3D bioprinting of BM-MSCs-loaded ECM biomimetic hydrogels for in vitro neocartilage formation

Marco Costantini , Joanna Idaszek , Krisztina Szöke , Jakub Jaroszewicz , Mariella Dentini , Andrea Barbetta , JE Brinchmann , Wojciech Święszkowski

Abstract

In this work we demonstrate how to print 3D biomimetic hydrogel scaffolds for cartilage tissue engineering with high cell density (>107 cells ml-1), high cell viability (85 90%) and high printing resolution (≈100 μm) through a two coaxial-needles system. The scaffolds were composed of modified biopolymers present in the extracellular matrix (ECM) of cartilage, namely gelatin methacrylamide (GelMA), chondroitin sulfate amino ethyl methacrylate (CS-AEMA) and hyaluronic acid methacrylate (HAMA). The polymers were used to prepare three photocurable bioinks with increasing degree of biomimicry: (i) GelMA, (ii) GelMA + CS-AEMA and (iii) GelMA + CS-AEMA + HAMA. Alginate was added to the bioinks as templating agent to form stable fibers during 3D printing. In all cases, bioink solutions were loaded with bone marrow-derived human mesenchymal stem cells (BM-MSCs). After printing, the samples were cultured in expansion (negative control) and chondrogenic media to evaluate the possible differentiating effect exerted by the biomimetic matrix or the synergistic effect of the matrix and chondrogenic supplements. After 7, 14, and 21 days, gene expression of the chondrogenic markers (COL2A1 and aggrecan), marker of osteogenesis (COL1A1) and marker of hypertrophy (COL10A1) were evaluated qualitatively by means of fluorescence immunocytochemistry and quantitatively by means of RT-qPCR. The observed enhanced viability and chondrogenic differentiation of BM-MSCs, as well as high robustness and accuracy of the employed deposition method, make the presented approach a valid candidate for advanced engineering of cartilage tissue.

Author Marco Costantini - Università Campus Bio-Medico di Roma, Rome, Italy [Warsaw University of Technology (PW)]
Marco Costantini,,
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- Politechnika Warszawska
, Joanna Idaszek (FMSE / DMD)
Joanna Idaszek,,
- Division of Materials Design
, Krisztina Szöke - [Oslo University Hospital]
Krisztina Szöke,,
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, Jakub Jaroszewicz (FMSE / DMD)
Jakub Jaroszewicz,,
- Division of Materials Design
, Mariella Dentini - Sapienza University of Rome [Universita degli Studi di Roma La Sapienza]
Mariella Dentini,,
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, Andrea Barbetta - Sapienza University of Rome [Universita degli Studi di Roma La Sapienza]
Andrea Barbetta,,
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, JE Brinchmann - [Oslo University Hospital]
JE Brinchmann,,
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, Wojciech Święszkowski (FMSE / DMD)
Wojciech Święszkowski,,
- Division of Materials Design
Journal seriesBiofabrication, ISSN 1758-5082
Issue year2016
Vol8
ASJC Classification2204 Biomedical Engineering; 2700 General Medicine; 2502 Biomaterials; 1303 Biochemistry; 1502 Bioengineering; 1305 Biotechnology
DOIDOI:10.1088/1758-5090/8/3/035002
Languageen angielski
Score (nominal)45
Score sourcejournalList
ScoreMinisterial score = 45.0, 25-06-2020, ArticleFromJournal
Ministerial score (2013-2016) = 45.0, 25-06-2020, ArticleFromJournal
Publication indicators Scopus Citations = 87; WoS Citations = 52; GS Citations = 98.0; Scopus SNIP (Source Normalised Impact per Paper): 2016 = 1.253; WoS Impact Factor: 2016 = 5.24 (2) - 2016=5.82 (5)
Citation count*98 (2020-06-25)
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* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.
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