The role of surface charge in cellular uptake and cytotoxicity of medical nanoparticles

Figures & data

Table 1 Overview of nanoparticle (NP) formulations in development for imaging and drug delivery with examples for approved drugs, with indication of the most important fields of application¹⁷²

Particle used	FDA-approved drugs
(A) Imaging
Iron oxide, gold NPs, gadolinium NPs, quantum dots	Endorem/feridex (superparamagnetic iron oxide associated with dextran, http://www.mr-tip.com/serv1.php?type=db1&dbs=Endorem)* Cliavist/resovist (superparamagnetic iron oxide, http://www.mr-tip.com/serv1.php?type=db1&dbs=resovist) Sinerem/combidex (ultrasmall superparamagnetic iron oxide, http://www.mr-tip.com/serv1.php?type=db1&dbs=sinerem)*
(B) Drug delivery
Polymer-based Polyacrylamide, polymethylmethacrylate, polyalkylcyanoacrylate, polylactate-co-glycolate, phosopholipid, pluronic, poly l-amino acid, cyclodextrins	Adagen (PEGylated adenosine deaminase, immune, http://www.rxlist.com/adagen-drug.htm) Cimzia (PEGylated fab fragments of human TNF-α, immune, http://www.rxlist.com/cimzia-drug.htm) Copaxone (amino acid polymer, MS, http://www.rxlist.com/copaxone-drug.htm) Eligard (leuprolide acetate/poly[d,l-lactide-co-glycolide] polymer, onco, http://www.rxlist.com/eligard-drug.htm) Genexol-PM (PEGylated poly[lactic acid] micelle formulation of paclitaxel, onco, http://pharmalicensing.com/public/outlicensing/view/4809/genexol-pm) Macugen (PEGylated anti-VEGF aptamer, ophth, http://www.rxlist.com/macugen-drug.htm) Mircera (methoxy PEG-epoetin beta, onco, http://www.rxlist.com/mircera-drug.htm) Neulasta (PEGylated filgrastin, onco, http://www.rxlist.com/neulasta-drug.htm) Oncaspar (PEGylated l-asparaginase, onco, http://www.rxlist.com/oncaspar-drug.htm) Pegasys (PEGylated interferon α-2a, immune, http://www.rxlist.com/pegasys-drug.htm) PegIntron (pegylated interferon α-2b, immune, http://www.rxlist.com/pegintron-and-rebetol-drug.htm) Renagel (polymeric amine, dialysis, http://www.rxlist.com/renagel-drug.htm) Somavert (PEGylated human hormone receptor antagonist, endo, http://www.rxlist.com/somavert-drug.htm) Taxotere (micellar docetaxel, onco, http://www.rxlist.com/taxotere-drug.htm)
Lipid-based Solid-lipid NPs, nanostructured lipid carriers	Abelcet (lipid complex of amphotericin B, infection, http://www.rxlist.com/abelcet-drug.htm) AmBisome (liposomal formulation of amphotericin B, infection, http://www.rxlist.com/ambisome-drug.htm) Amphocil/Amphotec (micellar amphotericin B, infection, http://dailymed.nlm.nih.gov/dailymed/drugInfo.cfm?id=6780) Estrasorb (micellar estradiol, endo, http://www.rxlist.com/script/main/rxlist.asp?drug=estrasorb&monotype=rx-desc&monopage=1) Definity (liposomal octofluoroproane, cardio, http://www.rxlist.com/definity-drug.htm) DaunoXome/Myocet (liposomal formulation of duanorubicin, onco, http://www.ema.europa.eu/ema/index.jsp?curl=pages/medicines/human/medicines/000297/human_med_000916.jsp&mid=WC0b01ac058001d124) DepoCyt (liposomal formulation of cytarabine, onco, http://www.rxlist.com/depocyt-drug.htm) DepoDur (liposomal formulation of morphine, onco, http://www.rxlist.com/depodur-drug.htm) Doxil/Caelyx (PEGylated liposomal formulation of doxorubicin, onco, http://www.rxlist.com/doxil-drug.htm) Mepact (liposomal mifamurtide, onco, http://www.cancer.gov/dictionary?cdrid=644758) Octocog alfa (liposomal factor VIII, hematology http://www.medicines.org.uk/guides/Octocog%20alfa/Haemophilia) Visudyne (liposomal formulation of verteporfin, ophth, http://www.rxlist.com/visudyne-drug.htm)
Protein-based Albumin	Abraxane (albumin-bound paclitaxel, onco, http://www.rxlist.com/abraxane-drug.htm)
Nanocrystals	Avinza (nanocrystal morphine, pain, http://www.rxlist.com/avinza-drug.htm) Cardizem (nanocrystal diltiazem, cardio, http://www.rxlist.com/cardizem-drug.htm) Emend (nanocrystal aprepitant, onco, http://www.rxlist.com/emend-drug.htm) Focalin (nanocrystal dexmethyl phenidate, ADHD, http://www.rxlist.com/focalin-drug.htm) Invega sustenna (nanocrystal paliperidone palmitate, schizophrenia, http://www.rxlist.com/invega-sustenna-drug.htm) Megace ES (nanocrystal megestrol, immune, http://www.rxlist.com/megace-drug.htm) Naprelan (nanocrystal naproxen, immune, http://www.rxlist.com/naprelan-drug.htm) Rapamune (nanocrystal sirolimus, immune, http://www.rxlist.com/rapamune-drug.htm) Ritalin (nanocrystal methyl phenidate, ADHD, http://www.rxlist.com/ritalin-drug.htm) Theo-Dur (nanocrystal theophylline, resp, http://www.rxmed.com/b.main/b2.pharmaceutical/b2.1.monographs/CPS-%20Monographs/CPS-%20%28General%20Monographs-%20T%29/THEO-dur.html) Tricor (nanocrystal fenofibrate, cardio, http://www.rxlist.com/tricor-drug.htm) Triglide (nanocrystal fenofibrate, cardio, http://www.rxlist.com/triglide-drug.htm) Verelan (nanocrystal verapamil, cardio, (http://www.rxlist.com/verelan-drug.htm)
Metallic NPs Gold, iron oxide, quantum dots	Feraheme (ferumoxytol, anemia, http://www.rxlist.com/feraheme-drug.htm)
Polysaccharide-based Chitosan, alginate	None
Dendrimers Poly(amido amine), poly(ethyleneimine)	None
Biological Retrovirus, lentivirus, adenovirus, herpes simplex virus -1, adeno-associated virus	Gardasil/Cevarix (human papilloma virus-like particles, vaccination, http://www.emea.europa.eu/ema/index.jsp?curl=pages/medicines/human/medicines/000721/human_med_000694.sjsp) Engerix/Recombivax (human hepatitis B virus-like particles, vaccination, http://www.ema.europa.eu/ema/index.jsp?curl=pages/medicines/human/referrals/Engerix_B/human_referral_000098.jsp&mid=WC0b01ac0580024e9a)
Others Fullerenes, ceramic (silica) NPs, titanium dioxide NPs,	None

