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3D bioprinting of multi-layered segments of a vessel-like structure with ECM and novel derived bioink
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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9437706/ https://www.ncbi.nlm.nih.gov/pubmed/36061430 http://dx.doi.org/10.3389/fbioe.2022.918690 |
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author | Potere, Federica Belgio, Beatrice Croci, Giorgio Alberto Tabano, Silvia Petrini, Paola Dubini, Gabriele Boschetti, Federica Mantero, Sara |
author_facet | Potere, Federica Belgio, Beatrice Croci, Giorgio Alberto Tabano, Silvia Petrini, Paola Dubini, Gabriele Boschetti, Federica Mantero, Sara |
author_sort | Potere, Federica |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9437706 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-94377062022-09-03 3D bioprinting of multi-layered segments of a vessel-like structure with ECM and novel derived bioink Potere, Federica Belgio, Beatrice Croci, Giorgio Alberto Tabano, Silvia Petrini, Paola Dubini, Gabriele Boschetti, Federica Mantero, Sara Front Bioeng Biotechnol Bioengineering and Biotechnology 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. Frontiers Media S.A. 2022-08-19 /pmc/articles/PMC9437706/ /pubmed/36061430 http://dx.doi.org/10.3389/fbioe.2022.918690 Text en Copyright © 2022 Potere, Belgio, Croci, Tabano, Petrini, Dubini, Boschetti and Mantero. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Bioengineering and Biotechnology Potere, Federica Belgio, Beatrice Croci, Giorgio Alberto Tabano, Silvia Petrini, Paola Dubini, Gabriele Boschetti, Federica Mantero, Sara 3D bioprinting of multi-layered segments of a vessel-like structure with ECM and novel derived bioink |
title | 3D bioprinting of multi-layered segments of a vessel-like structure with ECM and novel derived bioink |
title_full | 3D bioprinting of multi-layered segments of a vessel-like structure with ECM and novel derived bioink |
title_fullStr | 3D bioprinting of multi-layered segments of a vessel-like structure with ECM and novel derived bioink |
title_full_unstemmed | 3D bioprinting of multi-layered segments of a vessel-like structure with ECM and novel derived bioink |
title_short | 3D bioprinting of multi-layered segments of a vessel-like structure with ECM and novel derived bioink |
title_sort | 3d bioprinting of multi-layered segments of a vessel-like structure with ecm and novel derived bioink |
topic | Bioengineering and Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9437706/ https://www.ncbi.nlm.nih.gov/pubmed/36061430 http://dx.doi.org/10.3389/fbioe.2022.918690 |
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