Engineering meniscus structure and function via multi-layered mesenchymal stem cell-seeded nanofibrous scaffolds

Despite complex macroscopic and microscopic structural features of native tissue, including the circumferentially and radially aligned collagen bundles essential for mechanical function. To mimic this structural hierarchy, this study developed multi-lamellar mesenchymal stem cell (MSC)-seeded nanofibrous constructs. Bovine MSCs were seeded onto nanofibrous scaffolds comprised of poly(e-caprolactone) with fibers aligned in a single direction (0 degrees or 90 degrees to the scaffold long axis) or circumferentially aligned (C). Multi-layer groups (0 degrees/0 degrees/0 degrees, 90 degrees/90 degrees/90 degrees, 0 degrees/90 degrees/0 degrees, 90 degrees/0 degrees/90 degrees, and C/C/C) were created and cultured for a total of 6 weeks under conditions favoring fibrocartilaginous tissue formation. Tensile testing showed that 0 degrees and C single layer constructs had stiffness values several fold higher than 90 degrees constructs. For multi-layer groups, the stiffness of 0 degrees/0 degrees/0 degrees constructs was higher than all other groups, while 90 degrees/90 degrees/90 degrees constructs had the lowest values. Data for collagen content showed a general positive interactive effect for multi-layers relative to single layer constructs, while a positive interaction for stiffness was found only for the C/C/C group. Collagen content and cell infiltration occurred independent of scaffold alignment, and newly formed collagenous matrix followed the scaffold fiber direction. Structural hierarchies within multi-lamellar constructs dictated biomechanical properties, and only the C/C/C constructs with non-orthogonal alignment within layers featured positive mechanical reinforcement as a consequence of the layered construction. These multi-layer constructs may serve as functional substitutes for the meniscus as well as test beds to understand the complex mechanical principles that enable meniscus function. (C) 2015 Elsevier Ltd. All rights reserved.