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Clickable decellularized extracellular matrix as a new tool for building hybrid-hydrogels to model chronic fibrotic diseases in vitro

Petrou, Cassandra L. ; D'Ovidio, Tyler J. ; Bölükbas, Deniz LU ; Tas, Sinem LU ; Brown, R. Dale ; Allawzi, Ayed ; Lindstedt Ingemansson, Sandra LU ; Nozik-Grayck, Eva ; Stenmark, Kurt R. and Wagner, Darcy LU , et al. (2020) In Journal of Materials Chemistry B 8(31). p.6814-6826
Abstract

Fibrotic disorders account for over one third of mortalities worldwide. Despite great efforts to study the cellular and molecular processes underlying fibrosis, there are currently few effective therapies. Dual-stage polymerization reactions are an innovative tool for recreating heterogeneous increases in extracellular matrix (ECM) modulus, a hallmark of fibrotic diseases in vivo. Here, we present a clickable decellularized ECM (dECM) crosslinker incorporated into a dynamically responsive poly(ethylene glycol)-α-methacrylate (PEGαMA) hybrid-hydrogel to recreate ECM remodeling in vitro. An off-stoichiometry thiol-ene Michael addition between PEGαMA (8-arm, 10 kg mol-1) and the clickable dECM resulted in hydrogels with an elastic modulus... (More)

Fibrotic disorders account for over one third of mortalities worldwide. Despite great efforts to study the cellular and molecular processes underlying fibrosis, there are currently few effective therapies. Dual-stage polymerization reactions are an innovative tool for recreating heterogeneous increases in extracellular matrix (ECM) modulus, a hallmark of fibrotic diseases in vivo. Here, we present a clickable decellularized ECM (dECM) crosslinker incorporated into a dynamically responsive poly(ethylene glycol)-α-methacrylate (PEGαMA) hybrid-hydrogel to recreate ECM remodeling in vitro. An off-stoichiometry thiol-ene Michael addition between PEGαMA (8-arm, 10 kg mol-1) and the clickable dECM resulted in hydrogels with an elastic modulus of E = 3.6 ± 0.24 kPa, approximating healthy lung tissue (1-5 kPa). Next, residual αMA groups were reacted via a photo-initiated homopolymerization to increase modulus values to fibrotic levels (E = 13.4 ± 0.82 kPa) in situ. Hydrogels with increased elastic moduli, mimicking fibrotic ECM, induced a significant increase in the expression of myofibroblast transgenes. The proportion of primary fibroblasts from dual-reporter mouse lungs expressing collagen 1a1 and alpha-smooth muscle actin increased by approximately 60% when cultured on stiff and dynamically stiffened hybrid-hydrogels compared to soft. Likewise, fibroblasts expressed significantly increased levels of the collagen 1a1 transgene on stiff regions of spatially patterned hybrid-hydrogels compared to the soft areas. Collectively, these results indicate that hybrid-hydrogels are a new tool that can be implemented to spatiotemporally induce a phenotypic transition in primary murine fibroblasts in vitro.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Materials Chemistry B
volume
8
issue
31
pages
6814 - 6826
publisher
Royal Society of Chemistry
external identifiers
  • pmid:32343292
ISSN
2050-7518
DOI
10.1039/D0TB00613K
language
English
LU publication?
yes
id
c4c7e6dd-43c8-4285-9a41-9d35c62d10ba
date added to LUP
2020-04-23 20:31:21
date last changed
2020-09-02 13:56:11
@article{c4c7e6dd-43c8-4285-9a41-9d35c62d10ba,
  abstract     = {<p>Fibrotic disorders account for over one third of mortalities worldwide. Despite great efforts to study the cellular and molecular processes underlying fibrosis, there are currently few effective therapies. Dual-stage polymerization reactions are an innovative tool for recreating heterogeneous increases in extracellular matrix (ECM) modulus, a hallmark of fibrotic diseases in vivo. Here, we present a clickable decellularized ECM (dECM) crosslinker incorporated into a dynamically responsive poly(ethylene glycol)-α-methacrylate (PEGαMA) hybrid-hydrogel to recreate ECM remodeling in vitro. An off-stoichiometry thiol-ene Michael addition between PEGαMA (8-arm, 10 kg mol-1) and the clickable dECM resulted in hydrogels with an elastic modulus of E = 3.6 ± 0.24 kPa, approximating healthy lung tissue (1-5 kPa). Next, residual αMA groups were reacted via a photo-initiated homopolymerization to increase modulus values to fibrotic levels (E = 13.4 ± 0.82 kPa) in situ. Hydrogels with increased elastic moduli, mimicking fibrotic ECM, induced a significant increase in the expression of myofibroblast transgenes. The proportion of primary fibroblasts from dual-reporter mouse lungs expressing collagen 1a1 and alpha-smooth muscle actin increased by approximately 60% when cultured on stiff and dynamically stiffened hybrid-hydrogels compared to soft. Likewise, fibroblasts expressed significantly increased levels of the collagen 1a1 transgene on stiff regions of spatially patterned hybrid-hydrogels compared to the soft areas. Collectively, these results indicate that hybrid-hydrogels are a new tool that can be implemented to spatiotemporally induce a phenotypic transition in primary murine fibroblasts in vitro.</p>},
  author       = {Petrou, Cassandra L. and D'Ovidio, Tyler J. and Bölükbas, Deniz and Tas, Sinem and Brown, R. Dale and Allawzi, Ayed and Lindstedt Ingemansson, Sandra and Nozik-Grayck, Eva and Stenmark, Kurt R. and Wagner, Darcy and Magin, Chelsea M.},
  issn         = {2050-7518},
  language     = {eng},
  month        = {08},
  number       = {31},
  pages        = {6814--6826},
  publisher    = {Royal Society of Chemistry},
  series       = {Journal of Materials Chemistry B},
  title        = {Clickable decellularized extracellular matrix as a new tool for building hybrid-hydrogels to model chronic fibrotic diseases in vitro},
  url          = {http://dx.doi.org/10.1039/D0TB00613K},
  doi          = {10.1039/D0TB00613K},
  volume       = {8},
  year         = {2020},
}