Extruded bioreactor perfusion culture supports the chondrogenic differentiation of human mesenchymal stem/stromal cells in 3D porous poly(ɛ-caprolactone) scaffolds

Novel bioengineering strategies for the ex vivo fabrication of native-like tissue-engineered cartilage are crucial for the translation of these approaches to clinically manage highly prevalent and debilitating joint diseases. Bioreactors that provide different biophysical stimuli have been used in tissue engineering approaches aimed at enhancing the quality of the cartilage tissue generated. However, such systems are often highly complex, expensive, and not very versatile. In the current study, a novel, cost-effective, and customizable perfusion bioreactor totally fabricated by additive manufacturing (AM) is proposed for the study of the effect of fluid flow on the chondrogenic differentiation of human bone-marrow mesenchymal stem/stromal cells (hBMSCs) in 3D porous poly(ɛ-caprolactone) (PCL) scaffolds. hBMSCs are first seeded and grown on PCL scaffolds and hBMSC–PCL constructs are then transferred to 3D-extruded bioreactors for continuous perfusion culture under chondrogenic inductive conditions. Perfused constructs show similar cell metabolic activity and significantly higher sulfated glycosaminoglycan production (≈1.8-fold) in comparison to their non-perfused counterparts. Importantly, perfusion bioreactor culture significantly promoted the expression of chondrogenic marker genes while downregulating hypertrophy. This work highlights the potential of customizable AM platforms for the development of novel personalized repair strategies and more reliable in vitro models with a wide range of applications.

Description

The authors acknowledge financial support from Fundação para a Ciência e Tecnologia (FCT, Portugal) through UID/BIO/04565/2019, UID/Multi/04044/2013), Stimuli2BioScaffold grant (PTDC/EMESIS/ 32554/2017), and scholarships SFRH/BD/73970/2010, SFRH/BD/ 105771/2014, and SFRH/BSAB/128442/2017. Funding from Programa Operacional Regional de Lisboa 2020 (project no. 007317) and also through the projects PRECISE (PAC-PRECISE-LISBOA-01-0145-FEDER – 016394, SAICTPAC/0021/2015) and POCI-01-0145 FEDER-016800 is also acknowledged.

Keywords

Additive manufacturing Cartilage tissue engineering Extrusion-based perfusion bioreactor Mesenchymal stem/stromal cells Poly(ɛ-caprolactone) scaffolds

URI

http://hdl.handle.net/10400.8/8401

Citation

Silva, J. C., Moura, C. S., Borrecho, G., de Matos, A. P. A., da Silva, C. L., Cabral, J. M. S., Bártolo, P. J., Linhardt, R. J., & Ferreira, F. C. (2020). Extruded Bioreactor Perfusion Culture Supports the Chondrogenic Differentiation of Human Mesenchymal Stem/Stromal Cells in 3D Porous Poly(ɛ-Caprolactone) Scaffolds. Biotechnology Journal, 15(2). https://doi.org/10.1002/biot.201900078