3D-Reactive printing of engineered alginate inks

Sardelli, Lorenzo; Tunesi, Marta; Briatico‐Vangosa, Francesco; Petrini, Paola

doi:10.1039/d1sm00604e

Cited by 22 publications

(36 citation statements)

References 78 publications

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“…The presence of yielding points, defined as the point at which G″ starts to prevail over G′, in Hep3Gel, GEL, and ALG implies the existence of a shear stress level at which the hydrogels start behaving like viscous fluids, being thus able to be extruded. 31,82 The second test was showing that all of the hydrogels can recover their original viscoelastic properties after being extruded. In particular, the first interval aims to mimic the bioink in the cartridge before being printed.…”

Section: Characterization Of the Shape-shifting Behaviormentioning

confidence: 99%

“…With this test, it was possible to appreciate that despite changes in their rheological parameters due to the yielding occurring during extrusion, all the hydrogels were able to recover their original properties, thus highlighting the existence of some conditions in which the hydrogels are 3D printable. 31,82 The a priori knowledge of the yielding point of a bioink is crucial to reduce the trial-and-error procedure typical when studying the printability of a material. Actually, under the hypothesis that syringe extrusion is a process in which a high flow rate�associable with high-frequency deformations� dominates, the extrusion pressure needed to produce specific shear stresses inside the syringe can be estimated through a first-order mathematical model, as reported in eq 7.…”

Section: Characterization Of the Shape-shifting Behaviormentioning

confidence: 99%

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Hep3Gel: A Shape-Shifting Extracellular Matrix-Based, Three-Dimensional Liver Model Adaptable to Different Culture Systems

Guagliano

Volpini

Sardelli

et al. 2022

ACS Biomater. Sci. Eng.

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Drug-induced hepatotoxicity is a leading cause of clinical trial withdrawal. Therefore, in vitro modeling the hepatic behavior and functionalities is not only crucial to better understand physiological and pathological processes but also to support drug development with reliable high-throughput platforms. Different physiological and pathological models are currently under development and are commonly implemented both within platforms for standard 2D cultures and within tailor-made chambers. This paper introduces Hep3Gel: a hybrid alginate−extracellular matrix (ECM) hydrogel to produce 3D in vitro models of the liver, aiming to reproduce the hepatic chemomechanical niche, with the possibility of adapting its shape to different manufacturing techniques. The ECM, extracted and powdered from porcine livers by a specifically set-up procedure, preserved its crucial biological macromolecules and was embedded within alginate hydrogels prior to crosslinking. The viscoelastic behavior of Hep3Gel was tuned, reproducing the properties of a physiological organ, according to the available knowledge about hepatic biomechanics. By finely tuning the crosslinking kinetics of Hep3Gel, its dualistic nature can be exploited either by self-spreading or adapting its shape to different culture supports or retaining the imposed fiber shape during an extrusion-based 3D-bioprinting process, thus being a shape-shifter hydrogel. The self-spreading ability of Hep3Gel was characterized by combining empirical and numerical procedures, while its use as a bioink was experimentally characterized through rheological a priori printability evaluations and 3D printing tests. The effect of the addition of the ECM was evident after 4 days, doubling the survival rate of cells embedded within control hydrogels. This study represents a proof of concept of the applicability of Hep3Gel as a tool to develop 3D in vitro models of the liver.

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Section: Characterization Of the Shape-shifting Behaviormentioning

confidence: 99%

Section: Characterization Of the Shape-shifting Behaviormentioning

confidence: 99%

Hep3Gel: A Shape-Shifting Extracellular Matrix-Based, Three-Dimensional Liver Model Adaptable to Different Culture Systems

Guagliano

Volpini

Sardelli

et al. 2022

ACS Biomater. Sci. Eng.

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“…19 This approach has previously been quite extensively used to crosslink alginate hydrogels. 20 GdL has also been combined with calcium carbonate to form alginate/GG hybrids, although it is not clear whether the gellan gum was actually crosslinked via the same mechanism as the alginate in this case. 21 There has been some interest in incorporating CaCO 3 microparticles into GG gels – they can slowly release Ca 2+ even in the absence of an acid source and are also highly relevant in bone tissue engineering.…”

Section: Resultsmentioning

confidence: 99%

Combining gellan gum with a functional low-molecular-weight gelator to assemble stiff shaped hybrid hydrogels for stem cell growth

2022

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“…First, we optimized the decellularization protocol. The obtained dECM was then incorporated in a bioink whose formulation guaranteed its printability thus facing one of the main challenges with extrsuion-based 3D-bioprinting (Paxton et al, 2017) (Schwab et al, 2020) (Sardelli et al, 2021). Finally, we verified the bioink biocompatibility and cell infiltration monitoring cell growth in 3D printed constructs over time.…”

Section: Introductionmentioning

confidence: 84%

3D bioprinting of multi-layered segments of a vessel-like structure with ECM and novel derived bioink

Potere

Belgio

Croci

et al. 2022

Front. Bioeng. Biotechnol.

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3D-Bioprinting leads to the realization of tridimensional customized constructs to reproduce the biological structural complexity. The new technological challenge focuses on obtaining a 3D structure with several distinct layers to replicate the hierarchical organization of natural tissues. This work aims to reproduce large blood vessel substitutes compliant with the original tissue, combining the advantages of the 3D bioprinting, decellularization, and accounting for the presence of different cells. The decellularization process was performed on porcine aortas. Various decellularization protocols were tested and evaluated through DNA extraction, quantification, and amplification by PCR to define the adequate one. The decellularized extracellular matrix (dECM), lyophilized and solubilized, was combined with gelatin, alginate, and cells to obtain a novel bioink. Several solutions were tested, tuning the percentage of the components to obtain the adequate structural properties. The geometrical model of the large blood vessel constructs was designed with SolidWorks, and the construct slicing was done using the HeartWare software, which allowed generating the G-Code. The final constructs were 3D bioprinted with the Inkredible + using dual print heads. The composition of the bioink was tuned so that it could withstand the printing of a segment of a tubular construct up to 10 mm and reproduce the multicellular complexity. Among the several compositions tested, the suspension resulting from 8% w/v gelatin, 7% w/v alginate, and 3% w/v dECM, and cells successfully produced the designed structures. With this bioink, it was possible to print structures made up of 20 layers. The dimensions of the printed structures were consistent with the designed ones. We were able to avoid the double bioink overlap in the thickness, despite the increase in the number of layers during the printing process. The optimization of the parameters allowed the production of structures with a height of 20 layers corresponding to 9 mm. Theoretical and real structures were very close. The differences were 14% in height, 20% internal diameter, and 9% thickness. By tailoring the printing parameters and the amount of dECM, adequate mechanical properties could be met. In this study, we developed an innovative printable bioink able to finely reproduce the native complex structure of the large blood vessel.

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3D-Reactive printing of engineered alginate inks

Abstract: Alginate is a common component of bioinks due to its well-described ionic crosslinking mechanism and its tunable viscoelastic properties. The extrusion-based 3D-printing of alginate inks requires additives, such as gelatin...

Cited by 22 publications

References 78 publications

Hep3Gel: A Shape-Shifting Extracellular Matrix-Based, Three-Dimensional Liver Model Adaptable to Different Culture Systems

Hep3Gel: A Shape-Shifting Extracellular Matrix-Based, Three-Dimensional Liver Model Adaptable to Different Culture Systems

Combining gellan gum with a functional low-molecular-weight gelator to assemble stiff shaped hybrid hydrogels for stem cell growth

3D bioprinting of multi-layered segments of a vessel-like structure with ECM and novel derived bioink

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