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Deficient bone vasculature is a key component in pathological conditions ranging from developmental skeletal abnormalities to impaired bone repair. Vascularisation is dependent upon vascular endothelial growth factor (VEGF), which drives both angiogenesis and osteogenesis. The aim of this study was to examine the efficacy of blood vessel and bone formation following transfection with  VEGF  RNA or delivery of recombinant human VEGF 165  protein (rhVEGF 165 ) across  in vitro  and  in vivo  model systems. To quantify blood vessels within bone, an innovative approach was developed using high-resolution X-ray computed tomography (XCT) to generate quantifiable three-dimensional reconstructions. Application of rhVEGF 165  enhanced osteogenesis, as evidenced by increased human osteoblast-like MG-63 cell proliferation  in vitro  and calvarial bone thickness following  in vivo  administration. In contrast, transfection with  VEGF  RNA triggered angiogenic effects by promoting VEGF protein secretion from MG-63  VEGF 165  cells  in vitro , which resulted in significantly increased angiogenesis in the chorioallantoic (CAM) assay  in ovo . Furthermore, direct transfection of bone with  VEGF  RNA  in vivo  increased intraosseous vascular branching. This study demonstrates the importance of continuous supply as opposed to a single high dose of VEGF on angiogenesis and osteogenesis and, illustrates the potential of XCT in delineating in 3D, blood vessel connectivity in bone.

In vivo delivery of VEGF RNA and protein to increase osteogenesis and intraosseous angiogenesis