Interaction of polycationic polymers with supported lipid bilayers and cells: Nanoscale hole formation and enhanced membrane permeability

Seungpyo Hong, Pascale R. Leroueil, Elizabeth K. Janus, Jennifer L. Peters, Mary Margaret Kober, Mohammad T. Islam, Bradford G. Orr, James R. Baker, Mark M. Banaszak Holl

Research output: Contribution to journalArticle

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Abstract

Interactions of polycationic polymers with supported 1,2-dimyristoyl-sn- glycero-3-phosphocholine (DMPC) lipid bilayers and live cell membranes (KB and Rat2) have been investigated using atomic force microscopy (AFM), cytosolic enzyme assays, confocal laser scanning microscopy (CLSM), and a fluorescence-activated cell sorter (FACS). Polycationic polymers poly-L-lysine (PLL), polyethylenimine (PEI), and diethylaminoethyl-dextran (DEAE-DEX) and sphere-like poly(amidoamine) (PAMAM) dendrimers are employed because of their importance for gene and drug delivery. AFM studies indicate that all the polycationic polymers cause the formation and/or expansion of preexisting defects in supported DMPC bilayers in the concentration range of 1-3 μg/mL. By way of contrast, hydroxyl-containing neutral linear poly(ethylene glycol) (PEG) and poly(vinyl alcohol) (PVA) do not induce hole formation or expand the size of preexisting defects in the same concentration range. All polymers tested are not toxic to KB or Rat2 cells up to a 12 μg/mL concentration (XTT assay). In the concentration range of 6-12 μg/mL, however, significant amounts of the cytosolic enzymes lactate dehydrogenase (LDH) and luciferase (LUC) are released. PEI, which possesses the greatest density of charged groups on its chain, shows the most dramatic increase in membrane permeability. In addition, treatment with polycationic polymers allows the small dye molecules propidium idodide (PI) and fluorescein (FITC) to diffuse in and out of the cells. CLSM images also show internalization of PLL labeled with FITC dye. In contrast, controls of membrane permeability using the neutral linear polymers PEG and PVA show dramatically less LDH and LUC leakage and no enhanced dye diffusion. Taken together, these data are consistent with the hypothesis that polycationic polymers induce the formation of transient, nanoscale holes in living cells and that these holes allow a greatly enhanced exchange of materials across the cell membrane.

Original languageEnglish
Pages (from-to)728-734
Number of pages7
JournalBioconjugate Chemistry
Volume17
Issue number3
DOIs
Publication statusPublished - 2006 May 1

Fingerprint

Lipid bilayers
Lipid Bilayers
Permeability
Polymers
Membranes
Polyethylene glycols
Polyethyleneimine
Coloring Agents
Phosphorylcholine
Dyes
Fluorescein-5-isothiocyanate
Atomic Force Microscopy
Cell membranes
Luciferases
L-Lactate Dehydrogenase
Confocal Microscopy
Lysine
Assays
Atomic force microscopy
Microscopic examination

All Science Journal Classification (ASJC) codes

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

Cite this

Hong, Seungpyo ; Leroueil, Pascale R. ; Janus, Elizabeth K. ; Peters, Jennifer L. ; Kober, Mary Margaret ; Islam, Mohammad T. ; Orr, Bradford G. ; Baker, James R. ; Banaszak Holl, Mark M. / Interaction of polycationic polymers with supported lipid bilayers and cells : Nanoscale hole formation and enhanced membrane permeability. In: Bioconjugate Chemistry. 2006 ; Vol. 17, No. 3. pp. 728-734.
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abstract = "Interactions of polycationic polymers with supported 1,2-dimyristoyl-sn- glycero-3-phosphocholine (DMPC) lipid bilayers and live cell membranes (KB and Rat2) have been investigated using atomic force microscopy (AFM), cytosolic enzyme assays, confocal laser scanning microscopy (CLSM), and a fluorescence-activated cell sorter (FACS). Polycationic polymers poly-L-lysine (PLL), polyethylenimine (PEI), and diethylaminoethyl-dextran (DEAE-DEX) and sphere-like poly(amidoamine) (PAMAM) dendrimers are employed because of their importance for gene and drug delivery. AFM studies indicate that all the polycationic polymers cause the formation and/or expansion of preexisting defects in supported DMPC bilayers in the concentration range of 1-3 μg/mL. By way of contrast, hydroxyl-containing neutral linear poly(ethylene glycol) (PEG) and poly(vinyl alcohol) (PVA) do not induce hole formation or expand the size of preexisting defects in the same concentration range. All polymers tested are not toxic to KB or Rat2 cells up to a 12 μg/mL concentration (XTT assay). In the concentration range of 6-12 μg/mL, however, significant amounts of the cytosolic enzymes lactate dehydrogenase (LDH) and luciferase (LUC) are released. PEI, which possesses the greatest density of charged groups on its chain, shows the most dramatic increase in membrane permeability. In addition, treatment with polycationic polymers allows the small dye molecules propidium idodide (PI) and fluorescein (FITC) to diffuse in and out of the cells. CLSM images also show internalization of PLL labeled with FITC dye. In contrast, controls of membrane permeability using the neutral linear polymers PEG and PVA show dramatically less LDH and LUC leakage and no enhanced dye diffusion. Taken together, these data are consistent with the hypothesis that polycationic polymers induce the formation of transient, nanoscale holes in living cells and that these holes allow a greatly enhanced exchange of materials across the cell membrane.",
author = "Seungpyo Hong and Leroueil, {Pascale R.} and Janus, {Elizabeth K.} and Peters, {Jennifer L.} and Kober, {Mary Margaret} and Islam, {Mohammad T.} and Orr, {Bradford G.} and Baker, {James R.} and {Banaszak Holl}, {Mark M.}",
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Hong, S, Leroueil, PR, Janus, EK, Peters, JL, Kober, MM, Islam, MT, Orr, BG, Baker, JR & Banaszak Holl, MM 2006, 'Interaction of polycationic polymers with supported lipid bilayers and cells: Nanoscale hole formation and enhanced membrane permeability', Bioconjugate Chemistry, vol. 17, no. 3, pp. 728-734. https://doi.org/10.1021/bc060077y

