Oriented and sustained protein expression on biomimicking electrospun fibers for evaluating functionality of cells

Lin, Junquan, Chooi, Wai Hon, Ong, William, Zhang, Na, Bechler, Marie E., ffrench-Constant, Charles ORCID: https://orcid.org/0000-0002-5621-3377 and Chew, Sing Yian (2021) Oriented and sustained protein expression on biomimicking electrospun fibers for evaluating functionality of cells. Materials Science and Engineering C, 118. ISSN 0928-4931

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Abstract

A proper protein orientation is often required in order to achieve specific protein-receptor interaction to elicit a desired biological response. Here, we present a Protein A-based biomimicking platform that is capable of efficiently orienting proteins for evaluating cellular behaviour. By absorbing Protein A onto aligned bio-mimicking polycaprolactone (PCL) fibers, we demonstrate that protein binding could be retained on these fibers for at least 7 days under physiologically relevant conditions. We further show that Protein A served as a molecular orientor to arrange the recombinant proteins in similar orientations. Such protein-orienting scaffolds were further verified to be biologically functional by using sensitive primary rat cortical neurons (CNs) and oligodendrocyte progenitor cells (OPCs), as model neural cells for a stringent proof of concept. Specifically, CNs that were seeded on fibers coated with Protein A and a known enhancer of neurite growth (L1 Cell Adhesion Molecular L1CAM) displayed the longest total neurite length (462.77 ± 100.79 μm, p < 0.001) as compared to the controls. Besides that, OPCs seeded on fibers coated with Protein A and Neuregulin-1 Type III (Nrg1 type III) (myelin enhancer) produced the longest myelin ensheathment length (19.8 ± 11.69 μm). These results demonstrate the efficacy of this platform for protein screening applications.

Item Type: Article
Additional Information: Funding Information: This work is partially supported by the A*Star BMRC International Joint Grant - Singapore-China Joint Research Program ( 1610500024 ); the SingHealth-NTU-Research Collaborative Grant ( SHS-NTU/038/206 ); the Singapore National Research Foundation under its NMRC-CBRG grant ( NMRC/CBRG/0096/2015 ); and the Ministry of Education (Singapore) Tier 1 grant, ( RG38/19 ).
Uncontrolled Keywords: electrospinning,myelination,nerve regeneration,neurons,oligodendrocytes,protein a,materials science(all),condensed matter physics,mechanics of materials,mechanical engineering ,/dk/atira/pure/subjectarea/asjc/2500
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
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Depositing User: LivePure Connector
Date Deposited: 20 Jul 2022 11:30
Last Modified: 12 Aug 2022 05:36
URI: https://ueaeprints.uea.ac.uk/id/eprint/86722
DOI: 10.1016/j.msec.2020.111407

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