Introducing chemical functionality in fmoc-peptide gels for cell culture

V. Jayawarna, S.M. Richardson, A.R. Hirst, N.W. Hodson, A. Saiani, J.E. Gough, R.V. Ulijn

Research output: Contribution to journalArticle

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Abstract

Aromatic short peptide derivatives, i.e. peptides modified with aromatic groups such as 9-fluorenylmethoxycarbonyl (Fmoc), can self-assemble into self-supporting hydrogels. These hydrogels have some similarities to extracellular matrices due to their high hydration, relative stiffness and nanofibrous architecture. We previously demonstrated that Fmoc-diphenylalanine (Fmoc-F2) provides a suitable matrix for two-dimensional (2D) or three-dimensional (3D) culture of primary bovine chondrocytes. In this paper we investigate whether the introduction of chemical functionality, such as NH2, COOH or OH, enhances compatibility with different cell types. A series of hydrogel compositions consisting of combinations of Fmoc-F2 and n-protected Fmoc amino acids, lysine (K, with side chain R = (CH2)4NH2), glutamic acid (D, with side chain R = CH2COOH), and serine (S, with side chain R = CH2OH) were studied. All compositions produced fibrous scaffolds with fibre diameters in the range of 32-65 nm as assessed by cryo-scanning electron microscopy and atomic force microscopy. Fourier transform infrared spectroscopy analysis suggested that peptide segments adopt a predominantly antiparallel β-sheet conformation. Oscillatory rheology results show that all four hydrogels have mechanical profiles of soft viscoelastic materials with elastic moduli dependent on the chemical composition, ranging from 502 Pa (Fmoc-F2/D) to 21.2 KPa (Fmoc-F2). All gels supported the viability of bovine chondrocytes as assessed by a live-dead staining assay. Fmoc-F2/S and Fmoc-F2/D hydrogels in addition supported viability for human dermal fibroblasts (HDF) while Fmoc-F2/S hydrogel was the only gel type that supported viability for all three cell types tested. Fmoc-F2/S was therefore investigated further by studying cell proliferation, cytoskeletal organization and histological analysis in 2D culture. In addition, the Fmoc-F2/S gel was shown to support retention of cell morphology in 3D culture of bovine chondrocytes. These results demonstrate that introduction of chemical functionality into Fmoc-peptide scaffolds may provide gels with tunable chemical and mechanical properties for in vitro cell culture.
LanguageEnglish
Pages934-943
Number of pages9
JournalActa Biomaterialia
Volume5
Issue number3
DOIs
Publication statusPublished - Mar 2009

Fingerprint

Cell culture
Hydrogels
Peptides
Gels
Cell Culture Techniques
Chemical analysis
Chondrocytes
Cell proliferation
Fibroblasts
Hydrogel
Scaffolds (biology)
Rheology
Scaffolds
Hydration
Chemical properties
Fourier transform infrared spectroscopy
Conformations
9-fluorenylmethoxycarbonyl
Amino acids
Assays

Keywords

  • hydrogels
  • cell culture
  • self-assembly
  • biomolecule
  • peptides

Cite this

Jayawarna, V., Richardson, S. M., Hirst, A. R., Hodson, N. W., Saiani, A., Gough, J. E., & Ulijn, R. V. (2009). Introducing chemical functionality in fmoc-peptide gels for cell culture. Acta Biomaterialia, 5(3), 934-943. https://doi.org/10.1016/j.actbio.2009.01.006
Jayawarna, V. ; Richardson, S.M. ; Hirst, A.R. ; Hodson, N.W. ; Saiani, A. ; Gough, J.E. ; Ulijn, R.V. / Introducing chemical functionality in fmoc-peptide gels for cell culture. In: Acta Biomaterialia. 2009 ; Vol. 5, No. 3. pp. 934-943.
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Jayawarna, V, Richardson, SM, Hirst, AR, Hodson, NW, Saiani, A, Gough, JE & Ulijn, RV 2009, 'Introducing chemical functionality in fmoc-peptide gels for cell culture' Acta Biomaterialia, vol. 5, no. 3, pp. 934-943. https://doi.org/10.1016/j.actbio.2009.01.006

Introducing chemical functionality in fmoc-peptide gels for cell culture. / Jayawarna, V.; Richardson, S.M.; Hirst, A.R.; Hodson, N.W.; Saiani, A.; Gough, J.E.; Ulijn, R.V.

In: Acta Biomaterialia, Vol. 5, No. 3, 03.2009, p. 934-943.

Research output: Contribution to journalArticle

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T1 - Introducing chemical functionality in fmoc-peptide gels for cell culture

AU - Jayawarna, V.

AU - Richardson, S.M.

AU - Hirst, A.R.

AU - Hodson, N.W.

AU - Saiani, A.

AU - Gough, J.E.

