Investigation of the micro- and nano-structure of liver cells cultured on biodegradable silk fibroin-based scaffolds using scanning probe optical nanotomography

Abstract

Objective: to analyze the 3D micro- and nano-structure and quantitative morphological parameters of liver cellscultured on biodegradable silk fibroin-based film scaffolds. Materials and methods. Samples of biodegradable silk fibroin-based scaffolds with cultured Wistar rat liver cells were obtained for the study. The 3D structure of liver cells cultivated on the scaffolds was studied by scanning probe optical nanotomography using an experimental setup combining an ultramicrotome and a scanning probe microscope in correlation with fluorescence microscopy. Results. Nanoscale images and 3D nanotomographic reconstructions of rat liver cells cultured on scaffold were obtained. The morphological parameters of liver cells (average roughness, specific effective area) were determined. The average surface roughness of the liver cells Ra was found to be 124.8 ± 8.2 nm, while the effective surface area σ was 1.13 ± 0.02. Analysis of the volume distribution of lipid droplets showed that they occupy 28% of the cell volume. Conclusion. Scanning probe optical nanotomography can successfully analyze the nanostructure and quantify the nanomorphology of liver cells cultured on biodegradable scaffolds.

Keywords

• liver cells
• biodegradable scaffolds
• silk fibroin
• scanning probe microscopy
• fluorescence microscopy
• nanotomography