1. Oriented and sustained protein expression on biomimicking electrospun fibers for evaluating functionality of cells
- Author
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Na Zhang, Marie E. Bechler, William Ong, Junquan Lin, Wai Hon Chooi, Charles ffrench-Constant, Sing Yian Chew, School of Chemical and Biomedical Engineering, Interdisciplinary Graduate School (IGS), Lee Kong Chian School of Medicine (LKCMedicine), and NTU Institute for Health Technologies
- Subjects
Bioengineering [Engineering] ,Materials science ,Neurite ,L1 ,Bioengineering ,02 engineering and technology ,Plasma protein binding ,010402 general chemistry ,01 natural sciences ,law.invention ,Biomaterials ,chemistry.chemical_compound ,Myelination ,law ,Neurites ,Animals ,Enhancer ,Cell adhesion ,Cells, Cultured ,Neurons ,biology ,Electrospinning ,Oligodendrocytes ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,Nerve Regeneration ,Rats ,chemistry ,Mechanics of Materials ,Polycaprolactone ,Recombinant DNA ,Biophysics ,biology.protein ,Protein A ,0210 nano-technology - 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. Agency for Science, Technology and Research (A*STAR) Ministry of Education (MOE) Nanyang Technological University National Medical Research Council (NMRC) National Research Foundation (NRF) Submitted/Accepted version 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 NMRCCBRG grant (NMRC/CBRG/0096/2015); and the Ministry of Education (Singapore) Tier 1 grant, (RG38/19).
- Published
- 2020