Branched peptide-amphiphiles as self-assembling coatings for tissue engineering scaffolds.

An important challenge in regenerative medicine is the design of suitable bioactive scaffold materials that can act as artificial extracellular matrices. We reported previously on a family of peptide-amphiphile (PA) molecules that self-assemble into high-aspect ratio nanofibers under physiological conditions, and can display bioactive peptide epitopes along each nanofiber's periphery. One type of PA displays its epitope at a branched site using a lysine dendron, a molecular feature that improves epitope availability on the nanofiber surface. In this work, we describe the application of these branched PA (b-PA) systems as self-assembling coatings for fiber-bonded poly(glycolic acid) scaffolds. b-PAs bearing variations of the RGDS adhesion epitope from fibronectin were shown by elemental analysis to coat repeatably onto fiber scaffolds. The retention of supramolecular organization after coating on the scaffold was demonstrated through spectroscopic identification of beta-sheet structures and the close association of hydrophobic tails in a model pyrene-containing PA system. Primary human bladder smooth muscle cells demonstrated greater initial adhesion to b-PA-functionalized scaffolds than to bare scaffolds or to those coated with linear PAs. This strategy of molecular design and coating may have potential application in bladder tissue regeneration.

Supramolecular fluorophores for biological studies: phenylene vinylene-amino acid amphiphiles.

We report here on a family of self-assembling fluorescent organic amphiphiles with a biomolecular L-lysine hydrophile and a photonically active phenylene vinylene hydrophobe. Unlike conventional amphiphiles, these segmented dendrimers feature a rigid, branched hydrophobe, and have packing characteristics controlled by the ratio of cross-sectional areas of the hydrophobe and hydrophile. In dilute solution, the amphiphiles form supramolecular aggregates, which are easily taken in by cells through an endocytic pathway, and have no discernible effect on cell proliferation or morphology. An analogous pyrene-based amphiphile was cytotoxic, suggesting that cell survival may be linked either to the self-assembling nature of the amphiphiles, or to the specific properties of the phenylene vinylene segment. The combination of photonic and biological components in these amphiphiles provides great potential for applications in sensing or delivery of molecules to intracellular targets.

Probing the interior of peptide amphiphile supramolecular aggregates.

We present a study of the aqueous solvation within self-assembled structures formed from peptide amphiphiles. We have placed tryptophan and pyrene chromophores onto the peptide backbone to enable spectroscopic examinations of the interior of the resulting supramolecular objects. Self-assembly constrains the chromophores to a defined location within an aggregate, and they experience differing degrees of quencher penetration reflective of their depth within the nanostructure. Tryptophan fluorescence indicates that the interiors remain well-solvated, suggesting that the supramolecular aggregates maintain high degrees of free volume. The Stern-Volmer quenching constants and the fractional accessibility (of covalently bound pyrene) progressively increase as the chromophore is placed closer to the aggregate exterior. Furthermore, these aggregates encourage chromophore uptake from aqueous solution as evidenced by the solubilization of free pyrene chromophores. Our findings demonstrate that covalently bound fluorophores within an aggregate can interact with the external environment. Studies with small molecular probes indicate that these self-assembled architectures may represent viable vehicles to sequester hydrophobic, insoluble organic molecules (within the interior) and to present signaling protein epitopes to cells (on the periphery).

Modification of fibrous poly(L-lactic acid) scaffolds with self-assembling triblock molecules.

Molecular self-assembly offers an effective method to modify the surface properties of common biomaterials by presenting biologically relevant chemistry in a controlled, ordered fashion. This work reports on self-assembling triblock molecules containing rigid cholesteryl segments followed by flexible oligomers of L-(lactic acid) and second generation L-lysine dendrons. Second harmonic generation and small angle X-ray scattering indicate these molecules self-assemble into multilayer polar structures when cast from ethyl acetate solutions and segregate into polar polydomains when annealed. These self-assembled layers significantly improve water wettability when coated onto poly(L-lactic acid) fibers. Scaffolds formed from fibers modified by self-assembly enhance adhesion of 3T3 mouse calvaria cells and produce greater population growth rates. These results demonstrate the use of self-assembly to present biologically relevant chemistry on surfaces of biomaterials. Applications of this technology include the modification of substrates for cell culture, tissue engineering, and cell transplantation.

Aqueous self-assembly of unsymmetric Peptide bolaamphiphiles into nanofibers with hydrophilic cores and surfaces.

Unsymmetric peptide bolaamphiphiles that incorporate (l-glutamyl)3glycine at one terminus and either tetraethylene glycol or aspartic acid at the other were found to form hydrogels at low wt %, presumably by self-assembling into nanofibers presenting (l-glutamyl)3glycine at their surfaces and burying the second headgroup at their cores. Transmission electron microscopy measurements on 1 wt % gels negatively stained with phosphotungstic acid and positively stained with uranyl acetate show one-dimensional objects with diameters of 5 nm and lengths in excess of 1 mum. Circular dichroism and solid-state FTIR spectra indicate the adoption of beta-sheet structure within the nanofibers.