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Cell-laden alginate dialdehyde-gelatin hydrogels formed in 3D printed sacrificial gel.

ABSTRACT: Alginate dialdehyde-gelatin (ADA-GEL) hydrogels have been reported to be suitable matrices for cell encapsulation. In general, application of ADA-GEL as bioink has been limited to planar structures due to its low viscosity. In this work, ring shaped constructs of ADA-GEL hydrogel were fabricated by casting the hydrogel into sacrificial molds which were 3D printed from 9% methylcellulose and 5% gelatin. Dissolution of the supporting structure was observed during the 1^st week of sample incubation. In addition, the effect of different crosslinkers (Ba²⁺ and Ca²⁺) on the physicochemical properties of ADA-GEL and on the behavior of encapsulated MG-63 cells was investigated. It was found that Ba²⁺ crosslinked network had more than twice higher storage modulus, and mass decrease to 70% during incubation compared to 42% in case of hydrogels crosslinked with Ca²⁺. In addition, faster increase in cell viability during incubation and earlier cell network formation were observed after Ba²⁺ crosslinking. No negative effects on cell activity due to the use of sacrificial materials were observed. The approach presented here could be further developed for cell-laden ADA-GEL bioink printing into complex 3D structures.

SUBMITTER: Dranseikiene D

PROVIDER: S-EPMC7062650 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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Cell-laden alginate dialdehyde-gelatin hydrogels formed in 3D printed sacrificial gel.

Dranseikiene Dalia D Schrüfer Stefan S Schubert Dirk W DW Reakasame Supachai S Boccaccini Aldo R AR

Journal of materials science. Materials in medicine 20200309 3

Alginate dialdehyde-gelatin (ADA-GEL) hydrogels have been reported to be suitable matrices for cell encapsulation. In general, application of ADA-GEL as bioink has been limited to planar structures due to its low viscosity. In this work, ring shaped constructs of ADA-GEL hydrogel were fabricated by casting the hydrogel into sacrificial molds which were 3D printed from 9% methylcellulose and 5% gelatin. Dissolution of the supporting structure was observed during the 1<sup>st</sup> week of sample ...[more]

PMID: 32152812

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Project description:Background 3D bioprinting cardiac patches for epicardial transplantation are a promising approach for myocardial regeneration. Challenges remain such as quantifying printability, determining the ideal moment to transplant, and promoting vascularisation within bioprinted patches. We aimed to evaluate 3D bioprinted cardiac patches for printability, durability in culture, cell viability, and endothelial cell structural self-organisation into networks. Methods We evaluated 3D-bioprinted double-layer patches using alginate/gelatine (AlgGel) hydrogels and three extrusion bioprinters (REGEMAT3D, INVIVO, BIO X). Bioink contained either neonatal mouse cardiac cell spheroids or free (not-in-spheroid) human coronary artery endothelial cells with fibroblasts, mixed with AlgGel. To test the effects on durability, some patches were bioprinted as a single layer only, cultured under minimal movement conditions or had added fibroblast-derived extracellular matrix hydrogel (AlloECM). Controls included acellular AlgGel and gelatin methacryloyl (GELMA) patches. Results Printability was similar across bioprinters. For AlgGel compared to GELMA: resolutions were similar (200–700 ?m line diameters), printing accuracy was 45 and 25%, respectively (AlgGel was 1.7x more accurate; p < 0.05), and shape fidelity was 92% (AlgGel) and 96% (GELMA); p = 0.36. For durability, AlgGel patch median survival in culture was 14 days (IQR:10–27) overall which was not significantly affected by bioprinting system or cellular content in patches. We identified three factors which reduced durability in culture: (1) bioprinting one layer depth patches (instead of two layers); (2) movement disturbance to patches in media; and (3) the addition of AlloECM to AlgGel. Cells were viable after bioprinting followed by 28 days in culture, and all BIO X-bioprinted mouse cardiac cell spheroid patches presented contractile activity starting between day 7 and 13 after bioprinting. At day 28, endothelial cells in hydrogel displayed organisation into endothelial network-like structures. Conclusion AlgGel-based 3D bioprinted heart patches permit cardiomyocyte contractility and endothelial cell structural self-organisation. After bioprinting, a period of 2 weeks maturation in culture prior to transplantation may be optimal, allowing for a degree of tissue maturation but before many patches start to lose integrity. We quantify AlgGel printability and present novel factors which reduce AlgGel patch durability (layer number, movement, and the addition of AlloECM) and factors which had minimal effect on durability (bioprinting system and cellular patch content). Graphical Abstract

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Cell-laden alginate dialdehyde-gelatin hydrogels formed in 3D printed sacrificial gel.

Publications

Cell-laden alginate dialdehyde-gelatin hydrogels formed in 3D printed sacrificial gel.

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