acid) [2] have been widely applied for this purpose. Nevertheless, these materials present limitations related to mimicking the biological and/or biomechanical properties of natural bone apatite. For instance, although hydroxyapatite-based bone mimetics present chemical similarities with natural bone, they show poor resorbability and remain in the transplanted area even after 10 years. [1] Alternative therapeutics using growth factors (e.g., bone morphogenetic protein 2 and basic fibroblast growth factor), [3] or cells, such as osteoblasts or mesenchymal stem/progenitor cells (MSCs), have also been introduced. [4] These methods, however, are costly and time-consuming, because cell therapies usually involve the transplantation of mature osteoblasts, and osteoblastic differentiation requires ≈2-3 weeks. [5] Recent multidisciplinary approaches merging the knowledge in developmental biology with methods/techniques in tissue engineering have enabled the development of unprecedented technologies for cartilage and bone tissue engineering. [6] In this context, previous approaches for promoting bone regeneration were suboptimal as they were essentially based only on the concept of osteoblast-driven intramembranous ossification (IO).Bone is formed through two distinct processes: IO and endochondral ossification (EO). [7] Intramembranous ossification begins inside the osteoblast-secreted extracellular vesicles, i.e., matrix vesicles (MVs), and forms the flat bones (e.g., skull) and the cortical bone. [7][8][9] Matrix vesicles refer to small (20-200 nm) spherical bodies transported via lysosome and secreted by exocytosis from the cells, mainly osteoblasts. [10,11] On the other hand, in EO, bone replaces a cartilage intermediate. [12] Recently, EO was shown to initiate from chondrocyte-derived plasma membrane nanofragments (PMNFs), in the absence of osteoblasts and MVs. [8,13] Previous studies have shown that the membrane and enzymes constituting MVs are different from the parent cell membrane. [14,15] On the other hand, PMNFs are direct fragments of the parent cell membrane. Thus, MVs and PMNFs could be regarded to have different composition in phospholipids and enzymes, [14,16] and could be leading to the formation of apatite clusters with different structures. However, little is known about the exact morphology of bone apatite clusters formed from MVs (IO) and PMNFs (EO).