Interaction of poly(amidoamine) dendrimers with supported lipid bilayers and cells

Hole formation and the relation to transport

Seungpyo Hong, Anna U. Bielinska, Almut Mecke, Balazs Keszler, James L. Beals, Xiangyang Shi, Lajos Balogh, Bradford G. Orr, James R. Baker, Mark M. Banaszak Holl

Research output: Contribution to journalArticle

468 Citations (Scopus)

Abstract

We have investigated poly(amidoamine) (PAMAM) dendrimer interactions with supported 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) lipid bilayers and KB and Rat2 cell membranes using atomic force microscopy (AFM), enzyme assays, flow cell cytometry, and fluorescence microscopy. Amine-terminated generation 7 (G7) PAMAM dendrimers (10-100 nM) were observed to form holes of 15-40 nm in diameter in aqueous, supported lipid bilayers. G5 amine-terminated dendrimers did not initiate hole formation but expanded holes at existing defects. Acetamide-terminated G5 PAMAM dendrimers did not cause hole formation in this concentration range. The interactions between PAMAM dendrimers and cell membranes were studied in vitro using KB and Rat 2 cell lines. Neither G5 amine- nor acetamide-terminated PAMAM dendrimers were cytotoxic up to a 500 nM concentration. However, the dose dependent release of the cytoplasmic proteins lactate dehydrogenase (LDH) and luciferase (Luc) indicated that the presence of the amine-terminated G5 PAMAM dendrimer decreased the integrity of the cell membrane. In contrast, the presence of acetamide-terminated G5 PAMAM dendrimer had little effect on membrane integrity up to a 500 nM concentration. The induction of permeability caused by the amine-terminated dendrimers was not permanent, and leaking of cytosolic enzymes returned to normal levels upon removal of the dendrimers. The mechanism of how PAMAM dendrimers altered cells was investigated using fluorescence microscopy, LDH and Luc assays, and flow cytometry. This study revealed that (1) a hole formation mechanism is consistent with the observations of dendrimer internalization, (2) cytosolic proteins can diffuse out of the cell via these holes, and (3) dye molecules can be detected diffusing into the cell or out of the cell through the same membrane holes. Diffusion of dendrimers through holes is sufficient to explain the uptake of G5 amine-terminated PAMAM dendrimers into cells and is consistent with the lack of uptake of G5 acetamide-terminated PAMAM dendrimers.

Original languageEnglish
Pages (from-to)774-782
Number of pages9
JournalBioconjugate Chemistry
Volume15
Issue number4
DOIs
Publication statusPublished - 2004 Jul 1

Fingerprint

Dendrimers
Lipid bilayers
Lipid Bilayers
Amines
Cell membranes
Fluorescence microscopy
Cell Membrane
Luciferases
L-Lactate Dehydrogenase
Fluorescence Microscopy
Assays
Flow Cytometry
PAMAM Starburst
Poly(amidoamine)
Membranes
KB Cells
Phosphorylcholine
Flow cytometry
Atomic Force Microscopy
Enzyme Assays

