Atherosclerotic diseased murine models (The Jackson Laboratories, Bar Harbor, Maine, USA) were developed using angiotensin II infusion in both male and female ApoE−/− mice. ApoE−/− mice were switched to an atherogenic, Western diet (21% (wt/wt) fat, 0.2% (wt/wt) cholesterol, 19.5% (wt/wt) casein, Teklad, Indianapolis, IN, USA) at 24 weeks old for 6 weeks. At age 27 weeks, Alzet osmotic minipumps (Model 2004; ALZA Scientific Products, Mountain View, CA, USA) were implanted subcutaneously into ApoE−/− mice. Pumps were filled with angiotensin II (Bachem, Torrance, CA, USA) following the manufacturer’s protocol to deliver subcutaneous angiotensin II at a rate of 1.9 mg kg−1 d−1 for 28 days.[52 ] Mice were anesthetized using ketamine/xylazine and isoflurane at flow rate of 2%. Pumps were placed into the subcutaneous space behind the right shoulder of the mice through a small incision that was closed with surgical glue. Buprenorphine SR was also administered subcutaneously at a dose of 1.0 mg kg−1 as an analgesic.
One day after implantation of the osmotic pump, mice were injected with MC PAMs, SC PAMs (1000 μm , 100 μL, N = 4), or PBS (100 μL, N = 3) via tail vein IV injection. Subsequent injections were administered once a week over 3 weeks for a total of four injections. The last injection consisted of Cy7-labeled MCG or SCG PAMs for use as imaging contrast agents.
One day after implantation of the osmotic pump, mice were injected with MC PAMs, SC PAMs (1000 μ