Ex Vivo Generation and Characterization of Human Hyaline and Elastic Cartilaginous Microtissues for Tissue Engineering Applications.

Jan Vanfleteren,Jesús Chato-Astrain,Daniel Durand-Herrera,Ana B. Paes,José Darío Sánchez-López,Rik Verplancke,Víctor Carriel,David Sánchez-Porras,Óscar Darío García-García,Fernando Campos

doi:10.3390/biomedicines9030292

Jan Vanfleteren, Jesús Chato-Astrain

Open Access

https://doi.org/10.3390/biomedicines9030292

Copy DOI

Abstract

Considering the high prevalence of cartilage-associated pathologies, low self-repair capacity and limitations of current repair techniques, tissue engineering (TE) strategies have emerged as a promising alternative in this field. Three-dimensional culture techniques have gained attention in recent years, showing their ability to provide the most biomimetic environment for the cells under culture conditions, enabling the cells to fabricate natural, 3D functional microtissues (MTs). In this sense, the aim of this study was to generate, characterize and compare scaffold-free human hyaline and elastic cartilage-derived MTs (HC-MTs and EC-MTs, respectively) under expansion (EM) and chondrogenic media (CM). MTs were generated by using agarose microchips and evaluated ex vivo for 28 days. The MTs generated were subjected to morphometric assessment and cell viability, metabolic activity and histological analyses. Results suggest that the use of CM improves the biomimicry of the MTs obtained in terms of morphology, viability and extracellular matrix (ECM) synthesis with respect to the use of EM. Moreover, the overall results indicate a faster and more sensitive response of the EC-derived cells to the use of CM as compared to HC chondrocytes. Finally, future preclinical in vivo studies are still needed to determine the potential clinical usefulness of these novel advanced therapy products.

Highlights

It is possible to generate highly biomimetic functional units for tissue engineering (TE) by using the MT technique. This methodology has been used in different TE applications, demonstrating that cells can progressively produce extracellular matrix (ECM) molecules and establish different kinds of cell–cell or cell–ECM interactions when they are cultured in non-adherent molds, such as agarose microchips [4,31]
This study demonstrated that HC- and EC-derived chondrocytes cultured in agarose microchips were able to form stable, viable, metabolically active and ECM-rich MTs ex vivo
Histology demonstrated that MTforming chondrocytes expressed the S-100 marker over time and were able to synthesize diverse cartilage-specific ECM molecules

Summary

Introduction

Tissue engineering (TE) is an interdisciplinary field that applies the principles of engineering and life sciences toward to the development of bioengineered substitutes to restore or maintain the structure and function of tissues or organs [1,2]. To engineer living tissues in vitro, cultured cells are induced to grow within different kinds of scaffolds, or they are stimulated to create their own extracellular matrix (ECM). Both methods allow generating 3D bioartificial substitutes, with controlled biomechanical, structural and biological properties, to promote tissue regeneration in vivo [3,4,5].

Objectives

Discussion

Conclusion