Journal
ADVANCED FUNCTIONAL MATERIALS
Volume 32, Issue 3, Pages -Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/adfm.202104828
Keywords
cell-nanoneedle interfaces; endocytic pits; intracellular delivery; membrane deformations; mRNA transfection; polymeric nanoneedles
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Funding
- ARC CPPT, Motherson Innovation
- CSIRO Manufacturing
- Australian government (ARC DECRA) [DE170100021]
- Australian Research Council [DE170100021] Funding Source: Australian Research Council
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This study explores cell responses to substrate stiffness by engineering polymeric nanoneedle arrays of different stiffness, demonstrating potential intracellular delivery mechanisms of polymeric nanoneedles through interactions between cells and different polymeric substrates with varying stiffness.
Tunable vertically aligned nanostructures, usually fabricated using inorganic materials, are powerful nanoscale tools for advanced cellular manipulation. However, nanoscale precision typically requires advanced nanofabrication machinery and involves high manufacturing costs. By contrast, polymeric nanoneedles (NNs) of precise geometry can be produced by replica molding or nanoimprint lithography-rapid, simple, and cost-effective. Here, cytocompatible polymeric arrays of NNs are engineered with identical topographies but differing stiffness, using polystyrene (PS), SU8, and polydimethylsiloxane (PDMS). By interfacing the polymeric NN arrays with adherent and suspension mammalian cells, and comparing the cellular responses of each of the three polymeric substrates, the influence of substrate stiffness from topography on cell behavior is decoupled. Notably, the ability of PS, SU8, and PDMS NNs is demonstrated to facilitate mRNA delivery to GPE86 cells with 26.8% +/- 3.5%, 33.2% +/- 7.4%, and 30.1% +/- 4.1% average transfection efficiencies, respectively. Electron microscopy reveals the intricacy of the cell-NN interactions; and immunofluorescence imaging demonstrates that enhanced endocytosis is one of the mechanisms of PS NN-mediated intracellular delivery, involving the endocytic proteins caveolin-1 and clathrin heavy chain. The results provide insights into the interfacial interactions between cells and polymeric NNs, and their related intracellular delivery mechanisms.
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