Virginia Via Research Day Book 2026

Allyson Meador, Alina Xie, Claudia Ramos-Lopez, Alyssa Morrison, Claire Cho, Reece Girdharry, Ian Jeong, Braxton McDaniel, and Jia-Qiang He, PhD Corresponding author: allysonm24@vt.edu gavage (immediately after surgery, and then every 2 days for 1 week, and then 1x per week up to 4 weeks) of mEx loaded with or without α-CT1. Following the surgery, hindlimb blood flow will be evaluated weekly till the end of a 4-week period, when the animals will be euthanized, and the heart will be perfused to fix both limbs with 10% formalin. The gastrocnemius skeletal muscle and paw of both control and ischemic climb will be collected for post-histological analysis. We hope that our findings could justify progression toward pre clinical optimization and eventual clinical trials aimed at reducing the burden of PAD in human populations. 05 USING A HINDLIMB ISCHEMIA MODEL TO EVALUATE α -CT1-MODIFIED MILK EXOSOMES AS A POTENTIAL TREATMENT FOR PERIPHERAL ARTERIAL DISEASE Undergraduate Student Research Biomedical Department of Biomedical Sciences and Pathobiology, College of Veterinary Medicine, Virginia Tech, Blacksburg, Virginia

Peripheral Arterial Disease (PAD) results from partial or complete occlusion of distal arteries due to plaque accumulation, leading to impaired limb perfusion, ischemic pain, risk of necrosis, and even limb amputation. Current management strategies rely predominantly on surgical or endovascular interventions, motivating the development of less invasive therapeutic alternatives. This study aimed to 1) establish mouse hindlimb ischemia (to mimic human PAD), and 2) to evaluate therapeutic potential of milk-derived exosomes (mEx) loaded with the alpha-carboxyl terminus 1 (α-CT1) peptide. α-CT1 has previously demonstrated therapeutic benefit in mouse models of cardiac injury, and it is currently in Phase 3 clinical trial to treat skin injury. Briefly, murine hindlimb ischemia surgery was performed on a heated surgical bed, and the animal body temperature was maintained at 36.5 +/-0.5oC. Mice were anesthetized with 1–3% isoflurane delivered at a flow rate of 1 L/ min, initially in an induction chamber and subsequently maintained via continuous inhalation via a nose cone. Animals were positioned supine, secured via adhesive tapes, and the blood flow of both hindlimbs was

recorded using Laser Speckle imager before and after ligation. Prior to incision, hair was removed from the left hindlimb (termed ischemic limb, compared to the right limb, termed control limb). Under 10x to 40× magnification, a roughly 1 cm incision was made extending from the medial thigh to the knee. The exposure site was moistened with phosphate-buffered saline (PBS), which also aided in clearing subcutaneous adipose tissue. Sterilized ophthalmic forceps were applied transversely to dissect remaining connective tissue, exposing the femoral neurovascular bundle. The femoral artery, vein, and nerve were carefully separated. An 8-0 suture was passed beneath the distal femoral artery, proximal to the popliteal artery, and secured with double knots to achieve full occlusion. Following vessel occlusion, the vessel segments between two ligation sites were removed and the skin was closed using 5-0 suture. The results showed that a significant reduction of blood flow was identified by Laser Speckle in the experimental (i.e., ischemic) limb relative to its basal level (prior to surgery) and also to the control limb. In subsequent phases of this project, mice undergoing hindlimb ischemia will receive an oral

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126 Edward Via College of Osteopathic Medicine (VCOM)