Quantitation of angiogenesis in healing anastomoses of the rat colon

An accurate and reliable quantitation of angiogenesis is an essential requirement for a detailed study of new blood vessel growth during healing of colon anastomoses. In the present study we applied a computer-based digital image processing system for quantitative analysis of the anastomotic vascularization and perfusion. Rats underwent colonic resection (1 cm) followed by construction of an end-to-end anastomosis. The animals were killed 3 or 7 days after operation. The vascularization and perfusion were analysed in cyrostat sections using an anti-collagen type IV antibody and the fluorescent marker bisbenzimide H 33342, respectively. If compared with 3-day-old anastomoses, a significant increase in the median vascular and perfusion areas was found 7 days after operation. Within the same anastomosis, the vessel density and perfusion were lower in the central region than in the peripheral region of the wound. The present computerized digital image analysis system is a reliable technique that is suited for quantitative measurements of microvascular changes occurring in colon anastomoses during the first postoperative week.