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

Cite this

Hong, Seungpyo ; Bielinska, Anna U. ; Mecke, Almut ; Keszler, Balazs ; Beals, James L. ; Shi, Xiangyang ; Balogh, Lajos ; Orr, Bradford G. ; Baker, James R. ; Banaszak Holl, Mark M. / Interaction of poly(amidoamine) dendrimers with supported lipid bilayers and cells : Hole formation and the relation to transport. In: Bioconjugate Chemistry. 2004 ; Vol. 15, No. 4. pp. 774-782.
@article{fc6a1fd02b63436889596f753d16100b,
title = "Interaction of poly(amidoamine) dendrimers with supported lipid bilayers and cells: Hole formation and the relation to transport",
abstract = "We have investigated poly(amidoamine) (PAMAM) dendrimer interactions with supported 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) lipid bilayers and KB and Rat2 cell membranes using atomic force microscopy (AFM), enzyme assays, flow cell cytometry, and fluorescence microscopy. Amine-terminated generation 7 (G7) PAMAM dendrimers (10-100 nM) were observed to form holes of 15-40 nm in diameter in aqueous, supported lipid bilayers. G5 amine-terminated dendrimers did not initiate hole formation but expanded holes at existing defects. Acetamide-terminated G5 PAMAM dendrimers did not cause hole formation in this concentration range. The interactions between PAMAM dendrimers and cell membranes were studied in vitro using KB and Rat 2 cell lines. Neither G5 amine- nor acetamide-terminated PAMAM dendrimers were cytotoxic up to a 500 nM concentration. However, the dose dependent release of the cytoplasmic proteins lactate dehydrogenase (LDH) and luciferase (Luc) indicated that the presence of the amine-terminated G5 PAMAM dendrimer decreased the integrity of the cell membrane. In contrast, the presence of acetamide-terminated G5 PAMAM dendrimer had little effect on membrane integrity up to a 500 nM concentration. The induction of permeability caused by the amine-terminated dendrimers was not permanent, and leaking of cytosolic enzymes returned to normal levels upon removal of the dendrimers. The mechanism of how PAMAM dendrimers altered cells was investigated using fluorescence microscopy, LDH and Luc assays, and flow cytometry. This study revealed that (1) a hole formation mechanism is consistent with the observations of dendrimer internalization, (2) cytosolic proteins can diffuse out of the cell via these holes, and (3) dye molecules can be detected diffusing into the cell or out of the cell through the same membrane holes. Diffusion of dendrimers through holes is sufficient to explain the uptake of G5 amine-terminated PAMAM dendrimers into cells and is consistent with the lack of uptake of G5 acetamide-terminated PAMAM dendrimers.",
author = "Seungpyo Hong and Bielinska, {Anna U.} and Almut Mecke and Balazs Keszler and Beals, {James L.} and Xiangyang Shi and Lajos Balogh and Orr, {Bradford G.} and Baker, {James R.} and {Banaszak Holl}, {Mark M.}",
year = "2004",
month = "7",
day = "1",
doi = "10.1021/bc049962b",
language = "English",
volume = "15",
pages = "774--782",
journal = "Bioconjugate Chemistry",
issn = "1043-1802",
publisher = "American Chemical Society",
number = "4",

}

Hong, S, Bielinska, AU, Mecke, A, Keszler, B, Beals, JL, Shi, X, Balogh, L, Orr, BG, Baker, JR & Banaszak Holl, MM 2004, 'Interaction of poly(amidoamine) dendrimers with supported lipid bilayers and cells: Hole formation and the relation to transport', Bioconjugate Chemistry, vol. 15, no. 4, pp. 774-782. https://doi.org/10.1021/bc049962b

Interaction of poly(amidoamine) dendrimers with supported lipid bilayers and cells : Hole formation and the relation to transport. / Hong, Seungpyo; Bielinska, Anna U.; Mecke, Almut; Keszler, Balazs; Beals, James L.; Shi, Xiangyang; Balogh, Lajos; Orr, Bradford G.; Baker, James R.; Banaszak Holl, Mark M.

In: Bioconjugate Chemistry, Vol. 15, No. 4, 01.07.2004, p. 774-782.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Interaction of poly(amidoamine) dendrimers with supported lipid bilayers and cells

T2 - Hole formation and the relation to transport

AU - Hong, Seungpyo

AU - Bielinska, Anna U.

AU - Mecke, Almut

AU - Keszler, Balazs

AU - Beals, James L.

AU - Shi, Xiangyang

AU - Balogh, Lajos

AU - Orr, Bradford G.

AU - Baker, James R.

AU - Banaszak Holl, Mark M.

