Table 3.
Review of original articles assessing in vivo the toxicity of polymeric nanoparticles.
| Nanomaterial | Polymer characterization | Nanomaterial characterization | Testing method | Model | Administration route | Dose/concentration range | Results | References |
|---|---|---|---|---|---|---|---|---|
| Chitosan NPs | Chitosan hydrochloride salt (Protasan CL 110) | 289 nm + 36 Mv (phosphate buffer) |
In vivo exposure (acute toxicity) | New Zealand rabbits | Ocular | 30 μL of the 0.5 mg/mL CSNP formulation in the right eye every 30 min for 6 h | No signs of discomfort in rabbits eyes 24 h after the administration No histopathological changes in the eye compared to control |
de Salamanca A et al., 2006 |
| Insulin (ins) loaded alginate/chitosan (Alg/chi) NPs | Depolymerized chitosan (65 and 25 kDa, and 86% DDa) Alginate (M/Gb content 64.5/35.5%) |
3:1:1c 104 nm, + 4 mV 3:2:1c 157 nm, + 10 mV 3:3:1c 216 nm, + 16 mV |
In vivo exposure (acute toxicity) | Swiss albino mice | Oral | 150 mg/kg b.w. (ratio alg:chi:ins 3:1:1) | No mortality No change in biochemical or histopathological parameters No liver or renal toxicity |
Mukhopadhyay et al., 2015 |
| Eudragit® S100/alginate-enclosed chitosan-calcium phosphate-loaded lactoferrin nanocapsules | na | 240 nm −2.6 mV |
In vivo exposure (acute toxicity: 24 h) | Artemia salina (brine shrimp) | Oral (diluted in the water) | 20–5,000 μg/mL | No lethality | Leng et al., 2018 |
| Pluronic coated PLGA NPs | 75:25 Resomer® RG756 and Pluronic F68 | 240 nm −35 mV |
In vivo exposure (acute toxicity) | Balb/cJ mice | Intratracheal (nebulization) | 250 μg/50 μL in 5% glucose | Coated PLGA NPs did not induce an inflammatory response in mice, with no alterations of cellular population, protein quantity or expression of cytokines in BAL | Aragao-Santiago et al., 2015 |
| PVA coared PLGA NPs | 75:25 Resomer® RG756 and PVA (87–89% hydrolyzed, 30–70 kDa) | 220 nm −4 mV |
In vivo exposure (acute toxicity) | Balb/cJ mice | Intratracheal (nebulization) | 250 μg/50 μL in 5% glucose | Coated PLGA NPs did not induce an inflammatory response in mice, with no alterations of cellular population, protein quantity or expression of cytokines in BAL | Aragao-Santiago et al., 2015 |
| Chitosan coated PLGA NPs | 75:25 Resomer® RG756 and Protasan® UP CL113,75–90% deacetylation, 50–150 kDa | 200 nm + 18 mV |
In vivo exposure (acute toxicity) | Balb/cJ mice | Intratracheal (nebulization) | 250 μg/50 μL in 5% glucose | Coated PLGA NPs did not induce an inflammatory response in mice, with no alterations of cellular population, protein quantity or expression of cytokines in BAL | Aragao-Santiago et al., 2015 |
| Dissulfiram loaded PLGA nanoparticles, coated with PEG and functionalized with folate | PLGA (RG 504 H, acid terminated, lactide:glycolide 50:50, Mw: 38,000) and PEG-bis-amine (Mn: 10,000) | 204 nm −5.24 mV |
In vivo exposure (acute toxicity) | BALB/C mice | Intravenous | Equivalent to 120 and 60 mg/kg b.w. of dissulfiram | No lethality, no hematological parameters changes (2,000 mg/kg of loaded NPs ~100 mg/kg equivalent of disulfiram) |
Fasehee et al., 2016 |
| Dissulfiram loaded PLGA nanoparticles, coated with PEG and functionalized with folate | PLGA (RG 504 H, acid terminated, lactide:glycolide 50:50, Mw: 38,000) and PEG-bis-amine (Mn: 10,000) | 204 nm −5.24 mV |
