New experimental model of hind limb ischemia in pot-bellied pigs

The simulation of limb ischemia in large laboratory animals is a complex and currently topical task in experimental medicine. Meanwhile, there is a demand for a reliable and effective model of limb ischemia for further testing of medicines to stimulate circulation and induce angiogenesis, gene medicines in particular. Aim of this study was to develop and experimentally test an effective method of simulation of hind limb ischemia.

Female Vietnamese pot-bellied pigs were chosen as biological models. The reproduction of the pathology was evaluated using the following methods: laser doppler flowmetry, laboratory test of venous blood, immunohistochemical reaction with antibodies against CD31, a specific marker of endothelial cells, Van Gieson's staining of muscles for presence of connective tissue and clinical observation to detect the presence of lameness in pigs.

Laser doppler flowmetry recorded a significant decrease in the intensity of the blood circulation and a marked decrease in temperate in the operated limb. Increased lactate and creatine kinase were registered immediately after the surgery and were absent 3 or more days later. Clinical observation demonstrated presence of walking lameness. Histological and immunohistochemical methods revealed a credible increase in connective tissue area and a reduction in the number of blood vessels in the muscles, confirming the presence of ischemia.

An effective approach to modeling limb ischemia has been developed and experimentally tested. The proposed model may be used in cardiovascular surgery and will allow further testing of new medications designed to treat ischemia of hind limbs.