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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 <jats:italic>VEGF<\/jats:italic> RNA or delivery of recombinant human VEGF<jats:sub>165<\/jats:sub> protein (rhVEGF<jats:sub>165<\/jats:sub>) across <jats:italic>in vitro<\/jats:italic> and <jats:italic>in vivo<\/jats:italic> 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<jats:sub>165<\/jats:sub> enhanced osteogenesis, as evidenced by increased human osteoblast-like MG-63 cell proliferation <jats:italic>in vitro<\/jats:italic> and calvarial bone thickness following <jats:italic>in vivo<\/jats:italic> administration. In contrast, transfection with <jats:italic>VEGF<\/jats:italic> RNA triggered angiogenic effects by promoting VEGF protein secretion from MG-63<jats:sub><jats:italic>VEGF<\/jats:italic>165<\/jats:sub> cells <jats:italic>in vitro<\/jats:italic>, which resulted in significantly increased angiogenesis in the chorioallantoic (CAM) assay <jats:italic>in ovo<\/jats:italic>. Furthermore, direct transfection of bone with <jats:italic>VEGF<\/jats:italic> RNA <jats:italic>in vivo<\/jats:italic> 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.<\/jats:p>","DOI":"10.1038\/s41598-019-53249-4","type":"journal-article","created":{"date-parts":[[2019,11,28]],"date-time":"2019-11-28T11:03:21Z","timestamp":1574939001000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":33,"title":["In vivo delivery of VEGF RNA and protein to increase osteogenesis and intraosseous angiogenesis"],"prefix":"10.1038","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4266-5962","authenticated-orcid":false,"given":"Robin M. 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