Quantitative analysis of fibrosis formation on the microcapsule surface with the use of picro-sirius red staining, polarized light microscopy, and digital image analysis

Transplantation of microencapsulated cells shows potential to treat a variety of diseases. To improve the quality of microcapsules, efficient methods need to be established and standardized for the evaluation of biocompatibility of microcapsules. To this end, a microscopical method was developed to analyze quantitatively the degree of fibrotic overgrowth (FO), which serves as the index of biocompatibility. In this method, sections were stained with picro-sirius red and illuminated with a polarized light microscope. The images were digitized with the use of a computer-video system in which collagenous content was depicted by gray levels I to 255. Both the area of collagen and the thickness of FO were examined. The ratio of Type I/Type III collagen and the thickness were used as indexes of the degree of FO. The data showed that the ratio of Type I/Type III collagen and the thickness increased with time, and FO was not significant 4 weeks after transplantation. In comparison, this new method is more efficient to evaluate FO. (c) 2005 Wiley Periodicals, Inc