3D decellularized bovine meniscal sponge scaffold as a graft candidate: Comparative in vitro study with standard grafts

Background: Meniscal injury disrupts knee biomechanics and accelerates degeneration. Current treatments, including allografts, are limited by donor availability and biological constraints. Decellularized extracellular matrix (ECM) scaffolds offer a promising alternative. This study evaluated a three-dimensional (3D) decellularized bovine meniscal sponge scaffold and the effects of different crosslinking agents on its properties. Methods: An in vitro experimental study was conducted with four groups: human meniscus, non-crosslinked bovine scaffold, bovine scaffold treated with glutaraldehyde (GTA), and bovine scaffold treated with tannic acid. Scaffolds were characterized for morphology, ECM preservation, cytocompatibility, chemical properties, degradation, swelling, and mechanical performance using SEM, Masson’s trichrome staining, MTT assay, FTIR, and mechanical testing. Results: The optimized decellularization protocol effectively removed cellular components while preserving ECM structure. All scaffolds exhibited a porous 3D architecture with maintained collagen content, especially in tannic acid-treated groups. Crosslinking significantly improved mechanical properties, with the highest compressive strength in the tannic acid 4-h group (2.51 ± 1.02 MPa), followed by the GTA 24-h group (2.46 ± 1.15 MPa). Crosslinked scaffolds also showed enhanced degradation resistance while maintaining adequate swelling capacity. All groups exhibited cell viability above 70%, indicating acceptable cytocompatibility. Conclusion: The 3D decellularized bovine meniscal scaffold shows strong potential as a meniscal graft substitute. Tannic acid crosslinking, particularly at 4 h, provided the most balanced performance, supporting its potential for meniscus tissue engineering.