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Structural Reinforcement of Cell-Laden Hydrogels with Microfabricated Three Dimensional Scaffolds.

ABSTRACT: Hydrogels commonly used in tissue engineering are mechanically soft, thus often display structural weakness. Herein, we introduce a strategy for enhancing the structural integrity and fracture toughness of cell-laden hydrogels by incorporating a three-dimensional (3D) microfabricated scaffold as a structural element. A digital micromirror device projection printing (DMD-PP) system, a rapid prototyping technology which employs a layer-by-layer stereolithographic approach, was utilized to efficiently fabricate 3D scaffolds made from photocrosslinkable poly(ethylene glycol) diacrylate (PEGDA). The scaffold was incorporated into a photocrosslinkable gelatin hydrogel by placing it in a pre-gel solution, and inducing in situ hydrogel formation. The resulting scaffold-reinforced hydrogels demonstrated significant increase in ultimate stress and provided structural support for weak hydrogels. In addition, the scaffold did not affect the rigidity of hydrogels, as it was not involved in the crosslinking reaction to form the hydrogel. Therefore, the presented approach could avoid inadvertent and undesired changes in the hydrogel rigidity which is a known regulator of cellular activities. Furthermore, the biocompatibility of scaffold-reinforced hydrogels was confirmed by evaluating the viability and proliferation of encapsulated fibroblasts. Overall, the strategy of incorporating 3D scaffolds into hydrogels as structural reinforcements presented in this study will be highly useful for enhancing the mechanical toughness of hydrogels for various tissue engineering applications.

SUBMITTER: Cha C

PROVIDER: S-EPMC4000042 | biostudies-literature | 2014 May

REPOSITORIES: biostudies-literature

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Structural Reinforcement of Cell-Laden Hydrogels with Microfabricated Three Dimensional Scaffolds.

Cha Chaenyung C Soman Pranav P Zhu Wei W Nikkhah Mehdi M Camci-Unal Gulden G Chen Shaochen S Khademhosseini Ali A

Biomaterials science 20140501 5

Hydrogels commonly used in tissue engineering are mechanically soft, thus often display structural weakness. Herein, we introduce a strategy for enhancing the structural integrity and fracture toughness of cell-laden hydrogels by incorporating a three-dimensional (3D) microfabricated scaffold as a structural element. A digital micromirror device projection printing (DMD-PP) system, a rapid prototyping technology which employs a layer-by-layer stereolithographic approach, was utilized to efficien ...[more]

PMID: 24778793

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Project description:Three-dimensional (3D) cell culture systems, such as tumor organoids and multicellular tumor spheroids, have been developed in part as a result of major advances in tissue engineering and biofabrication techniques. 3D cell culture offers great capabilities in drug development, screening, testing, and precision medicine owing to its physiological accuracy. However, since the inception of 3D systems, few methods have been reported to successfully analyze cell viability quantitatively within hydrogel constructs. In this study, we describe and compare commercially available viability assays developed for two-dimensional (2D) applications for use in 3D constructs composed of organic, synthetic, or hybrid hydrogel formulations. We utilized Promega's CellTiter-Glo®, CellTiter-Glo 3D, and CellTiter 96® MTS Assay along with Thermo Fisher's PrestoBlue™ assay to determine if these assays can be used accurately in 3D systems. Compared with direct cell viability commonly used in 2D cell culture, our results show cellular health output inaccuracies among each assay in differing hydrogel formulations. Our results should inform researchers of potential errors when using cell viability measurements in 3D cultures and conclude that microscopic imaging should be used, in combination, for validation. Impact statement Three-dimensional (3D) tissue organoids models are a valuable tool not only for studying drug toxicity but also for understanding human embryonic development, intra-tissue morphogenesis, and mechanisms of disease. In cancer organoids, such 3D models may be used for preclinical chemotherapy screening and for understanding cell death and viability mechanisms under physiologically relevant conditions. Cell viability assays are necessary for assessing the effect of biological reagents on cellular health and have been used on in vitro cell cultures for many years. With the increase of 3D systems in cellular biology research to determine therapeutic efficacy, two-dimensional assays that measure cell viability are being used outside their intended use on 3D constructs. In this study, we assess the accuracy of using various commercially available cell viability assays on different 3D hydrogel constructs to help researchers understand expected variability in their experimentation along microscopic imaging validation.

Dataset Information

Structural Reinforcement of Cell-Laden Hydrogels with Microfabricated Three Dimensional Scaffolds.

Publications

Structural Reinforcement of Cell-Laden Hydrogels with Microfabricated Three Dimensional Scaffolds.

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