Accelerated development of supramolecular corneal stromal-like assemblies from corneal fibroblasts in the presence of macromolecular crowders.
Tissue Engineering Part C: Methods, December 2014
Pramod Kumar, Abhigyan Satyam, Xingliang Fan, Yury Rochev, Brian J Rodriguez, Alexander Gorelov, Lokesh Joshi, Michael Raghunath, Abhay Pandit, Dimitrios I Zeugolis, Kumar P, Satyam A, Fan X, Rochev Y, Rodriguez BJ, Gorelov A, Joshi L, Raghunath M, Pandit A, Zeugolis DI, Kumar, Pramod, Satyam, Abhigyan, Fan, Xingliang, Rochev, Yury, Rodriguez, Brian J., Gorelov, Alexander, Joshi, Lokesh, Raghunath, Michael, Pandit, Abhay, Zeugolis, Dimitrios I.
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 setting, the major obstacle for wide acceptance of this technology remains the tardy extracellular matrix formation. Herein, we assessed the influence of macromolecular crowding / excluding volume effect, a biophysical phenomenon that accelerates thermodynamic activities and biological processes by several orders of magnitude, in human corneal fibroblast culture. Our data indicate that the addition of negatively charged galactose derivative (carrageenan) in human corneal fibroblast culture, even at 0.5% serum, increases by 12-fold tissue-specific matrix deposition, whilst maintaining physiological cell morphology and protein / gene expression. Gene analysis indicates that a glucose derivative (dextran sulphate) may drive corneal fibroblasts towards a myofibroblast lineage. Collectively, these results indicate that macromolecular crowding may be suitable not only for clinical translation and commercialisation of tissue engineering by self-assembly therapies, but also for the development of in vitro pathophysiology models.
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