Interaction of polycationic polymers with supported lipid bilayers and cells : Nanoscale hole formation and enhanced membrane permeability. / Hong, Seungpyo; Leroueil, Pascale R.; Janus, Elizabeth K.; Peters, Jennifer L.; Kober, Mary Margaret; Islam, Mohammad T.; Orr, Bradford G.; Baker, James R.; Banaszak Holl, Mark M.

In: Bioconjugate Chemistry, Vol. 17, No. 3, 01.05.2006, p. 728-734.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Interaction of polycationic polymers with supported lipid bilayers and cells

T2 - Nanoscale hole formation and enhanced membrane permeability

AU - Hong, Seungpyo

AU - Leroueil, Pascale R.

AU - Janus, Elizabeth K.

AU - Peters, Jennifer L.

AU - Kober, Mary Margaret

AU - Islam, Mohammad T.

AU - Orr, Bradford G.

AU - Baker, James R.

AU - Banaszak Holl, Mark M.

PY - 2006/5/1

Y1 - 2006/5/1

N2 - Interactions of polycationic polymers with supported 1,2-dimyristoyl-sn- glycero-3-phosphocholine (DMPC) lipid bilayers and live cell membranes (KB and Rat2) have been investigated using atomic force microscopy (AFM), cytosolic enzyme assays, confocal laser scanning microscopy (CLSM), and a fluorescence-activated cell sorter (FACS). Polycationic polymers poly-L-lysine (PLL), polyethylenimine (PEI), and diethylaminoethyl-dextran (DEAE-DEX) and sphere-like poly(amidoamine) (PAMAM) dendrimers are employed because of their importance for gene and drug delivery. AFM studies indicate that all the polycationic polymers cause the formation and/or expansion of preexisting defects in supported DMPC bilayers in the concentration range of 1-3 μg/mL. By way of contrast, hydroxyl-containing neutral linear poly(ethylene glycol) (PEG) and poly(vinyl alcohol) (PVA) do not induce hole formation or expand the size of preexisting defects in the same concentration range. All polymers tested are not toxic to KB or Rat2 cells up to a 12 μg/mL concentration (XTT assay). In the concentration range of 6-12 μg/mL, however, significant amounts of the cytosolic enzymes lactate dehydrogenase (LDH) and luciferase (LUC) are released. PEI, which possesses the greatest density of charged groups on its chain, shows the most dramatic increase in membrane permeability. In addition, treatment with polycationic polymers allows the small dye molecules propidium idodide (PI) and fluorescein (FITC) to diffuse in and out of the cells. CLSM images also show internalization of PLL labeled with FITC dye. In contrast, controls of membrane permeability using the neutral linear polymers PEG and PVA show dramatically less LDH and LUC leakage and no enhanced dye diffusion. Taken together, these data are consistent with the hypothesis that polycationic polymers induce the formation of transient, nanoscale holes in living cells and that these holes allow a greatly enhanced exchange of materials across the cell membrane.

AB - Interactions of polycationic polymers with supported 1,2-dimyristoyl-sn- glycero-3-phosphocholine (DMPC) lipid bilayers and live cell membranes (KB and Rat2) have been investigated using atomic force microscopy (AFM), cytosolic enzyme assays, confocal laser scanning microscopy (CLSM), and a fluorescence-activated cell sorter (FACS). Polycationic polymers poly-L-lysine (PLL), polyethylenimine (PEI), and diethylaminoethyl-dextran (DEAE-DEX) and sphere-like poly(amidoamine) (PAMAM) dendrimers are employed because of their importance for gene and drug delivery. AFM studies indicate that all the polycationic polymers cause the formation and/or expansion of preexisting defects in supported DMPC bilayers in the concentration range of 1-3 μg/mL. By way of contrast, hydroxyl-containing neutral linear poly(ethylene glycol) (PEG) and poly(vinyl alcohol) (PVA) do not induce hole formation or expand the size of preexisting defects in the same concentration range. All polymers tested are not toxic to KB or Rat2 cells up to a 12 μg/mL concentration (XTT assay). In the concentration range of 6-12 μg/mL, however, significant amounts of the cytosolic enzymes lactate dehydrogenase (LDH) and luciferase (LUC) are released. PEI, which possesses the greatest density of charged groups on its chain, shows the most dramatic increase in membrane permeability. In addition, treatment with polycationic polymers allows the small dye molecules propidium idodide (PI) and fluorescein (FITC) to diffuse in and out of the cells. CLSM images also show internalization of PLL labeled with FITC dye. In contrast, controls of membrane permeability using the neutral linear polymers PEG and PVA show dramatically less LDH and LUC leakage and no enhanced dye diffusion. Taken together, these data are consistent with the hypothesis that polycationic polymers induce the formation of transient, nanoscale holes in living cells and that these holes allow a greatly enhanced exchange of materials across the cell membrane.

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