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      Aspiration-assisted bioprinting for precise positioning of biologics

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          Abstract

          Aspiration-assisted bioprinting enables precise positioning of viscoelastic spheroids in both scaffold-based and free manner.

          Abstract

          Three-dimensional (3D) bioprinting is an appealing approach for building tissues; however, bioprinting of mini-tissue blocks (i.e., spheroids) with precise control on their positioning in 3D space has been a major obstacle. Here, we unveil “aspiration-assisted bioprinting (AAB),” which enables picking and bioprinting biologics in 3D through harnessing the power of aspiration forces, and when coupled with microvalve bioprinting, it facilitated different biofabrication schemes including scaffold-based or scaffold-free bioprinting at an unprecedented placement precision, ~11% with respect to the spheroid size. We studied the underlying physical mechanism of AAB to understand interactions between aspirated viscoelastic spheroids and physical governing forces during aspiration and bioprinting. We bioprinted a wide range of biologics with dimensions in an order-of-magnitude range including tissue spheroids (80 to 600 μm), tissue strands (~800 μm), or single cells (electrocytes, ~400 μm), and as applications, we illustrated the patterning of angiogenic sprouting spheroids and self-assembly of osteogenic spheroids.

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          Most cited references46

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          A Computational Tool for Quantitative Analysis of Vascular Networks

          Angiogenesis is the generation of mature vascular networks from pre-existing vessels. Angiogenesis is crucial during the organism' development, for wound healing and for the female reproductive cycle. Several murine experimental systems are well suited for studying developmental and pathological angiogenesis. They include the embryonic hindbrain, the post-natal retina and allantois explants. In these systems vascular networks are visualised by appropriate staining procedures followed by microscopical analysis. Nevertheless, quantitative assessment of angiogenesis is hampered by the lack of readily available, standardized metrics and software analysis tools. Non-automated protocols are being used widely and they are, in general, time - and labour intensive, prone to human error and do not permit computation of complex spatial metrics. We have developed a light-weight, user friendly software, AngioTool, which allows for quick, hands-off and reproducible quantification of vascular networks in microscopic images. AngioTool computes several morphological and spatial parameters including the area covered by a vascular network, the number of vessels, vessel length, vascular density and lacunarity. In addition, AngioTool calculates the so-called “branching index” (branch points / unit area), providing a measurement of the sprouting activity of a specimen of interest. We have validated AngioTool using images of embryonic murine hindbrains, post-natal retinas and allantois explants. AngioTool is open source and can be downloaded free of charge.
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            Organ printing: tissue spheroids as building blocks.

            Organ printing can be defined as layer-by-layer additive robotic biofabrication of three-dimensional functional living macrotissues and organ constructs using tissue spheroids as building blocks. The microtissues and tissue spheroids are living materials with certain measurable, evolving and potentially controllable composition, material and biological properties. Closely placed tissue spheroids undergo tissue fusion - a process that represents a fundamental biological and biophysical principle of developmental biology-inspired directed tissue self-assembly. It is possible to engineer small segments of an intraorgan branched vascular tree by using solid and lumenized vascular tissue spheroids. Organ printing could dramatically enhance and transform the field of tissue engineering by enabling large-scale industrial robotic biofabrication of living human organ constructs with "built-in" perfusable intraorgan branched vascular tree. Thus, organ printing is a new emerging enabling technology paradigm which represents a developmental biology-inspired alternative to classic biodegradable solid scaffold-based approaches in tissue engineering.
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              Spheroid culture as a tool for creating 3D complex tissues.

              3D cell culture methods confer a high degree of clinical and biological relevance to in vitro models. This is specifically the case with the spheroid culture, where a small aggregate of cells grows free of foreign materials. In spheroid cultures, cells secrete the extracellular matrix (ECM) in which they reside, and they can interact with cells from their original microenvironment. The value of spheroid cultures is increasing quickly due to novel microfabricated platforms amenable to high-throughput screening (HTS) and advances in cell culture. Here, we review new possibilities that combine the strengths of spheroid culture with new microenvironment fabrication methods that allow for the creation of large numbers of highly reproducible, complex tissues. Copyright © 2013. Published by Elsevier Ltd.
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                Author and article information

                Journal
                Sci Adv
                Sci Adv
                SciAdv
                advances
                Science Advances
                American Association for the Advancement of Science
                2375-2548
                March 2020
                06 March 2020
                : 6
                : 10
                : eaaw5111
                Affiliations
                [1 ]Engineering Science and Mechanics Department, Penn State University, University Park, PA 16802, USA.
                [2 ]The Huck Institutes of the Life Sciences, Penn State University, University Park, PA 16802, USA.
                [3 ]Department of Dental Materials, School of Dentistry, Kyung Hee University, Seoul 02447, Republic of Korea.
                [4 ]Department of Chemistry, Penn State University, University Park, PA 16802, USA.
                [5 ]Biomedical Engineering Department, Penn State University, University Park, PA 16802, USA.
                [6 ]Materials Research Institute, Penn State University, University Park, PA 16802, USA.
                Author notes
                [*]

                These authors contributed equally to this work.

                []Corresponding author. Email: ito1@ 123456psu.edu
                Author information
                http://orcid.org/0000-0003-4614-263X
                http://orcid.org/0000-0002-7717-7184
                http://orcid.org/0000-0003-1577-3316
                http://orcid.org/0000-0002-9523-8083
                http://orcid.org/0000-0002-0012-3006
                http://orcid.org/0000-0002-6128-8285
                http://orcid.org/0000-0001-8328-4528
                Article
                aaw5111
                10.1126/sciadv.aaw5111
                7060055
                32181332
                86eaddd7-4d91-461d-854c-70b49c6fb973
                Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

                This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license, which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited.

                History
                : 31 January 2019
                : 13 December 2019
                Funding
                Funded by: doi http://dx.doi.org/10.13039/100000001, National Science Foundation;
                Award ID: 1624515
                Funded by: doi http://dx.doi.org/10.13039/100000001, National Science Foundation;
                Award ID: 1914885
                Funded by: doi http://dx.doi.org/10.13039/100000002, National Institutes of Health;
                Award ID: R21 CA224422 01A1
                Funded by: doi http://dx.doi.org/10.13039/100011612, Materials Research Institute, Pennsylvania State University;
                Categories
                Research Article
                Research Articles
                SciAdv r-articles
                Engineering
                Life Sciences
                Engineering
                Custom metadata
                Fritzie Benzon

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