Meniscus tissue engineering using a novel combination of electrospun scaffolds and human meniscus cells embedded within an extracellular matrix hydrogel

Abstract

Meniscus injury and degeneration have been linked to the development of secondary osteoarthritis (OA). Therapies that successfully repair or replace the meniscus are therefore likely to prevent or delay OA progression. We investigated the novel approach of building layers of aligned polylactic acid (PLA) electrospun (ES) scaffolds with human meniscus cells embedded in extracellular matrix (ECM) hydrogel to lead to formation of neotissues that resemble meniscus-like tissue. PLA ES scaffolds with randomly oriented or aligned fibers were seeded with human meniscus cells derived from vascular or avascular regions. Cell viability, cell morphology, and gene expression profiles were monitored via confocal microscopy, scanning electron microscopy (SEM), and real-time PCR, respectively. Seeded scaffolds were used to produce multilayered constructs and were examined via histology and immunohistochemistry. Morphology and mechanical properties of PLA scaffolds (with and without cells) were influenced by fiber direction of the scaffolds. Both PLA scaffolds supported meniscus tissue formation with increased COL1A1, SOX9, COMP, yet no difference in gene expression was found between random and aligned PLA scaffolds. Overall, ES materials, which possess mechanical strength of meniscus and can support neotissue formation, show potential for use in cell-based meniscus regeneration strategies.

Effect of scaffold morphology and cell co-culture on tenogenic differentiation of HADMSC on centrifugal melt electrospun poly (L‑lactic acid) fibrous meshes

“…In this study, we fabricated PLLA fibrous meshes with controlled fiber diameter, orientation, and thickness by the adjustment of the appropriate spinning parameters. PLLA was chosen as the biomaterial for the tendon regeneration application because it is a fully biocompatible, biodegradable, synthetic material which can undergo remodeling and ultimately be replaced by regenerated tissue in vivo [43, 44]. In addition, PLLA is thermoplastics and it can be heated to its melting point, cooled, and reheated again without significant degradation [45].…”

Anatomical region-dependent enhancement of 3-dimensional chondrogenic differentiation of human mesenchymal stem cells by soluble meniscus extracellular matrix

“…Baek et al [35] fabricated multilayered scaffolds with alternating layers of electrospun nanofibers and MSC-seeded alginate hydrogels, which allowed tunable tensile anisotropy depending on fiber orientation. However, compressive mechanical properties were not measured.…”

Structured Macroporous Hydrogels: Progress, Challenges, and Opportunities

“…[143] Lee et al used a wet spinning process with a rotating collector to directly fabricate aligned alginate microribbon hydrogels for cell encapsulation applications, [144] while both Baek et al and Yang et al have fabricated aligned electrospun polylactic acid (PLA) fibers using rotating drum and static parallel bar collectors respectively. [145,146] These PLA fibers can then be used as templates for gel channels aiming to dictate the growth and orientation of cells within a collagen-based hydrogel. [145,146] Tonsomboon and Oyen demonstrated a comparable system using a rotating mandrel collector with electrospun gelatin fibers subsequently used to reinforce alginate hydrogels, showing control over hydrogel mechanical properties through fiber alignment.…”

“…[145,146] These PLA fibers can then be used as templates for gel channels aiming to dictate the growth and orientation of cells within a collagen-based hydrogel. [145,146] Tonsomboon and Oyen demonstrated a comparable system using a rotating mandrel collector with electrospun gelatin fibers subsequently used to reinforce alginate hydrogels, showing control over hydrogel mechanical properties through fiber alignment. [147] Directed phase separation has also been used to fabricate anisotropic macroporous hydrogels, although the consistency of the anisotropy resulting from such processes is typically lower than achieved using other approaches.…”