Praveesuda L. Michael, Miguel Santos, Elysse C. Filipe, Alex H.P. Chan, Juichien Hung, Bob S.L. Lee, Richard Tan, Minh Huynh, Clare Hawkins, Anna Waterhouse, Marcela M.M. Bilek, and Steven G. Wise*.

 

Applied Materials Group, The Heart Research Institute
Sydney, NSW 2042, Australia

 

The achievement of multiple functionalities on a single nanocarrier holds significant promise for improving efficiency by combining targeting, imaging and drug delivery on a single platform. However, with current technology, the addition of each new functionality adds substantial complexity including additional steps of synthesis, purification and characterisation resulting in escalating costs, regulatory burden and reduced yield. Multifunctional carbon-based nanoparticles (nanoP3) are a new platform that achieves multiple functionalities in a single, simple step enabling delivery of multiple molecular cargos into cells without cell penetrating agents.

nanoP3 were manufactured and characterized in-house using a custom-built plasma chamber. Size, roughness, surface charge and chemical composition were readily controlled in the system, giving rise to a versatile nanoparticle platform. Unfunctionalized nanoP3 also penetrated the membrane of human coronary artery endothelial cells (hCAECs) and accumulated in the cytoplasm, observed using 3View cross-sectional SEM. Incubation with hCAECs, epithelial MCF10A or human breast adenocarcinoma cells (MCF7) for up to 5 days did not significantly affect cell viability or morphology, with up to 3.1x108 particles per mm2. The presence of long-lived free radicals, formed during synthesis, within nanoP3 suggests the potential for direct covalent immobilization of biomolecules by simple incubation. To realize this potential, we immobilized three different IgG antibodies tagged with 40, 20 and 10 nm gold labels respectively to demonstrate single, double and triple functionalization on a single nanoparticle, following simple incubation (Figure. 1). To exemplify one possible functional utility of nanoP3, we delivered functional small interfering RNA (siRNA) targeted to the vascular endothelial growth factor (VEGF) gene in HUVECs, impairing the formation of tubules in an established Matrigel assay.

Conclusions:  To our knowledge this is the first demonstration of the co-delivery of three different surface bound cargos into multiple cell systems following one-step co-incubation with the nanoparticle platform and without chemical pre-functionalization steps. This approach eliminates the trade-off between additional functionality and complexity and could facilitate the upscaling of nanoparticle-based therapies into the clinic.

Biographic Details
Name: Steven G Wise
Title: Dr
Affiliation, Country: The Heart Research Institute, Sydney, Australia
Phone: +612 8208 8928 E-mail: steve.wise@hri.org.au
Research interests: biomaterials, device engineering, nanomedicine
 

Venue

Room: 
AEB Auditorium