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Tissue Eng Part C Methods. 2015 Jul;21(7):660-70. doi: 10.1089/ten.TEC.2014.0387. Epub 2015 Mar 12.

Accelerated Development of Supramolecular Corneal Stromal-Like Assemblies from Corneal Fibroblasts in the Presence of Macromolecular Crowders.

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1 Network of Excellence for Functional Biomaterials (NFB), Bioscience Research Building, National University of Ireland Galway (NUI Galway) , Galway, Ireland .
2 Conway Institute of Biomolecular & Biomedical Research, University College Dublin , Dublin, Ireland .
3 School of Chemistry & Chemical Biology, University College Dublin , Dublin, Ireland .
4 Alimentary Glycoscience Research Cluster, NUI Galway , Galway, Ireland .
5 Department of Bioengineering, Faculty of Engineering, National University of Singapore , Singapore, Singapore .
6 Tissue Engineering Programme, Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore , Singapore, Singapore .


Tissue engineering by self-assembly uses the cells' secretome as a regeneration template and biological factory of trophic factors. Despite the several advantages that have been witnessed in preclinical and clinical settings, the major obstacle for wide acceptance of this technology remains the tardy extracellular matrix formation. In this study, we assessed the influence of macromolecular crowding (MMC)/excluding volume effect, a biophysical phenomenon that accelerates thermodynamic activities and biological processes by several orders of magnitude, in human corneal fibroblast (HCF) culture. Our data indicate that the addition of negatively charged galactose derivative (carrageenan) in HCF culture, even at 0.5% serum, increases by 12-fold tissue-specific matrix deposition, while maintaining physiological cell morphology and protein/gene expression. Gene analysis indicates that a glucose derivative (dextran sulfate) may drive corneal fibroblasts toward a myofibroblast lineage. Collectively, these results indicate that MMC may be suitable not only for clinical translation and commercialization of tissue engineering by self-assembly therapies, but also for the development of in vitro pathophysiology models.

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