Delivery of proteins encapsulated in chitosan-tripolyphosphate nanoparticles to human skin melanoma cells

Mai Bay Stie, Henrik Seir Thoke, Olaf-Georg Issinger, Jennifer Hochscherf, Barbara Guerra, Lars Folke Olsen

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

We have successfully encapsulated two proteins, bovine serum albumin (BSA) and p53, in chitosan-tripolyphosphate (TPP) nanoparticles at various pH values from 5.5 to 6.5 and delivered the particles to human melanoma cells. The particles have diameters ranging from 180 nm to 280 nm and a zeta potential of +15 to + 40 mV. Cellular uptake of the particles by human skin melanoma cells was evaluated by: (i) fluorescence microscopy and (ii) gel electrophoresis showing that FITC-labeled BSA and p53 could be recovered in the soluble cell fraction after lysis of the cells. Our data also show that the highest cellular uptake takes place at the lowest pH as the particles have the highest positive charge under these conditions. The method we describe appears to be a general method for delivery of proteins to cells using chitosan-TPP nanoparticles as a drug delivery system, since structurally unrelated proteins such as BSA and p53 with different isoelectrical points can be encapsulated in the chitosan-TPP nanoparticles and be effectively internalized by the cells.

OriginalsprogEngelsk
TidsskriftColloids and Surfaces B: Biointerfaces
Vol/bind174
Sider (fra-til)216-223
ISSN0927-7765
DOI
StatusUdgivet - feb. 2019

Fingeraftryk

Chitosan
Bovine Serum Albumin
Melanoma
delivery
Skin
Nanoparticles
proteins
Proteins
nanoparticles
cells
albumins
serums
Fluorescein-5-isothiocyanate
Fluorescence microscopy
Zeta potential
Electrophoresis
Gels
electrophoresis
Fluorescence Microscopy
triphosphoric acid

Citer dette

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title = "Delivery of proteins encapsulated in chitosan-tripolyphosphate nanoparticles to human skin melanoma cells",
abstract = "We have successfully encapsulated two proteins, bovine serum albumin (BSA) and p53, in chitosan-tripolyphosphate (TPP) nanoparticles at various pH values from 5.5 to 6.5 and delivered the particles to human melanoma cells. The particles have diameters ranging from 180 nm to 280 nm and a zeta potential of +15 to + 40 mV. Cellular uptake of the particles by human skin melanoma cells was evaluated by: (i) fluorescence microscopy and (ii) gel electrophoresis showing that FITC-labeled BSA and p53 could be recovered in the soluble cell fraction after lysis of the cells. Our data also show that the highest cellular uptake takes place at the lowest pH as the particles have the highest positive charge under these conditions. The method we describe appears to be a general method for delivery of proteins to cells using chitosan-TPP nanoparticles as a drug delivery system, since structurally unrelated proteins such as BSA and p53 with different isoelectrical points can be encapsulated in the chitosan-TPP nanoparticles and be effectively internalized by the cells.",
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Delivery of proteins encapsulated in chitosan-tripolyphosphate nanoparticles to human skin melanoma cells. / Stie, Mai Bay; Thoke, Henrik Seir; Issinger, Olaf-Georg; Hochscherf, Jennifer; Guerra, Barbara; Olsen, Lars Folke.

I: Colloids and Surfaces B: Biointerfaces, Bind 174, 02.2019, s. 216-223.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Delivery of proteins encapsulated in chitosan-tripolyphosphate nanoparticles to human skin melanoma cells

AU - Stie, Mai Bay

AU - Thoke, Henrik Seir

AU - Issinger, Olaf-Georg

AU - Hochscherf, Jennifer

AU - Guerra, Barbara

AU - Olsen, Lars Folke

PY - 2019/2

Y1 - 2019/2

N2 - We have successfully encapsulated two proteins, bovine serum albumin (BSA) and p53, in chitosan-tripolyphosphate (TPP) nanoparticles at various pH values from 5.5 to 6.5 and delivered the particles to human melanoma cells. The particles have diameters ranging from 180 nm to 280 nm and a zeta potential of +15 to + 40 mV. Cellular uptake of the particles by human skin melanoma cells was evaluated by: (i) fluorescence microscopy and (ii) gel electrophoresis showing that FITC-labeled BSA and p53 could be recovered in the soluble cell fraction after lysis of the cells. Our data also show that the highest cellular uptake takes place at the lowest pH as the particles have the highest positive charge under these conditions. The method we describe appears to be a general method for delivery of proteins to cells using chitosan-TPP nanoparticles as a drug delivery system, since structurally unrelated proteins such as BSA and p53 with different isoelectrical points can be encapsulated in the chitosan-TPP nanoparticles and be effectively internalized by the cells.

AB - We have successfully encapsulated two proteins, bovine serum albumin (BSA) and p53, in chitosan-tripolyphosphate (TPP) nanoparticles at various pH values from 5.5 to 6.5 and delivered the particles to human melanoma cells. The particles have diameters ranging from 180 nm to 280 nm and a zeta potential of +15 to + 40 mV. Cellular uptake of the particles by human skin melanoma cells was evaluated by: (i) fluorescence microscopy and (ii) gel electrophoresis showing that FITC-labeled BSA and p53 could be recovered in the soluble cell fraction after lysis of the cells. Our data also show that the highest cellular uptake takes place at the lowest pH as the particles have the highest positive charge under these conditions. The method we describe appears to be a general method for delivery of proteins to cells using chitosan-TPP nanoparticles as a drug delivery system, since structurally unrelated proteins such as BSA and p53 with different isoelectrical points can be encapsulated in the chitosan-TPP nanoparticles and be effectively internalized by the cells.

KW - Chitosan

KW - Nanoparticles

KW - Protein

KW - SK-mel 28

KW - p53

U2 - 10.1016/j.colsurfb.2018.11.005

DO - 10.1016/j.colsurfb.2018.11.005

M3 - Journal article

VL - 174

SP - 216

EP - 223

JO - Colloids and Surfaces B: Biointerfaces

JF - Colloids and Surfaces B: Biointerfaces

SN - 0927-7765

ER -