A Reversible and Repeatable Thiol–Ene Bioconjugation
for Dynamic Patterning of Signaling Proteins in Hydrogels
Version 3 2018-07-25, 04:04
Version 2 2018-07-12, 12:24
Version 1 2018-07-11, 21:14
Posted on 2018-07-25 - 04:04
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.
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Grim, Joseph
C.; Brown, Tobin
E.; Aguado, Brian
A.; Chapnick, Douglas
A.; Viert, Alexandrea
L.; Liu, Xuedong; et al. (2018). A Reversible and Repeatable Thiol–Ene Bioconjugation
for Dynamic Patterning of Signaling Proteins in Hydrogels. ACS Publications. Collection. https://doi.org/10.1021/acscentsci.8b00325
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AUTHORS (7)
JG
Joseph
C. Grim
TB
Tobin
E. Brown
BA
Brian
A. Aguado
DC
Douglas
A. Chapnick
AV
Alexandrea
L. Viert
XL
Xuedong Liu
KA
Kristi S. Anseth
KEYWORDS
protein patterningprotein concentrationsHydrogels Biomolecule-functionalized hydrogelsrepeatable tetheringcell culture conditionshydrogel backboneSignaling Proteinsregenerative medicine applicationsfunctionalize hydrogelsbiomolecule compositionextracellular nichethiolated proteinDynamic Patterningcell culture platformsPEG thiolrepeatable patterningallyl sulfide moietyallyl sulfide chain-transfer agentbioconjugation reaction