Toxicity assessment of superparamagnetic iron oxide nanoparticles in different tissues

Figures & data

Table 1. Some of the commercialized SPIONs which are used for different diagnostic and therapeutic applications.

Brand name	Core material	Surface	Biological target/applications	Clinical relevance	Refs.
Ferumoxides (Feridex IV^®, Endorem™)	Fe3O4–γ–Fe2O3	Dextran	Reticuloendothelial system/liver stem cell labeling	Used and discontinued	[6]
Ferucarbotran (Resovist)	Fe3O4	Carboxydextran	Blood pool/stem cell labeling	Approved	[7]
Ferumoxtran 10 (Combidex) Ferumoxytol	Fe3O4	Dextran Polyglucose sorbitol	Noncancerous lymphatic tissue/tissue imaging	Clinicaltrials	[8]
(Feraheme/Rienso) Ferumoxsil (Lumirem)	Fe3O4	Carboxymethyl ether	Brain/tumor imaging	Clinical trials	[9]
GastroMARK, Oral-SPIO)	Fe3O4–γ–Fe2O3	Silicon metallic iron and activated	Liver imaging	Used and discontinued	[10]
SPION-epirubicin	γ-Fe2O3	Carbon	Drug targeting	Clinical trials (terminated)	[11]
NanoTherm	Not available	Aminosilane	Hyperthermia on brain tumors	Approved/in use in Europe	[12]

Figure 1. Distinguished in vitro assays that probe the impact of nanoparticles on cells.

Table 2. Cytotoxicity of Fe₃O₄ nanoparticles in different cell lines.

Cell line	Surface coating	NP concentration (average size)	Test	Exposure duration	Toxicity	Year	Refs.
EC219 cell and N9 & N11 cells	PVA, amino PVA, Carboxyl PVA, Thiol PVA	2.5 µL NPs/mL, 11.3 mg iron/mL (core = 9–10 nm, d = 30 nm)	MTT	48 h	Uptake of aminoPVA-SPION by N11	2006	[30]
BE-2-C cell	–COOH, –NH2, bare	(2–32 mmol/mL), size of -COOH, -NH2, Bare, respectively, 13.8 ± 2.1, 17.8 ± 2.6, 13.7 ± 2.1	MTT	24 h	Toxicity amounts: bare SPIONs > positive SPIONs > negative particles	2011	[25]
BE-2-C, A172 cells	Bare, CES-grafted, PEGylated, APTES-grafted	(2–32 mM), d = 13.762.1, 13.862.1, 14.961.8, 17.862.6 for Bare, CES-grafted, PEGylated, APTES-grafted, respectively.	MTT and XTT	24 h	Significant toxicity on both neuronal and glial cells	2012	[28]
NIH 3T3	CTAB BSA- and HSA-coated NPs	60 mg/mL (core = 7–9 nm, d = 50–70 nm)	MTT, LDH, and DCFDA	244,872 h	Cellular viability for protein-coated NPs >95%, for CTAB-coated NPs decrease to 90%, for IONPs a clear decline	2014	[35]
hTERT-BJ1	MA-PEG	0–1000 mg/mL (d = 50 nm)	MTT, live/dead	24 h	For 250 mg/mL: 25–50% viable for bare SPIONs; for 1 mg/mL: 99% viable for PEG-coated NPs	2004	[36]
L929	poly(ethylene glycol)-co-fumarate	0.4, 0.8, and 1.6 mg/mL, d = 82 ± 12	MTT	24, 48 and 72 h	Lower toxicity for PEGF-coated NPs compare to bare one	2009	[37]
L929	Bare, CES-grafted, PEGylated, APTES-grafted	(2–32 mM), d = 13.762.1, 13.862.1, 14.961.8, 17.862.6 for Bare, CES-grafted, PEGylated, APTES-grafted, respectively	MTT and XTT	24h	Very little toxicity	2012	[28]
Hepa1-6	DMSA	50 and 100 μg/mL (core= 11 ± 1.24 nm, d = 32 nm)	GeneChip analysis	24 h	Overall significant toxicity, repression of Id3 expression	2015	[38]
BRL 3A	None	0–250 mg mL^–1 (d = 30, 47 nm)	LDH, MTT, GSH	24 h	EC50 > 250 mg/mL	2005	[46]
Hep G2	Bare, CES-grafted, PEGylated, APTES-grafted	(2–32 mM), d = 13.762.1, 13.862.1, 14.961.8, 17.862.6 for Bare, CES-grafted, PEGylated, APTES-grafted, respectively	MTT and XTT	24 h	Very little toxicity	2012	[28]
LLC	Poly(TMSMA-r-PEGMA)	From 1 to 100 µg iron per well, core = 4–8 nm, d = 16 nm).	MTT	12 h	No toxicity even at relatively high concentrations of the SPION	2006	[40]
A549	Without coating	0.2–10 μm, 2–3 μm, 0.5–1.0 μm, 20–60 nm	DCFH-DA, mmp Comet assay, cytokinesis block-micronucleus test	24 h	Genotoxicity of magnetite in A549 cells	2011	[41]
A549	TEPSA, TPED	1 × 10^5 cells per well in to in a 12-well plate. ( core= 9.3 nm, d = 10.1 ± 1.3, and 10.4 ± 1.6 nm for Fe3O4@NH2, and Fe3O4@COOH	DCFH-DA, MTS assay and etc.	24 h	No cell deaths, proliferation reduction is dose dependent and highest for bare SPIONs. Negatively charged NPs are most biocompatible	2015	[42]
A549	Bare, CES-grafted, PEGylated, APTES-grafted	(2–32 mM), d = 13.762.1, 13.862.1, 14.961.8, 17.862.6 for Bare, CES-grafted, PEGylated, APTES-grafted, respectively.	MTT and XTT	24 h	Significant toxicity	2012	[28]
HCM	–COOH, –NH2, bare	(2–32 mmol/mL), size of -COOH, -NH2, Bare NPs, respectively, 13.8 ± 2.1, 17.8 ± 2.6, 13.7 ± 2.1	MTT	24 h	Toxicity amounts: bare SPIONs > positive SPIONs > negative particles SPIONs-NH2	2011	[25]
HCM	Bare, CES-grafted, PEGylated, APTES-grafted	(2–32 mM), d = 13.762.1, 13.862.1, 14.961.8, 17.862.6 for Bare, CES-grafted, PEGylated, APTES-grafted, respectively.	MTT and XTT	24 h	Very little toxicity on the Human cardiac myocytes	2012	[28]
Human MSCs	Resovist SPIONs, Endorem SPIONs	25–1000, and 25–2000 for Resovist and Endorem, respectively.	MTT	24 h	No significant cytotoxicity	2012	[13]

