Applying Fractal Dimension and Image Analysis to Quantify Fibrotic Collagen Deposition and Organization in the Normal and Hypertensive Heart

Hearts of mice with reduction of function mutation in STAT3 (SA/SA) develop fibrotic collagen foci and reduced systolic function with hypertension. This model was used to determine if fractal dimension and image analysis can provide a quantitative description of myocardial fibrosis using routinely prepared trichome-stained material. Collagen was characterized by relative density [integrated optical density/area (IOD/A)] and fractal dimension (D), an index of complexity. IOD/A of collagen in wild type mice increased with hypertension while D decreased, suggesting tighter collagen packing that could eventually stiffen the myocardium as in diastolic heart failure. Reduced STAT3 function caused modest collagen fibrosis with increased IOD/A and D, indicating more tightly packed, but more disorganized collagen than normotensive and hypertensive controls. Hypertension in SA/SA mice resulted in large regions where myocytes were lost and replaced by fibrotic collagen characterized by decreased density and increased disorder. This indicates that collagen associated with reparative fibrosis in SA/SA hearts experiencing hypertension was highly disorganized and more space filling. Loss of myocytes and their replacement by disordered collagen fibers may further weaken the myocardium leading to systolic heart failure. Our findings highlight the utility of image analysis in revealing importance of a cellular protein for normal and reparative extracellular matrix deposition.