title: Characterisation of minimalist co-assembled fluorenylmethyloxycarbonyl self-assembling peptide systems for presentation of multiple bioactive peptides.
abstract: The nanofibrillar structures that underpin self-assembling peptide (SAP) hydrogels offer great potential for the development of finely tuned cellular microenvironments suitable for tissue engineering. However, biofunctionalisation without disruption of the assembly remains a key issue. SAPS present the peptide sequence within their structure, and studies to date have typically focused on including a single biological motif, resulting in chemically and biologically homogenous scaffolds. This limits the utility of these systems, as they cannot effectively mimic the complexity of the multicomponent extracellular matrix (ECM). In this work, we demonstrate the first successful co-assembly of two biologically active SAPs to form a coassembled scaffold of distinct two-component nanofibrils, and demonstrate that this approach is more bioactive than either of the individual systems alone. Here, we use two bioinspired SAPs from two key ECM proteins: Fmoc-FRGDF containing the RGD sequence from fibronectin and Fmoc-DIKVAV containing the IKVAV sequence from laminin. Our results demonstrate that these SAPs are able to co-assemble to form stable hybrid nanofibres containing dual epitopes. Comparison of the co-assembled SAP system to the individual SAP hydrogels and to a mixed system (composed of the two hydrogels mixed together post-assembly) demonstrates its superior stable, transparent, shear-thinning hydrogels at biological pH, ideal characteristics for tissue engineering applications. Importantly, we show that only the coassembled hydrogel is able to induce in vitro multinucleate myotube formation with C2C12 cells. This work illustrates the importance of tissue engineering scaffold functionalisation and the need to develop increasingly advanced multicomponent systems for effective ECM mimicry.
  
  
  
  Successful control of stem cell fate in tissue engineering applications requires the use of sophisticated scaffolds that deliver biological signals to guide growth and differentiation. The complexity of such processes necessitates the presentation of multiple signals in order to effectively mimic the native extracellular matrix (ECM). Here, we establish the use of two biofunctional, minimalist self-assembling peptides (SAPs) to construct the first co-assembled SAP scaffold. Our work characterises this construct, demonstrating that the physical, chemical, and biological properties of the peptides are maintained during the co-assembly process. Importantly, the coassembled system demonstrates superior biological performance relative to the individual SAPs, highlighting the importance of complex ECM mimicry. This work has important implications for future tissue engineering studies.
authors.affiliation: Research School of Engineering, The Australian National University, Canberra 2601, Australia.
authors.forename: Conor C
authors.initials: CC
authors.lastname: Horgan
authors.affiliation: Research School of Engineering, The Australian National University, Canberra 2601, Australia.
authors.forename: Alexandra L
authors.initials: AL
authors.lastname: Rodriguez
authors.affiliation: Centre for Chemistry and Biotechnology, Deakin University, Waurn Ponds 3217, Australia.
authors.forename: Rui
authors.initials: R
authors.lastname: Li
authors.affiliation: Research School of Engineering, The Australian National University, Canberra 2601, Australia.
authors.forename: Kiara F
authors.initials: KF
authors.lastname: Bruggeman
authors.affiliation: School of Medicine, Deakin University, Waurn Ponds 3217, Australia.
authors.forename: Nicole
authors.initials: N
authors.lastname: Stupka
authors.affiliation: CSIRO Food and Nutritional Sciences, Werribee 3030, Australia.
authors.forename: Jared K
authors.initials: JK
authors.lastname: Raynes
authors.affiliation: CSIRO Food and Nutritional Sciences, Werribee 3030, Australia; AgResearch Ltd, Grasslands Research Centre, Palmerston North 4442, New Zealand.
authors.forename: Li
authors.initials: L
authors.lastname: Day
authors.affiliation: Research School of Chemistry, The Australian National University, Canberra 2601, Australia.
authors.forename: John W
authors.initials: JW
authors.lastname: White
authors.affiliation: School of Aerospace, Mechanical and Manufacturing Engineering, RMIT University, Melbourne 3001, Australia. Electronic address: richard.williams@rmit.edu.au.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Williams
authors.affiliation: Research School of Engineering, The Australian National University, Canberra 2601, Australia. Electronic address: david.nisbet@anu.edu.au.
authors.forename: David R
authors.initials: DR
authors.lastname: Nisbet
mesh_terms: D005109:Extracellular Matrix / Q000737:chemistry*; D005449:Fluorenes / Q000737:chemistry*; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry*
chemical_list: D005449:Fluorenes; D010455:Peptides
keywords: Co-assembly; Fmoc; Hydrogels; Self-assembling peptides; Tissue engineering