Multiphoton microscopy of engineered dermal substitutes: assessment of 3D collagen matrix remodeling induced by fibroblasts contraction

A.-M. Pena; C. Olive; J.-F. Michelet; J.-B. Galey; D. Fagot; F. Leroy; J.-L. Martin; A. Colonna; M.-C. Schanne-Klein

doi:10.1117/12.839947

2 March 2010 Multiphoton microscopy of engineered dermal substitutes: assessment of 3D collagen matrix remodeling induced by fibroblasts contraction

A.-M. Pena, C. Olive, J.-F. Michelet, J.-B. Galey, D. Fagot, F. Leroy, J.-L. Martin, A. Colonna, M.-C. Schanne-Klein

Author Affiliations +

Proceedings Volume 7548, Photonic Therapeutics and Diagnostics VI; 754802 (2010) https://doi.org/10.1117/12.839947
Event: SPIE BiOS, 2010, San Francisco, California, United States

Abstract

One of the main functions of dermal fibroblasts is the generation of mechanical forces within their surrounding extracellular matrix. Investigating molecules that could modulate fibroblast contraction and act as potent anti aging ingredients requires the development of three-dimensional in situ imaging methodologies for dermal substitute analysis. Here we use multiphoton microscopy in order to investigate the fibroblast-induced collagen matrix reorganization in engineered dermal tissue and to evaluate the effect of Y27632, a RhoA kinase inhibitor on dermal substitutes contraction. We observe that collagen fibrils rearrange around fibroblast with increasing density in control samples, whereas collagen fibrils show no remodeling in the samples containing the RhoA kinase inhibitor. Moreover, when the culture medium containing the inhibitor was replaced with a control medium, the dermal substitutes presented the same 3D reorganization as the control samples, which indicates that the inhibitory effects are reversible. In conclusion, our study demonstrates the relevance of multiphoton microscopy to visualize three-dimensional remodeling of the matrix induced by fibroblast contraction.

Citation Download Citation

A.-M. Pena, C. Olive, J.-F. Michelet, J.-B. Galey, D. Fagot, F. Leroy, J.-L. Martin, A. Colonna, and M.-C. Schanne-Klein "Multiphoton microscopy of engineered dermal substitutes: assessment of 3D collagen matrix remodeling induced by fibroblasts contraction", Proc. SPIE 7548, Photonic Therapeutics and Diagnostics VI, 754802 (2 March 2010); https://doi.org/10.1117/12.839947

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