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    Fabrication and Characterization of Recombinant Silk-elastinlike Protein Fibers for Tissue Engineering Applications

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    Author
    Qiu, Weiguo
    Issue Date
    2011
    Keywords
    Silk-elastinlike protein
    Tissue engineering
    Mechanical Engineering
    Biomaterial
    Scaffold
    Advisor
    Wu, Xiaoyi
    
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    Publisher
    The University of Arizona.
    Rights
    Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
    Abstract
    The integration of functional and structural properties makes genetically engineered proteins appealing in tissue engineering. Silk-elastinlike proteins (SELPs), containing tandemly repeated polypeptide sequence derived from natural silk and elastin, are recently under active study due to the interesting structure. The biological, chemical, physical properties of SELPs have been extensively investigated for their possible applications in drug/gene delivery, surgical tissue sealing and spine repair surgery. However, the mechanical aspect has rarely been looked into. Moreover, many other biomaterials have been fabricated into fibers in micrometer and nanometer scale to build extracellular matrix-mimic scaffolds for tissue regeneration, but many have one or mixed defects such as: poor strength, mild toxicity or immune repulsion etc. The SELP fibers, with the intrinsic primary structures, have novel mechanical properties that can make them defects-minimized scaffolds in tissue engineering.In this study, one SELP (SELP-47K) was fabricated into microfibers and nanofibers by the techniques of wet-spinning and electrospinning. Microfibers of meters long were formed and collected from a methanol coagulation bath, and later were crosslinked by glutaraldehyde (GTA) vapor. The resultant microfibers displayed higher tensile strength up to 20 MPa and higher deformability as high as 700% when tested in hydrated state. Electrospinnig of SELP-47K in formic acid and water resulted in rod-like and ribbon-like nanofibrous scaffolds correspondingly. Both chemical (methanol and/or GTA) and physical (autoclaving) crosslinking methods were utilized to stabilize the scaffolds. The chemical crosslinked hydrated scaffolds exhibit elastic moduli of 3.4-13.2 MPa, ultimate tensile strength of 5.7-13.5 MPa, and deformability of 100-130%, closely matching or exceeding the native aortic elastin; while the autoclaved one had lower numbers: 1.0 MPa elastic modulus, 0.3 MPa ultimate strength and 29% deformation. However, the resilience was all above 80%, beyond the aortic elastin, which is 77%. Additionally, Fourier transform infrared spectra showed clear secondary structure transition after crosslinking, explaining the phenomenon of scaffold water-insolubility from structural perspective and showed a direct relationship with the mechanical performance. Furthermore, the in vitro biocompatibility of SELP-47K nanofibrous scaffolds were verified through the culture of NIH 3T3 mouse embryonic fibroblast cells.
    Type
    text
    Electronic Dissertation
    Degree Name
    Ph.D.
    Degree Level
    doctoral
    Degree Program
    Graduate College
    Mechanical Engineering
    Degree Grantor
    University of Arizona
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