title: Design and synthesis of elastin-like polypeptides for an ideal nerve conduit in peripheral nerve regeneration.
abstract: The study involves design and synthesis of three different elastin like polypeptide (ELP) gene monomers namely ELP1, ELP2 and ELP3 that encode for ELP proteins. The formed ELPs were assessed as an ideal nerve conduit for peripheral nerve regeneration. ELP1 was constructed with a small elongated pentapeptide carrying VPGVG sequence to mimic the natural polypeptide ELP. The ELP2 was designed by the incorporation of 4-penta peptide chains to improve the biocompatibility and mechanical strength. Thus, the third position in unique VPGVG was replaced with alanine to VPAVG and in a similar way modified to VPGKG, VPGEG and VPGIG with the substitution of lysine, glutamic acid and isoleucine. In ELP3, fibronectin C5 domain endowed with REDV sequence was introduced to improve the cell attachment. The ELP1, ELP2 and ELP3 proteins expressed by Escherichia coli were purified by inverse transition cycling (ITC). The purified ELPs were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting. The Schwann cell (SC) morphology and cell adhesion were assessed by fabrication of ELP membrane cross-linked with glutaraledhyde. The Schwann cell proliferation was measured by WST-1 assay. Immunofluorostaining of Schwann cells was accomplished with SC specific phenotypic marker, S100.
authors.affiliation: Institute of Biomedical Engineering, College of Engineering, National Taiwan University, Taipei 100, Taiwan; Institute of Biomedical Engineering, College of Medicine, National Taiwan University, Taipei 100, Taiwan.
authors.forename: Yu-Sheng
authors.initials: YS
authors.lastname: Hsueh
authors.affiliation: Institute of Biomedical Engineering, College of Engineering, National Taiwan University, Taipei 100, Taiwan; Department of Biotechnology, Sree Sastha Institute of Engineering and Technology, Chennai, India; Institute of Biomedical Engineering, College of Medicine, National Taiwan University, Taipei 100, Taiwan.
authors.forename: S
authors.initials: S
authors.lastname: Savitha
authors.affiliation: Division of Biomedical Engineering and Nanomedicine Research, National Health Research Institutes, Miaoli 350, Taiwan.
authors.forename: S
authors.initials: S
authors.lastname: Sadhasivam
authors.affiliation: Institute of Biomedical Engineering, College of Engineering, National Taiwan University, Taipei 100, Taiwan; Division of Biomedical Engineering and Nanomedicine Research, National Health Research Institutes, Miaoli 350, Taiwan; Institute of Biomedical Engineering, College of Medicine, National Taiwan University, Taipei 100, Taiwan. Electronic address: double@ntu.edu.tw.
authors.forename: Feng-Huei
authors.initials: FH
authors.lastname: Lin
authors.affiliation: Institute of Biomedical Engineering, College of Engineering, National Taiwan University, Taipei 100, Taiwan; College of Medicine, National Taiwan University Hospital, Taipei 100, Taiwan; Institute of Biomedical Engineering, College of Medicine, National Taiwan University, Taipei 100, Taiwan.
authors.forename: Ming-Jium
authors.initials: MJ
authors.lastname: Shieh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D004549:Elastin / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D051379:Mice; D008969:Molecular Sequence Data; D009416:Nerve Regeneration / Q000187:drug effects*; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D010525:Peripheral Nerves / Q000187:drug effects / Q000502:physiology*; D010641:Phenotype; D009418:S100 Proteins / Q000378:metabolism; D012583:Schwann Cells / Q000166:cytology; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D015415:Biomarkers; D010455:Peptides; D009418:S100 Proteins; D004549:Elastin
keywords: ELP; Inverse transition cycling; Peripheral nerve injury; Schwann cells; VPGVG