Collagen Type II enhances chondrogenic differentiation in agarose-based modular microtissues

Abstract

Background Cell-based therapies have made an impact on the treatment of osteoarthritis, however the repair and regeneration of thick cartilage defects is an important and growing clinical problem. Next-generation therapies that combine cells with biomaterials may provide improved outcomes. We have developed modular microenvironments that mimic the composition of articular cartilage as a delivery system for consistently differentiated cells. Methods Human bone marrow-derived mesenchymal stem cells (MSC) were embedded in modular microbeads consisting of agarose (AG) supplemented with 0%, 10%, and 20% collagen Type II (COL-II) using a water-in-oil emulsion technique. AG and AG/COL-II microbeads were characterized in terms of their structural integrity, size distribution, and protein content. The viability of embedded MSC and their ability to differentiate into osteogenic, adipogenic and chondrogenic lineages over three weeks in culture were also assessed. Results Microbeads made with <20% COL-II were robust, generally spheroidal in shape and 80 ± 10 µm in diameter. MSC viability in microbeads was consistently high over a week in culture, while viability in corresponding bulk hydrogels decreased with increasing COL-II content. Osteogenic differentiation of MSC was modestly supported in both AG and AG/COL-II microbeads, while adipogenic differentiation was strongly inhibited in COL-II containing microbeads. Chondrogenic differentiation of MSC was clearly promoted in microbeads containing COL-II, compared to pure AG matrices. Conclusions Inclusion of collagen Type II in agarose matrices in microbead format can potentiate chondrogenic differentiation of human MSC. Such compositionally tailored microtissues may find utility for cell delivery in next-generation cartilage repair therapies.

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

“…In total, we provide further evidence that ACPCs can be regarded as a promising alternative cell source for cartilage engineering. Moreover, taken together with previous studies [ 21 , 44 , 45 ], the results underscore the importance of the choice of an appropriate hydrogel for each cell type. Agarose appeared to be a suitable hydrogel for ACPCs, and thus may also be utilized in future studies when further investigating layered zonal constructs, while other types of gels may be considered for layers in which MSCs are applied.…”

“…Agarose has been shown to support the native phenotype and morphology of chondrocytes [ 22 , 23 ], and appears to be a suitable environment for chondrocytes and their subpopulation, ACPCs. Taken as a whole, these results support that the selected hydrogel can largely influence the performance of the cells, and the choice of the hydrogel and cell type must be well matched to achieve the desired results [ 21 , 44 , 45 , 46 ].…”

Plant-Derived Biomaterials: A Review of 3D Bioprinting and Biomedical Applications

“…It has however been noted to be adversely mechanically affected by the presence of cells: cells within agarose diminish the gel strength due to interference with the hydrogen bonding required for crosslinking and gelation (Shoichet et al, 1996). This limitation can be somewhat surmounted through combination with other polymers and proteins such as collagen, chitosan, and cellulose to increase cell affinity (Annamalai et al, 2016;Awadhiya et al, 2017). Research into the role of agarose composites has been explored for tissue engineering purposes for neural, vascular, bone, and pancreatic tissue (Bhatnagar et al, 2016;Zarrintaj et al, 2018).…”

Core-binding factor beta is required for osteoblast differentiation during fibula fracture healing

“…Collagen II, known to regulate chondrogenesis of mesenchymal stem cells, has been demonstrated to have the potential of facilitating osteogenesis and suppressing adipogenesis during early stage of mesenchymal stem cell differentiation [18]. Stegemann et al also revealed Collagen II to be a key factor in enhancing chondrogenic differentiation in agarose-based modular microtissues [19]. PCNA is widely accepted to be involved in distinct pathways of DNA postreplication repair [20].…”