3D Culturing and differentiation of SH-SY5Y neuroblastoma cells on bacterial nanocellulose scaffolds

Abstract

A new in vitro model, mimicking the complexity of nerve tissue, was developed based on a bacterial nanocellulose (BNC) scaffold that supports 3D culturing of neuronal cells. BNC is extracellularly excreted by Gluconacetobacter xylinus (G. xylinus) in the shape of long non-aggregated nanofibrils. The cellulose network created by G. xylinus has good mechanical properties, 99% water content, and the ability to be shaped into 3D structures by culturing in different molds. Surface modification with trimethyl ammonium beta-hydroxypropyl (TMAHP) to induce a positive surface charge, followed by collagen I coating, has been used to improve cell adhesion, growth, and differentiation on the scaffold. In the present study, we used SH-SY5Y neuroblastoma cells as a neuronal model. These cells attached and proliferated well on the BNC scaffold, as demonstrated by scanning electron microscopy (SEM) and the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) assay. Following neuronal differentiation, we demonstrated functional action potentials (APs) by electrophysiological recordings, indicating the presence of mature neurons on the scaffolds. In conclusion, we have demonstrated for the first time that neurons can attach, proliferate, and differentiate on BNC. This 3D model based on BNC scaffolds could possibly be used for developing in vitro disease models, when combined with human induced pluripotent stem (iPS) cells (derived from diseased patients) for detailed investigations of neurodegenerative disease mechanisms and in the search for new therapeutics.

Gold nano‐decorated aligned polyurethane nanofibers for enhancement of neurite outgrowth and elongation

“…Neurite outgrowth and alignment are two important factors in neural regeneration in order to reach to the degenerating distal stump and recreate the lost network. Several nanofibrous polymeric based scaffolds were developed to support neural regeneration . Biazar et al generated a porous poly(3‐hydroxy butyric acid‐ co ‐3‐hydroxy valeric acid nerve guidance conduit which was fabricated by electrospinning.…”

Sitagliptin promotes mitochondrial biogenesis in human SH‐SY5Y cells by increasing the expression of PGC‐1α/NRF1/TFAM

“…Human SH-SY5Y neuronal cells were cultured in DMEM/F12 cell culture medium supplemented with 10% fetal bovine serum (FBS) and 1% antibiotics including penicillin and streptomycin (Life Technologies). 14 To determine the effect of sitagliptin, cells were incubated with 50, 100, 200 nM sitagliptin or DMSO for 48 hr. To determine the possible involvement of CREB, cells were stimulated with 100 nM sitagliptin in the presence or absence of the specific inhibitor H89 (10 μM) for 48 hr.…”

In Vitro Modeling of Tissue-Specific 3D Microenvironments and Possibile Application to Pediatric Cancer Research

“…Besides the common matrices, new biomolecules have been explored, such as the bacterial nanocellulose, excreted by Gluconacetobacter xylinus [310], or synthesized materials such as poly (L-lactic acid) PLL [311], photocurable polylactide (PLA) resin [312], poly(DL-lactide-co-glycolide) acid [313], carbon structures [314] or polymeric microspheres [315]. However, the main applicative field, by using these new materials, is not the NBL cancer research, but the neural tissue engineering, thanks to the properties of NBL cells to form neuron-like tissues.…”