A true biomimetic of the cartilage extracellular matrix (ECM) could greatly contribute to our ability to regenerate this tissue in a mechanically demanding, often inflamed environment. Articular cartilage is a composite tissue made of cells and fibrillar proteins embedded in a hydrophilic polymeric meshwork. Here, a polyanionic functionalized alginate is used to mimic the glycosaminoglycan component of the native ECM. To create the fibrillar component, cryoelectrospinning of poly(ε-caprolactone) on a -78 °C mandrel, subsequently treated by O plasma, is used to create a stable, ultraporous and hydrophillic nanofiber network. In this study, cell-laden, fiber-reinforced composite scaffolds thicker than 1.5 mm can be created by infiltrating a chondrocyte/alginate solution into the fiber mesh, which is then physically cross-linked. The fibrillar component significantly reinforces the chondroinductive, but mechanically weak sulfated alginate hydrogels. This allows the production of a glycosaminoglycan- and collagen type II-rich matrix by the chondrocytes as well as survival of the composite in vivo. To further enhance the system, the electrospun component is loaded with dexamethasone, which protected the cells from an IL-1β-mediated inflammatory insult.

Enzymatic crosslinking has immense appeal for tissue engineers as one of the most biocompatible methods of hydrogel crosslinking. Sortase A has a number of unique advantages over previous systems. We show an impressive and tunable range of crosslinking kinetics, from almost instantaneous gelation to several minutes. We also demonstrate that Sortase A crosslinked hydrogels have good cytocompatibility and cause no immune reaction when implanted in vivo. With its additional benefits of excellent stability in solution and easy large-scale synthesis available to any lab, we believe this novel crosslinking modality will find multiple applications in high throughput screening, tissue engineering, and biofabrication.

Intra-articular administration of glucocorticoids such as dexamethasone is a common treatment for osteoarthritic inflammation and pain. Despite its potent anti-inflammatory properties, multiple barriers hinder the drug's effectiveness in the articular space. In particular, the high turnover rate of the synovial fluid and the dense cartilage extracellular matrix (ECM) lead to poor drug penetration into cartilage. In order to increase the infiltration and retention time, two dexamethasone prodrugs were developed. Firstly, dexamethasone was conjugated to polycationic chitosan, which led to deep and sustained infiltration of the drug into full thickness cartilage, due to its strong electrostatic interactions with the high negative fixed charge of the cartilage ECM. Secondly, dexamethasone was conjugated to a collagen type II-binding peptide, WYRGRL, and this prodrug was shown to be retained in the deep zones of cartilage through specific interactions with cartilage-specific collagen type II bundles. In both cases, active dexamethasone was released from the carrier by ester linkage hydrolysis. Complexing dexamethasone with either chitosan or collagen type II-affinity carriers increased its binding and therapeutic efficacy inside cartilage, compared to free drug. Both dexamethasone conjugates significantly reduced levels of inflammatory markers and slowed the loss of glycosaminoglycans in an ex vivo model. A single dose of a cartilage-targeting dexamethasone prodrug represents a promising alternative to the repetitive glucocorticoid injections needed to compensate for its rapid clearance from the joint cavity.

The goal of this study was to evaluate human epiphyseal chondroprogenitor cells (ECPs) as a potential new cell source for cartilage regeneration. ECPs were compared to human bone marrow stromal cells (MSCs) and human adult articular chondrocytes (ACs) for their chondrogenic potential and phenotypic stability in vitro and in vivo. The cells were seeded in Optimaix-3D scaffolds at 5 × 10 cells/mm and gene expression, matrix production and mechanical properties were analysed up to 6 weeks. In vitro, ECPs synthesized consistently high collagen 2 and low collagen 10. AC-seeded constructs exhibited high donor variability in GAG/DNA values as well as in collagen 2 staining, but showed low collagen 10 production. MSCs, on the other hand, expressed high levels of collagen 2 but also of collagens 1 and 10, and were therefore not considered further. In vivo, there was considerable loss of matrix proteins in ECPs compared to in vitro cultured samples. To overcome this, a second implantation study investigated the effect of mixing cells with alginate prior to seeding in the scaffold. ECPs in alginate maintained their cartilage matrix and resisted mineralization and vessel infiltration better 6 weeks after subcutaneous implantation, whereas ACs lost their chondrogenic matrix completely. This study shows the great potential of ECPs as an off-the-shelf, highly chondrogenic cell type that produces stable cartilage in vivo. Copyright © 2016 John Wiley & Sons, Ltd.

Highlights  Chondrogenesis of a tissue-engineered cartilage graft is feasible in immunocompetent small animals  Immunocompetent and immunodeficient animals lead to analogous results in terms of chondrogenesis, as long as the implanted cells are shielded from the host by a biomaterial  Subcutaneous implantation in small animals with a complete and human immune system could help to predict the outcome of engineered grafts for cartilage applications