Transport of supramolecular drugs across the cell membrane by calcium phosphate nanoparticles

Rotan, Olga; Sokolova, Viktoriya; Gilles, Patrick; Hu, Wenbin; Dutt, Som; Schräder, Thomas; Epple, Matthias

doi:10.1002/mawe.201300085

Cited by 10 publications

(25 citation statements)

References 18 publications

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“…The protein was not able to cross the cell membrane alone for any cell line, corroborating earlier results on the nanoparticle-mediated uptake of (bio)molecules by cells [3, 11, 29, 40]. The nanoparticle-treated cells did not exhibit any adverse effects due to the presence of calcium or polyethylenimine (PEI), probably due to the low dose (about 1.1 g μL -1 calcium phosphate in the well).…”

Section: Resultssupporting

confidence: 86%

Delivery of the autofluorescent protein R-phycoerythrin by calcium phosphate nanoparticles into four different eukaryotic cell lines (HeLa, HEK293T, MG-63, MC3T3): Highly efficient, but leading to endolysosomal proteolysis in HeLa and MC3T3 cells

et al. 2017

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Nanoparticles can be used as carriers to transport biomolecules like proteins and synthetic molecules across the cell membrane because many molecules are not able to cross the cell membrane on their own. The uptake of nanoparticles together with their cargo typically occurs via endocytosis, raising concerns about the possible degradation of the cargo in the endolysosomal system. As the tracking of a dye-labelled protein during cellular uptake and processing is not indicative of the presence of the protein itself but only for the fluorescent label, a label-free tracking was performed with the red-fluorescing model protein R-phycoerythrin (R-PE). Four different eukaryotic cell lines were investigated: HeLa, HEK293T, MG-63, and MC3T3. Alone, the protein was not taken up by any cell line; only with the help of calcium phosphate nanoparticles, an efficient uptake occurred. After the uptake into HeLa cells, the protein was found in early endosomes (shown by the marker EEA1) and lysosomes (shown by the marker Lamp1). There, it was still intact and functional (i.e. properly folded) as its red fluorescence was detected. However, a few hours after the uptake, proteolysis started as indicated by the decreasing red fluorescence intensity in the case of HeLa and MC3T3 cells. 12 h after the uptake, the protein was almost completely degraded in HeLa cells and MC3T3 cells. In HEK293T cells and MG-63 cells, no degradation of the protein was observed. In the presence of Bafilomycin A1, an inhibitor of acidification and protein degradation in lysosomes, the fluorescence of R-PE remained intact over the whole observation period in the four cell lines. These results indicate that despite an efficient nanoparticle-mediated uptake of proteins by cells, a rapid endolysosomal degradation may prevent the desired (e.g. therapeutic) effect of a protein inside a cell.

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Section: Resultssupporting

confidence: 86%

Delivery of the autofluorescent protein R-phycoerythrin by calcium phosphate nanoparticles into four different eukaryotic cell lines (HeLa, HEK293T, MG-63, MC3T3): Highly efficient, but leading to endolysosomal proteolysis in HeLa and MC3T3 cells

et al. 2017

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“…Click Approach to the Preparation of Dendrimer-Type Multi(thia)calix [4]arenes (CH Ar ), 136.5 (CH Ar ), 143.5 (C i,Ar-t-Bu ), 143.8 (C i,Ar-t-Bu ), 148.9 (C i,Ar-t-Bu ), 155.9 (C i,Ar-O ), 156.5 (C i,Ar-O ), 157.6 (C i,Ar-O ).…”

Section: Synthesis Of Compoundsmentioning

confidence: 99%

“…A promising scaffold for the design of dendrimer structures is represented by macrocycles, in particular, thiacalixarenes, which offer conformational diversity (that can be frozen in case bulky substituents are introduced), variety of reaction centers and their relatively facile modification giving almost unlimited possibilities towards the design of a specific receptor. [2] According to published data, multicalixarenes are capable of self-association, [3] they are promising supramolecular drugs, [4] and they can bind large biological objects such as DNA. [5] The complexation of multicalixarenes with metal ions was also studied.…”

