A Photo‐Crosslinkable Kidney ECM‐Derived Bioink Accelerates Renal Tissue Formation

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Abstract

3D bioprinting strategies in tissue engineering aim to fabricate clinically applicable tissue constructs that can replace the damaged or diseased tissues and organs. One of the main prerequisites in 3D bioprinting is finding an appropriate bioink that provides a tissue-specific microenvironment supporting the cellular growth and maturation. In this respect, decellularized extracellular matrix (dECM)-derived hydrogels have been considered as bioinks for the cell-based bioprinting due to their capability to inherit the intrinsic cues from native ECM. Herein, a photo-crosslinkable kidney ECM-derived bioink (KdECMMA) is developed that could provide a kidney-specific microenvironment for renal tissue bioprinting. Porcine whole kidneys are decellularized through a perfusion method, dissolved in an acid solution, and chemically modified by methacrylation. A KdECMMA-based bioink is formulated and evaluated for rheological properties and printability for the printing process. The results show that the bioprinted human kidney cells in the KdECMMA bioink are highly viable and mature with time. Moreover, the bioprinted renal constructs exhibit the structural and functional characteristics of the native renal tissue. The potential of the tissue-specific ECM-derived bioink is demonstrated for cell-based bioprinting that could enhance the cellular maturation and eventually tissue formation.

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Peter M Crapo, Thomas W Gilbert, Stephen F Badylak (2011)

Biologic scaffold materials composed of extracellular matrix (ECM) are typically derived by processes that involve decellularization of tissues or organs. Preservation of the complex composition and three-dimensional ultrastructure of the ECM is highly desirable but it is recognized that all methods of decellularization result in disruption of the architecture and potential loss of surface structure and composition. Physical methods and chemical and biologic agents are used in combination to lyse cells, followed by rinsing to remove cell remnants. Effective decellularization methodology is dictated by factors such as tissue density and organization, geometric and biologic properties desired for the end product, and the targeted clinical application. Tissue decellularization with preservation of ECM integrity and bioactivity can be optimized by making educated decisions regarding the agents and techniques utilized during processing. An overview of decellularization methods, their effect upon resulting ECM structure and composition, and recently described perfusion techniques for whole organ decellularization techniques are presented herein. Copyright © 2011 Elsevier Ltd. All rights reserved.

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Brian Derby (2012)

New manufacturing technologies under the banner of rapid prototyping enable the fabrication of structures close in architecture to biological tissue. In their simplest form, these technologies allow the manufacture of scaffolds upon which cells can grow for later implantation into the body. A more exciting prospect is the printing and patterning in three dimensions of all the components that make up a tissue (cells and matrix materials) to generate structures analogous to tissues; this has been termed bioprinting. Such techniques have opened new areas of research in tissue engineering and regenerative medicine.

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Effect of bioink properties on printability and cell viability for 3D bioplotting of embryonic stem cells.

Liliang Ouyang, Rui Yao, Yu ZHAO … (2016)

3D cell printing is an emerging technology for fabricating complex cell-laden constructs with precise and pre-designed geometry, structure and composition to overcome the limitations of 2D cell culture and conventional tissue engineering scaffold technology. This technology enables spatial manipulation of cells and biomaterials, also referred to as 'bioink', and thus allows study of cellular interactions in a 3D microenvironment and/or in the formation of functional tissues and organs. Recently, many efforts have been made to develop new bioinks and to apply more cell sources for better biocompatibility and biofunctionality. However, the influences of printing parameters on the shape fidelity of 3D constructs as well as on cell viability after the cell printing process have been poorly characterized. Furthermore, parameter optimization based on a specific cell type might not be suitable for other types of cells, especially cells with high sensibility. In this study, we systematically studied the influence of bioink properties and printing parameters on bioink printability and embryonic stem cell (ESC) viability in the process of extrusion-based cell printing, also known as bioplotting. A novel method was established to determine suitable conditions for bioplotting ESCs to achieve both good printability and high cell viability. The rheological properties of gelatin/alginate bioinks were evaluated to determine the gelation properties under different bioink compositions, printing temperatures and holding times. The bioink printability was characterized by a newly developed semi-quantitative method. The results demonstrated that bioinks with longer gelation times would result in poorer printability. The live/dead assay showed that ESC viability increased with higher printing temperatures and lower gelatin concentrations. Furthermore, an exponential relationship was obtained between ESC viability and induced shear stress. By defining the proper printability and acceptable viability ranges, a combined parameters region was obtained. This study provides guidance for parameter optimization and the fine-tuning of 3D cell printing processes regarding both bioink printability and cell viability after bioplotting, especially for easily damaged cells, like ESCs.