3D Scaffolds of Polycaprolactone/Copper-Doped Bioactive Glass: Architecture Engineering with Additive Manufacturing and Cellular Assessments in a Coculture of Bone Marrow Stem Cells and Endothelial Cells.

The local delivery of Cu from copper-doped bioactive glass (Cu-BaG) was combined with 3D printing of polycaprolactone (PCL) scaffolds for its potent angiogenic effect in bone tissue engineering. PCL and Cu-BaG were, respectively, dissolved and dispersed in acetone to formulate a moderately homogeneous ink. The PCL/Cu-BaG scaffolds were fabricated via direct ink writing into a cold ethanol bath. The architecture of the printed scaffolds, including strut diameter, strut spacing, and porosity, were investigated and characterized. The PCL/Cu-BaG scaffolds showed a Cu-BaG content-dependent mechanical property, as the compressive Young's modulus ranged from 7 to 13 MPa at an apparent porosity of 60%. The ion dissolution behavior in simulated body fluid was evaluated, and the hydroxyapatite-like precipitation on the strut surface was confirmed. Furthermore, the cytocompatibility of the PCL/Cu-BaG scaffolds was assessed in human bone marrow stem cell (hBMSC) culture, and a dose-dependent cytotoxicity of Cu was observed. Here, the PCL/BaG scaffold induced the higher expression of late osteogenic genes and in comparison to the PCL scaffold. The doping of Cu in BaG elicited higher expression of the early osteogenic marker gene but decreased the expression of late osteogenic marker genes and in comparison to the PCL/BaG scaffold, demonstrating the suppressing effect of Cu on osteogenic differentiation of hBMSCs. In a coculture of hBMSCs and human umbilical vein endothelial cells, both the PCL/BaG and PCL/Cu-BaG scaffolds stimulated the formation of a denser tubule network, compared to the PCL scaffold. Meanwhile, only slightly higher gene expression of was observed with the PCL/Cu-BaG scaffold than with the PCL/BaG scaffold, indicating the potent angiogenic effect of the released Cu.