Fabrication of vascularized and scaffold-free bone tissue using endothelial and osteogenic cells differentiated from bone marrow derived mesenchymal stem cells

Abstract

Over-dependence on existing synthetic scaffolds and insufficient vascularization limit the development of tissue engineered bone (TEB). The purpose of this study is to fabricate vascularized and scaffold-free bone tissue using cell sheet technology and to assess its feasibility to repair critical-sized calvarial defects in rats. Firstly, the pre-vascularized cell sheet was formed by seeding BMSC-derived endothelial cells (ECs) on an undifferentiated BMSCs cell sheet layer in vitro. After 3 days of co-culture, ECs migrated and rearranged to form lumens on the BMSC sheet. Secondly, osteogenic cell sheet was formed by inducing osteogenic differentiation of high density BMSCs. Then, the pre-vascularized cell sheet was stacked on BMSC-derived osteogenic cell sheet to fabricate a scaffold-free construct for bone regeneration. Finally, the scaffold-free construct with both angiogenic and osteogenic potential was implanted into critical-sized calvarial defects in adult Wistar rats. Results showed that more functional perfused blood vessels and new bone tissue formed in the pre-vascularized group than that in the controls (both empty and non-pre-vascularized cell sheet group). This study indicates a new promising strategy for bone tissue regeneration.

Keywords: Bone marrow derived mesenchymal stem cell; Cell sheet; Rat skull critical defect; Tissue engineering; pre-vascularization.