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Materials (Basel). 2018 May 30;11(6). pii: E927. doi: 10.3390/ma11060927.

A Single-Step Surface Modification of Electrospun Silica Nanofibers Using a Silica Binding Protein Fused with an RGD Motif for Enhanced PC12 Cell Growth and Differentiation.

Author information

1
Department of Chemistry, Center for Nanotechnology, Center for Biomedical Technology, Chung Yuan Christian University, Chung Li 32023, Taiwan. allanson92@yahoo.com.tw.
2
Department of Chemistry, Center for Nanotechnology, Center for Biomedical Technology, Chung Yuan Christian University, Chung Li 32023, Taiwan. sunny_day80917@yahoo.com.tw.
3
Department of Science, Concordia University Saint Paul, Saint Paul, MN 55104, USA. amasroujeh@gmail.com.
4
Department of Science, Concordia University Saint Paul, Saint Paul, MN 55104, USA. aaugustine@atsu.edu.
5
Department of Chemistry, Center for Nanotechnology, Center for Biomedical Technology, Chung Yuan Christian University, Chung Li 32023, Taiwan. chengkang20071994@gmail.com.
6
Department of Bioscience Technology, Chung Yuan Christian University, Chung Li, 32023, Taiwan. tychin@cycu.edu.tw.
7
Department of Chemistry, Center for Nanotechnology, Center for Biomedical Technology, Chung Yuan Christian University, Chung Li 32023, Taiwan. yuiwhei@cycu.edu.tw.
8
Department of Science, Concordia University Saint Paul, Saint Paul, MN 55104, USA. myang2@csp.edu.

Abstract

In this study, a previously known high-affinity silica binding protein (SB) was genetically engineered to fuse with an integrin-binding peptide (RGD) to create a recombinant protein (SB-RGD). SB-RGD was successfully expressed in Escherichia coli and purified using silica beads through a simple and fast centrifugation method. A further functionality assay showed that SB-RGD bound to the silica surface with an extremely high affinity that required 2 M MgCl₂ for elution. Through a single-step incubation, the purified SB-RGD proteins were noncovalently coated onto an electrospun silica nanofiber (SNF) substrate to fabricate the SNF-SB-RGD substrate. SNF-SB-RGD was characterized by a combination of scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, and immunostaining fluorescence microscopy. As PC12 cells were seeded onto the SNF-SB-RGD surface, significantly higher cell viability and longer neurite extensions were observed when compared to those on the control surfaces. These results indicated that SB-RGD could serve as a noncovalent coating biologic to support and promote neuron growth and differentiation on silica-based substrates for neuronal tissue engineering. It also provides proof of concept for the possibility to genetically engineer protein-based signaling molecules to noncovalently modify silica-based substrates as bioinspired material.

KEYWORDS:

RGD; electrospun silica nanofibers; neuronal tissue engineering; silica binding protein

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