Note:

* Withdrawn from the market.

Abbreviations: PEG, polyethylene glycol; TNF, tumor necrosis factor; MS, multiple sclerosis; VEGF, vascular endothelial growth factor; onco, oncology; immune, modulation of immune system; endo, endocrine; cardio, cardiology; resp, respiratory problems; ophth, ophthalmology; ADHD, attention deficit hyperactivity disorder.

Figure 1 Targets for cytotoxicity of nanoparticles (NPs).

Notes: NPs may act through extracellular generation of reactive oxygen species (ROS) (1), they may physically damage the plasma membrane by causing holes (2) or bind to membrane proteins like nicotinamide adenine dinucleotide phosphate-oxidase (3), Ca²⁺ channels (4), and membrane receptors (5), thereby inducing oxidative signaling, increasing intracellular Ca²⁺ levels and activating second-messenger cascades. Inside the cells, NPs may interfere with mitochondrial metabolism (6), causing generation of radicals and induction of apoptosis. Intracellular ROS generation by NPs or by metals from lysosomal degradation (7) as well as lysosomal disruption (8) and direct binding to components of the cytoskeleton (9) and the induction of structural alterations of proteins (10) are additional modes of toxic actions. In the nucleus, interference with the transcription machinery and oxidative damage of the DNA (11) may occur.

Figure 2 Simplified representation of active uptake mechanisms in nonphagocytic cells.

