title: The in vivo performance of an enzyme-assisted self-assembled peptide/protein hydrogel.
abstract: We demonstrate the distribution of the important extracellular matrix protein laminin in a novel biomaterial consisting of a hydrogel underpinned by nanofibrillar networks. These are formed by the immobilised enzyme mediated self-assembly of fmoc-L(3) (9-fluorenylmethoxycarbonyl-tri-leucine). The peptide assembly yields nanofibrils formed of β-sheets that are locked together via π-stacking interactions. This ordering allows the localisation of the peptide sidechains on the surface, creating a hydrophobic environment. This induces the formation of bundles of these nanofibrils producing a clear hydrogel. This mechanism enables the three dimensional distribution of laminin throughout the network via supramolecular interactions. These forces favour the formation and improve the order of the network itself, as observed by spectroscopic and mechanical testing. In order to test the stability and suitability of this class of material for in vivo applications, we utilise microinjection to deliver the biomaterial under fine spatial control into a dystrophic zebrafish model organism, which lacks laminin as a result of a genetic mutation. Using confocal and transmission electron microscopy, we confirm that the biomaterial remains stable structurally, and is confined spatially to the site of injection.
authors.affiliation: CSIRO Materials Science and Engineering, Ian Wark Laboratories, Bag 10, Clayton South, VIC 3169, Australia. Richard.Williams@CSIRO.au
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Williams
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Hall
authors.forename: Veronica
authors.initials: V
authors.lastname: Glattauer
authors.forename: Jacinta
authors.initials: J
authors.lastname: White
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Pasic
authors.forename: Anders B
authors.initials: AB
authors.lastname: Sorensen
authors.forename: Lynne
authors.initials: L
authors.lastname: Waddington
authors.forename: Keith M
authors.initials: KM
authors.lastname: McLean
authors.forename: Peter D
authors.initials: PD
authors.lastname: Currie
authors.forename: Patrick G
authors.initials: PG
authors.lastname: Hartley
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D005449:Fluorenes / Q000737:chemistry; D020100:Hydrogels / Q000138:chemical synthesis / Q000737:chemistry*; D007797:Laminin / Q000235:genetics / Q000378:metabolism; D007930:Leucine / Q000737:chemistry; D008422:Materials Testing; D015394:Molecular Structure; D057139:Nanofibers / Q000737:chemistry / Q000648:ultrastructure; D010455:Peptides / Q000737:chemistry*; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D015027:Zebrafish / Q000033:anatomy & histology / Q000235:genetics; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: C496789:9-fluorenylmethoxycarbonyl; D005449:Fluorenes; D020100:Hydrogels; D007797:Laminin; D010455:Peptides; D011506:Proteins; D029961:Zebrafish Proteins; D007930:Leucine