Enhanced osteogenesis in co-cultures with human mesenchymal stem cells and endothelial cells on polymeric microfiber scaffolds

In this work, human mesenchymal stem cells (hMSCs) and their osteogenically precultured derivatives were directly co-cultured with human umbilical vein endothelial cells (HUVECs) on electrospun 3D poly(-caprolactone) microfiber scaffolds in order to evaluate the co-culture’s effect on the generation of osteogenic constructs. Specifically, cells were cultured on scaffolds for up to three weeks, and the cellularity, alkaline phosphatase activity (ALP), and bone-like matrix formation were assessed. Constructs with co-cultures and monocultures had almost identical cellularity after the first week, however lower cellularity was observed in co-cultures compared to monocultures during the subsequent two weeks of culture. Scaffolds with co-cultures showed significantly higher ALP activity, glycosaminoglycan and collagen production, as well as greater calcium deposition over the course of study compared to monocultures of hMSCs. Furthermore, the osteogenic outcome was equally robust in co-cultures containing osteogenically precultured and non-precultured hMSCs. The results demonstrate that the combination of MSC and HUVEC populations within a porous scaffold material under osteogenic culture conditions is an effective strategy to promote osteogenesis