AU - Ulijn, R.V.

PY - 2009/3

Y1 - 2009/3

N2 - Aromatic short peptide derivatives, i.e. peptides modified with aromatic groups such as 9-fluorenylmethoxycarbonyl (Fmoc), can self-assemble into self-supporting hydrogels. These hydrogels have some similarities to extracellular matrices due to their high hydration, relative stiffness and nanofibrous architecture. We previously demonstrated that Fmoc-diphenylalanine (Fmoc-F2) provides a suitable matrix for two-dimensional (2D) or three-dimensional (3D) culture of primary bovine chondrocytes. In this paper we investigate whether the introduction of chemical functionality, such as NH2, COOH or OH, enhances compatibility with different cell types. A series of hydrogel compositions consisting of combinations of Fmoc-F2 and n-protected Fmoc amino acids, lysine (K, with side chain R = (CH2)4NH2), glutamic acid (D, with side chain R = CH2COOH), and serine (S, with side chain R = CH2OH) were studied. All compositions produced fibrous scaffolds with fibre diameters in the range of 32-65 nm as assessed by cryo-scanning electron microscopy and atomic force microscopy. Fourier transform infrared spectroscopy analysis suggested that peptide segments adopt a predominantly antiparallel β-sheet conformation. Oscillatory rheology results show that all four hydrogels have mechanical profiles of soft viscoelastic materials with elastic moduli dependent on the chemical composition, ranging from 502 Pa (Fmoc-F2/D) to 21.2 KPa (Fmoc-F2). All gels supported the viability of bovine chondrocytes as assessed by a live-dead staining assay. Fmoc-F2/S and Fmoc-F2/D hydrogels in addition supported viability for human dermal fibroblasts (HDF) while Fmoc-F2/S hydrogel was the only gel type that supported viability for all three cell types tested. Fmoc-F2/S was therefore investigated further by studying cell proliferation, cytoskeletal organization and histological analysis in 2D culture. In addition, the Fmoc-F2/S gel was shown to support retention of cell morphology in 3D culture of bovine chondrocytes. These results demonstrate that introduction of chemical functionality into Fmoc-peptide scaffolds may provide gels with tunable chemical and mechanical properties for in vitro cell culture.

AB - Aromatic short peptide derivatives, i.e. peptides modified with aromatic groups such as 9-fluorenylmethoxycarbonyl (Fmoc), can self-assemble into self-supporting hydrogels. These hydrogels have some similarities to extracellular matrices due to their high hydration, relative stiffness and nanofibrous architecture. We previously demonstrated that Fmoc-diphenylalanine (Fmoc-F2) provides a suitable matrix for two-dimensional (2D) or three-dimensional (3D) culture of primary bovine chondrocytes. In this paper we investigate whether the introduction of chemical functionality, such as NH2, COOH or OH, enhances compatibility with different cell types. A series of hydrogel compositions consisting of combinations of Fmoc-F2 and n-protected Fmoc amino acids, lysine (K, with side chain R = (CH2)4NH2), glutamic acid (D, with side chain R = CH2COOH), and serine (S, with side chain R = CH2OH) were studied. All compositions produced fibrous scaffolds with fibre diameters in the range of 32-65 nm as assessed by cryo-scanning electron microscopy and atomic force microscopy. Fourier transform infrared spectroscopy analysis suggested that peptide segments adopt a predominantly antiparallel β-sheet conformation. Oscillatory rheology results show that all four hydrogels have mechanical profiles of soft viscoelastic materials with elastic moduli dependent on the chemical composition, ranging from 502 Pa (Fmoc-F2/D) to 21.2 KPa (Fmoc-F2). All gels supported the viability of bovine chondrocytes as assessed by a live-dead staining assay. Fmoc-F2/S and Fmoc-F2/D hydrogels in addition supported viability for human dermal fibroblasts (HDF) while Fmoc-F2/S hydrogel was the only gel type that supported viability for all three cell types tested. Fmoc-F2/S was therefore investigated further by studying cell proliferation, cytoskeletal organization and histological analysis in 2D culture. In addition, the Fmoc-F2/S gel was shown to support retention of cell morphology in 3D culture of bovine chondrocytes. These results demonstrate that introduction of chemical functionality into Fmoc-peptide scaffolds may provide gels with tunable chemical and mechanical properties for in vitro cell culture.

KW - hydrogels

KW - cell culture

KW - self-assembly

KW - biomolecule

KW - peptides

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JF - Acta Biomaterialia

SN - 1742-7061

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Jayawarna V, Richardson SM, Hirst AR, Hodson NW, Saiani A, Gough JE et al. Introducing chemical functionality in fmoc-peptide gels for cell culture. Acta Biomaterialia. 2009 Mar;5(3):934-943. https://doi.org/10.1016/j.actbio.2009.01.006