Aligned Nanofibers for Regenerating Arteries, Nerves, and Muscles

Cells are the fundamental unit of the human body, and therefore the ability to control cell behavior is the most important challenge in regenerative medicine. Peptides are the language of biology which is why synthetic peptide amphiphile (PA) molecules hold great potential as a biomaterial. The work presented in this dissertation explores a variety of liquid crystalline PA nanofibers as a means for directing cell growth. Shaping the alignment of these nanofiber networks requires a deep understanding of their rheological properties which presents a difficult challenge as they exist in complex solid and liquid environments. Using PA molecules that self-assemble into high aspect ratio nanofibers and liquid crystalline solutions, this work investigates the influence of shear flow on macroscopic and microscopic nanofiber alignment. To this end, a shear force applied to PA solutions was systematically varied while the alignment was probed using small angle x-ray scattering. Nanofibers were found to respond to shear flow by aligning parallel to the flow direction. By changing pH and PA chemical sequence it was observed that increasing the interfiber electrostatic repulsive interactions resulted in a greater dependence on shear rate. Nanofiber solutions having greater repulsion did not drastically increase in alignment when the applied strain was increased by two orders of magnitude (1 s -1 to 100 s-1), while solutions with nanofibers having less repulsion increased there alignment four fold with the same strain increase. say exactly what you mean by resulted in greater dependence: did it result in fibers aligning under lower shear rates or higher rates--give the results Anionic PA solutions typically used to encapsulate living cells at neutral pH were found to require minimal shear rates, <1s-1, to achieve significant nanofiber alignment. In an effort to produce tubular hydrogels composed of circumferentially aligned nanofibers, a procedure was designed that used an annular gap containing PA solution with a rotating rod. Using the shear aligning properties of PA solutions this rotating surface in contact with the PA solution induced a high degree of alignment in the nanofibers which was subsequently locked in place by introducing gelating calcium ions. again say something about what this fabrication procedure entails Cells encapsulated within these tubes responded to the alignment by extending in the circumferential direction mimicking the same cellular alignment observed in native arteries. A similar design strategy was also used to align nanofibers within the core of biopolymer nerve conduits, and these scaffolds were implanted in a rat sciatic nerve model. Histological and behavioral observations confirmed that PA implants sustained regeneration rates comparable to autologous grafts and significantly better than empty biopolymer grafts. Furthermore, these nanofiber gels were used as a vehicle to deliver stem cells into muscle tissue. A specialized injector was designed to introduce aligned PA gels into mouse leg muscles in a 1cm long channel. Bioluminescence and histology showed that stem cell engraftment into myofibers was greatly enhanced when delivered by PA gels compared to saline solution. The final section of this thesis describes a new series of PA molecules designed to degrade upon exposure to UV lightstate here why is this of interest in the context of the work described in the thesis. This was done to understand the degradation behavior of PA nanofibers and provide a controlled approach to changing the rheological properties post gelation.The three PA molecules in this series contained the same peptide sequence V3A3E3, while varying the location of a nitrobenzyl UV-reactive group along the backbone of the molecule. This system allowed for a quick reaction that cleaves the molecule at the reactive nitrobenzyl site without introducing any other reactive molecules. While all three molecules produced nanofibers that remained intact upon UV exposure, the PA having its cleavage point nearest to the hydrophobic core resulted in the most dramatic drop in storage modulus. This work has demonstrated the control of alignment, macroscopic shape, and rheological properties of nanofiber gels tailored to assist in the regeneration of tissues with orientational order such as blood vessels, peripheral nerves, and muscle tissue.