Ultrasound and Microbubble-Targeted Delivery of Macromolecules Is Regulated by Induction of Endocytosis and Pore Formation

Autor: René J. P. Musters, Inge S. Zuhorn, Lynda J. M. Juffermans, Klazina Kooiman, Nico de Jong, Amanda M. G. Versteilen, Walter Paulus, Robert H. Henning, Annemieke van Wamel, Otto Kamp, Wiek H. van Gilst, M. Emmer, Leo E. Deelman, Bernadet D. M. Meijering
Přispěvatelé: Cardiology, Physiology, ICaR - Heartfailure and pulmonary arterial hypertension, Physics of Fluids, Center for Liver, Digestive and Metabolic Diseases (CLDM), Nanotechnology and Biophysics in Medicine (NANOBIOMED), Cardiovascular Centre (CVC), Groningen Kidney Center (GKC), Vascular Ageing Programme (VAP), Groningen Institute for Organ Transplantation (GIOT)
Rok vydání: 2009
Předmět:
Time Factors
Physiology
Caveolin 1
Contrast Media
CONTRAST-AGENT
ultrasound microbubble targeted delivery
cell membrane pore
Bulk endocytosis
GENE DELIVERY
ACTIVATION
chemistry.chemical_compound
Adenosine Triphosphate
Cytosol
Drug Delivery Systems
Ultrasonics
DRUG-DELIVERY
Infusions
Intravenous
Cells
Cultured
Phospholipids
Microbubbles
SHEAR-STRESS
biology
Chemistry
Vesicle
Pinocytosis
Dextrans
AORTIC ENDOTHELIAL-CELLS
MEMBRANE POROSITY
endothelial cells
Femoral Artery
Dextran
Biochemistry
dextran
Wortmannin
Cardiology and Cardiovascular Medicine
Chlorpromazine
Sulfur Hexafluoride
Gene delivery
Caveolae
Endocytosis
Clathrin
Imaging
Three-Dimensional
HYDROGEN-PEROXIDE
Pressure
Animals
endocytosis
Filipin
Rats
Wistar
Transport Vesicles
Fluorescent Dyes
Biological Transport
Receptor-mediated endocytosis
Rats
Androstadienes
Molecular Weight
Microscopy
Fluorescence
PLASMA-MEMBRANE
biology.protein
Biophysics
Cattle
EXPOSED MICROBUBBLES
Zdroj: Circulation Research, 104(5), 679-687. Lippincott Williams and Wilkins
Circulation research, 104(5), 679-687. Lippincott Williams & Wilkins
Circulation research, 104(5), 679-U226. LIPPINCOTT WILLIAMS & WILKINS
Circulation Research, 104(5), 679-U226. Lippincott Williams & Wilkins
Meijering, B D M, Juffermans, L J M, van Wamel, A, Henning, R H, Zuhorn, I S, Emmer, M, Versteilen, A M G, Paulus, W J, van Gilst, W H, Kooiman, K, de Jong, N, Musters, R J P, Deelman, L E & Kamp, O 2009, ' Ultrasound and Microbubble-Targeted Delivery of Macromolecules Is Regulated by Induction of Endocytosis and Pore Formation ', Circulation Research, vol. 104, no. 5, pp. 679-687 . https://doi.org/10.1161/CIRCRESAHA.108.183806
ISSN: 1524-4571
0009-7330
DOI: 10.1161/circresaha.108.183806
Popis: Contrast microbubbles in combination with ultrasound (US) are promising vehicles for local drug and gene delivery. However, the exact mechanisms behind intracellular delivery of therapeutic compounds remain to be resolved. We hypothesized that endocytosis and pore formation are involved during US and microbubble targeted delivery (UMTD) of therapeutic compounds. Therefore, primary endothelial cells were subjected to UMTD of fluorescent dextrans (4.4 to 500 kDa) using 1 MHz pulsed US with 0.22-MPa peak-negative pressure, during 30 seconds. Fluorescence microscopy showed homogeneous distribution of 4.4-and 70-kDa dextrans through the cytosol, and localization of 155-and 500-kDa dextrans in distinct vesicles after UMTD. After ATP depletion, reduced uptake of 4.4-kDa dextran and no uptake of 500-kDa dextran was observed after UMTD. Independently inhibiting clathrin-and caveolae-mediated endocytosis, as well as macropinocytosis significantly decreased intracellular delivery of 4.4-to 500-kDa dextrans. Furthermore, 3D fluorescence microscopy demonstrated dextran vesicles (500 kDa) to colocalize with caveolin-1 and especially clathrin. Finally, after UMTD of dextran (500 kDa) into rat femoral artery endothelium in vivo, dextran molecules were again localized in vesicles that partially colocalized with caveolin-1 and clathrin. Together, these data indicated uptake of molecules via endocytosis after UMTD. In addition to triggering endocytosis, UMTD also evoked transient pore formation, as demonstrated by the influx of calcium ions and cellular release of preloaded dextrans after US and microbubble exposure. In conclusion, these data demonstrate that endocytosis is a key mechanism in UMTD besides transient pore formation, with the contribution of endocytosis being dependent on molecular size. (Circ Res. 2009; 104: 679-687.)
Databáze: OpenAIRE
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