BSA: bovine serum albumin, CTAB: cetyl trimethylammonium bromide, DMSA: dimercaptosuccinic acid, HSA : human serum albumin, poly(TMSMA-r-PEGMA): poly(trimethoxysilyl)propyl methacrylate and PEG methacrylate, TPED: N-[3-(trimethoxysilyl) propyl]ethylenediamine, TEPSA: 3-(triethoxysilyl)-propyl succinic anhydride, MSCs: mesenchymal stem cells. Id3: can be used as a nanotoxicity biomarker for iron nanoparticles.

Table 3. Toxicity results of SPIONs on various tissues; in vivo studies.

SPION used	Dose (mg/kg)	Animal model	Administration route	Results	Refs.
γ-Fe2O3	0.8	Wistar rats	I.V	Showing harmful properties	[46]
Oleic acid-pluronic coated iron oxide NPs	10	Sprague–Dawley rats	I.V	Safe for drug delivery and imaging applications	[47]
Ferumoxtran-10, Ferumoxides, Ferumoxytol,	5 or 25	Long Evans rats	I.V	Safe as MRI contrast agents in the brain	[48]
Fe3O4	0.4, 2, and 10 (µg/kg)	BALB/c mice	I.V	Thrombosis and cardiac oxidative stress, and DNA damage	[49]
SPIONs	100	KM mice	S.I	low hazard potential	[50]
PEGylated SPIONs	350–750	BALB/c mice	I.V	Dose-dependent toxicity	[51]
Ferumoxtran-10	5 or 25	Normal rats, Long Evans rats	I.I in normal rats, I.V in nude rats	Safe as MRI contrast agents in the brain	[52]
Bare, Dex, and PEG-coated SPIONs	5	Wistar rats	I.V	Showing harmful properties in vivo	[53]

I.V: intravenous injection, S.I: subcutaneous injection, I.I: intracerebral injection, PEG: polyethylene glycol, Dex: dextran.

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