In vivo exposure (acute toxicity) | BALB/C mice | Intraperitoneal | Equivalent to 2,000 and 225 mg/kg b.w. of dissulfiram | No lethality, hematological parameters altered (2,000 mg/kg of loaded NPs ~100 mg/kg equivalent of disulfiram) |
Fasehee et al., 2016 |
| Poly(ε-caprolactone)-poly(ethylene glycol)-poly(ε-caprolactone) (PCEC) nanoparticles | PCEC copolymer with a molecular weight of 17,500 (1H NMR spectrum) | 40 nm | In vivo exposure (acute toxicity) | Sprague-Dawley rats | Intravenous | 2.4 g/kg (divided in 2 administration within 12 h) | No clinical symptoms 14-days post-injection No histopathological findings after animal's sacrifice |
Huang et al., 2010 |
| Paclitaxel loaded PLA NPs | Inherent viscosity 0.55–0.75 dL/g and average molecular weight 75,000–1,20,000 | 150–175 nm, and zeta potentials lower than −15 mV | In vivo exposure (acute toxicity) | Wistar rats | Intravenous | 10 mg/kg b.w.of paclitaxel | No induction of histopathological alterations (number, arrangement and architecture of cells) of the heart, lungs, liver, spleen, kidney, and brain Blank nanoparticles (unspecified dose) did not cause any toxicity as well |
VasanthaKumar et al., 2014 |
| Paclitaxel loaded PLGA NPs | Lactide:glycolide 50/50 and average molecular weight 5000–1,5000 | 150–175 nm < -15 mV |
In vivo exposure (acute toxicity) | Wistar rats | Intravenous | 10 mg/kg b.w.of paclitaxel | No induction of histopathological alterations (number, arrangement and architecture of cells) of the heart, lungs, liver, spleen, kidney, and brain Blank nanoparticles (unspecified dose) did not cause any toxicity as well |
VasanthaKumar et al., 2014 |
| Paclitaxel loaded PCL NPs | Average molecular weight 14,000 and average molecular number 10,000 | 150–175 nm, and zeta potentials lower than −15 mV | In vivo exposure (acute toxicity) | Wistar rats | Intravenous | 10 mg/kg b.w.of paclitaxel | No induction of histopathological alterations (number, arrangement and architecture of cells) of the heart, lungs, liver, spleen, kidney, and brain Blank nanoparticles (unspecified dose) did not cause any toxicity as well |
VasanthaKumar et al., 2014 |
| Danorubicin loaded polyethylene glycol-poly L-lysine-poly lactic-co-glycolic acid (PEG-PLL-PLGA) NPs | na | 229 nm −20 mV |
In vivo exposure (Acute toxicity) | Kunming mice | Intravenous | 40, 30, 22, 17, and 13 mg/kg b.w.of Danunorubicin (DNR) loaded in the particles | LD50: 464.4 mg/kg b.w.(23.22 mg/kg b.w.of DNR) 95% confidence interval: 399–542 mg/kg b.w.(20–27 mg/kg b.w.OF DNR) No significant pathological changes of organizational structure and cell morphology |
Guo et al., 2015 |
| Danorubicin loaded polyethylene glycol-poly L-lysine-poly lactic-co-glycolic acid (PEG-PLL-PLGA) NPs | na | 229 nm −20 mV |
In vivo exposure (Acute toxicity) | Kunming mice | Intravenous | 200 mg/kg b.w.of DNR loaded in the particles | No lethality No physical signs of toxicity No changes in hepatic or renal markers |
Guo et al., 2015 |