PY - 2004/7/1

Y1 - 2004/7/1

N2 - We have investigated poly(amidoamine) (PAMAM) dendrimer interactions with supported 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) lipid bilayers and KB and Rat2 cell membranes using atomic force microscopy (AFM), enzyme assays, flow cell cytometry, and fluorescence microscopy. Amine-terminated generation 7 (G7) PAMAM dendrimers (10-100 nM) were observed to form holes of 15-40 nm in diameter in aqueous, supported lipid bilayers. G5 amine-terminated dendrimers did not initiate hole formation but expanded holes at existing defects. Acetamide-terminated G5 PAMAM dendrimers did not cause hole formation in this concentration range. The interactions between PAMAM dendrimers and cell membranes were studied in vitro using KB and Rat 2 cell lines. Neither G5 amine- nor acetamide-terminated PAMAM dendrimers were cytotoxic up to a 500 nM concentration. However, the dose dependent release of the cytoplasmic proteins lactate dehydrogenase (LDH) and luciferase (Luc) indicated that the presence of the amine-terminated G5 PAMAM dendrimer decreased the integrity of the cell membrane. In contrast, the presence of acetamide-terminated G5 PAMAM dendrimer had little effect on membrane integrity up to a 500 nM concentration. The induction of permeability caused by the amine-terminated dendrimers was not permanent, and leaking of cytosolic enzymes returned to normal levels upon removal of the dendrimers. The mechanism of how PAMAM dendrimers altered cells was investigated using fluorescence microscopy, LDH and Luc assays, and flow cytometry. This study revealed that (1) a hole formation mechanism is consistent with the observations of dendrimer internalization, (2) cytosolic proteins can diffuse out of the cell via these holes, and (3) dye molecules can be detected diffusing into the cell or out of the cell through the same membrane holes. Diffusion of dendrimers through holes is sufficient to explain the uptake of G5 amine-terminated PAMAM dendrimers into cells and is consistent with the lack of uptake of G5 acetamide-terminated PAMAM dendrimers.

AB - We have investigated poly(amidoamine) (PAMAM) dendrimer interactions with supported 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) lipid bilayers and KB and Rat2 cell membranes using atomic force microscopy (AFM), enzyme assays, flow cell cytometry, and fluorescence microscopy. Amine-terminated generation 7 (G7) PAMAM dendrimers (10-100 nM) were observed to form holes of 15-40 nm in diameter in aqueous, supported lipid bilayers. G5 amine-terminated dendrimers did not initiate hole formation but expanded holes at existing defects. Acetamide-terminated G5 PAMAM dendrimers did not cause hole formation in this concentration range. The interactions between PAMAM dendrimers and cell membranes were studied in vitro using KB and Rat 2 cell lines. Neither G5 amine- nor acetamide-terminated PAMAM dendrimers were cytotoxic up to a 500 nM concentration. However, the dose dependent release of the cytoplasmic proteins lactate dehydrogenase (LDH) and luciferase (Luc) indicated that the presence of the amine-terminated G5 PAMAM dendrimer decreased the integrity of the cell membrane. In contrast, the presence of acetamide-terminated G5 PAMAM dendrimer had little effect on membrane integrity up to a 500 nM concentration. The induction of permeability caused by the amine-terminated dendrimers was not permanent, and leaking of cytosolic enzymes returned to normal levels upon removal of the dendrimers. The mechanism of how PAMAM dendrimers altered cells was investigated using fluorescence microscopy, LDH and Luc assays, and flow cytometry. This study revealed that (1) a hole formation mechanism is consistent with the observations of dendrimer internalization, (2) cytosolic proteins can diffuse out of the cell via these holes, and (3) dye molecules can be detected diffusing into the cell or out of the cell through the same membrane holes. Diffusion of dendrimers through holes is sufficient to explain the uptake of G5 amine-terminated PAMAM dendrimers into cells and is consistent with the lack of uptake of G5 acetamide-terminated PAMAM dendrimers.

UR - http://www.scopus.com/inward/record.url?scp=3242748278&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3242748278&partnerID=8YFLogxK

U2 - 10.1021/bc049962b

DO - 10.1021/bc049962b

M3 - Article

VL - 15

SP - 774

EP - 782

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

SN - 1043-1802

IS - 4

ER -