Research sites – SELF-ASSEMBLING NANOEMULSIONS -- data collected mostly at following institutions:

1) Research School of Physical SciencesAustralian National Univ.; Canberra, Australia

2) Dept. of Surgery, Div. of NeurosurgeryUniv. of Connecticut Health Ctr.; Farmington,

CT

3) Dept. of Neurosurgery, Hartford Hospital; CT

( chronology same as “slide order” below ):

set-up for

Langmuir trough set-up for lipid monolayer analysis ( Stable Nanoemulsions, 2010, 3rd edition, Elsevier, -- [ Fig. 6.1 ] )

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Surface pressure-area curves for a microbubble-surfactant monolayer spread at air/water interface

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Surface pressure-area curves for microbubble surfactant monolayers spread on aqueous phases

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ARTIFICIAL “MIXED-LIPID” NANOEMULSIONS:

• Self-assembly using ONLY NONionic lipids;

• Contains both submicron-sized1) lipid-coated microbubbles, and2) liquid-crystal microparticles

predominantly( and both labeled “LCM” in slides below);

• Lipids employed consist of glycerides, cholesterol, and cholesterol esters (but NOT phospholipids).

Lipid-coated microbubble histograms (from flow cytometer): A) saturated solution of FilmixTM lipid mixture; B) distilled water alone; C) computed “difference histogram” of LCM.

Relative size distribution of LCM: Unstirred medium.(Light-scatter signals collected during 1,000-sec period.)

Relative size distribution of LCM: Stirred medium.(Light-scatter signals collected during 1,000-sec. period.)

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Direct optical imaging of lipid-coated microbubbles (LCM) by phase-measurement interferometric microscopy.

TARGETED DRUG DELIVERY using a STABLE LIPID NANOEMULSION vehicle:

• Lipid nanoemulsion targets certain “lipoprotein receptors”;

• Drug delivery to target cells is by “ACTIVE uptake” process -- i.e., by ENDOCYTOSIS;

• LCM uptake by target cells is rapid (-- often 2 minutes after i.v. injection in rats).

Microbubble count contour map. Lipid-coated microbubble (LCM) distribution in tumor -- represented by lines of microbubble isodensity in contour map (bottom right).

Tumor targeting ability of LCM, after i.v. injection, in rat with liver tumor. (Stained LCM appear here as solid black discs.)

Confinement of LCM to 9L-gliosarcoma (top left) and to C6-glioma (top right) in rats. Bottom panels show 9L tumor with diO-labeled LCM, in rat, by confocal laser microscopy.

Interactions of LCM with cultured tumor cells. C6 cells with diO-LCM (panels A,B) or diO alone (panels C,D)

Kinetics of LCM uptake by tumor cells in culture.( C6 glioma cells incubated with diO-LCM at 3 temper-

atures for periods ranging from 5 to 60 minutes. )

Localization of diO-LCM in intracellular acidic com-partments. ( C6 tumor cell seen using dual-mode recording for diO [A] and TR stains [B]. Bar = 10 µm)

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Effects of paclitaxel-loaded LCM on the C6-tumor cell morphology. (Control culture [top panel]; paclitaxel [bottom left panel]; paclitaxel-LCM [bottom right].)

Photomicrograph demonstrating tumor morphology of Sprague-Dawley rats bearing C6 glioma treated with paclitaxel-CRE (A), and paclitaxel- LCM (B, C).

Photomicrograph demonstrating tumor morphologyof Fischer 344 rats bearing 9L gliosarcomas treatedwith cremophor-LCM (A), and paclitaxel-LCM (B,C).

Summary of findings on STABLE, MIXED-LIPID,“LCM/nanoparticle-derived” NANOEMULSIONS:

Self-assemble readily from nonionic lipids; Display marked capability for rapid (actively)

targeted chemotherapy; Selective uptake by target cells occurs via

“lipoprotein receptor”-mediated endocyto-sis (particularly via scavenger receptors);

Long-lived (liquid-crystalline) lipid nanostruc-tures predominate in the submicron

range[ cf. particle-size-distribution slides below ].