Accelerated Aneurysmal Dilation Associated with Apoptosis and Inflammation in a Newly Developed Calcium Phosphate Rodent Abdominal Aortic Aneurysm Model

Objective The calcium chloride (CaCl 2 ) model is a widely accepted rodent model for abdominal aortic aneurysms (AAAs). Calcium deposition, mainly consisting of calcium phosphate (CaPO 4 ) crystals, has been reported to exist in human and experimental aneurysms. CaPO 4 crystals have been used for in vitro DNA transfection by mixing CaCl 2 and phosphate-buffered saline (PBS). Here, we describe accelerated aneurysm formation resulting from a modification of the CaCl 2 model. Methods A modified CaCl 2 model, the CaPO 4 model, was created by applying PBS onto the mouse infrarenal aorta after CaCl 2 treatment. Morphologic, histologic, and immunohistochemical analyses were performed on arteries treated with the CaPO 4 model and the conventional CaCl 2 model as the control. In vitro methods were performed using a mixture of CaCl 2 and PBS to create CaPO 4 crystals. CaPO 4 - induced apoptosis of primary cultured mouse vascular smooth muscle cells (VSMCs) was measured by DNA fragmentation enzyme-linked immunosorbent assay. Results The CaPO 4 model produces AAA, defined as an increase of ≥50% in the diameter of the aorta, faster than in the CaCl 2 model. The CaPO 4 model showed significantly larger aneurysmal dilation at 7, 28, and 42 days, as reflected by a maximum diameter (measured in mm) fold-change of 1.69 ± 0.07, 1.99 ± 0.14, and 2.13 ± 0.09 vs 1.22 ± 0.04, 1.48 ± 0.07, and 1.68 ± 0.06 in a CaCl 2 model, respectively (n = 6; P 4 model compared with the CaCl 2 model (2.7 ± 0.2 vs 1.5 ± 0.2; n=6; P 4 model (apoptosis index at 1, 2, and 3 days postsurgery: 0.26 ± 0.14, 0.37 ± 0.14, and 0.33 ± 0.08 vs 0.012 ± 0.10, 0.15 ± 0.02, and 0.12 ± 0.05 in the conventional CaCl 2 model; n=3; P 4 model. CaPO 4 induced approximately 3.7 times more apoptosis in VSMCs than a mixture of CaCl 2 (n = 4; P Conclusions The CaPO 4 model accelerates aneurysm formation with the enhancement of apoptosis, macrophage infiltration, and calcium deposition. This modified model, with its rapid and robust dilation, can be used as a new model for AAAs.