Graphene-Based MicroRNA Transfection Blocks Preosteoclast Fusion to Increase Bone Formation and Vascularization

The objective of this study is to design a graphene-based miRNA transfection drug delivery system for antiresorptive therapy. An efficient nonviral gene delivery system is developed using polyethylenimine (PEI) functionalized graphene oxide (GO) complex loaded with miR-7b overexpression plasmid. GO-PEI complex exhibits excellent transfection efficiency within the acceptable range of cytotoxicity. The overexpression of miR-7b after GO-PEI-miR-7b transfection significantly abrogates osteoclast (OC) fusion and bone resorption activity by hampering the expression of an essential fusogenic molecule dendritic cell-specific transmembrane protein. However, osteoclastogenesis occurs without cell-cell fusion and preosteoclast (POC) is preserved. Through preservation of POC, GO-PEI-miR-7b transfection promotes mesenchymal stem cell osteogenesis and endothelial progenitor cells angiogenesis in the coculture system. Platelet-derived growth factor-BB secreted by POC is increased by GO-PEI-miR-7b both in vitro and in vivo. In treating osteoporotic ovariectomized mice, GO-PEI-miR-7b significantly enhances bone mineral density, bone volume as well as bone vascularization through increasing CD31^hiEmcn^hi cell number. This study provides a cell-cell fusion targeted miRNA transfection drug delivery strategy in treating bone disorders with excessive osteoclastic bone resorption.