Issue 2, 2010

Investigation of microsphere-mediated cellular delivery by chemical, microscopic and gene expression analysis

Abstract

Amino functionalised cross-linked polystyrene microspheres of well defined sizes (0.2–2 μm) have been prepared and shown to be efficient and controllable delivery devices, capable of transporting anything from small dye molecules to bulky proteins into cells. However, the specific mechanism of cellular entry is largely unknown and widely variant from study to study. As such, chemical, biological and microscopic methods are used to elucidate the mechanism of cellular uptake for polystyrene microspheres of 0.2, 0.5 and 2 μm in mouse melanoma cells. Uptake is found to be wholly unreliant upon energetic processes, while lysosomal and endosomal tracking agents failed to show co-localisation with lysosomes/endosomes, suggesting a non-endocytic uptake pathway. To further explore the consequences of microsphere uptake, gene expression profiling is used to determine if there is a transcriptional response to “beadfection” in both murine and human cells. None of the common transcriptional responses to enhanced endocytosis are observed in beadfected cells, further supporting a non-endocytic uptake mechanism. Furthermore, the microspheres are noted to have a limited interaction with cells at a transcriptional level, supporting them as a non-toxic delivery vehicle.

Graphical abstract: Investigation of microsphere-mediated cellular delivery by chemical, microscopic and gene expression analysis

Supplementary files

Article information

Article type
Paper
Submitted
20 Jul 2009
Accepted
22 Oct 2009
First published
26 Nov 2009

Mol. BioSyst., 2010,6, 399-409

Investigation of microsphere-mediated cellular delivery by chemical, microscopic and gene expression analysis

L. M. Alexander, S. Pernagallo, A. Livigni, R. M. Sánchez-Martín, J. M. Brickman and M. Bradley, Mol. BioSyst., 2010, 6, 399 DOI: 10.1039/B914428E

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