Notes: Nanoparticle (●) uptake has been evaluated mainly according to macropinocytosis, represented here as only one route, through macropinosome (MP), clathrin-mediated uptake by clathrin-coated pits (CC), and caveolae-dependent uptake by caveosomes (Cav). Uptake by clathrin-independent caveolae-independent endocytosis, which includes flotillin-, Arf6-, Cdc42-, and RhoA-dependent uptake, is also presented as only one route. Fluid-phase endocytosis, which mainly uses the clathrin-coated pits, is not depicted as a separate route. All pathways deliver their content to endosomes (E), late endosomes (LE) and lysosomes (L); the content of caveolosomes may also be delivered to the endoplasmic reticulum (ER) and the Golgi apparatus. Vesicular transport through the cell occurs through transcytotic vesicles (TV). © 2012, Elsevier. Reproduced with permission from Fröhlich E, Roblegg E. Models for oral uptake of nanoparticles in consumer products. Toxicology. 2012;291(1–3):8.¹⁷⁹

Table 2 Routes of endocytic uptake in nonphagocytic cells

Particle	Charge	Uptake route	References
Quantum dots	Anionic	Clathrin	^Citation173
	Anionic	Clathrin-independent, caveolae-independent	^Citation174
Polystyrene	Anionic	Clathrin	^Citation119
	Plain	Macropinocytosis	^Citation119
	Cationic	Clathrin	^Citation175
	Plain	Clathrin-independent	^Citation175
	Anionic	Clathrin (43 nm)	^Citation176
		Clathrin-independent, caveolae-independent (24 nm)	^Citation176
	Cationic	Macropinocytosis	^Citation177
		Caveolin	^Citation178
Chitosan	Anionic	Clathrin	^Citation159
	Cationic	Clathrin	^Citation159
Poly(lactic acid)	Cationic	Clathrin	^Citation129,Citation160
	Anionic	Clathrin, clathrinin-dependent	^Citation160

LedetGMandalTNanomedicine: emerging therapeutics for the 21st centuryUS Pharm2012373711

 Google Scholar

FröhlichERobleggEModels for oral uptake of nanoparticles in consumer productsToxicology20122911–38

 Web of Science ®Google Scholar

XiaoYForrySPGaoXHolbrookRDTelfordWGTonaADynamics and mechanisms of quantum dot nanoparticle cellular uptakeJ Nanobiotechnol2010813

 PubMedGoogle Scholar

ZhangLWMonteiro-RiviereNAMechanisms of quantum dot nanoparticle cellular uptakeToxicol Sci2009110113815519414515

 PubMed Web of Science ®Google Scholar

JiangXMusyanovychARockerCLandfesterKMailanderVNienhausGUSpecific effects of surface carboxyl groups on anionic polystyrene particles in their interactions with mesenchymal stem cellsNanoscale2011352028203521409242

 PubMed Web of Science ®Google Scholar

JiangXDausendJHafnerMSpecific effects of surface amines on polystyrene nanoparticles in their interactions with mesenchymal stem cellsBiomacromolecules201011374875320166675

 PubMed Web of Science ®Google Scholar

LaiSKHidaKManSTPrivileged delivery of polymer nanoparticles to the perinuclear region of live cells via a nonclathrin, non-degradative pathwayBiomaterials200728182876288417363053

 PubMed Web of Science ®Google Scholar

DausendJMusyanovychADassMUptake mechanism of oppositely charged fluorescent nanoparticles in HeLa cellsMacromol Biosci20088121135114318698581

 PubMed Web of Science ®Google Scholar

XiaTKovochichMLiongMZinkJINelAECationic polystyrene nanosphere toxicity depends on cell-specific endocytic and mitochondrial injury pathwaysACS Nano200821859619206551

 PubMed Web of Science ®Google Scholar

ZakiNMNastiATirelliNNanocarriers for cytoplasmic delivery: cellular uptake and intracellular fate of chitosan and hyaluronic acid-coated chitosan nanoparticles in a phagocytic cell modelMacromol Biosci201111121747176021954171

 PubMed Web of Science ®Google Scholar

Harush-FrenkelORozenturEBenitaSAltschulerYSurface charge of nanoparticles determines their endocytic and transcytotic pathway in polarized MDCK cellsBiomacromolecules20089243544318189360

 PubMed Web of Science ®Google Scholar

Harush-FrenkelODebottonNBenitaSAltschulerYTargeting of nanoparticles to the clathrin-mediated endocytic pathwayBiochem Biophys Res Commun20073531263217184736

 PubMed Web of Science ®Google Scholar