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The development of  in vitro  3D models to get insights into the mechanisms of bone regeneration could accelerate the translation of experimental findings to the clinic, reducing costs and duration of experiments. This work explores the design and manufacturing of multi-compartments structures in poly(ε-caprolactone) (PCL) 3D-printed by Fused Filament Fabrication technique. The construct was designed with interconnected stalls to host stem cells and endothelial cells. Cells were encapsulated within an optimised gellan gum (GG)-based hydrogel matrix, crosslinked using strontium (Sr 2+ ) ions to exploit its bioactivity and finally, assembled within compartments with different sizes. Calcium (Ca 2+ )-crosslinked gels were also used as control for comparison of Sr 2+  osteogenic effect. The results obtained demonstrated that Sr 2+  ions were successfully diffused within the hydrogel matrix and increased the hydrogel matrix strength properties under compressive load. The  in vitro  co-culture of human-TERT mesenchymal stem cells (TERT- hMSCs) and human umbilical vein endothelial cells (HUVECs), encapsulated within Sr 2+  ions containing GG-hydrogels and inter-connected by compartmentalised scaffolds under osteogenic conditions, enhanced cell viability and supported osteogenesis, with a significant increase of alkaline phosphatase activity, osteopontin and osteocalcin respect with the Ca 2+ -crosslinked GG-PCL scaffolds. These outcomes demonstrate that the design and manufacturing of compartmentalised co-culture of TERT-hMSCs and HUVEC populations enables an effective system to study and promote osteogenesis.

Multi-compartment scaffold fabricated via 3D-printing as in vitro co-culture osteogenic model