Three-dimensional cell culture of chondrocytes on modified di-phenylalanine scaffolds.
The design of self-assembled peptide-based structures for three-dimensional cell culture and tissue repair has been a key objective in biomaterials science for decades. In search of the simplest possible peptide system that can self-assemble, we discovered that combinations of di-peptides that are modified with aromatic stacking ligands could form nanometre-sized fibres when exposed to physiological conditions. For example, we demonstrated that a number of Fmoc (fluoren-9-ylmethyloxycarbonyl) modified di- and tri-peptides form highly ordered hydrogels via hydrogen-bonding and pi-pi interactions from the fluorenyl rings. These highly hydrated gels allowed for cell proliferation of chondrocytes in three dimensions [Jayawarna, Ali, Jowitt, Miller, Saiani, Gough and Ulijn (2006) Adv. Mater. 18, 611-614]. We demonstrated that fibrous architecture and physical properties of the resulting materials were dictated by the nature of the amino acid building blocks. Here, we report the self-assembly process of three di-phenylalanine analogues, Fmoc-Phe-Phe-OH, Nap (naphthalene)-Phe-Phe-OH and Cbz (benzyloxycarbonyl)-Phe-Phe-OH, to compare and contrast the self-assembly properties and cell culture conditions attributable to their protecting group difference. Fibre morphology analysis of the three structures using cryo-SEM (scanning electron microscopy) and TEM (transmission electron microscopy) suggested fibrous structures with dramatically varying fibril dimensions, depending on the aromatic ligand used. CD and FTIR (Fourier-transform IR) data confirmed beta-sheet arrangements in all three samples in the gel state. The ability of these three new hydrogels to support cell proliferation of chondrocytes was confirmed for all three materials.
Jayawarna
V
V
School of Materials, The University of Manchester, PO Box 88, Manchester M60 1QD, UK. V.Jayawarna@postgrad.manchester.ac.uk
Smith
A
A
Gough
J E
JE
Ulijn
R V
RV
D001672:Biocompatible Materials / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology*; D004151:Dipeptides; D005449:Fluorenes / Q000737:chemistry; D020100:Hydrogels; D008024:Ligands; D008422:Materials Testing; D008854:Microscopy, Electron; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000737:chemistry; D023822:Tissue Engineering / Q000379:methods
D001672:Biocompatible Materials; D004151:Dipeptides; D005449:Fluorenes; D020100:Hydrogels; D008024:Ligands; C026650:phenylalanylphenylalanine; D010649:Phenylalanine