Section: Introductionmentioning

confidence: 99%

“…The aim of this work was to synthesize dendrimershaped multicalixarenes with triazole fragments on the lower rim (Scheme 1). The proposed strategy is focused on the conjugation of tetrasubstituted thiacalix [4]arene in 1,3-alternate stereoisomeric form as a core with coneshaped monosubstituted thiacalix [4]arenes as a monodendron by CuAAC reaction. In this case, azido or ethynyl groups can be located on the core or dendron.…”

Section: Introductionmentioning

confidence: 99%

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Azide–Akyne Click Approach to the Preparation of Dendrimer–Type Multi(thia)calix[4]arenes with Triazole Linkers

Muravev¹,

Laishevtsev²,

Galieva³

et al. 2017

MHC

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The design of polymacrocyclic structures is of high importance for the improvement of recognition ability to various guests. We present a convenient approach towards two types of dendrimer-shaped pentakis-thiacalix[4]arenes with triazolyl linkers using the click reaction of tetraazidoalkoxycalixarenes with monoalkynyloxy derivatives, as well as tetraalkynyloxycalixarenes with monoazidoalkoxy derivatives. Mitsunobu alkylation of hydroxyl groups has been employed to afford tetrasubstituted derivatives in 1,3-alternate configuration. A facile procedure for the synthesis of monosubstituted cone precursors in high yields by the hydrolysis of one ether fragment in disubstituted counterparts under basic conditions (n-BuNH 2 or NaN 3 in DMF) has been suggested and optimized. CuAAC reaction has been conducted in microwave reactor and on a hot plate. It has been found that microwave irradiation promotes the reaction and the yields of products are as high as 40-80 %.

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“…[14][15][16][17][18][19][20] In addition, calcium phosphate nanoparticles are efficiently taken up by cells and subsequently dissolved in lysosomes at a pH below 5. [21][22][23][24][25][26] However, due to the negative surface charge (negative zeta potential) and limited protection from nucleases (e.g. RNases), the transfection efficiency of such "single shell" calcium phosphate nanoparticles is typically low.…”

Section: Introductionmentioning

confidence: 99%

A pH-sensitive poly(methyl methacrylate) copolymer for efficient drug and gene delivery across the cell membrane

2014

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A versatile drug delivery system based on calcium phosphate/Eudragit®-E100 nanoparticles with a diameter below 200 nm was realized by a water-in-oil-in-water (W 1 /O/W 2) emulsion solvent evaporation technique. Hydrophilic drugs (siRNA for gene silencing and bovine serum albumin as model protein) and hydrophobic drugs (5,10,15,20-tetrakis(4-hydroxyphenyl)-21H,23H-porphine, THPP, a photosensitizer for photodynamic therapy) were encapsulated as model drugs. Eudragit®-E100 is a poly(methyl methacrylate) copolymer with a low solubility at neutral pH and a high solubility at low pH. Thus, the particles are stable in cell culture media and rapidly dissolved in the lysosome after cellular uptake and delivery of their cargo into the cell. The particles had a positive charge (zeta potential +49 mV) and were taken up very well by epithelial cells (HeLa) as shown by fluorescence microscopy and confocal laser scanning microscopy. Gene-silencing experiments on HeLa-eGFP cells gave knockdown efficiencies of 39% with no toxic effects. The particles can be freeze-dried without cryoprotectant and easily redispersed in water, thus making their transport and storage convenient.

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Transport of supramolecular drugs across the cell membrane by calcium phosphate nanoparticles

Cited by 10 publications

References 18 publications

Delivery of the autofluorescent protein R-phycoerythrin by calcium phosphate nanoparticles into four different eukaryotic cell lines (HeLa, HEK293T, MG-63, MC3T3): Highly efficient, but leading to endolysosomal proteolysis in HeLa and MC3T3 cells

Delivery of the autofluorescent protein R-phycoerythrin by calcium phosphate nanoparticles into four different eukaryotic cell lines (HeLa, HEK293T, MG-63, MC3T3): Highly efficient, but leading to endolysosomal proteolysis in HeLa and MC3T3 cells

Azide–Akyne Click Approach to the Preparation of Dendrimer–Type Multi(thia)calix[4]arenes with Triazole Linkers

A pH-sensitive poly(methyl methacrylate) copolymer for efficient drug and gene delivery across the cell membrane

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