Neuronal Networks on Nanocellulose Scaffolds
Journal article, 2015

Proliferation, integration, and neurite extension of PC12 cells, a widely used culture model for cholinergic neurons, were studied in nanocellulose scaffolds biosynthesized by Gluconacetobacter xylinus to allow a three-dimensional (3D) extension of neurites better mimicking neuronal networks in tissue. The interaction with control scaffolds was compared with cationized nanocellulose (trimethyl ammonium betahydroxy propyl [TMAHP] cellulose) to investigate the impact of surface charges on the cell interaction mechanisms. Furthermore, coatings with extracellular matrix proteins (collagen, fibronectin, and laminin) were investigated to determine the importance of integrin-mediated cell attachment. Cell proliferation was evaluated by a cellular proliferation assay, while cell integration and neurite propagation were studied by simultaneous label-free Coherent anti-Stokes Raman Scattering and second harmonic generation microscopy, providing 3D images of PC12 cells and arrangement of nanocellulose fibrils, respectively. Cell attachment and proliferation were enhanced by TMAHP modification, but not by protein coating. Protein coating instead promoted active interaction between the cells and the scaffold, hence lateral cell migration and integration. Irrespective of surface modification, deepest cell integration measured was one to two cell layers, whereas neurites have a capacity to integrate deeper than the cell bodies in the scaffold due to their fine dimensions and amoeba-like migration pattern. Neurites with lengths of >50 μm were observed, successfully connecting individual cells and cell clusters. In conclusion, TMAHP-modified nanocellulose scaffolds promote initial cellular scaffold adhesion, which combined with additional cell-scaffold treatments enables further formation of 3D neuronal networks.

Author

M. Jonsson

Chalmers, Biology and Biological Engineering

Christian Brackmann

Chalmers, Biology and Biological Engineering

Maja Puchades

Chalmers, Biology and Biological Engineering

Karoline E Brattås

Chalmers, Biology and Biological Engineering

Andrew Ewing

Chalmers, Chemistry and Chemical Engineering, Chemistry and Biochemistry, Analytical Chemistry

Paul Gatenholm

Chalmers, Chemistry and Chemical Engineering, Applied Chemistry, Polymer Technology

Annika Enejder

Chalmers, Biology and Biological Engineering, Chemical Biology

Tissue Engineering - Part C: Methods

1937-3384 (ISSN)

Vol. 21 11 1162-1170

Subject Categories

Biochemistry and Molecular Biology

DOI

10.1089/ten.tec.2014.0602

PubMed

26398224

More information

Latest update

11/23/2018