| Amphotericin loaded PEG-PLGA nanoparticles | Copolymer produced with 6,000 Da PLGA (lactic to glycolic acid molar ratio of 1:1) and 15% PEG | 25 nm | In vivo exposure (acute toxicity) | Albino Sprague-Dawley rats | Intravenous | Equivalent to 1 mg/kg of amphotericin and blank NPs | No nephrotoxicity (evaluated by renal injury biomarkers BUN and PCr) Although described no results presented for blank nanoparticles group |
Radwan et al., 2017a |
| Angiopoietin-2 (Ang2) small interfering (si)RNA plasmid chitosan magnetic nanoparticles (CMNPs) | Chitosan polysaccharides (Mwd 1,38,0000, 90% DD) | nae | In vivo exposure (acute toxicity) | Kunming mice | Intravenous | 92, 153, 255, 424, and 707 mg/kg b.w. | All doses: no mortality, no changes in b.w. Higher doses: short-term staggering, reduced activities and accelerated breathing, as well as transient reduction of eating, lung uneven dark red coloring and particles aggregated inside the lungs Based on the conversion method of equivalent dose co efficient, the non-toxic dose in humans should be <222 mg/kg per day for 14 day, overall a total of 3117 mg/kg, which is significantly higher compared with the quantity required clinically |
Shan et al., 2017 |
| Tween 80 modified chitosan nanoparticles (TmCS-NPs) | Chitosan (100 kDa, 85% DD) | 251 nm +26.5 mV |
In vivo exposure (7 days) |
Sprague-Dawley rats | Intravenous | 3, 10, and 30 mg/kg b.w. | Body weight of rats remarkably decreased dose-dependently Dose-dependent neuron apoptosis and slight inflammatory response in the frontal cortex, and downregulation of GFAP expression in the cerebellum Study aim: neurotoxicity |
Yuan et al., 2015 |
| Chitosan/alginate (Chi/alg) NPs | Chitosan (Mvf of 1,10,000–1,50,000) Sodium alginate (very low viscosity) |
1:10g 300 nm, −30 mV (water) 900 nm, −25 mV (cell culture medium) 10:1g 500 nm, + 30 mV (water) 1,100 nm, + 10 mV (cell culture medium) |
In vivo exposure (14 days) | Wistar albino rats | Oral | 9 mg/kg b.w. (in 0.5 ml/100 g b.w.) | No mortality No behavioral changes No changes in body weight or relative liver weight No changes in MDA levels GSH levels decreased for the 10:1 (chit:alg) ratio No hematological parameters altered |
Aluani et al., 2017 |
| Chitosan/alginate (Chi/alg) NPs | Chitosan (low molecular weight; 200 cp viscosity) Sodium Alginate (low viscosity −0.02 Pa.s) |
1:9g 254 nm, −35 mV |
In vivo exposure (14 days) | Wistar albino rats | Oral | 24.5 mg (in 2 mL) | No mortality No adverse reaction in the condition of the eye, nose and motor activity No histopathological alteration in animal's organs Normal feed intake and weight gain |
Radwan et al., 2017b |
| pH sensitive chitosan/poly-γ-glutamic acid (Chi/PGA) NPs | Chitosan (80 kDa, 85% DD) γ -PGA (60 kDa) |
218 nm +25.3 mV |
In vivo exposure (14 days) | ICR mice | Oral | 100 mg/kg b.w. | No clinical signs or weight loss No change in hematological or biochemical parameters No pathological changes in liver, kidney and intestinal segments The dose (100 mg/kg) was 18 times higher than the dose they used in the pharmacokinetic study of insulin-loaded nanoparticles (5.5 mg/kg) |
Sonaje et al., 2009 |
| α-tocopherol succinate-grafted carboxymethyl chitosan polymeric micelles | low molecular weight chitosan: 22 kDa | 114–187 nm −20 to −22 mV |
In vivo exposure (14 days) | Sprague Dawley rats | Oral | 500 mg/kg b.w. | No mortality Normal weight gain Normal red blood cells morphology No pathological changes in the liver, kidney, and intestine |
