Induction of zonal-specific cellular morphology and matrix synthesis for biomimetic cartilage regeneration using hybrid scaffolds

Abstract

<jats:p>Cartilage is anisotropic in nature and organized into distinct zones. Our goal was to develop zonal-specific three-dimensional hybrid scaffolds which could induce the generation of zonal-specific cellular morphology and extracellular matrix (ECM) composition. The superficial and middle zones comprised two layers of hyaluronic acid (HA) hydrogel which enveloped specifically orientated or randomly arranged polylactic acid nanofibre meshes. The deep zone comprised a HA hydrogel with multiple vertical channels. Primary bovine chondrocytes were seeded into the individual zonal scaffolds, cultured for 14 days and then the ECM was analysed. The aligned nanofibre mesh used in the superficial zone induced an elongated cell morphology, lower glycosaminoglycan (GAG) and collagen II production, and higher cell proliferation and collagen I production than the cells in the middle zone scaffold. Within the middle zone scaffold, which comprised a randomly orientated nanofibre mesh, the cells were clustered and expressed more collagen II. The deep zone scaffold induced the highest GAG production, the lowest cell proliferation and the lowest collagen I expression of the three zones. Assembling the three zones and stabilizing the arrangement with a HA hydrogel generated aligned, randomly aggregated and columnar cells in the superficial, middle and deep zones. This study presents a method to induce zonal-specific chondrocyte morphology and ECM production.</jats:p>

Differential Production of Cartilage ECM in 3D Agarose Constructs by Equine Articular Cartilage Progenitor Cells and Mesenchymal Stromal Cells

“…The zonal architecture is regarded as another potentially important factor when considering the development of functional cartilage [ 47 ], and mimicking cartilage zones in tissue engineering may be approached by using different cell types or materials for the different layers [ 20 , 26 , 29 , 48 , 49 ]. As it is not possible today to produce cartilage tissue in vitro the same way it develops in the body, these different approaches are employed to recapitulate a zonal architecture that is similar to the native zonal structure and has the potential to mimic some of the properties that render native zonal cartilage more stable than non-zonal repair tissue [ 48 ].…”

“…In the field of cartilage tissue engineering, several different approaches exist that try to produce cartilage constructs with zones similar to the native structure. Such zones are, for example, defined by the use of different cells (zone-specific chondrocytes, MSCs) [ 26 , 27 , 28 ] or different material properties (stiffness gradients, different hydrogels) [ 29 , 30 , 31 , 32 ]. The combination of ACPCs in a superficial layer and MSCs in a bottom layer has been tested in other hydrogels with encouraging results [ 18 , 20 , 21 ].…”

Maintenance and Acceleration of Pericellular Matrix Formation within 3D Cartilage Cell Culture Models

“…The hydrated HA was dialyzed against deionized water and PBS to remove any residual BDDE and the resulting gel was adjusted with PBS to a concentration of 20 mg/mL. 29 The hydrated HA was further processed to allow the reconstruction of gels into different size. Briefly, the crosslinked HA was pulverized with a homogenizer to obtain mini viscous gel particles of 0 to 400 µm.…”

Isotropic and Anisotropic Scaffolds for Tissue Engineering: Collagen, Conventional, and Textile Fabrication Technologies and Properties

“…However, in contrast to the hypothesis, a long-term study of 12 months failed to show any effect of fiber orientation on tissue formation (analysis based on collagen orientation) [210]. Owida et al prepared a three-layered electrospun polylactide nanofiber scaffold resembling the layered structure of natural articular cartilage of the knee [211]. After culturing chondrocytes in each layer for 14 days, both orientation of the cells and orientation of the newly formed ECM resembled the orientation of the respective layer.…”