Intramural Stress and Inflammation in Arterial Branches: A Histology-Based Approach

Abstract

Hypertension is a major risk factor for coronary artery disease, stroke, and kidney disease. Many studies suggest that elevated intramural stresses caused by hypertension may stimulate inflammatory changes, but little has been done to ascertain whether inflammation and stress are spatially correlated. Such correlations are a first step in identifying the mechanisms that may relate intramural stress to disease so that more effective clinical treatments may be developed. Arterial branches exhibit local stress peaks and are focal points for the onset of disease. They are thus a logical place to examine whether high stresses spatially correlate with increased inflammation. This research seeks to 1) develop a histology-based method to reconstruct small arterial branches; 2) use finite element analysis to evaluate intramural stresses where experimental testing is of limited use; 3) quantify biological measures of inflammation; and 4) visually and statistically compare the distribution of stress with the distribution of inflammation. Hypertension was induced in Sprague-Dawley rats by implanting Angiotensin II pumps for 7 days or 21 days. Normotensive rats were used as controls. To preserve morphology the mesentery was pressure-fixed in situ, harvested, processed, and embedded in glycol methacrylate resin. Branch geometry was reconstituted from serial sections. This involved: correcting deformations caused by sectioning; aligning sections into an image stack; identifying vessel boundaries; creating a surface suitable for finite element analysis; reducing the branch geometry to a midplane surface; and using Ansys (Ansys, Inc.) to mesh the midplane surface with a variable-thickness shell element. The pattern of inflammation was characterized by measuring the local density of monocytes and macrophages. Cell density was expressed as a distribution on the branch surface, which simplified visualization and facilitated statistical comparisons of inflammation with stress. Both intramural stresses and inflammation were greater near branches during hypertension. In most cases, however, high stresses and high cell density were not spatially collocated. The onset of an adaptive response may reduce the strength of this correlation. Maximal wall tension, defined as the maximal midplane stress multiplied by the wall thickness, was elevated near branches and strongly correlated with cell density.