Coherent anti-Stokes Raman scattering microscopy of human smooth muscle cells in bioengineered tissue scaffolds

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 CH(2)-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.

Synergistic roles for lipids and proteins in the permanent adhesive of barnacle larvae

“…Coherent anti-Stokes Raman scattering 47 is used as an analytical tool and an imaging contrast mechanism in the realms of materials and biological sciences [47][48][49][50] . BCARS contributes to the suite of traditional CARS capabilities by providing improved chemical selectivity and sensitivity over a 3,000 cm À 1 spectral range.…”

Mechanical evaluation of bacterial nanocellulose as an implant material for ear cartilage replacement

“…It has been shown that BNC supports adhesion and proliferation of different cell types (Andersson et al, 2010;Backdahl et al, 2006;Bodin et al, 2010;Brackmann et al, 2011;Svensson et al, 2005), in particular chondrocytes (Andersson et al, 2010;Svensson et al, 2005). This ability to induce chondrocyte proliferation makes BNC a promising scaffold material for ear cartilage TE.…”

Neuronal Networks on Nanocellulose Scaffolds

“…CARS microscopy has been used to visualize the cell body and neurites of the PC12 cells and has also given good specific contrast for other cell types in nanocellulose scaffolds. 20,27,28 The nanocellulose fibrils in the support network are simultaneously visualized by filtering out the scattered second harmonic (SH) of the probe light. 27,29 Together with conventional assay measurements of the overall cell proliferation, this provides detailed insights in neuronal cell growth in a 3D tissue matrix.…”