Cell-instructive surface gradients of photo-responsive amyloid-like fibrils

06 July 2021, Version 1
This content is a preprint and has not undergone peer review at the time of posting.

Abstract

Gradients of bioactive molecules play a crucial role in various biological processes like vascularization, tissue regeneration or cell migration. In order to study these complex biological systems, it is necessary to control the concentration of bioactive molecules on their substrates. Here, we created a photochemical strategy to generate gradients using amyloid-like fibrils as scaffolds functionalized with a model epitope, i.e. the integrin-binding peptide RGD, to modulate cell adhesion. The self-assembling -sheet forming peptide (CKFKFQF) was connected to the RGD epitope via a photo-sensitive nitrobenzyl linker and assembled into photo-responsive nanofibrils. The fibrils were spray-coated on glass substrates and macroscopic gradients were generated by UV-light over a cm-scale. We confirmed the gradient formation using matrix assisted laser desorption ionization mass spectroscopy imaging (MALDI-MSI), which directly visualizes the molecular species on the surface. The RGD gradient was used to instruct cells. In consequence, A549 adapted their adhesion properties in dependence of the RGD-epitope density

Keywords

Peptide amyloid fiber
MALDI-MSI
bioactive gradient
photo-controlled cell adhesion

Supplementary materials

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Title
Cell-instructive surface gradients of photo-responsive amyloid-like fibrils
Description
Gradients of bioactive molecules play a crucial role in various biological processes like vascularization, tissue regeneration or cell migration. To study these complex biological systems, it is necessary to control the concentration of bioactive molecules on their substrates. Here, we created a photochemical strategy to generate gradients using amyloid-like fibrils as scaffolds functionalized with a model epitope, i.e. the integrin-binding peptide RGD, to modulate cell adhesion. The self-assembling -sheet forming peptide (CKFKFQF) was connected to the RGD epitope via a photo-sensitive nitrobenzyl linker and assembled into photo-responsive nanofibrils. The fibrils were spray-coated on glass substrates and macroscopic gradients were generated by UV-light over a cm-scale. We confirmed the gradient formation using matrix assisted laser desorption ionization mass spectroscopy imaging (MALDI-MSI), which directly visualizes the molecular species on the surface. The RGD gradient was used to instruct cells. In consequence, A549 adapted their adhesion properties in dependence of the RGD-epitope density
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