Dose response of angiogenesis to basic fibroblast growth factor in rat corneal pocket assay: I. Experimental characterizations

Understanding mechanisms of formation of vascular networks under different experimental conditions is essential for improving treatment of angiogenesis-dependent diseases. To this end, we investigated the dose response of angiogenesis to basic fibroblast growth factor (bFGF) using the rat corneal pocket assay. The response was quantified, in terms of (i) the migration distance of vascular networks, (ii) the total vessel length, (iii) the distribution of the projected width of vessels, (iv) the distribution of the number of vessels, and (v) the distribution of vessel diameters. The quantification was based on new image analysis methods developed in the study. It was observed that the migration distance and the total vessel length increased by 82% and 199%, respectively, when the dose of bFGF was increased from 5 ng to 50 ng. The number and the diameter of vessels increased with the dose of bFGF as well. However, the last two parameters at a given dose of bFGF were approximately independent of the location in the middle region between the pellet and the limbus. These results provided useful information for understanding mechanisms of angiogenesis induced by bFGF and important data for validating a mathematical model of angiogenesis described in the second part of the study.