Optimization of swim bladder decellularization with sodium dodecyl sulfate, triton X-100, and aminosulfabetaine-16 for use as a biomaterial

Swim bladders are biomaterials with numerous potential applications, but effective decellularization is essential to maximize retention of the extracellular matrix (ECM) and minimize immunogenicity. However, systematic optimization of detergent-based decellularization protocols for swim bladder remains limited, particularly with respect to balancing efficient nuclear removal with preservation of ECM, mechanics, and anti-calcification properties. We performed a systematic evaluation of the efficacy of three detergents-sodium dodecyl sulfate (SDS), Triton X-100 (TX-100), and aminosulfobetaine-16 (ASB-16)-for decellularizing<i>Hypophthalmichthys nobilis</i>swim bladders to develop an optimized decellularization protocol. Swim bladders were decellularized using varying concentrations of each detergent for varying lengths of time. Nuclear removal was first assessed by hematoxylin and eosin staining and DNA quantification, and cytotoxicity, hemocompatibility, mechanical properties, ECM preservation, microstructure, anti-calcification potential, and immunogenicity were subsequently evaluated. Selective detergent combinations were then performed based on their effects on the swim bladder tissues to optimize the decellularization protocol. SDS achieved the best nuclear removal, but compromised ECM integrity and mechanical properties, with some calcification observed. TX-100 demonstrated limited decellularization efficacy and negatively impacted the ECM. ASB-16 effectively removed nuclei while preserving ECM and mechanical properties, exhibiting good biocompatibility and anti-calcification properties. Cytotoxicity testing showed cell viability above 70% for all groups, and hemolysis rates were below 2%. Based on these findings, a combined decellularization protocol was developed, incubating swim bladders in 0.2% SDS for 2 h, followed by 0.5% ASB-16 for 12 h. This demonstrated a high decellularization efficiency, good biocompatibility and hemocompatibility, preservation of ECM and mechanical properties, and minimization of immunogenicity and calcification. This protocol can yield a biomaterial with numerous<i>in vivo</i>uses, including potential cardiovascular applications, and provides a practical framework for rational optimization of swim bladder decellularization.