Fibrinogen-Based Bioink for Application in Skin Equivalent 3D Bioprinting

Three-dimensional bioprinting has emerged as an attractive technology due to its ability to mimic native tissue architecture using different cell types and biomaterials. Nowadays, cell-laden bioink development or skin tissue equivalents are still at an early stage. The aim of the study is to propose...

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Glavni autori: Aida Cavallo, Tamer Al Kayal, Angelica Mero, Andrea Mezzetta, Lorenzo Guazzelli, Giorgio Soldani, Paola Losi
Format: Članak
Jezik:English
Izdano: MDPI AG 2023-09-01
Serija:Journal of Functional Biomaterials
Teme:
Online pristup:https://www.mdpi.com/2079-4983/14/9/459
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author Aida Cavallo
Tamer Al Kayal
Angelica Mero
Andrea Mezzetta
Lorenzo Guazzelli
Giorgio Soldani
Paola Losi
author_facet Aida Cavallo
Tamer Al Kayal
Angelica Mero
Andrea Mezzetta
Lorenzo Guazzelli
Giorgio Soldani
Paola Losi
author_sort Aida Cavallo
collection DOAJ
description Three-dimensional bioprinting has emerged as an attractive technology due to its ability to mimic native tissue architecture using different cell types and biomaterials. Nowadays, cell-laden bioink development or skin tissue equivalents are still at an early stage. The aim of the study is to propose a bioink to be used in skin bioprinting based on a blend of fibrinogen and alginate to form a hydrogel by enzymatic polymerization with thrombin and by ionic crosslinking with divalent calcium ions. The biomaterial ink formulation, composed of 30 mg/mL of fibrinogen, 6% of alginate, and 25 mM of CaCl<sub>2</sub>, was characterized in terms of homogeneity, rheological properties, printability, mechanical properties, degradation rate, water uptake, and biocompatibility by the indirect method using L929 mouse fibroblasts. The proposed bioink is a homogeneous blend with a shear thinning behavior, excellent printability, adequate mechanical stiffness, porosity, biodegradability, and water uptake, and it is in vitro biocompatible. The fibrinogen-based bioink was used for the 3D bioprinting of the dermal layer of the skin equivalent. Three different normal human dermal fibroblast (NHDF) densities were tested, and better results in terms of viability, spreading, and proliferation were obtained with 4 × 10<sup>6</sup> cell/mL. The skin equivalent was bioprinted, adding human keratinocytes (HaCaT) through bioprinting on the top surface of the dermal layer. A skin equivalent stained by live/dead and histological analysis immediately after printing and at days 7 and 14 of culture showed a tissuelike structure with two distinct layers characterized by the presence of viable and proliferating cells. This bioprinted skin equivalent showed a similar native skin architecture, paving the way for its use as a skin substitute for wound healing applications.
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spelling doaj.art-06f86926c99e4461b73eb7eb7b905a822023-11-19T11:23:23ZengMDPI AGJournal of Functional Biomaterials2079-49832023-09-0114945910.3390/jfb14090459Fibrinogen-Based Bioink for Application in Skin Equivalent 3D BioprintingAida Cavallo0Tamer Al Kayal1Angelica Mero2Andrea Mezzetta3Lorenzo Guazzelli4Giorgio Soldani5Paola Losi6Institute of Life Sciences, Scuola Superiore Sant’Anna, 56127 Pisa, ItalyInstitute of Clinical Physiology, National Research Council, 54100 Massa, ItalyDepartment of Pharmacy, University of Pisa, 56126 Pisa, ItalyDepartment of Pharmacy, University of Pisa, 56126 Pisa, ItalyDepartment of Pharmacy, University of Pisa, 56126 Pisa, ItalyInstitute of Clinical Physiology, National Research Council, 54100 Massa, ItalyInstitute of Clinical Physiology, National Research Council, 54100 Massa, ItalyThree-dimensional bioprinting has emerged as an attractive technology due to its ability to mimic native tissue architecture using different cell types and biomaterials. Nowadays, cell-laden bioink development or skin tissue equivalents are still at an early stage. The aim of the study is to propose a bioink to be used in skin bioprinting based on a blend of fibrinogen and alginate to form a hydrogel by enzymatic polymerization with thrombin and by ionic crosslinking with divalent calcium ions. The biomaterial ink formulation, composed of 30 mg/mL of fibrinogen, 6% of alginate, and 25 mM of CaCl<sub>2</sub>, was characterized in terms of homogeneity, rheological properties, printability, mechanical properties, degradation rate, water uptake, and biocompatibility by the indirect method using L929 mouse fibroblasts. The proposed bioink is a homogeneous blend with a shear thinning behavior, excellent printability, adequate mechanical stiffness, porosity, biodegradability, and water uptake, and it is in vitro biocompatible. The fibrinogen-based bioink was used for the 3D bioprinting of the dermal layer of the skin equivalent. Three different normal human dermal fibroblast (NHDF) densities were tested, and better results in terms of viability, spreading, and proliferation were obtained with 4 × 10<sup>6</sup> cell/mL. The skin equivalent was bioprinted, adding human keratinocytes (HaCaT) through bioprinting on the top surface of the dermal layer. A skin equivalent stained by live/dead and histological analysis immediately after printing and at days 7 and 14 of culture showed a tissuelike structure with two distinct layers characterized by the presence of viable and proliferating cells. This bioprinted skin equivalent showed a similar native skin architecture, paving the way for its use as a skin substitute for wound healing applications.https://www.mdpi.com/2079-4983/14/9/459fibrinogenalginatebioink3D bioprintingskin equivalent
spellingShingle Aida Cavallo
Tamer Al Kayal
Angelica Mero
Andrea Mezzetta
Lorenzo Guazzelli
Giorgio Soldani
Paola Losi
Fibrinogen-Based Bioink for Application in Skin Equivalent 3D Bioprinting
Journal of Functional Biomaterials
fibrinogen
alginate
bioink
3D bioprinting
skin equivalent
title Fibrinogen-Based Bioink for Application in Skin Equivalent 3D Bioprinting
title_full Fibrinogen-Based Bioink for Application in Skin Equivalent 3D Bioprinting
title_fullStr Fibrinogen-Based Bioink for Application in Skin Equivalent 3D Bioprinting
title_full_unstemmed Fibrinogen-Based Bioink for Application in Skin Equivalent 3D Bioprinting
title_short Fibrinogen-Based Bioink for Application in Skin Equivalent 3D Bioprinting
title_sort fibrinogen based bioink for application in skin equivalent 3d bioprinting
topic fibrinogen
alginate
bioink
3D bioprinting
skin equivalent
url https://www.mdpi.com/2079-4983/14/9/459
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