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Biological Atomic Force Microscopy for Imaging Gold-Labeled Liposomes on Human Coronary Artery Endothelial Cells.

Zaske AM, Danila D, Queen MC, Golunski E, Conyers JL - J Pharm (Cairo) (2013)

Bottom Line: Dynasore monohydrate, an inhibitor of endocytosis, obstructed the internalization of the gold-liposomes.This study showed the versatility of the AFM technique, combined with fluorescent microscopy, for investigating liposome uptake by endothelial cells.The 90 nm colloidal gold nanoparticles proved to be a noninvasive contrast agent that efficiently improves AFM imaging during the investigation of biological nanoprocesses.

View Article: PubMed Central - PubMed

Affiliation: Division of Cardiology, Department of Internal Medicine, The University of Texas Health Science Center at Houston, 1881 East Road, Houston, TX 77054, USA.

ABSTRACT
Although atomic force microscopy (AFM) has been used extensively to characterize cell membrane structure and cellular processes such as endocytosis and exocytosis, the corrugated surface of the cell membrane hinders the visualization of extracellular entities, such as liposomes, that may interact with the cell. To overcome this barrier, we used 90 nm nanogold particles to label FITC liposomes and monitor their endocytosis on human coronary artery endothelial cells (HCAECs) in vitro. We were able to study the internalization process of gold-coupled liposomes on endothelial cells, by using AFM. We found that the gold-liposomes attached to the HCAEC cell membrane during the first 15-30 min of incubation, liposome cell internalization occurred from 30 to 60 min, and most of the gold-labeled liposomes had invaginated after 2 hr of incubation. Liposomal uptake took place most commonly at the periphery of the nuclear zone. Dynasore monohydrate, an inhibitor of endocytosis, obstructed the internalization of the gold-liposomes. This study showed the versatility of the AFM technique, combined with fluorescent microscopy, for investigating liposome uptake by endothelial cells. The 90 nm colloidal gold nanoparticles proved to be a noninvasive contrast agent that efficiently improves AFM imaging during the investigation of biological nanoprocesses.

No MeSH data available.


Related in: MedlinePlus

The hydrodynamic radii of noncoupled (a) and gold-coupled liposomes (b), estimated by DLS. Uncoupled liposomes (a) observed a mean diameter value of 129 ± 1.3 nm. The size distribution for gold-labeled liposomes (b) described a major peak for particles about 285 ± 5.3 nm in size. DLS results represent the averages of 13 runs at three different measurements per sample.
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fig1: The hydrodynamic radii of noncoupled (a) and gold-coupled liposomes (b), estimated by DLS. Uncoupled liposomes (a) observed a mean diameter value of 129 ± 1.3 nm. The size distribution for gold-labeled liposomes (b) described a major peak for particles about 285 ± 5.3 nm in size. DLS results represent the averages of 13 runs at three different measurements per sample.

Mentions: The average diameter of the non-gold-coupled liposomes, as determined by DLS, was 129 ± 1.3 nm (Figure 1(a)), and a polydispersity index (PDI) of 0.126. The DLS results represent the averages of three different measurements (13 runs each) per sample. For the gold-coupled liposomes (Figure 1(b)), the DLS results showed a major peak at 285 ± 5.3 nm. The PDI for the gold-liposomes was 0.689.


Biological Atomic Force Microscopy for Imaging Gold-Labeled Liposomes on Human Coronary Artery Endothelial Cells.

Zaske AM, Danila D, Queen MC, Golunski E, Conyers JL - J Pharm (Cairo) (2013)

The hydrodynamic radii of noncoupled (a) and gold-coupled liposomes (b), estimated by DLS. Uncoupled liposomes (a) observed a mean diameter value of 129 ± 1.3 nm. The size distribution for gold-labeled liposomes (b) described a major peak for particles about 285 ± 5.3 nm in size. DLS results represent the averages of 13 runs at three different measurements per sample.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4590807&req=5

fig1: The hydrodynamic radii of noncoupled (a) and gold-coupled liposomes (b), estimated by DLS. Uncoupled liposomes (a) observed a mean diameter value of 129 ± 1.3 nm. The size distribution for gold-labeled liposomes (b) described a major peak for particles about 285 ± 5.3 nm in size. DLS results represent the averages of 13 runs at three different measurements per sample.
Mentions: The average diameter of the non-gold-coupled liposomes, as determined by DLS, was 129 ± 1.3 nm (Figure 1(a)), and a polydispersity index (PDI) of 0.126. The DLS results represent the averages of three different measurements (13 runs each) per sample. For the gold-coupled liposomes (Figure 1(b)), the DLS results showed a major peak at 285 ± 5.3 nm. The PDI for the gold-liposomes was 0.689.

Bottom Line: Dynasore monohydrate, an inhibitor of endocytosis, obstructed the internalization of the gold-liposomes.This study showed the versatility of the AFM technique, combined with fluorescent microscopy, for investigating liposome uptake by endothelial cells.The 90 nm colloidal gold nanoparticles proved to be a noninvasive contrast agent that efficiently improves AFM imaging during the investigation of biological nanoprocesses.

View Article: PubMed Central - PubMed

Affiliation: Division of Cardiology, Department of Internal Medicine, The University of Texas Health Science Center at Houston, 1881 East Road, Houston, TX 77054, USA.

ABSTRACT
Although atomic force microscopy (AFM) has been used extensively to characterize cell membrane structure and cellular processes such as endocytosis and exocytosis, the corrugated surface of the cell membrane hinders the visualization of extracellular entities, such as liposomes, that may interact with the cell. To overcome this barrier, we used 90 nm nanogold particles to label FITC liposomes and monitor their endocytosis on human coronary artery endothelial cells (HCAECs) in vitro. We were able to study the internalization process of gold-coupled liposomes on endothelial cells, by using AFM. We found that the gold-liposomes attached to the HCAEC cell membrane during the first 15-30 min of incubation, liposome cell internalization occurred from 30 to 60 min, and most of the gold-labeled liposomes had invaginated after 2 hr of incubation. Liposomal uptake took place most commonly at the periphery of the nuclear zone. Dynasore monohydrate, an inhibitor of endocytosis, obstructed the internalization of the gold-liposomes. This study showed the versatility of the AFM technique, combined with fluorescent microscopy, for investigating liposome uptake by endothelial cells. The 90 nm colloidal gold nanoparticles proved to be a noninvasive contrast agent that efficiently improves AFM imaging during the investigation of biological nanoprocesses.

No MeSH data available.


Related in: MedlinePlus