Prechondrogenic ATDC5 Cell Attachment and Differentiation on Graphene Foam; Modulation by Surface Functionalization with Fibronectin

Stephanie M. Frahs, Jonathon C. Reeck, Katie M. Yocham, Anders Frederiksen, Kiyo Fujimoto, Crystal M. Scott, Richard S. Beard, Raquel J. Brown, Trevor J. Lujan, Ilia A. Solov'yov, David Estrada, Julia Thom Oxford*

*Kontaktforfatter for dette arbejde

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Graphene foam holds promise for tissue engineering applications. In this study, graphene foam was used as a three-dimension scaffold to evaluate cell attachment, cell morphology, and molecular markers of early differentiation. The aim of this study was to determine if cell attachment and elaboration of an extracellular matrix would be modulated by functionalization of graphene foam with fibronectin, an extracellular matrix protein that cells adhere well to, prior to the establishment of three-dimensional cell culture. The molecular dynamic simulation demonstrated that the fibronectin-graphene interaction was stabilized predominantly through interaction between the graphene and arginine side chains of the protein. Quasi-static and dynamic mechanical testing indicated that fibronectin functionalization of graphene altered the mechanical properties of graphene foam. The elastic strength of the scaffold increased due to fibronectin, but the viscoelastic mechanical behavior remained unchanged. An additive effect was observed in the mechanical stiffness when the graphene foam was both coated with fibronectin and cultured with cells for 28 days. Cytoskeletal organization assessed by fluorescence microscopy demonstrated a fibronectin-dependent reorganization of the actin cytoskeleton and an increase in actin stress fibers. Gene expression assessed by quantitative real-time polymerase chain reaction of 9 genes encoding cell attachment proteins (Cd44, Ctnna1, Ctnnb1, Itga3, Itga5, Itgav, Itgb1, Ncam1, Sgce), 16 genes encoding extracellular matrix proteins (Col1a1, Col2a1, Col3a1, Col5a1, Col6a1, Ecm1, Emilin1, Fn1, Hapln1, Lamb3, Postn, Sparc, Spp1, Thbs1, Thbs2, Tnc), and 9 genes encoding modulators of remodeling (Adamts1, Adamts2, Ctgf, Mmp14, Mmp2, Tgfbi, Timp1, Timp2, Timp3) indicated that graphene foam provided a microenvironment conducive to expression of genes that are important in early chondrogenesis. Functionalization of graphene foam with fibronectin modified the cellular response to graphene foam, demonstrated by decreases in relative gene expression levels. These findings illustrate the combinatorial factors of microscale materials properties and nanoscale molecular features to consider in the design of three-dimensional graphene scaffolds for tissue engineering applications. ©

OriginalsprogEngelsk
TidsskriftACS Applied Materials and Interfaces
Vol/bind11
Udgave nummer45
Sider (fra-til)41906-41924
Antal sider19
ISSN1944-8244
DOI
StatusUdgivet - 13. nov. 2019
Udgivet eksterntJa

Fingeraftryk

Graphite
Fibronectins
Graphene
Foams
Modulation
Gene encoding
Proteins
Extracellular Matrix Proteins
Tissue engineering
Gene expression
Scaffolds
Actins
Arginine
Mechanical testing
Fluorescence microscopy
Polymerase chain reaction
Scaffolds (biology)
Cell culture
Modulators
Molecular dynamics

