Self-assembled organoid-tissue modules for scalable organoid engineering: Application to chondrogenic regeneration

Abstract

Tissue engineering has made significant strides in creating biomimetic grafts for the repair and regeneration of damaged tissues; however, the scalability of engineered tissue constructs remains a major technical hurdle. This study introduces a method for generating organoid-tissue modules (Organoid-TMs) through scaffold-free self-assembly of microblocks (MiBs) derived from adipose-derived mesenchymal stem cells (ADMSCs). The key parameters influencing Organoid-TM formation were identified as the density of MiBs and the controlled mixing ratio of large and small MiBs. The resulting Organoid-TM exhibited a distinctive cup-shaped morphology, a millimeter-scale structure with enhanced nutrient and oxygen diffusion compared to conventional spherical aggregates. Despite their larger size, Organoid-TMs maintained ADMSC stemness and differentiation potential, while stemness and differentiation were halted during fabrication. Organoid-TMs receiving chondrogenic cues during fabrication were transplanted into cartilage defect sites in animal models, demonstrating cartilage regeneration efficacy in a scaffold-independent and xeno-free manner. This fabrication method represents a highly reproducible and consistent process for developing spheroids or organoids, offering a robust platform for regenerative medicine applications. Specifically, Organoid-TMs provide a foundational framework for therapeutic strategies targeting cartilage defects and osteoarthritis, paving the way for advancements in tissue-engineered therapeutics. STATEMENT OF SIGNIFICANCE: This study introduces a distinct approach in tissue engineering, utilizing self-assembled Organoid-Tissue Modules (Organoid-TMs) to address persistent challenges in scalable organoid production and cartilage regeneration. By leveraging adipose-derived mesenchymal stem cells (ADMSCs) and carefully optimizing the size, ratio, and spatial organization of microblocks (MiBs), we successfully generated millimeter-scale Organoid-TMs. The distinctive cup-shaped architecture of these Organoid-TMs enhances oxygen and nutrient diffusion, effectively overcoming limitations such as core necrosis typically encountered in large-scale organoid culture. This system demonstrated substantial regenerative potential, particularly in chondrogenic differentiation and cartilage repair in both rabbit and pig models, without the use of artificial scaffolds or xenogenic materials.

Keywords: Chondrogenesis; Mesenchymal stem cell; Organoid; Regeneration; Self-organization; Tissue engineering.