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

被引:223
作者
Costantini, Marco [1 ,2 ]
Idaszek, Joanna [1 ]
Szoke, Krisztina [3 ,4 ]
Jaroszewicz, Jakub [1 ]
Dentini, Mariella [2 ]
Barbetta, Andrea [2 ]
Brinchmann, Jan E. [3 ,4 ,5 ]
Swieszkowski, Wojciech [1 ]
机构
[1] Warsaw Univ Technol, Fac Mat Sci & Engn, PL-02507 Warsaw, Poland
[2] Sapienza Univ Rome, Dept Chem, I-00185 Rome, Italy
[3] Oslo Univ Hosp, Dept Immunol, Rikshosp, NO-0424 Oslo, Norway
[4] Oslo Univ Hosp, Norwegian Ctr Stem Cell Res, Rikshosp, NO-0424 Oslo, Norway
[5] Univ Oslo, Dept Mol Med, Inst Basic Med Sci, NO-0372 Oslo, Norway
关键词
3D bioprinting; cartilage regeneration; mesenchymal stem cells; hydrogels; bioink formulation; MESENCHYMAL STEM-CELLS; HYALURONIC-ACID; CHONDROGENIC DIFFERENTIATION; GENE-EXPRESSION; BONE-MARROW; CARTILAGE; SCAFFOLDS; GELATIN; CHONDROCYTES; TISSUES;
D O I
10.1088/1758-5090/8/3/035002
中图分类号
R318 [生物医学工程];
学科分类号
100103 [病原生物学];
摘要
In this work we demonstrate how to print 3D biomimetic hydrogel scaffolds for cartilage tissue engineering with high cell density (> 10(7) cells ml(-1)), high cell viability (85 divided by 90%) and high printing resolution (approximate to 100 mu m) 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.
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页数:13
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