Citer dette

Frahs, S. M., Reeck, J. C., Yocham, K. M., Frederiksen, A., Fujimoto, K., Scott, C. M., ... Oxford, J. T. (2019). Prechondrogenic ATDC5 Cell Attachment and Differentiation on Graphene Foam; Modulation by Surface Functionalization with Fibronectin. ACS Applied Materials and Interfaces, 11(45), 41906-41924. https://doi.org/10.1021/acsami.9b14670
Frahs, Stephanie M. ; Reeck, Jonathon C. ; Yocham, Katie M. ; Frederiksen, Anders ; Fujimoto, Kiyo ; Scott, Crystal M. ; Beard, Richard S. ; Brown, Raquel J. ; Lujan, Trevor J. ; Solov'yov, Ilia A. ; Estrada, David ; Oxford, Julia Thom. / Prechondrogenic ATDC5 Cell Attachment and Differentiation on Graphene Foam; Modulation by Surface Functionalization with Fibronectin. I: ACS Applied Materials and Interfaces. 2019 ; Bind 11, Nr. 45. s. 41906-41924.
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title = "Prechondrogenic ATDC5 Cell Attachment and Differentiation on Graphene Foam; Modulation by Surface Functionalization with Fibronectin",
abstract = "Graphene foam holds promise for tissue engineering applications. In this study, graphene foam was used as a three-dimension scaffold to evaluate cell attachment, cell morphology, and molecular markers of early differentiation. The aim of this study was to determine if cell attachment and elaboration of an extracellular matrix would be modulated by functionalization of graphene foam with fibronectin, an extracellular matrix protein that cells adhere well to, prior to the establishment of three-dimensional cell culture. The molecular dynamic simulation demonstrated that the fibronectin-graphene interaction was stabilized predominantly through interaction between the graphene and arginine side chains of the protein. Quasi-static and dynamic mechanical testing indicated that fibronectin functionalization of graphene altered the mechanical properties of graphene foam. The elastic strength of the scaffold increased due to fibronectin, but the viscoelastic mechanical behavior remained unchanged. An additive effect was observed in the mechanical stiffness when the graphene foam was both coated with fibronectin and cultured with cells for 28 days. Cytoskeletal organization assessed by fluorescence microscopy demonstrated a fibronectin-dependent reorganization of the actin cytoskeleton and an increase in actin stress fibers. Gene expression assessed by quantitative real-time polymerase chain reaction of 9 genes encoding cell attachment proteins (Cd44, Ctnna1, Ctnnb1, Itga3, Itga5, Itgav, Itgb1, Ncam1, Sgce), 16 genes encoding extracellular matrix proteins (Col1a1, Col2a1, Col3a1, Col5a1, Col6a1, Ecm1, Emilin1, Fn1, Hapln1, Lamb3, Postn, Sparc, Spp1, Thbs1, Thbs2, Tnc), and 9 genes encoding modulators of remodeling (Adamts1, Adamts2, Ctgf, Mmp14, Mmp2, Tgfbi, Timp1, Timp2, Timp3) indicated that graphene foam provided a microenvironment conducive to expression of genes that are important in early chondrogenesis. Functionalization of graphene foam with fibronectin modified the cellular response to graphene foam, demonstrated by decreases in relative gene expression levels. These findings illustrate the combinatorial factors of microscale materials properties and nanoscale molecular features to consider in the design of three-dimensional graphene scaffolds for tissue engineering applications. {\circledC}",
keywords = "ATDC5, bioscaffold, chondroprogenitor cells, differentiation, dynamic mechanical analysis, extracellular matrix, fibronectin, graphene foam, molecular dynamic simulation, three-dimensional cell culture, tissue engineering",
author = "Frahs, {Stephanie M.} and Reeck, {Jonathon C.} and Yocham, {Katie M.} and Anders Frederiksen and Kiyo Fujimoto and Scott, {Crystal M.} and Beard, {Richard S.} and Brown, {Raquel J.} and Lujan, {Trevor J.} and Solov'yov, {Ilia A.} and David Estrada and Oxford, {Julia Thom}",
year = "2019",
month = "11",
day = "13",
doi = "10.1021/acsami.9b14670",
language = "English",
volume = "11",
pages = "41906--41924",
journal = "A C S Applied Materials and Interfaces",
issn = "1944-8244",
publisher = "American Chemical Society",
number = "45",

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Frahs, SM, Reeck, JC, Yocham, KM, Frederiksen, A, Fujimoto, K, Scott, CM, Beard, RS, Brown, RJ, Lujan, TJ, Solov'yov, IA, Estrada, D & Oxford, JT 2019, 'Prechondrogenic ATDC5 Cell Attachment and Differentiation on Graphene Foam; Modulation by Surface Functionalization with Fibronectin', ACS Applied Materials and Interfaces, bind 11, nr. 45, s. 41906-41924. https://doi.org/10.1021/acsami.9b14670

Prechondrogenic ATDC5 Cell Attachment and Differentiation on Graphene Foam; Modulation by Surface Functionalization with Fibronectin. / Frahs, Stephanie M.; Reeck, Jonathon C.; Yocham, Katie M.; Frederiksen, Anders; Fujimoto, Kiyo; Scott, Crystal M.; Beard, Richard S.; Brown, Raquel J.; Lujan, Trevor J.; Solov'yov, Ilia A.; Estrada, David; Oxford, Julia Thom.

I: ACS Applied Materials and Interfaces, Bind 11, Nr. 45, 13.11.2019, s. 41906-41924.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Prechondrogenic ATDC5 Cell Attachment and Differentiation on Graphene Foam; Modulation by Surface Functionalization with Fibronectin

AU - Frahs, Stephanie M.

AU - Reeck, Jonathon C.

AU - Yocham, Katie M.

AU - Frederiksen, Anders

AU - Fujimoto, Kiyo

AU - Scott, Crystal M.

AU - Beard, Richard S.

AU - Brown, Raquel J.

AU - Lujan, Trevor J.

AU - Solov'yov, Ilia A.

