Ricerc@Sapienza

Vascularization is a crucial factor when approaching any engineered tissue. Vascular
wall–mesenchymal stem cells are an excellent in vitro model to study vascular remodeling due to
their strong angiogenic attitude. This study aimed to demonstrate the angiogenic potential of
experimental highly porous scaffolds based on polylactic acid (PLA) or poly‐e‐caprolactone (PCL)
doped with calcium silicates (CaSi) and dicalcium phosphate dihydrate (DCPD), namely PLA‐
10CaSi‐10DCPD and PCL‐10CaSi‐10DCPD, designed for the regeneration of bone defects. Vascular
wall–mesenchymal stem cells (VW‐MSCs) derived from pig thoracic aorta were seeded on the
scaffolds and the expression of angiogenic markers, i.e. CD90 (mesenchymal stem/stromal cell
surface marker), pericyte genes α‐SMA (alpha smooth muscle actin), PDGFR‐β (platelet‐derived
growth factor receptor‐β), and NG2 (neuron‐glial antigen 2) was evaluated. Pure PLA and pure PCL
scaffolds and cell culture plastic were used as controls (3D in vitro model vs. 2D in vitro model).
The results clearly demonstrated that the vascular wall mesenchymal cells colonized the scaffolds
and were metabolically active. Cells, grown in these 3D systems, showed the typical gene expression
profile they have in control 2D culture, although with some main quantitative differences. DNA
staining and immunofluorescence assay for alpha‐tubulin confirmed a cellular presence on both
scaffolds. However, VW‐MSCs cultured on PLA‐10CaSi‐10DCPD showed an individual cells
growth, whilst on PCL‐10CaSi‐10DCPD scaffolds VW‐MSCs grew in spherical clusters. In
conclusion, vascular wall mesenchymal stem cells demonstrated the ability to colonize PLA and
PCL scaffolds doped with CaSi‐DCPD for new vessels formation and a potential for tissue
regeneration.