Use of a novel Screen–Enrich–Combine(-biomaterials) Circulating System to fill a 3D-printed open Ti6Al4V frame with mesenchymal stem cells/β-tricalcium phosphate to repair complex anatomical bone defects in load-bearing areas

BACKGROUND: Repairing complex anatomical load-bearing bone defects is difficult because it requires the restoration of the load-bearing function, reconstructing the anatomical shape, and repair by regenerated bone. We previously developed a Screen–Enrich–Combine(-biomaterials) Circulating System (SECCS) for rapid intraoperative enrichment of autologous bone marrow mesenchymal stem cells (MSCs) to enhance the osteogenic ability of porous bone substitutes. In this study, we prepared a 3D-printed Ti6A14V macroporous frame matching the defect shape to provide early load-bearing support and evaluated the efficacy of filling the frame with SECCS-processed MSCs/beta tricalcium phosphate (β-TCP) for long-term bone growth. METHODS: Fifteen 2-year-old goats were involved in this study, and the lateral part of their distal femur was removed by an electric saw and was fitted by a matching electron beam melting technology-prepared (EBM) Ti6Al4V frame. Three types of frames, filled with nothing, pure porous β-TCP, or SECCS-processed MSCs/β-TCP, were fixed onto the defect site. Repair efficacy was evaluated by X-ray radiography, computed tomography (CT), histology, and histomorphometry. RESULTS: In the basic regular hexagon printing unit, the combined side width (w) and the inscribed circle diameter (d) determines the printing frame’s mechanical strength. The compressive load was significantly higher for w=1.9 mm, d=4.4 mm than for w=1.7 mm, d=4.0 mm or w=2.0 mm, d=5.0 mm (P<0.05). The EBM-prepared Ti6Al4V defect-matched frame was well maintained 9 months after implantation. The MSCs successfully adhered to the wall of the porous β-TCP in the SECCS-processed group and had spread fully in the test samples. Each goat in the MSCs/β-TCP–the filled group, had approximately 31,321.7±22,554.7 of MSCs and a larger area of new bone growth inside the frame than the control and blank areas groups. CONCLUSIONS: Filling the 3D-printed Ti6Al4V large-aperture frame with osteogenic materials achieved biological reconstruction over a larger area of regenerated bone to repair complex anatomical weight-bearing bone defects under the condition of early frame-supported load bearing. MSCs/β-TCP prepared by SECCS can be used as a filling material for this type of bone defect to obtain more efficacious bone repair.