AU - Estrada, David

AU - Oxford, Julia Thom

PY - 2019/11/13

Y1 - 2019/11/13

N2 - Graphene foam holds promise for tissue engineering applications. In this study, graphene foam was used as a three-dimension scaffold to evaluate cell attachment, cell morphology, and molecular markers of early differentiation. The aim of this study was to determine if cell attachment and elaboration of an extracellular matrix would be modulated by functionalization of graphene foam with fibronectin, an extracellular matrix protein that cells adhere well to, prior to the establishment of three-dimensional cell culture. The molecular dynamic simulation demonstrated that the fibronectin-graphene interaction was stabilized predominantly through interaction between the graphene and arginine side chains of the protein. Quasi-static and dynamic mechanical testing indicated that fibronectin functionalization of graphene altered the mechanical properties of graphene foam. The elastic strength of the scaffold increased due to fibronectin, but the viscoelastic mechanical behavior remained unchanged. An additive effect was observed in the mechanical stiffness when the graphene foam was both coated with fibronectin and cultured with cells for 28 days. Cytoskeletal organization assessed by fluorescence microscopy demonstrated a fibronectin-dependent reorganization of the actin cytoskeleton and an increase in actin stress fibers. Gene expression assessed by quantitative real-time polymerase chain reaction of 9 genes encoding cell attachment proteins (Cd44, Ctnna1, Ctnnb1, Itga3, Itga5, Itgav, Itgb1, Ncam1, Sgce), 16 genes encoding extracellular matrix proteins (Col1a1, Col2a1, Col3a1, Col5a1, Col6a1, Ecm1, Emilin1, Fn1, Hapln1, Lamb3, Postn, Sparc, Spp1, Thbs1, Thbs2, Tnc), and 9 genes encoding modulators of remodeling (Adamts1, Adamts2, Ctgf, Mmp14, Mmp2, Tgfbi, Timp1, Timp2, Timp3) indicated that graphene foam provided a microenvironment conducive to expression of genes that are important in early chondrogenesis. Functionalization of graphene foam with fibronectin modified the cellular response to graphene foam, demonstrated by decreases in relative gene expression levels. These findings illustrate the combinatorial factors of microscale materials properties and nanoscale molecular features to consider in the design of three-dimensional graphene scaffolds for tissue engineering applications. ©

AB - Graphene foam holds promise for tissue engineering applications. In this study, graphene foam was used as a three-dimension scaffold to evaluate cell attachment, cell morphology, and molecular markers of early differentiation. The aim of this study was to determine if cell attachment and elaboration of an extracellular matrix would be modulated by functionalization of graphene foam with fibronectin, an extracellular matrix protein that cells adhere well to, prior to the establishment of three-dimensional cell culture. The molecular dynamic simulation demonstrated that the fibronectin-graphene interaction was stabilized predominantly through interaction between the graphene and arginine side chains of the protein. Quasi-static and dynamic mechanical testing indicated that fibronectin functionalization of graphene altered the mechanical properties of graphene foam. The elastic strength of the scaffold increased due to fibronectin, but the viscoelastic mechanical behavior remained unchanged. An additive effect was observed in the mechanical stiffness when the graphene foam was both coated with fibronectin and cultured with cells for 28 days. Cytoskeletal organization assessed by fluorescence microscopy demonstrated a fibronectin-dependent reorganization of the actin cytoskeleton and an increase in actin stress fibers. Gene expression assessed by quantitative real-time polymerase chain reaction of 9 genes encoding cell attachment proteins (Cd44, Ctnna1, Ctnnb1, Itga3, Itga5, Itgav, Itgb1, Ncam1, Sgce), 16 genes encoding extracellular matrix proteins (Col1a1, Col2a1, Col3a1, Col5a1, Col6a1, Ecm1, Emilin1, Fn1, Hapln1, Lamb3, Postn, Sparc, Spp1, Thbs1, Thbs2, Tnc), and 9 genes encoding modulators of remodeling (Adamts1, Adamts2, Ctgf, Mmp14, Mmp2, Tgfbi, Timp1, Timp2, Timp3) indicated that graphene foam provided a microenvironment conducive to expression of genes that are important in early chondrogenesis. Functionalization of graphene foam with fibronectin modified the cellular response to graphene foam, demonstrated by decreases in relative gene expression levels. These findings illustrate the combinatorial factors of microscale materials properties and nanoscale molecular features to consider in the design of three-dimensional graphene scaffolds for tissue engineering applications. ©

KW - ATDC5

KW - bioscaffold

KW - chondroprogenitor cells

KW - differentiation

KW - dynamic mechanical analysis

KW - extracellular matrix

KW - fibronectin

KW - graphene foam

KW - molecular dynamic simulation

KW - three-dimensional cell culture

KW - tissue engineering

U2 - 10.1021/acsami.9b14670

DO - 10.1021/acsami.9b14670

M3 - Journal article

C2 - 31639302

AN - SCOPUS:85074779934

VL - 11

SP - 41906

EP - 41924

JO - A C S Applied Materials and Interfaces

JF - A C S Applied Materials and Interfaces

SN - 1944-8244

IS - 45

ER -