Introduction: Lack of sufficient vascularization to support cell viability, growth and function in scaffold guided tissue regeneration (SGTR) is a prevalent challenge facing tissue engineering today. Matching axial vascularization is a regenerative therapeutic approach which incorporates the benefits of flap-based techniques for neo-vascularization to further aid tissue regeneration. Methodology: An ovine critical-size tibial defect model (Medium defect volume=9.5 cm3) was undertaken in eight sheep to evaluate the novel tissue engineering approach involving a 3D-printed medical-grade ε-polycaprolactone b-tricalcium phosphate (mPCL-TCP) scaffold with a cortico-periosteal flap (CPF). Biomechanical, radiological, histological and immunohistochemical analysis confirmed functional bone regeneration comparable to the clinical gold standard control (autologous bone graft) and was superior to a scaffold control group (mPCL-TCP only). A pilot study was performed on two sheep where the defect volume was doubled to 19 cm3 (X-Large defect volume) to represent the most challenging clinical situation. Results: Positive results of both the M and XL defect volume study supported clinical translation. A 27-year-old adult male underwent reconstruction of a 36 cm near-total intercalary defect of the tibia secondary to osteomyelitis using this original scaffold guided bone regeneration concept. Bone regeneration was confirmed postoperatively both radiologically and histologically with complete independent weight bearing achieved within 24 months. Conclusion: This article represents the widely advocated and seldomly accomplished concept of “bench-to-bedside” research. The presented original scaffold-guided treatment concept to reconstruct major bone loss in load-bearing limbs will have significant implications for reconstructive surgery and regenerative medicine more generally.