Jena and Sangamwar, 2016 |
| Alginate coated CS core-shell NPs | Sodium alginate (ALG) of low viscosity, ~50 kDa Low molecular weight CS (25 kDa, DDA 82%) |
216 nm −36 mV (with naringenin encapsulated) |
In vivo exposure (19 days) | Wistar rats | Oral | 50 mg/kg b.w. (blank NPs) | No significant differences in hair texture or color, water and food intake No hepatic toxicity No abnormalities found in the hepatic or intestinal tissues No hematological parameters change (glucose and lipids) |
Maity et al., 2017 |
| Oleoyl-carboxymethyl-chitosan (OCMCS) nanoparticles | 170 kDa chitosan, 92.56% DD modified with chloroactic acid and oleoyl chloride | 171 nm + 19 mV |
In vivo exposure (7 days) |
Carp | Oral (catheter) | 2 mg/mL (500 μL) | No lethality or histopathological signs of inflammation (liver, spleen, kidneys) | Liu et al., 2013 |
| Amphotericin loaded PEG-PLGA NPs | PLGA lactic to glycolic acid 50:50 with 40–75 KDa and PEG with 10 KDa | 170 nm |
In vivo exposure (7 days) |
Wistar rats | Intraperitoneal and oral | Equivalent to 10 mg/kg b.w.of amphotericin | No lethality, no body weight loss, no hematological parameters alterations, no histopathological changes in liver, and kidneys | Moraes Moreira Carraro et al., 2017 |
| Amphotericin loaded PLGA NPs | PLGA lactic to glycolic acid 50:50 with 40–75 KDa | 190 nm | ||||||
| Chitosan/alginate (Chi/alg) NPs | Chitosan (Mvh of 1,10,000–1,50,000) Sodium alginate (very low viscosity) |
1:10i 300 nm, −30 mV (water) 900 nm, −25 mV (cell culture medium) 10:1i 500 nm, + 30 mV (water) 1,100 nm, + 10 mV (cell culture medium) |
In vivo exposure (14 days) | Wistar albino rats | Oral | 9 mg/kg b.w. (in 0.5 ml/100 g b.w.) | No mortality No behavioral changes No changes in body weight or relative liver weight No changes in MDA levels GSH levels decreased for the 10:1 (chit:alg) ratio No hematological parameters altered |
Aluani et al., 2017 |
| Chitosan/alginate (Chi/alg) NPs | Chitosan (low molecular weight; 200 cp viscosity) Sodium Alginate (low viscosity −0.02 Pa.s) |
1:9i254 nm, −35 mV | In vivo exposure (14 days) | Wistar albino rats | Oral | 24.5 mg (in 2 mL) | No mortality No adverse reaction in the condition of the eye, nose, and motor activity No histopathological alteration in animal's organs Normal feed intake and weight gain |
Radwan et al., 2017b |
| pH sensitive chitosan/poly-γ-glutamic acid (Chi/PGA) NPs | Chitosan (80 kDa, 85% DD) γ -PGA (60 kDa) |
218 nm +25.3 mV |
In vivo exposure (14 days) | ICR mice | Oral | 100 mg/kg b.w. | No clinical signs or weight loss No change in hematological or biochemical parameters No pathological changes in liver, kidney, and intestinal segments The dose (100 mg/kg) was 18 times higher than the dose they used in the pharmacokinetic study of insulin-loaded nanoparticles (5.5 mg/kg) |
Sonaje et al., 2009 |
| Dissulfiram loaded PLGA nanoparticles, coated with PEG and functionalized with folate | PLGA (RG 504 H, acid terminated, lactide:glycolide 50:50, Mw: 38,000) and PEG-bis-amine (Mn: 10,000) | 204 nm −5.24 mV |
In vivo exposure (7 days) |
BALB/C mice | Intravenous | Equivalent to 120, 60, 30, and 15 mg/kg of dissulfiram 120 mg/kg b.w. blank nanoparticles |
