Nanoscale molecular quantification of stem cell-hydrogel interactions
File(s)
Published version
Author(s)
Type
Journal Article
Abstract
A common approach to tailoring synthetic hydrogels for regenerative medicine applications involves incorporating RGD cell adhesion peptides, yet assessing the cellular response to engineered microenvironments at the nanoscale remains challenging. To date, no study has demonstrated how RGD concentration in hydrogels affects the presentation of individual cell surface receptors. Here we studied the interaction between human mesenchymal stem cells (hMSCs) and RGD-functionalized poly(ethylene glycol) hydrogels, by correlating macro- and nanoscale single-cell interfacial quantification techniques. We quantified RGD unbinding forces on a synthetic hydrogel using single cell atomic force spectroscopy, revealing that short-term binding of hMSCs was sensitive to RGD concentration. We also performed direct stochastic optical reconstruction microscopy (dSTORM) to quantify the molecular interactions between integrin α5β1 and a biomaterial, unexpectedly revealing that increased integrin clustering at the hydrogel-cell interface correlated with fewer available RGD binding sites. Our complementary, quantitative approach uncovered mechanistic insights into specific stem cell-hydrogel interactions, where dSTORM provides nanoscale sensitivity to RGD-dependent differences in cell surface localization of integrin α5β1. Our findings reveal that it is possible to precisely determine how peptide-functionalized hydrogels interact with cells at the molecular scale, thus providing a basis to fine-tune the spatial presentation of bioactive ligands.
Date Issued
2020-12-22
Date Acceptance
2020-11-17
Citation
ACS Nano, 2020, 14 (12), pp.17321-17332
ISSN
1936-0851
Publisher
American Chemical Society
Start Page
17321
End Page
17332
Journal / Book Title
ACS Nano
Volume
14
Issue
12
Copyright Statement
© 2020 American Chemical Society. This is an open access article published under a Creative Commons Attribution (CC-BY)License, which permits unrestricted use, distribution and reproduction in any medium,provided the author and source are cited
License URL
Sponsor
Commission of the European Communities
Medical Research Council (MRC)
Wellcome Trust
Engineering & Physical Science Research Council (EPSRC)
Engineering and Physical Sciences Research Council
Commission of the European Communities
Grant Number
701664
MR/R015651/1
098411/Z/12/Z
EP/M020398/1
EP/R512540/1
660757
Subjects
Science & Technology
Physical Sciences
Technology
Chemistry, Multidisciplinary
Chemistry, Physical
Nanoscience & Nanotechnology
Materials Science, Multidisciplinary
Chemistry
Science & Technology - Other Topics
Materials Science
AFM
single cell force spectroscopy
dSTORM
RGD
PEG hydrogel
integrin alpha 5 beta 1
INTEGRIN ALPHA(5)BETA(1)
TISSUE REGENERATION
FORCE
ADHESION
FIBRONECTIN
ACTIVATION
BETA
DIFFERENTIATION
SPECIFICITY
MODULATION
AFM
PEG hydrogel
RGD
dSTORM
integrin α5β1
single cell force spectroscopy
Nanoscience & Nanotechnology
Publication Status
Published
Date Publish Online
2020-11-20