Modulating Temporospatial Phosphate Equilibrium by Nanoparticulate Mineralized Collagen Materials Induces Osteogenesis via PiT‐1 and PiT‐2

The temporospatial equilibrium of phosphate contributes to physiological bone development and fracture healing, yet optimal control of phosphate content has not been explored in skeletal regenerative materials. Nanoparticulate mineralized collagen glycosaminoglycan (MC‐GAG) is a synthetic, tunable material that promotes in vivo skull regeneration. In this work, the effects of MC‐GAG phosphate content on the surrounding microenvironment and osteoprogenitor differentiation are investigated. This study finds that MC‐GAG exhibits a temporal relationship with soluble phosphate with elution early in culture shifting to absorption with or without differentiating primary bone marrow‐derived human mesenchymal stem cells (hMSCs). The intrinsic phosphate content of MC‐GAG is sufficient to stimulate osteogenic differentiation of hMSCs in basal growth media without the addition of exogenous phosphate in a manner that can be severely reduced, but not eliminated, by knockdown of the sodium phosphate transporters PiT‐1 or PiT‐2. The contributions of PiT‐1 and PiT‐2 to MC‐GAG‐mediated osteogenesis are nonredundant but also nonadditive, suggestive that the heterodimeric form is essential to its activity. These findings indicate that the mineral content of MC‐GAG alters phosphate concentrations within a local microenvironment resulting in osteogenic differentiation of progenitor cells via both PiT‐1 and PiT‐2. Dynamic phosphate fluxes influence physiological bone development and healing; however, temporospatial phosphate control has not been explored in skeletal regenerative materials. This work describes the microenvironmental changes in phosphate equilibrium induced by a nanoparticulate mineralized collagen glycosaminoglycan material (MC‐GAG). The function of phosphate intrinsic to MC‐GAG in promoting osteoprogenitor differentiation via sodium phosphate cotransporters, PiT‐1 and PiT‐2, is characterized.