Osteoblast‐Derived Mitochondria Formulated with Cationic Liposome Guide Mesenchymal Stem Cells into Osteogenic Differentiation

Abstract While mitochondria are known to be essential for intracellular energy production and overall function, emerging evidence highlights their role in influencing cell behavior through mitochondrial transfer. This phenomenon provides a potential basis for the development of treatment strategies for tissue damage and degeneration. This study aims to evaluate whether mitochondria isolated from osteoblasts can promote osteogenic differentiation in mesenchymal stem cells (MSCs). Mitochondria from MSCs, which primarily utilize glycolysis, are compared with those from MG63 cells, which depend on oxidative phosphorylation. Mitochondria from both cell types are then encapsulated in cationic liposomes and transferred to MSCs, and their impact on differentiation is assessed. Mitochondria delivery from MG63 cells to MSCs grown in both two‐ and three‐dimensional cultures results in increased expression of osteogenic markers, including Runt‐related transcription factor 2, Osterix, and Osteopontin, and upregulation of genes involved in Bone morphogenetic protein 2 signaling and calcium import. This is accompanied by increased calcium influx and regulated by the Wnt/β‐catenin signaling pathway. Transplantation of spheroids containing MSCs with MG63‐derived mitochondria in bone defect animal models improves bone regeneration. The results suggest that delivery of MG63‐derived mitochondria effectively guides MSCs toward osteogenesis, paving the way for the development of mitochondria‐transplantation therapies.