Given their ability to dictate initial cell alignment and subsequent matrix organization, aligned electrospun scaffolds are a fitting means for engineering fiber-reinforced, anisotropic tissues such as tendon, ligament, the knee meniscus, and the annulus fibrosus [1–3]. However, one commonly observed limitation of such scaffolds is the relatively slow infiltration rates of surface-seeded cells, where the central thicknesses of constructs cultured for 10 weeks remain devoid of cells [2]. This limitation arises from the tight packing of fibers which yields small pore sizes, thereby hampering cell migration. Towards accelerating cell ingress, we have recently reported on two-polymer composite scaffolds containing both slow eroding poly(ε-caprolactone) (PCL) fibers as well as water-soluble poly(ethylene oxide) (PEO) fibers that serve as space holders during scaffold formation [4]. Removal of these PEO fibers prior to seeding resulted in improved cell infiltration after 3 weeks, but the long term maturation of such constructs has yet to be characterized. To assess the effect of sacrificial PEO fiber content on construct growth, a triple-jet electrospinning device was employed to generate PCL/PEO scaffolds with PEO fiber fractions ranging from 0 to 60%. After seeding with mesenchymal stem cells (MSCs), constructs were clamped in custom grips to maintain strip morphology. The mechanical and biochemical maturation of constructs was assessed over 9 weeks of free swelling culture in a chemically defined medium (CDM), along with cell infiltration and matrix distribution. We hypothesized that enhanced pore size in dual-fiber constructs would lead to not only a better distribution of cells, but also larger increases in stiffness resulting from enhanced matrix production and distribution.
An inverse small molecule screen to design a chemically defined medium supporting long-term growth of Drosophila cell lines
