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Journal of Biomedical Optics • Open Access

Multiphoton microscopy of engineered dermal substitutes: assessment of 3-D collagen matrix remodeling induced by fibroblast contraction
Author(s): Ana-Maria Pena; Dominique Fagot; Christian Olive; Jean-François Michelet; Jean-Baptiste Galey; Frederic Leroy; Emmanuel Beaurepaire; Jean-Louis Martin; Anne Colonna; Marie-Claire Schanne-Klein

Paper Abstract

Dermal fibroblasts are responsible for the generation of mechanical forces within their surrounding extracellular matrix and can be potentially targeted by anti-aging ingredients. Investigation of the modulation of fibroblast contraction by these ingredients requires the implementation of three-dimensional in situ imaging methodologies. We use multiphoton microscopy to visualize unstained engineered dermal tissue by combining second-harmonic generation that reveals specifically fibrillar collagen and two-photon excited fluorescence from endogenous cellular chromophores. We study the fibroblast-induced reorganization of the collagen matrix and quantitatively evaluate the effect of Y-27632, a RhoA-kinase inhibitor, on dermal substitute contraction. We observe that collagen fibrils rearrange around fibroblasts with increasing density in control samples, whereas collagen fibrils show no remodeling in the samples containing the RhoA-kinase inhibitor. Moreover, we show that the inhibitory effects are reversible. Our study demonstrates the relevance of multiphoton microscopy to visualize three-dimensional remodeling of the extracellular matrix induced by fibroblast contraction or other processes.

Paper Details

PDF: 7 pages
J. Biomed. Opt. 15(5) 056018 doi: 10.1117/1.3503411
Published in: Journal of Biomedical Optics Volume 15, Issue 5, September 2010
Show Author Affiliations
Ana-Maria Pena, L'Oréal Recherche et Innovation (France)
Dominique Fagot, L'Oréal Recherche et Innovation (France)
Christian Olive, L'Oréal Recherche et Innovation (France)
Jean-François Michelet, L'Oréal Recherche et Innovation (France)
Jean-Baptiste Galey, L'Oréal Recherche et Innovation (France)
Frederic Leroy, L'Oreal (France)
Emmanuel Beaurepaire, Ecole Polytechnique (France)
Jean-Louis Martin, Institut d'Optique Graduate School (France)
Anne Colonna, L'Oréal Recherche et Innovation (France)
Marie-Claire Schanne-Klein, Lab. d'Optique et Biosciences (France)

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