Development and characterization of decellularized meniscus-derived bioscaffolds

This study developed and validated a porcine meniscus decellularization protocol designed to avoid harsh anionic detergents while preserving key extracellular matrix (ECM) constituents for downstream tissue-engineering and bioink applications. Inner and outer meniscus regions were processed separately, and the new protocol (freeze–thaw + hypotonic lysis + Triton X-100/KCl extraction + DNase/RNase digestion) achieved near-complete cell removal (reported ~99% dsDNA reduction) while retaining collagen architecture and region-dependent ECM markers (collagen I/II/IV/VI, fibronectin, laminin) by histology, immunostaining, and SEM. Decellularized inner (DIM) and outer (DOM) meniscus ECM was then pepsin-digested and incorporated into a proprietary alginate formulation to create composite beads intended as a cell-instructive hydrogel/bioink platform. Bead size, sol content, swelling, and initial compressive stiffness were similar across formulations, indicating ECM peptides did not disrupt alginate crosslinking or bulk mechanics. In long-term 3D culture, human adipose-derived stromal cells (ASCs) encapsulated in ECM-containing beads remained highly viable for 28 days; viable cell density increased significantly in ALG+DIM beads and was higher than alginate-only controls at day 28. Under chondrogenic differentiation conditions, ECM-containing beads showed markedly enhanced fibrochondrogenic ECM marker accumulation (collagen I, collagen II, fibronectin), supporting meniscus-derived ECM peptides as bioactive cues for meniscus tissue engineering and region-informed bioink design.