore, the current study describes how to induce deep vein thrombosis in mice using the inferior vena cava stasis model and how to monitor it using high frequency ultrasound.

Introduction

Venous thromboembolism (VTE), which is comprised of deep venous thrombosis (DVT) and pulmonary embolism, is the third leading cause of cardiovascular death after myocardial infarction and stroke. It is a common condition affecting 1 - 2% of the population, with an annual incidence of 1 in 500¹. VTE can lead to: 1) death through pulmonary embolism; 2) post-thrombotic syndrome, characterized by chronic leg pain, swelling, and ulceration; or 3) chronic pulmonary hypertension resulting in significant chronic respiratory compromise. VTE is a multi-factorial disease and may result from stasis of blood flow, damage to vessel walls, and/or hypercoagula....