The use of collagen in the biomedical device industry has led to major advances in soft tissue repair. This is attributed largely to the favourable biological and physiochemical properties of collagen. Regenerative medicine and tissue engineering favoured the use of this biomaterial and various commercial products have become available in the past few decades. This study aims to develop a collagen and chondroitin-6-sulphate dermal regeneration scaffold with enhanced resistance against enzymatic degradation. Frozen slurries (0.5% collagen) were dried under vacuum, coated with silicone, crosslinked and then thoroughly rinsed. The scaffolds were subjected to a range of quantitative and qualitative tests that included: scanning electron microscopy analysis, collagenase enzymatic degradation, and cytotoxicity assessment. Scaffold resistance to enzymatic degradation was manipulated after dehydrothermal treatment by employing combinations of crosslinking agents, such as glutaraldehyde and / or carbodiimide, with or without the presence of L-lysine. Results indicate that highly porous (mean pore diameter of 87.3 µm), bioactive, non-cytotoxic tissue engineering matrices were obtained. Enhanced stability of these scaffolds was achieved through extensive crosslinking and suggests the potential to prevent in vivo wound contraction sufficiently.