Abstract

Tissue engineering techniques for the regeneration of large bone defects require sufficient vascularisation of the applied constructs to ensure a sufficient supply of oxygen and nutrients. In our previous work, prevascularised 3D scaffolds have been successfully established by coculture of bone marrow derived stem cells (MSCs) and endothelial progenitor cells (EPCs). We identified stabilising pericytes (PCs) as part of newly formed capillary-like structures. In the present study, we report preliminary data on the interactions between MSCs and EPCs, leading to the differentiation of pericyte-like cells. MSCs and EPCs were seeded in transwell cultures, direct cocultures, and single cultures. Cells were cultured for 10 days in IMDM 10% FCS or IMDM 5% FCS 5% platelet lysate medium. Gene expression of PC markers, CD146, NG2, αSMA, and PDGFR-β, was analysed using RT-PCR at days 0, 3, 7, and 10. The upregulation of CD146, NG2, and αSMA in MSCs in direct coculture with EPCs advocates the MSCs' differentiation towards a pericyte-like phenotype in vitro. These results suggest that pericyte-like cells derive from MSCs and that cell-cell contact with EPCs is an important factor for this differentiation process. These findings emphasise the concept of coculture strategies to promote angiogenesis for cell-based tissue engineered bone grafts.

Highlights

  • The remarkable regeneration potential of bone tissue is based on the presence of a highly branched vessel network, providing oxygen, nutrients, growth factors, and precursor cells to the injured tissue [1]

  • The upregulation of CD146, NG2, and αSMA in marrow derived stem cells (MSCs) in direct coculture with endothelial progenitor cells (EPCs) advocates the MSCs’ differentiation towards a pericyte-like phenotype in vitro. These results suggest that pericyte-like cells derive from MSCs and that cell-cell contact with EPCs is an important factor for this differentiation process

  • We developed an in vitro prevascularised 3D polyurethane (PU) bone implant seeded with EPCs (CD34+/ CD133+) and MSCs, which showed the formation of luminal tubular structures after 7 days of coculture [10]

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Summary

Introduction

The remarkable regeneration potential of bone tissue is based on the presence of a highly branched vessel network, providing oxygen, nutrients, growth factors, and precursor cells to the injured tissue [1]. The capacity of regeneration is limited and fails in large bone defects. Cell-based tissue engineering strategies have been developed and are accepted since a few years to address the challenge of bone repair. The approach of engineering bone tissue depends on the presence of osteogenic cells at the healing site and requires an adequate vascularisation of the applied biomaterial [2]. Bone-forming osteoblasts and endothelial cells (or their progenitors) are playing a crucial role for successful engraftment of cell seeded biomaterials [3]

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