Collagen-based vascular substitutes represent in VTE a valid alternative for the replacement of diseased small-calibre blood vessels. In this study, collagen gel-based scaffolds were crosslinked combining modulation of pH and UV-C radiation. The effects on the mechanical properties, on the molecular structure and on cell viability and morphology were investigated. The mechanical response increased as a function of pH or UV-C dose and strongly depended on the test speed. Collagen molecular conformation resulted only slightly modified. While cell adhesion was not significantly altered, cell proliferation partially decreased in function of pH and UV-C. These findings suggest that UV-C treated collagen gels can represent an adequate substrate for VTE applications.