Construction of multifunctional cell aggregates in angiogenesis and osteogenesis through incorporating hVE-cad-Fc-modified PLGA/β-TCP microparticles for enhancing bone regeneration.

Multicellular aggregates have been widely utilized for regenerative medicine; however, the heterogeneous structure and undesired bioactivity of cell-only aggregates hinder their clinical translation. In this study, we fabricated an innovative kind of microparticle-integrated cellular aggregate with multifunctional activities in angiogenesis and osteogenesis, by combining stem cells from human exfoliated deciduous teeth (SHEDs) and bioactive composite microparticles. The poly(lactide-co-glycolide) (PLGA)-based bioactive microparticles (PTV microparticles) were ∼15 μm in diameter, with dispersed β-tricalcium phosphate (β-TCP) nanoparticles and surface-modified vascular endothelialcadherin fusion protein (hVE-cad-Fc). After co-culturing with microparticles in U-bottomed culture plates, SHEDs could firmly attach to the microparticles with a homogeneous distribution. The PTV microparticle-integrated SHED aggregates (PTV/SHED aggregates) showed significant positive CD31 and ALP expression, as well as the significantly upregulated osteogenesis makers (Runx2, ALP, and OCN) and angiogenesis makers (Ang-1 and CD31), compared with PLGA, PLGA/β-TCP (PT) and PLGA/hVE-cad-Fc (PV) microparticle-integrated SHED aggregates. Finally, in mice, 3 mm calvarial defects filled with the PTV microparticle-integrated SHED aggregates achieved abundant vascularized neo-bone regeneration within 4 weeks. Overall, we believe that these multifunctional PTV/SHED aggregates could be used as modules for bottom-up regenerative medicine, and provide a promising method for vascularized bone regeneration.