Sterilization is a mandatory requirement for the manufacture and clinical translation of biomaterials and medical devices intended to contact with cells and tissues. Conventional sterilization methods, such as heat sterilization, gamma irradiation and ethylene oxide treatment, often compromise the structural and functional integrity of chemically sensitive materials, demonstrating the need for alternative approaches specifically tailored to sensitive biomaterials. Supercritical carbon dioxide (scCO₂) has emerged as a promising candidate owing to its low operating temperature, chemical inertness, non-toxicity, and tunable solvent properties. This study assessed the efficacy of scCO₂ sterilization applied to Gelatin methacryloyl (GelMA) and amphiphilic poly(ether urethane)s (PEUs), a natural-derived and a synthetic polymeric system, respectively, employed for the preparation of hydrogel systems with application in tissue engineering and regenerative medicine.