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A Reversible and Repeatable Thiol–Ene Bioconjugation for Dynamic Patterning of Signaling Proteins in Hydrogels
[Image: see text] Biomolecule-functionalized hydrogels have emerged as valuable cell culture platforms to recapitulate the mechanical and biochemical properties of the extracellular niche. The typical strategy to functionalize hydrogels with biomolecules involves directly tethering them to the hydro...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2018
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6062832/ https://www.ncbi.nlm.nih.gov/pubmed/30062120 http://dx.doi.org/10.1021/acscentsci.8b00325 |
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author | Grim, Joseph C. Brown, Tobin E. Aguado, Brian A. Chapnick, Douglas A. Viert, Alexandrea L. Liu, Xuedong Anseth, Kristi S. |
author_facet | Grim, Joseph C. Brown, Tobin E. Aguado, Brian A. Chapnick, Douglas A. Viert, Alexandrea L. Liu, Xuedong Anseth, Kristi S. |
author_sort | Grim, Joseph C. |
collection | PubMed |
description | [Image: see text] Biomolecule-functionalized hydrogels have emerged as valuable cell culture platforms to recapitulate the mechanical and biochemical properties of the extracellular niche. The typical strategy to functionalize hydrogels with biomolecules involves directly tethering them to the hydrogel backbone resulting in a static material. Thus, this approach fails to capture the dynamic changes in biomolecule composition that occur during biological processes or that may be required for regenerative medicine applications. Moreover, it also limits the scope of biomolecules to simple peptides, as signaling proteins generally have poor stability under cell culture conditions and lose their bioactivity over time. To that end, we sought to develop a bioconjugation reaction that would enable reversible and repeatable tethering of signaling proteins to hydrogels, so that spent protein could be released on-demand and replaced with fresh protein as needed. Specifically, we designed an allyl sulfide chain-transfer agent that enables a reversible, photomediated, thiol–ene bioconjugation of signaling proteins to hydrogels. Upon addition of a thiolated protein to the allyl sulfide moiety, the previously tethered protein is released, and the “ene” functionality is regenerated. Using this approach, we demonstrate that protein patterning can be achieved in hydrogels through a thiol–ene reaction, and the patterned protein can then be released through a subsequent thiol–ene reaction of a PEG thiol. Importantly, this process is repeatable through multiple iterations and proceeds at physiologically relevant signaling protein concentrations. Finally, we demonstrate that whole signaling proteins can be patterned and released in the presence of cells, and that cells respond to their presentation with spatial fidelity. Combined, these data represent the first example of a methodology that enables fully reversible and repeatable patterning and release of signaling proteins from hydrogels. |
format | Online Article Text |
id | pubmed-6062832 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-60628322018-07-30 A Reversible and Repeatable Thiol–Ene Bioconjugation for Dynamic Patterning of Signaling Proteins in Hydrogels Grim, Joseph C. Brown, Tobin E. Aguado, Brian A. Chapnick, Douglas A. Viert, Alexandrea L. Liu, Xuedong Anseth, Kristi S. ACS Cent Sci [Image: see text] Biomolecule-functionalized hydrogels have emerged as valuable cell culture platforms to recapitulate the mechanical and biochemical properties of the extracellular niche. The typical strategy to functionalize hydrogels with biomolecules involves directly tethering them to the hydrogel backbone resulting in a static material. Thus, this approach fails to capture the dynamic changes in biomolecule composition that occur during biological processes or that may be required for regenerative medicine applications. Moreover, it also limits the scope of biomolecules to simple peptides, as signaling proteins generally have poor stability under cell culture conditions and lose their bioactivity over time. To that end, we sought to develop a bioconjugation reaction that would enable reversible and repeatable tethering of signaling proteins to hydrogels, so that spent protein could be released on-demand and replaced with fresh protein as needed. Specifically, we designed an allyl sulfide chain-transfer agent that enables a reversible, photomediated, thiol–ene bioconjugation of signaling proteins to hydrogels. Upon addition of a thiolated protein to the allyl sulfide moiety, the previously tethered protein is released, and the “ene” functionality is regenerated. Using this approach, we demonstrate that protein patterning can be achieved in hydrogels through a thiol–ene reaction, and the patterned protein can then be released through a subsequent thiol–ene reaction of a PEG thiol. Importantly, this process is repeatable through multiple iterations and proceeds at physiologically relevant signaling protein concentrations. Finally, we demonstrate that whole signaling proteins can be patterned and released in the presence of cells, and that cells respond to their presentation with spatial fidelity. Combined, these data represent the first example of a methodology that enables fully reversible and repeatable patterning and release of signaling proteins from hydrogels. American Chemical Society 2018-07-12 2018-07-25 /pmc/articles/PMC6062832/ /pubmed/30062120 http://dx.doi.org/10.1021/acscentsci.8b00325 Text en Copyright © 2018 American Chemical Society This is an open access article published under an ACS AuthorChoice License (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) , which permits copying and redistribution of the article or any adaptations for non-commercial purposes. |
spellingShingle | Grim, Joseph C. Brown, Tobin E. Aguado, Brian A. Chapnick, Douglas A. Viert, Alexandrea L. Liu, Xuedong Anseth, Kristi S. A Reversible and Repeatable Thiol–Ene Bioconjugation for Dynamic Patterning of Signaling Proteins in Hydrogels |
title | A Reversible and Repeatable Thiol–Ene Bioconjugation
for Dynamic Patterning of Signaling Proteins in Hydrogels |
title_full | A Reversible and Repeatable Thiol–Ene Bioconjugation
for Dynamic Patterning of Signaling Proteins in Hydrogels |
title_fullStr | A Reversible and Repeatable Thiol–Ene Bioconjugation
for Dynamic Patterning of Signaling Proteins in Hydrogels |
title_full_unstemmed | A Reversible and Repeatable Thiol–Ene Bioconjugation
for Dynamic Patterning of Signaling Proteins in Hydrogels |
title_short | A Reversible and Repeatable Thiol–Ene Bioconjugation
for Dynamic Patterning of Signaling Proteins in Hydrogels |
title_sort | reversible and repeatable thiol–ene bioconjugation
for dynamic patterning of signaling proteins in hydrogels |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6062832/ https://www.ncbi.nlm.nih.gov/pubmed/30062120 http://dx.doi.org/10.1021/acscentsci.8b00325 |
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