No lethality, no hematological parameters changes (2,000 mg/kg of loaded NPs ~100 mg/kg equivalent of disulfiram) |
Fasehee et al., 2016 |
| Polyphenolic bio-enhancers with oleanolic acid in chitosan coated PLGA NPs (CH-OA-B-PLGA NPs) | chitosan (molecular weight 150 kDa, deacetylation degree 85%), Poly (lactide-coglycolide) (PLGA) 50:50, mw 40–75 kDa | 342 nm + 34 mV |
In vivo exposure (15 days) | Sprague Dawley rats | Oral | 100 mg/kg b.w. of OA | No mortality No histopathological changes No abnormal behavior (100 mg/kg is the double of the OA effective dose) |
Sharma et al., 2017 |
| Polyphenolic bio-enhancers with oleanolic acid in PLGA NPs (OA-B-PLGA NPs) | chitosan (molecular weight 150 kDa, deacetylation degree 85%), Poly (lactide-coglycolide) (PLGA) 50:50, mw 40–75 kDa | 221 nm −19 mV |
In vivo exposure (15 days) | Sprague Dawley rats | Oral | 100 mg/kg b.w. of OA | No mortality No histopathological changes No abnormal behavior (100 mg/kg is the double of the OA effective dose) |
Sharma et al., 2017 |
| Amphotericin loaded PEG-PLGA nanoparticles | Copolymer produced with 6,000 Da PLGA (lactic to glycolic acid molar ratio of 1:1) and 15% PEG | 25 nm |
In vivo exposure (7 days) |
Albino Sprague-Dawley rats | Intravenous | Equivalent to 1 mg/kg of amphotericin and blank NPs | No nephrotoxicity (evaluated by renal injury biomarkers BUN and PCr) No histopathological damage of the kidney Although described no results presented for blank nanoparticles group |
Radwan et al., 2017a |
| Paclitaxel loaded monomethoxypoly (ethylene glycol)-b-poly(lactic acid) (mPEG-PLA) polymeric micelles |
mPEG-PLA copolymer (40/60) with a number average molecular weight of 4488.4 | (40/60): 37 nm After incubation with BSA: 40 nm (50/50): 44 nm After ncubation with BSA: 71 nm |
In vivo exposure (4 weeks, 1 injection per week) | Beagle dogs | Injection in the foreleg (intravenous) | Equivalent to 0.5 mg/mL of paclitaxel | mPEG-PLA (40/60): no sign of pathological changes except the lung congestion. mPEG-PLA (50/50): liver index was higher and the thymus index was lower;pylorus and small intestine congestion were also observed The toxicity of paclitaxel loaded mPEG-PLA (40/60) polymeric micelles was significantly lower than those of mPEG-PLA (50/50) |
Li et al., 2014 |
| Angiopoietin-2 (Ang2) small interfering (si)RNA plasmid chitosan magnetic nanoparticles (CMNPs) | Chitosan polysaccharides (Mwj 13,80,000, 90% DD) | nae | In vivo exposure (14 days) | Sprague-Dawley rats | Intravenous | 35, 70, and 353 mg/kg b.w. | Higher doses: chronic pulmonary congestion in Sprague-Dawley rats, as well as simultaneous pulmonary inflammation and partial fibrosis All doses: total number of white blood was significantly higher Based on the conversion method of equivalent dose co-efficient, the non-toxic dose in humans should be <222 mg/kg per day for 14 day, overall a total of 3,117 mg/kg, which is significantly higher compared with the quantity required clinically |
Shan et al., 2017 |
a
DD, deacetylation degree.
b
M/G, β-D-mannuronic acid/α-L-guluronic acid.
c
Ratio alg:chi:ins.
d
Mw, molecular weight number.
e
na, not available.
f
Mv, viscosity molecular weight.
g
Ratio chi:alg.