Collagen is a potential candidate for the preparation of biomimetic materials able to stimulate bone mineralization and cartilage regeneration. The human cartilage has a low ability of regeneration due to its avascularity. Therefore tissue engineering aims at obtaining biomimetic materials with controlled biodegradability rates and able to improve specific local cell stimulation on site of defects. Chondroitin sulfate is one of the main glycosaminoglycan component of cartilage and it can connect or retains growth factors. Carrageenan has chemical and structural characteristics similar to chondroitin sulfate, so it is a candidate as a substitute of this glycosaminoglycan. Furthermore, carrageenan has better rheological properties compared to chondroitin sulfate and can be obtained from renewable sources. This study aimed at comparing mechanical properties of type II collagen containing either kappa-carrageenan or chondroitin sulfate. Type II collagen was isolated from pig’s ear at a concentration of 1.7 mg/mL. After concentration by tangential flow filtration, a solution containing 5 mg/mL of type II collagen was used to prepare the scaffolds. The tertiary helical structure of the collagen triple-helix was confirmed by circular dichroism spectrum. The molecular weight of collagen was obtained by SDS-page electrophoresis and it was found bands associated to the 3 α1 polypeptide chains (192.5, 156.6 and 114.9 kDa). The scaffolds were prepared and characterized. Bands associated to the chemical structure of collagen, kappa-carrageenan and chondroitin sulfate at 1540, 1225 and 1640 cm−1 were detected by FTIR. The modification of size distribution and organization of collagen fibrils caused by the addition of kappa-carrageenan and chondroitin sulfate was observed by SEM images, which indicates the potential of the substitute kappa-carrageenan associated with collagen for cartilage regeneration.