Neuronal Networks on Nanocellulose Scaffolds

Area of Science:

• Biomaterials Science
• Neuroscience
• Cell Biology

Background:

• PC12 cells, a model for cholinergic neurons, require suitable scaffolds for three-dimensional (3D) network formation.
• Nanocellulose scaffolds offer a promising platform for neuronal tissue engineering.
• Surface modifications can influence cell-scaffold interactions and neuronal network development.

Purpose of the Study:

• To investigate the proliferation, integration, and neurite extension of PC12 cells in nanocellulose scaffolds.
• To evaluate the impact of surface charge (cationized TMAHP-cellulose) and extracellular matrix protein coatings on cell behavior.
• To understand the mechanisms of cell attachment and 3D network formation in engineered neuronal tissues.

Main Methods:

• PC12 cells cultured on native and modified nanocellulose scaffolds (TMAHP-cellulose, protein coatings).
• Cell proliferation assessed using a cellular proliferation assay.
• 3D cell integration and neurite propagation visualized using label-free Coherent anti-Stokes Raman Scattering and second harmonic generation microscopy.

Main Results:

• Cationized TMAHP-nanocellulose enhanced PC12 cell attachment and proliferation compared to control scaffolds.
• Extracellular matrix protein coatings promoted cell migration and integration but not initial attachment or proliferation.
• Neurites extended deeper (>50 μm) into the scaffold than cell bodies, forming connections and contributing to 3D network formation.

Conclusions:

• TMAHP-modified nanocellulose scaffolds improve initial cell adhesion and proliferation for PC12 cells.
• Combined scaffold modifications (surface charge and protein coating) facilitate deeper cell integration and neurite extension.
• These engineered nanocellulose scaffolds show potential for developing functional 3D neuronal networks in vitro.