Pro-angiogenic Activity Assay of Chondroitin Sulfate and Glucosamine Sulfate on Vascular Network of Mouse and of Chick Embryo Chorioallantoic Membrane

Abstract

Objective: Target of this study was to test the capacity of chondroitin sulfate (CS) and glucosamine sulfate (GS) to induce in vivo angiogenesis. Methods: The proangiogenic activity of these compounds was analyzed through the assays in chorioallantoic membrane (CAM) of chick embryo and dorsal skin vascularization in mice, but before was realized a cell viability assay with human umbilical veins endothelial cells (HUVEC). Results: In the viability assay, concentrations tested between 30 and 3000 µg/ml showed a reduction of viable HUVEC number. In the CAM assay, CS and GS in an amount 2.0 mg/implant increased the vessels number as compared to control (phosphate buffered saline-PBS). In the assay of the dorsal skin vascularization of adult Swiss mice, the groups treated with CS (2 mg/implant; Gelfoam plug) exhibited an increase in the vessels number into plugs (0.52 ± 0.08 g/dl; measured as plug-hemoglobin content), a similar effect to that promoted by fibroblast growth factor-2 (FGF-2; 50 ng/implant) (0.53 ± 0.1 g/dl). However the group treated with GS did not exhibit significant effect on mice skin vascularization. Conclusion: CS was capable to promote angiogenesis on CAM and dorsal skin vascularization, but GS only had pro-angiogenic activity in CAM vascular network.