Nanofibrous PLGA electrospun scaffolds modified with type I collagen influence hepatocyte function and support viability in vitro

A major challenge of maintaining primary hepatocytes in vitro is progressive loss of hepatocyte-specific functions, such as protein synthesis and cytochrome P450 (CYP450) catalytic activity. We developed a three-dimensional (3D) nanofibrous scaffold made from poly(l-lactide-co-glycolide) (PLGA) poly...

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Main Authors: Brown, Jessica H., Das, Prativa, DiVito, Michael D., Ivancic, David, Tan, Lay Poh, Wertheim, Jason A.
Other Authors: School of Materials Science & Engineering
Format: Article
Language:English
Published: 2019
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Online Access:https://hdl.handle.net/10356/90298
http://hdl.handle.net/10220/48478
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Institution: Nanyang Technological University
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spelling sg-ntu-dr.10356-902982020-11-01T04:44:19Z Nanofibrous PLGA electrospun scaffolds modified with type I collagen influence hepatocyte function and support viability in vitro Brown, Jessica H. Das, Prativa DiVito, Michael D. Ivancic, David Tan, Lay Poh Wertheim, Jason A. School of Materials Science & Engineering Interdisciplinary Graduate School (IGS) Tissue Engineering Electrospun Nanofibers DRNTU::Engineering::Materials A major challenge of maintaining primary hepatocytes in vitro is progressive loss of hepatocyte-specific functions, such as protein synthesis and cytochrome P450 (CYP450) catalytic activity. We developed a three-dimensional (3D) nanofibrous scaffold made from poly(l-lactide-co-glycolide) (PLGA) polymer using a newly optimized wet electrospinning technique that resulted in a highly porous structure that accommodated inclusion of primary human hepatocytes. Extracellular matrix (ECM) proteins (type I collagen or fibronectin) at varying concentrations were chemically linked to electrospun PLGA using amine coupling to develop an in vitro culture system containing the minimal essential ECM components of the liver micro-environment that preserve hepatocyte function in vitro. Cell-laden nanofiber scaffolds were tested in vitro to maintain hepatocyte function over a two-week period. Incorporation of type I collagen onto PLGA scaffolds (PLGA-Chigh: 100 µg/mL) led to 10-fold greater albumin secretion, 4-fold higher urea synthesis, and elevated transcription of hepatocyte-specific CYP450 genes (CYP3A4, 3.5-fold increase and CYP2C9, 3-fold increase) in primary human hepatocytes compared to the same cells grown within unmodified PLGA scaffolds over two weeks. These indices, measured using collagen-bonded scaffolds, were also higher than scaffolds coupled to fibronectin or an ECM control sandwich culture composed of type I collagen and Matrigel. Induction of CYP2C9 activity was also higher in these same type I collagen PLGA scaffolds compared to other ECM-modified or unmodified PLGA constructs and was equivalent to the ECM control at 7 days. Together, we demonstrate a minimalist ECM-based 3D synthetic scaffold that accommodates primary human hepatocyte inclusion into the matrix, maintains long-term in vitro survival and stimulates function, which can be attributed to coupling of type I collagen. Accepted version 2019-05-30T02:46:28Z 2019-12-06T17:45:10Z 2019-05-30T02:46:28Z 2019-12-06T17:45:10Z 2018 Journal Article Brown, J. H., Das, P., DiVito, M. D., Ivancic, D., Tan, L. P., & Wertheim, J. A. (2018). Nanofibrous PLGA electrospun scaffolds modified with type I collagen influence hepatocyte function and support viability in vitro. Acta Biomaterialia, 73, 217-227. doi:10.1016/j.actbio.2018.02.009 1742-7061 https://hdl.handle.net/10356/90298 http://hdl.handle.net/10220/48478 10.1016/j.actbio.2018.02.009 en Acta Biomaterialia © 2018 Acta Materialia Inc. All rights reserved. This paper was published by Elsevier Ltd in Acta Biomaterialia and is made available with permission of Acta Materialia Inc. 44 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic Tissue Engineering
Electrospun Nanofibers
DRNTU::Engineering::Materials
spellingShingle Tissue Engineering
Electrospun Nanofibers
DRNTU::Engineering::Materials
Brown, Jessica H.
Das, Prativa
DiVito, Michael D.
Ivancic, David
Tan, Lay Poh
Wertheim, Jason A.
Nanofibrous PLGA electrospun scaffolds modified with type I collagen influence hepatocyte function and support viability in vitro
description A major challenge of maintaining primary hepatocytes in vitro is progressive loss of hepatocyte-specific functions, such as protein synthesis and cytochrome P450 (CYP450) catalytic activity. We developed a three-dimensional (3D) nanofibrous scaffold made from poly(l-lactide-co-glycolide) (PLGA) polymer using a newly optimized wet electrospinning technique that resulted in a highly porous structure that accommodated inclusion of primary human hepatocytes. Extracellular matrix (ECM) proteins (type I collagen or fibronectin) at varying concentrations were chemically linked to electrospun PLGA using amine coupling to develop an in vitro culture system containing the minimal essential ECM components of the liver micro-environment that preserve hepatocyte function in vitro. Cell-laden nanofiber scaffolds were tested in vitro to maintain hepatocyte function over a two-week period. Incorporation of type I collagen onto PLGA scaffolds (PLGA-Chigh: 100 µg/mL) led to 10-fold greater albumin secretion, 4-fold higher urea synthesis, and elevated transcription of hepatocyte-specific CYP450 genes (CYP3A4, 3.5-fold increase and CYP2C9, 3-fold increase) in primary human hepatocytes compared to the same cells grown within unmodified PLGA scaffolds over two weeks. These indices, measured using collagen-bonded scaffolds, were also higher than scaffolds coupled to fibronectin or an ECM control sandwich culture composed of type I collagen and Matrigel. Induction of CYP2C9 activity was also higher in these same type I collagen PLGA scaffolds compared to other ECM-modified or unmodified PLGA constructs and was equivalent to the ECM control at 7 days. Together, we demonstrate a minimalist ECM-based 3D synthetic scaffold that accommodates primary human hepatocyte inclusion into the matrix, maintains long-term in vitro survival and stimulates function, which can be attributed to coupling of type I collagen.
author2 School of Materials Science & Engineering
author_facet School of Materials Science & Engineering
Brown, Jessica H.
Das, Prativa
DiVito, Michael D.
Ivancic, David
Tan, Lay Poh
Wertheim, Jason A.
format Article
author Brown, Jessica H.
Das, Prativa
DiVito, Michael D.
Ivancic, David
Tan, Lay Poh
Wertheim, Jason A.
author_sort Brown, Jessica H.
title Nanofibrous PLGA electrospun scaffolds modified with type I collagen influence hepatocyte function and support viability in vitro
title_short Nanofibrous PLGA electrospun scaffolds modified with type I collagen influence hepatocyte function and support viability in vitro
title_full Nanofibrous PLGA electrospun scaffolds modified with type I collagen influence hepatocyte function and support viability in vitro
title_fullStr Nanofibrous PLGA electrospun scaffolds modified with type I collagen influence hepatocyte function and support viability in vitro
title_full_unstemmed Nanofibrous PLGA electrospun scaffolds modified with type I collagen influence hepatocyte function and support viability in vitro
title_sort nanofibrous plga electrospun scaffolds modified with type i collagen influence hepatocyte function and support viability in vitro
publishDate 2019
url https://hdl.handle.net/10356/90298
http://hdl.handle.net/10220/48478
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