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Rapid and adequate vascularization is vital to the long-term success of porous orbital enucleation implants. In this study, porous hydroxyapatite (HA) scaffolds coated with vascular endothelial growth factor (VEGF)-functionalized collagen (COL)/heparin (HEP) multilayers (porosity 75%, pore size 316.8 ± 77.1 μm, VEGF dose 3.39 ng/mm 3 ) were fabricated to enhance vascularization by inducing the differentiation of mesenchymal stem cells (MSCs) to endothelial cells. The  in vitro  immunofluorescence staining, quantitative real-time polymerase chain reaction (qRT-PCR), and western blotting results demonstrated that the expression of the endothelial differentiation markers CD31, Flk-1, and von Willebrand factor (vWF) was significantly increased in the HA/(COL/HEP) 5 /VEGF/MSCs group compared with the HA/VEGF/MSCs group. Moreover, the HA/(COL/HEP) 5  scaffolds showed a better entrapment of the MSCs and accelerated cell proliferation. The  in vivo  assays showed that the number of newly formed vessels within the constructs after 28 d was significantly higher in the HA/(COL/HEP) 5 /VEGF/MSCs group (51.9 ± 6.3/mm 2 ) than in the HA (26.7 ± 2.3/mm 2 ) and HA/VEGF/MSCs (38.2 ± 2.4/mm 2 ) groups. The qRT-PCR and western blotting results demonstrated that the HA/(COL/HEP) 5 /VEGF/MSCs group also had the highest expression of CD31, Flk-1, and vWF at both the mRNA and protein levels.

In vivo vascularization of MSC-loaded porous hydroxyapatite constructs coated with VEGF-functionalized collagen/heparin multilayers