Differentiation of BMSCs into Nerve Precursor Cells on Fiber-Foam Constructs for Peripheral Nerve Tissue Engineering

Abstract

Bone marrow stem cells (BMSCs) are frequently used in nerve tissue engineering studies due to ease of their isolation and high potential for differentiation into nerve cells. A bilayer fiber-foam construct containing nanofibrous elements to house and guide BMSCs was designed as a model to study the regeneration of damaged peripheral nerve tissue and eventually serve as a nerve guide. The construct consisted of a) a macroporous bottom layer to serve as the backing and support, and for nutrient transport, and b) an electrospun, fibrous upper layer for cell attachment and guidance. Porosity and pore sizes of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) bottom layer were 85% and 5-200 µm, respectively, suitable for cell attachment and growth. Alignment of the cells is essential for cell-to-cell contact and the degree of alignment of electrospun PHBV/Collagen fibers was 11° when a frame type collector was used, while it was much higher (53°) for random fibers produced on an ordinary aluminum sheet collector. When the fibers were electrospun directly onto a PHBV foam attached on the frame type collector to create the bilayer, the degree of alignment of fibers decreased, alignment angle increased from 11° to 44°. This value did not change when the fibers were electrospun directly on the foams on the aluminum collector (53° vs 55°). A new media was designed to achieve comparable differentiation with the commercial media. It was found that the commercial Mesenchymal Stem Cell Neurogenic Differentiation Medium (PromoCell, Germany) was the better in terms of the expressions of neuronal markers nestin and β-III tubulin and the medium made in the lab with known constituents led to neuronal marker expressions very close to that with the commercial