Development of porcine skeletal muscle extracellular matrix-derived hydrogels with improved properties and low immunogenicity

• 成果类型: Article
• 署名作者: Barajaa, Mohammed A.; Otsuka, Takayoshi; Ghosh, Debolina; Kan, Ho-Man; Laurencin, Cato T.
• 署名单位: Imam Abdulrahman Bin Faisal University; University of Connecticut; University of Connecticut; University of Connecticut; University of Connecticut; University of Connecticut
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-14191
• DOI: 10.1073/pnas.2322822121
• 发表日期: 2024-05-07
• 关键词: immune-response; decellularization process; collagen fibrillogenesis; macrophage phenotype; biologic scaffolds; antigen removal; neural tissue; xenotransplantation; cells; pigs

摘要

Hydrogels derived from decellularized extracellular matrices (ECM) of animal origin show immense potential for regenerative applications due to their excellent cytocompatibility and biomimetic properties. Despite these benefits, the impact of decellularization protocols on the properties and immunogenicity of these hydrogels remains relatively unexplored. In this study, porcine skeletal muscle ECM (smECM) underwent decellularization using mechanical disruption (MD) and two commonly employed decellularization detergents, sodium deoxycholate (SDC) or Triton X - 100. To mitigate immunogenicity associated with animal - derived ECM, all decellularized tissues were enzymatically treated with alpha- galactosidase to cleave the primary xenoantigen-the alpha- Gal antigen. Subsequently, the impact of the different decellularization protocols on the resultant hydrogels was thoroughly investigated. All methods significantly reduced total DNA content in hydrogels. Moreover, alpha- galactosidase treatment was crucial for cleaving alpha- Gal antigens, suggesting that conventional decellularization methods alone are insufficient. MD preserved total protein, collagen, sulfated glycosaminoglycan, laminin, fibronectin, and growth factors more efficiently than other protocols. The decellularization method impacted hydrogel gelation kinetics and ultrastructure, as confirmed by turbidimetric and scanning electron microscopy analyses. MD hydrogels demonstrated high cytocompatibility, supporting satellite stem cell recruitment, growth, and differentiation into multinucleated myofibers. In contrast, the SDC and Triton X - 100 protocols exhibited cytotoxicity. Comprehensive in vivo immunogenicity assessments in a subcutaneous xenotransplantation model revealed MD hydrogels' biocompatibility and low immunogenicity. These findings highlight the significant influence of the decellularization protocol on hydrogel properties. Our results suggest that combining MD with alpha- galactosidase treatment is an efficient method for preparing low - immunogenic smECM - derived hydrogels with enhanced properties for skeletal muscle regenerative engineering and clinical applications.