Rapid formation of 3D: Decellularized extracellular matrix spheroids for enhancing bone formation

Bone marrow mesenchymal stem cell sheet-derived spheroids (BMSCs spheroids) have been widely studied as native bioactive scaffolds. However, the abundant cells in BMSCs spheroids cause immunogenicity and make them difficult to store. This paper aimed to construct a new bioactive scaffold called 3D-decellularized extracellular matrix spheroids (ECM spheroids) via decellularization of BMSCs spheroids to enhance bone formation. Hematoxylin and eosin staining (HE), nuclear and cytoskeletal fluorescence, immunofluorescence (IF), and scanning electron microscopy (SEM) were utilized to detect the characteristics and components of ECM spheroids. Furthermore, the biological properties of migration, adhesion, and recellularization of cells in ECM spheroids were assessed in vitro, and bone formation was evaluated in rat calvarial defects. The results showed that both the nuclei and cytoskeleton in ECM spheroids were greatly altered and one of the major components of FN was intact. The migration, adhesion, and recellularization potential were improved in vitro. Meanwhile, ECM spheroids promoted osteogenesis in rat skull defects after 3 months (p < .01). In conclusion, ECM spheroids were successfully prepared and proven to promote cell migration, adhesion, and proliferation. Bone formation in vivo was also accelerated. We believe that ECM spheroids can be used as bioactive and biocompatible 3D scaffolds in the future.