Effect of In Vitro Culture Length on the Bone-Forming Capacity of Osteoblast-Derived Decellularized Extracellular Matrix Melt Electrowritten Scaffolds

Recent advancements in decellularization have seen the development of extracellular matrix (ECM)-decorated scaffolds for bone regeneration; however, little is understood of the impact of in vitro culture prior to decellularization on the performances of these constructs. Therefore, this study investigated the effect of in vitro culture on ECM-decorated melt electrowritten polycaprolactone scaffold bioactivity. The scaffolds were seeded with osteoblasts and cultured for 1, 2, or 4 weeks to facilitate bone-specific ECM deposition and subsequently decellularized to form an acellular ECM-decorated scaffold. The utilization of mild chemicals and DNase was highly efficient in removing DNA while preserving ECM structure and composition. ECM decoration of the melt electrowritten fibers was observed within the first week of culture, with increased ECM at 2 and 4 week culture periods. Infiltration of re-seeded cells as well as overall bone regeneration in a rodent calvarial model was impeded by a longer culture period. Thus, it was demonstrated that the length of culture has a key influence on the osteogenic properties of decellularized ECM-decorated scaffolds, with long-term culture (2+ weeks) causing pore obstruction and creating a physical barrier which interfered with bone formation. These findings have important implications for the development of effective ECM-decorated scaffolds for bone regeneration.