Abstract
Many bioactive molecules have intracellular targets, but have difficulty crossing the cell membrane to reach those targets. To address this difficulty, we fabricated arrays of nanoneedles to gently and simultaneously puncture 105 cells and thereby provide transient pathways for transport of molecules into the cells. The nanoneedles were microfabricated by etching silicon to create arrays of nanoneedles measuring 12 μm in height, tapering to a sharp tip less than 30 nm wide to facilitate puncture into cells and spaced 10 μm apart in order to have at least one nanoneedle puncture each cell in a confluent monolayer. These nanoneedles were used for intracellular delivery in two ways: puncture loading, in which nanoneedle arrays were pressed into cell monolayers, and centrifuge loading, in which cells in suspension were spun down onto nanoneedle arrays. The effects on intracellular uptake and cell viability were determined as a function of nanoneedle length and sharpness, puncture force and duration, and molecular weight of the molecule delivered. Under optimal conditions, intracellular uptake was seen in approximately 50 % of cells while maintaining high cell viability. Overall, this study provides a comparative analysis of intracellular delivery using nanoneedle arrays by two different loading methods over a range of operating parameters.
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Acknowledgments
We thank Dr. Vladimir Zarnitsyn and Dr. Xin Dong Guo for their guidance on preliminary experiments, our collaborators in the laboratories of Dr. Wilbur Lam and Dr. Gang Bao at Georgia Institute of Technology, Dr. William Dynan at Emory University and Dr. Matthew Porteus at Stanford University for helpful discussions and resources, and Donna Bondy for administrative support. This work was carried out in the Institute for Bioengineering and Biosciences at Georgia Tech and was supported by a grant from the National Institutes of Health. Mark Prausnitz is an inventor of patents that have been or may be licensed to companies developing microneedle-based products, a paid advisor to companies developing microneedle-based products and is a founder/shareholder of companies developing microneedle-based products. The resulting potential conflict of interest has been disclosed and is managed by the Georgia Institute of Technology and Emory University.
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Park, S., Choi, SO., Paik, Sj. et al. Intracellular delivery of molecules using microfabricated nanoneedle arrays. Biomed Microdevices 18, 10 (2016). https://doi.org/10.1007/s10544-016-0038-2
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DOI: https://doi.org/10.1007/s10544-016-0038-2