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Journal of Biomedical Optics • Open Access

Coherent anti-Stokes Raman scattering microscopy of human smooth muscle cells in bioengineered tissue scaffolds
Author(s): Christian Brackmann; Annika M. Enejder; Maricris Esguerra; Daniel Olausson; Dick Delbro; Alexandra Krettek; Paul Gatenholm

Paper Abstract

The integration of living, human smooth muscle cells in biosynthesized cellulose scaffolds was monitored by nonlinear microscopy toward contractile artificial blood vessels. Combined coherent anti-Stokes Raman scattering (CARS) and second harmonic generation (SHG) microscopy was applied for studies of the cell interaction with the biopolymer network. CARS microscopy probing CH2-groups at 2845 cm−1 permitted three-dimensional imaging of the cells with high contrast for lipid-rich intracellular structures. SHG microscopy visualized the fibers of the cellulose scaffold, together with a small signal obtained from the cytoplasmic myosin of the muscle cells. From the overlay images we conclude a close interaction between cells and cellulose fibers. We followed the cell migration into the three-dimensional structure, illustrating that while the cells submerge into the scaffold they extrude filopodia on top of the surface. A comparison between compact and porous scaffolds reveals a migration depth of <10 μm for the former, whereas the porous type shows cells further submerged into the cellulose. Thus, the scaffold architecture determines the degree of cell integration. We conclude that the unique ability of nonlinear microscopy to visualize the three-dimensional composition of living, soft matter makes it an ideal instrument within tissue engineering.

Paper Details

Date Published: 1 February 2011
PDF: 6 pages
J. Biomed. Opt. 16(2) 021115 doi: 10.1117/1.3534782
Published in: Journal of Biomedical Optics Volume 16, Issue 2
Show Author Affiliations
Christian Brackmann, Chalmers Univ. of Technology (Sweden)
Annika M. Enejder, Chalmers Univ. of Technology (Sweden)
Maricris Esguerra, Sahlgrenska Univ. Hospital (Sweden)
Daniel Olausson, Sahlgrenska Univ. Hospital (Sweden)
Dick Delbro, Sahlgrenska Univ. Hospital (Sweden)
Alexandra Krettek, Sahlgrenska Univ. Hospital (Sweden)
Paul Gatenholm, Chalmers Univ. of Technology (Sweden)

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