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.
Highlights
Osteoarthritis is a highly prevalent degenerative osteoarticular disease that causes serious debilitating condition and the interest of the scientific community is oriented to the development of new Polymers 2018, 10, 738; doi:10.3390/polym10070738 www.mdpi.com/journal/polymersPolymers 2018, 10, 738 cost-effective therapies [1]
In cartilage tissue engineering field, a critical factor is the achievement of an adequate number of functional chondrocytes for the complete colonization of scaffolds; in this context, the main limit is represented by the low number of cells obtained from donor tissues and by their easy loss of phenotype that causes the loss of functionality
We considered articular (ACs) and nasal septum (NCs) chondrocytes to evaluate the optimal cell source for the production of engineered cartilage tissue in a short time
Summary
Osteoarthritis is a highly prevalent degenerative osteoarticular disease that causes serious debilitating condition and the interest of the scientific community is oriented to the development of new Polymers 2018, 10, 738; doi:10.3390/polym10070738 www.mdpi.com/journal/polymersPolymers 2018, 10, 738 cost-effective therapies [1]. Animals must be replaced by alternative methods (Replacement), their number must be reduced to a minimum (Reduction) and, where not possible, procedures should minimize the suffering of the same (Refinement). In this contest, 3D cell cultures have been proposed to obtain predictive in vitro models of numerous pathologies; in particular, during the screening of new drugs, 3D models can be used to test a large number of bioactive compounds reducing the cost of the early stages of drug discovery [3,4,5]. 3D cell cultures have been proposed to obtain predictive in vitro models of numerous pathologies; in particular, during the screening of new drugs, 3D models can be used to test a large number of bioactive compounds reducing the cost of the early stages of drug discovery [3,4,5]. 3D models can not totally substitute animal testing, but could be considered as a valid support to obtain more information about the efficacy of bioactive compounds [5]
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