Macromolecularly crowded in vitro microenvironments accelerate the production of extracellular matrix-rich supramolecular assemblies.
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.
Network of Excellence for Functional Biomaterials (NFB), National University of Ireland Galway (NUI Galway), Bioscience Research Building, Galway, Ireland.
Pramod
P
Kumar
Network of Excellence for Functional Biomaterials (NFB), National University of Ireland Galway (NUI Galway), Bioscience Research Building, Galway, Ireland.
Abhigyan
A
Satyam
Network of Excellence for Functional Biomaterials (NFB), National University of Ireland Galway (NUI Galway), Bioscience Research Building, Galway, Ireland.
Xingliang
X
Fan
Network of Excellence for Functional Biomaterials (NFB), National University of Ireland Galway (NUI Galway), Bioscience Research Building, Galway, Ireland.
Estelle
E
Collin
Network of Excellence for Functional Biomaterials (NFB), National University of Ireland Galway (NUI Galway), Bioscience Research Building, Galway, Ireland.
Yury
Y
Rochev
Conway Institute of Biomolecular &Biomedical Research, University College Dublin, Dublin, Ireland.
Brian J
BJ
Rodriguez
School of Chemistry &Chemical Biology, University College Dublin, Dublin, Ireland.
Alexander
A
Gorelov
BIDMC Genomics, Proteomics, Bioinformatics and Systems Biology Center, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, USA.
Simon
S
Dillon
Alimentary Glycoscience Research Cluster, NUI Galway, Galway, Ireland.
Lokesh
L
Joshi
Department of Bioengineering, Faculty of Engineering, National University of Singapore Tissue Engineering Programme, Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
Michael
M
Raghunath
Network of Excellence for Functional Biomaterials (NFB), National University of Ireland Galway (NUI Galway), Bioscience Research Building, Galway, Ireland.
Abhay
A
Pandit
Network of Excellence for Functional Biomaterials (NFB), National University of Ireland Galway (NUI Galway), Bioscience Research Building, Galway, Ireland.
Dimitrios I
DI
Zeugolis
D000818:Animals; D000831:Animals, Newborn; D002417:Cattle; D018929:Cell Culture Techniques; D002468:Cell Physiological Phenomena; D002478:Cells, Cultured; D060833:Cellular Microenvironment / Q000187:drug effects / Q000502:physiology*; D003094:Collagen / Q000378:metabolism; D003315:Cornea / Q000166:cytology; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D046911:Macromolecular Substances / Q000378:metabolism*; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D018625:Microscopy, Atomic Force; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D008954:Models, Biological; D011347:Procollagen / Q000378:metabolism; D044967:Serum / Q000737:chemistry; D059008:Time-Lapse Imaging; D023822:Tissue Engineering / Q000379:methods
D003470:Culture Media; D046911:Macromolecular Substances; D011347:Procollagen; D003094:Collagen; D020778:Matrix Metalloproteinase 2