Density of connective and white and red pulp tissue of spleens of adult working horses determined by stereological and morphometric methods.

摘要

The spleen is organized in arterial branches where smaller arterioles end in a venous sinus system surrounded by a fibrous capsule of connective tissue and trabeculae to sustain greater vascularization. Authors quantify volume densities of connectiveand white and red pulp tissue of spleen of 10 adult working horses by histomorphometric and stereological techniques. Tissues were stained with hematoxylin & eosin, Picrosirius Red, Gomori and Azan trichrome and then photomicrographed. Images fields werecaptured by Olympus BX51 microscope analyzed with Image-Pro Plus~R software to measure and individualize tissues in 5 different fields per slide. Furthermore, the STEPanizer~R software was also used for the point system analysis to compare the proportions of red and white pulps and connective tissue. Normalized data was subjected to Pearson correlation at P <0.05 significance. Mean + SD density of spleens was 0.99 g/ml ± 0.068. Connective tissue densities were 24.34±7.12 with Picrosirius Red, 6.95 ±3.78 with Gomori, and 8.41 ± 3.75% with Azan trichrome staining. However, with hematoxylin & eosin stain, volume density of connective tissue was 8.64% ± 8.27, white pulp 3.70 ± 3.41%, and red pulp 66.66% ± 11.36. We assume picrosirius red stained splenic musculature whereas Gomori and Azan Trichrome differentiated the connective tissue. Immunohistochemical analysis of spleen allowed us to determine percentage of connective tissue in each spleen to establish a possible relationship with its contractile function. Images of spleen with stains are shown.