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Development of Gelatin-Coated Microspheres for Novel Bioink Design.

ABSTRACT: A major challenge in tissue engineering is the formation of vasculature in tissue and organs. Recent studies have shown that positively charged microspheres promote vascularization, while also supporting the controlled release of bioactive molecules. This study investigated the development of gelatin-coated pectin microspheres for incorporation into a novel bioink. Electrospray was used to produce the microspheres. The process was optimized using Design-Expert® software. Microspheres underwent gelatin coating and EDC catalysis modifications. The results showed that the concentration of pectin solution impacted roundness and uniformity primarily, while flow rate affected size most significantly. The optimal gelatin concentration for microsphere coating was determined to be 0.75%, and gelatin coating led to a positively charged surface. When incorporated into bioink, the microspheres did not significantly alter viscosity, and they distributed evenly in bioink. These microspheres show great promise for incorporation into bioink for tissue engineering applications.

SUBMITTER: Kanungo M

PROVIDER: S-EPMC8512326 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature

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Development of Gelatin-Coated Microspheres for Novel Bioink Design.

Kanungo Muskan M Wang Yale Y Hutchinson Noah N Kroll Emma E DeBruine Anna A Kumpaty Subha S Ren Lixia L Wu Yuelin Y Hua Xiaolin X Zhang Wujie W

Polymers 20210929 19

A major challenge in tissue engineering is the formation of vasculature in tissue and organs. Recent studies have shown that positively charged microspheres promote vascularization, while also supporting the controlled release of bioactive molecules. This study investigated the development of gelatin-coated pectin microspheres for incorporation into a novel bioink. Electrospray was used to produce the microspheres. The process was optimized using Design-Expert<sup>®</sup> software. Microspheres ...[more]

PMID: 34641153

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Project description:Three-dimensional (3D)in vitrotumor models that can capture the pathophysiology of human tumors are essential for cancer biology and drug development. However, simulating the tumor microenvironment is still challenging because it consists of a heterogeneous mixture of various cellular components and biological factors. In this regard, current extracellular matrix (ECM)-mimicking hydrogels used in tumor tissue engineering lack physical interactions that can keep biological factors released by encapsulated cells within the hydrogel and improve paracrine interactions. Here, we developed a nanoengineered ion-covalent cross-linkable bioink to construct 3D bioprinted organotypic tumor models. The bioink was designed to implement the tumor ECM by creating an interpenetrating network composed of gelatin methacryloyl (GelMA), a light cross-linkable polymer, and synthetic nanosilicate (Laponite) that exhibits a unique ionic charge to improve retention of biological factors released by the encapsulated cells and assist in paracrine signals. The physical properties related to printability were evaluated to analyze the effect of Laponite hydrogel on bioink. Low GelMA (5%) with high Laponite (2.5%-3.5%) composite hydrogels and high GelMA (10%) with low Laponite (1.0%-2.0%) composite hydrogels showed acceptable mechanical properties for 3D printing. However, a low GelMA composite hydrogel with a high Laponite content could not provide acceptable cell viability. Fluorescent cell labeling studies showed that as the proportion of Laponite increased, the cells became more aggregated to form larger 3D tumor structures. Reverse transcription-polymerase chain reaction (RT-qPCR) and western blot experiments showed that an increase in the Laponite ratio induces upregulation of growth factor and tissue remodeling-related genes and proteins in tumor cells. In contrast, cell cycle and proliferation-related genes were downregulated. On the other hand, concerning fibroblasts, the increase in the Laponite ratio indicated an overall upregulation of the mesenchymal phenotype-related genes and proteins. Our study may provide a rationale for using Laponite-based hydrogels in 3D cancer modeling.

Project description:Three-dimensional bioprinting has emerged as a promising technique in tissue engineering applications through the precise deposition of cells and biomaterials in a layer-by-layer fashion. However, the limited availability of hydrogel bioinks is frequently cited as a major issue for the advancement of cell-based extrusion bioprinting technologies. It is well known that highly viscous materials maintain their structure better, but also have decreased cell viability due to the higher forces which are required for extrusion. However, little is known about the effect of the two distinct components of dynamic modulus of viscoelastic materials, storage modulus (G') and loss modulus (G?), on the printability of hydrogel-based bioinks. Additionally, 'printability' has been poorly defined in the literature, mostly consisting of gross qualitative measures which do not allow for direct comparison of bioinks. This study developed a framework for evaluating printability and investigated the effect of dynamic modulus, including storage modulus (G'), loss modulus (G?), and loss tangent (G?/G') on the printing outcome. Gelatin and alginate as model hydrogels were mixed at various concentrations to obtain hydrogel formulations with a wide range of storage and loss moduli. These formulations were then evaluated for the quantitatively defined values of extrudability, extrusion uniformity, and structural integrity. For extrudability, increasing either the loss or storage modulus increased the pressure required to extrude the bioink. A mathematical model relating the G' and G? to the required extrusion pressure was derived based on the data. A lower loss tangent was correlated with increased structural integrity while a higher loss tangent correlated with increased extrusion uniformity. Gelatin-alginate composite hydrogels with a loss tangent in the range of 0.25-0.45 exhibited an excellent compromise between structural integrity and extrusion uniformity. In addition to the characterization of a common bioink, the methodology introduced in this paper could also be used to evaluate the printability of other bioinks in the future.

Dataset Information

Development of Gelatin-Coated Microspheres for Novel Bioink Design.

Publications

Development of Gelatin-Coated Microspheres for Novel Bioink Design.

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