Human Engineered Cartilage and Decellularized Matrix as an Alternative to Animal Osteoarthritis Model

Abstract

(1) Objective: to obtain a reproducible, robust, well-defined, and cost-affordable in vitro model of human cartilage degeneration, suitable for drug screening; (2) Methods: we proposed 3D models of engineered cartilage, considering two human chondrocyte sources (articular/nasal) and five culture methods (pellet, alginate beads, silk/alginate microcarriers, and decellularized cartilage). Engineered cartilages were treated with pro-inflammatory cytokine IL-1β to promote cartilage degradation; (3) Results: articular chondrocytes have been rejected since they exhibit low cellular doubling with respect to nasal cells, with longer culture time for cell expansion; furthermore, pellet and alginate bead cultures lead to insufficient cartilage matrix production. Decellularized cartilage resulted as good support for degeneration model, but long culture time and high cell amount are required to obtain the adequate scaffold colonization. Here, we proposed, for the first time, the combined use of decellularized cartilage, as aggrecanase substrate, with pellet, alginate beads, or silk/alginate microcarriers, as polymeric scaffolds for chondrocyte cultures. This approach enables the development of suitable models of cartilaginous pathology. The results obtained after cryopreservation also demonstrated that beads and microcarriers are able to preserve chondrocyte functionality and metabolic activity; (4) Conclusions: alginate and silk/alginate-based scaffolds can be easily produced and cryopreserved to obtain a cost-affordable and ready-to-use polymer-based product for the subsequent screening of anti-inflammatory drugs for cartilage diseases.

Models of Osteoarthritis: Relevance and New Insights

“…May be included in a co-culture system, facilitates investigation into cell-cell and cell-ECM interactions under disease phenotype. Has potential to allow for investigating the effects of loading on cells in vitro Can be costly, 3D culture is still in infancy, cell proliferation is decreased, a broad range of model systems and raw materials exist which can influence cell activity necessitating additional optimization for in vitro model development [38,45,47,49,52,58,61,64] on a flat surface in polystyrene culture flasks, which exposes the cultured cells to an equal volume of the surrounding media containing the various nutrients and growth factors essential for cell development and proliferation [18,19]. 2D in vitro models have routinely allowed for the screening of chondroprotective compounds to attenuate the catabolic factors involved in AC degradation.…”

Equine Mesenchymal Stem/Stromal Cells Freeze-Dried Secretome (Lyosecretome) for the Treatment of Musculoskeletal Diseases: Production Process Validation and Batch Release Test for Clinical Use

“…Musculoskeletal disorders (MSDs) have an extensive and growing impact representing a major health problem worldwide. Despite years of research, the understanding of these diseases and the control on their progression has not yet been achieved; indeed, different therapeutic approaches are being investigated to prevent the disorders or to promote recovery and/or regeneration of the musculoskeletal system, including muscles, bones, and joints [1,2]. MSDs often result from traumatic injuries due, for example, to vigorous physical activity leading to excessive muscle fatigue, damage of joint structures (articular cartilage, subchondral bone, synovial membrane), and inflammation or lesion of the tendon/ligament structure.…”

Silk/Fibroin Microcarriers for Mesenchymal Stem Cell Delivery: Optimization of Cell Seeding by the Design of Experiment

“…Fibroin-coated alginate microcarriers (FAMs) were fabricated as previously described [36,40,41,42]. Briefly, sodium alginate (1% w / v , medium viscosity, Sigma-Aldrich, St. Louis, MO, USA) was solubilized in distilled water and then the solution was added dropwise into an aqueous solution containing calcium chloride (Sigma-Aldrich) 100 mM under magnetic stirring using a bead generator (Encapsulator VAR V1, Nisco Engineering AG, Zurich, Switzerland) to obtain alginate microcarriers (AMs).…”