Abstract
A major limitation of the 2D culture systems is that they fail to recapitulate the in vivo 3D cellular microenvironment whereby cell–cell and cell–extracellular matrix (ECM) interactions occur. In this paper, a biomaterial scaffold that mimics the structure of collagen fibers was produced by jet-spraying. This micro-fiber polycaprolactone (PCL) scaffold was evaluated for 3D culture of human bone marrow mesenchymal stromal cells (MSCs) in comparison with a commercially available electrospun scaffold. The jet-sprayed scaffolds had larger pore diameters, greater porosity, smaller diameter fibers, and more heterogeneous fiber diameter size distribution compared to the electrospun scaffolds. Cells on jet-sprayed constructs exhibited spread morphology with abundant cytoskeleton staining, whereas MSCs on electrospun scaffolds appeared less extended with fewer actin filaments. MSC proliferation and cell infiltration occurred at a faster rate on jet-sprayed compared to electrospun scaffolds. Osteogenic differentiation of MSCs and ECM production as measured by ALP, collagen and calcium deposition was superior on jet-sprayed compared to electrospun scaffolds. The jet-sprayed scaffold which mimics the native ECM and permits homogeneous cell infiltration is important for 3D in vitro applications such as bone cellular interaction studies or drug testing, as well as bone tissue engineering strategies.
Highlights
Bone cells in vivo reside in a 3D environment within an extracellular matrix (ECM) which acts as a structural and biochemical support
Further supporting evidence for the importance of 3D cultures for drug testing comes from several studies which demonstrate that drugs have dramatically different effects on cells when cultured in 3D compared to in 2D [10, 11]
In terms of osteogenic differentiation and ECM production, while both scaffolds induced similar quantities of collagen production, Alkaline phosphatase (ALP) and calcium production were higher on jet-sprayed scaffolds compared to electrospun scaffolds
Summary
Bone cells in vivo reside in a 3D environment within an extracellular matrix (ECM) which acts as a structural and biochemical support. A major limitation of the 2D culture systems is that they fail to recapitulate the in vivo 3D cellular microenvironment where cell–cell, cell–ECM interactions, as well as biochemical and mechanical signalling occur. The attrition rate for new drug therapies after entering clinical development is approximately 90% despite the enormous investment (~1 billion dollars/drug) in development and screening [2, 3]. This failure is attributed to the inadequate 2D in vitro systems employed. Further supporting evidence for the importance of 3D cultures for drug testing comes from several studies which demonstrate that drugs have dramatically different effects on cells when cultured in 3D compared to in 2D [10, 11]
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