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Optimizing Bioink Composition for Human Chondrocyte Expression of Lubricin

The surface zone of articular cartilage is the first area impacted by cartilage defects, commonly resulting in osteoarthritis. Chondrocytes in the surface zone of articular cartilage synthesize and secrete lubricin, a proteoglycan that functions as a lubricant protecting the deeper layers from shear...

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Autores principales: Martyniak, Kari, Kennedy, Sean, Karimzadeh, Makan, Cruz, Maria A., Jeon, Oju, Alsberg, Eben, Kean, Thomas J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10526043/
https://www.ncbi.nlm.nih.gov/pubmed/37760099
http://dx.doi.org/10.3390/bioengineering10090997
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author Martyniak, Kari
Kennedy, Sean
Karimzadeh, Makan
Cruz, Maria A.
Jeon, Oju
Alsberg, Eben
Kean, Thomas J.
author_facet Martyniak, Kari
Kennedy, Sean
Karimzadeh, Makan
Cruz, Maria A.
Jeon, Oju
Alsberg, Eben
Kean, Thomas J.
author_sort Martyniak, Kari
collection PubMed
description The surface zone of articular cartilage is the first area impacted by cartilage defects, commonly resulting in osteoarthritis. Chondrocytes in the surface zone of articular cartilage synthesize and secrete lubricin, a proteoglycan that functions as a lubricant protecting the deeper layers from shear stress. Notably, 3D bioprinting is a tissue engineering technique that uses cells encapsulated in biomaterials to fabricate 3D constructs. Gelatin methacrylate (GelMA) is a frequently used biomaterial for 3D bioprinting cartilage. Oxidized methacrylated alginate (OMA) is a chemically modified alginate designed for its tunable degradation rate and mechanical properties. To determine an optimal combination of GelMA and OMA for lubricin expression, we used our novel high-throughput human articular chondrocyte reporter system. Primary human chondrocytes were transduced with PRG4 (lubricin) promoter-driven Gaussia luciferase, allowing for temporal assessment of lubricin expression. A lubricin expression-driven Design of Experiment screen and subsequent validation identified 14% GelMA/2% OMA for further study. Therefore, DoE optimized 14% GelMA/2% OMA, 14% GelMA control, and 16% GelMA (total solid content control) were 3D bioprinted. The combination of lubricin protein expression and shape retention over the 22 days in culture, successfully determined the 14% GelMA/2%OMA to be the optimal formulation for lubricin secretion. This strategy allows for rapid analysis of the role(s) of biomaterial composition, stiffness or other cell manipulations on lubricin expression by chondrocytes, which may improve therapeutic strategies for cartilage regeneration.
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spelling pubmed-105260432023-09-28 Optimizing Bioink Composition for Human Chondrocyte Expression of Lubricin Martyniak, Kari Kennedy, Sean Karimzadeh, Makan Cruz, Maria A. Jeon, Oju Alsberg, Eben Kean, Thomas J. Bioengineering (Basel) Article The surface zone of articular cartilage is the first area impacted by cartilage defects, commonly resulting in osteoarthritis. Chondrocytes in the surface zone of articular cartilage synthesize and secrete lubricin, a proteoglycan that functions as a lubricant protecting the deeper layers from shear stress. Notably, 3D bioprinting is a tissue engineering technique that uses cells encapsulated in biomaterials to fabricate 3D constructs. Gelatin methacrylate (GelMA) is a frequently used biomaterial for 3D bioprinting cartilage. Oxidized methacrylated alginate (OMA) is a chemically modified alginate designed for its tunable degradation rate and mechanical properties. To determine an optimal combination of GelMA and OMA for lubricin expression, we used our novel high-throughput human articular chondrocyte reporter system. Primary human chondrocytes were transduced with PRG4 (lubricin) promoter-driven Gaussia luciferase, allowing for temporal assessment of lubricin expression. A lubricin expression-driven Design of Experiment screen and subsequent validation identified 14% GelMA/2% OMA for further study. Therefore, DoE optimized 14% GelMA/2% OMA, 14% GelMA control, and 16% GelMA (total solid content control) were 3D bioprinted. The combination of lubricin protein expression and shape retention over the 22 days in culture, successfully determined the 14% GelMA/2%OMA to be the optimal formulation for lubricin secretion. This strategy allows for rapid analysis of the role(s) of biomaterial composition, stiffness or other cell manipulations on lubricin expression by chondrocytes, which may improve therapeutic strategies for cartilage regeneration. MDPI 2023-08-23 /pmc/articles/PMC10526043/ /pubmed/37760099 http://dx.doi.org/10.3390/bioengineering10090997 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Martyniak, Kari
Kennedy, Sean
Karimzadeh, Makan
Cruz, Maria A.
Jeon, Oju
Alsberg, Eben
Kean, Thomas J.
Optimizing Bioink Composition for Human Chondrocyte Expression of Lubricin
title Optimizing Bioink Composition for Human Chondrocyte Expression of Lubricin
title_full Optimizing Bioink Composition for Human Chondrocyte Expression of Lubricin
title_fullStr Optimizing Bioink Composition for Human Chondrocyte Expression of Lubricin
title_full_unstemmed Optimizing Bioink Composition for Human Chondrocyte Expression of Lubricin
title_short Optimizing Bioink Composition for Human Chondrocyte Expression of Lubricin
title_sort optimizing bioink composition for human chondrocyte expression of lubricin
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10526043/
https://www.ncbi.nlm.nih.gov/pubmed/37760099
http://dx.doi.org/10.3390/bioengineering10090997
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