Fibrocartilage decellularized extracellular matrix (dECM) is a promising alternative material for damaged fibrocartilage repair and replacement due to its biomimetic gross morphology and internal microstructure. However, the alterations in the microstructure and micromechanical properties of fibrocartilage after decellularization interfere with the macroscopic functional application of the scaffold. Therefore, this study aims to present an analytical atlas of the microstructure and micromechanics of the fibrocartilaginous dECM scaffold to elucidate the effect of decellularization treatment on the macroscopic function of the scaffold. The fibrocartilage dECM was prepared using the temporomandibular joint (TMJ) disc as the model, and its durability was evaluated under three functional states (physiological, physiological limit, and beyond the limit). The macroscopic function of different fibrocartilage dECM exhibits notable differences, which are attributed to the destruction of the multilevel collagen structure. This process involves unwinding triple-helix tropocollagen molecules, destroying collagen fibril D-periodicity, expanding collagen fiber bundle curling, and loosening of the collagen fiber network. The impairment of multiscale collagen structures degrades the cross-scale mechanical modulus and energy dissipation of dECM from the triple helix molecules to the fibril level to the fiber bundle that extends to the fiber network. This study provides important data for further optimizing decellularized fibrocartilage scaffolds and evaluating their translational potential.