Development of a two-stage limb ischemia model to better simulate human peripheral artery disease

Abstract

peripheral arterial disease (pAD) develops due to the narrowing or blockage of arteries supplying blood to the lower limbs. Surgical and endovascular interventions are the main treatments for advanced pAD but alternative and adjunctive medical therapies are needed. currently the main preclinical experimental model employed in pAD research is based on induction of acute hind limb ischemia (HLI) by a 1-stage procedure. Since there are concerns regarding the ability to translate findings from this animal model to patients, we aimed to develop a novel clinically relevant animal model of pAD. HLi was induced in male Apolipoprotein e (ApoE −/− ) deficient mice by a 2-stage procedure of initial gradual femoral artery occlusion by ameroid constrictors for 14 days and subsequent excision of the femoral artery. This 2-stage HLI model was compared to the classical 1-stage HLI model and sham controls. ischemia severity was assessed using Laser Doppler perfusion imaging (LDpi). Ambulatory ability was assessed using an open field test, a treadmill test and using established scoring scales. Molecular markers of angiogenesis and shear stress were assessed within gastrocnemius muscle tissue samples using quantitative polymerase chain reaction. HLI was more severe in mice receiving the 2-stage compared to the 1-stage ischemia induction procedure as assessed by LDPI (p = 0.014), and reflected in a higher ischemic score (p = 0.004) and lower average distance travelled on a treadmill test (p = 0.045). Mice undergoing the 2-stage HLI also had lower expression of angiogenesis markers (vascular endothelial growth factor, p = 0.004; vascular endothelial growth factor-receptor 2, p = 0.008) and shear stress response mechano-transducer transient receptor potential vanilloid 4 (p = 0.041) within gastrocnemius muscle samples, compared to animals having the 1-stage HLI procedure. Mice subjected to the 2-stage HLI receiving an exercise program showed significantly greater improvement in their ambulatory ability on a treadmill test than a sedentary control group. this study describes a novel model of HLi which leads to more severe and sustained ischemia than the conventionally used model. Exercise therapy, which has established efficacy in PAD patients, was also effective in this new model. this new model maybe useful in the evaluation of potential novel pAD therapies.Peripheral arterial disease (PAD) leads to impaired lower limb blood supply usually as a result of atherosclerosis and associated thrombosis 1 . In 2010 an estimated 200 million people worldwide were living with PAD, an increase of ~30% since 2000 which has been referred to as a PAD pandemic 2,3 . The burden of PAD is increasing worldwide and particularly within low and middle-income countries 4 . PAD is associated with significant morbidity including intense leg pain during walking (intermittent claudication, IC), impaired walking ability, poor health-related quality of life and risk of serious complications such as major leg amputation and death 1 . Medical management of PAD is ...

Comparative analysis of mouse bone marrow and adipose tissue mesenchymal stem cells for critical limb ischemia cell therapy

“…We used a model of limb ischemia that was induced by the complete removal of the femoral artery and the closing of its branches. CLIinduced mice were evaluated for functional recovery and ischemia using the Tarlov, ischemia, modified ischemia, function, and the grade of limb necrosis scoring system at days 3, 7, 14, 21, and 28 after cell transplantation [44,45]. The results showed that BM-MSC transplantation improved the treatment efficacy of hindlimb ischemia compared to AT-MSCs and control groups.…”

Interleukin 35 Delays Hindlimb Ischemia-Induced Angiogenesis Through Regulating ROS-Extracellular Matrix but Spares Later Regenerative Angiogenesis

“…Third , there are variabilities in the angiogenic processes in different mouse strains ( 78 ). Fourth , we acknowledge that current one stage HLI model ( 81 ) would not allow us to examine the regenerative angiogenesis in details. Thus, anti-inflammatory treatment has been presumed a promising therapy to control the local inflammation and promote desired neovascularization.…”

Impact of aerobic exercise type on blood flow, muscle energy metabolism, and mitochondrial biogenesis in experimental lower extremity artery disease

“…To our knowledge, we show for the first time that improvement in voluntary endurance exercise performance following ET is not associated with enhanced blood flow, oxygenation and/or vascularization in a mouse model of LEAD. During the revision process of our manuscript, Krishna et al 25 reported that 4 weeks of running wheel exercise (30-45 min per day,average distance covered per day: 80-200 m) increased functional capacity (i.e. total distance walked by mice until exhaustion) but not limb perfusion in ApoE −/− mice subjected to a two-stage hindlimb ischemia (i.e.…”

“…total distance walked by mice until exhaustion) but not limb perfusion in ApoE −/− mice subjected to a two-stage hindlimb ischemia (i.e. initial gradual femoral artery occlusion by ameroid constrictors for 14 days and subsequent excision of the femoral artery) 25 . These findings are directly in line with our observations.…”