Macromolecularly crowded in vitro microenvironments accelerate the production of extracellular matrix-rich supramolecular assemblies

Scientific Reports
Pramod KumarDimitrios Zeugolis

Abstract

Therapeutic strategies based on the principles of tissue engineering by self-assembly put forward the notion that functional regeneration can be achieved by utilising the inherent capacity of cells to create highly sophisticated supramolecular assemblies. However, in dilute ex vivo microenvironments, prolonged culture time is required to develop an extracellular matrix-rich implantable device. Herein, we assessed the influence of macromolecular crowding, a biophysical phenomenon that regulates intra- and extra-cellular activities in multicellular organisms, in human corneal fibroblast culture. In the presence of macromolecules, abundant extracellular matrix deposition was evidenced as fast as 48 h in culture, even at low serum concentration. Temperature responsive copolymers allowed the detachment of dense and cohesive supramolecularly assembled living substitutes within 6 days in culture. Morphological, histological, gene and protein analysis assays demonstrated maintenance of tissue-specific function. Macromolecular crowding opens new avenues for a more rational design in engineering of clinically relevant tissue modules in vitro.

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Citations

Oct 13, 2015·ACS Applied Materials & Interfaces·Akash BachhukaK Vasilev
Oct 27, 2015·Biotechnology and Bioengineering·Dilip ThomasDimitrios I Zeugolis
Aug 28, 2016·Drug Discovery Today·Diana Gaspar, Dimitrios I Zeugolis
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Sep 7, 2016·Journal of Tissue Engineering and Regenerative Medicine·Pramod KumarDimitrios I Zeugolis
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Oct 18, 2018·Journal of Wound Care·Majana Tomic-CanicHans Smola
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Jun 28, 2019·The Journal of the American Academy of Orthopaedic Surgeons·Dimitrios Tsiapalis, Dimitrios I Zeugolis
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Feb 2, 2021·Frontiers in Medicine·Nataly Puerta CavanzoRuud A Bank
Mar 4, 2021·American Journal of Physiology. Cell Physiology·Dimitrios I Zeugolis
Feb 27, 2021·Journal of Tissue Engineering·Yoann TorresNicolas L'Heureux
Mar 31, 2021·Npj Regenerative Medicine·Andrea De PieriDimitrios I Zeugolis
Apr 15, 2021·Advanced Biology·Andrea De PieriKarin Wuertz-Kozak
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May 15, 2015·Bioconjugate Chemistry·Christina N M RyanDimitrios I Zeugolis
Sep 9, 2021·ACS Applied Materials & Interfaces·Gerard Rubí-SansElisabeth Engel

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Methods Mentioned

BETA
electrophoresis
atomic force microscopy
SMA
zymography
phase contrast microscopy
PCR
proteomic profiling
reverse phase chromatography
AFM

Software Mentioned

Scope
Pro Plus
Protein Pilot
GeneTools

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