Table 4.
Review of original articles assessing oxidative stress induction by polymeric nanoparticles.
| Nanomaterial | Polymer characterization | Nanomaterial characterization | Testing method | Cellular model | Dose/concentration range | Results | Observations | References |
|---|---|---|---|---|---|---|---|---|
| Chitosan NPs | Low molecular weight chitosan (50–190 kDa, 75–85% DDa) | 92 nm +32 mV |
2′,7′-dichlorodihydro- fluorescein diacetate (H2DCF-DA) probe (72 h incubation) |
HeLa, MDA-MB-231 and THP-1 cells | 1% | Significant reduction in the generation of reactive oxygen species when compared to control | Similar results for plasmid loaded chitosan NPs | Bor et al., 2016 |
| Chitosan NPs | 80% DD 400 kDa |
100 nm + 19 mV |
Dichlorofluorescin diacetate (DCFH-DA) probe (6/12/24 h incubation) | Hela and SMMC-7721 cells | 10; 100 μg/mL | Chitosan NPs increase ROS production in a concentration-dependent manner | – | Wang et al., 2018 |
| Chitosan NPs | Low molecular weight chitosan (85% DD) | ≤ 100 nm + 40 mV |
Dichlorofluorescin diacetate (DCFH-DA) probe (unknown h incubation) | BCL2(AAA) Jurkat cells | 10–50 μg/mL | All concentrations induced ROS production (concentration dependent manner) | Bulk chitosan was tested at the same concentrations. ROS production was concentration dependent but lower than with chitosan NPs | Sarangapani et al., 2018 |
| Chitosan NPs | na | 164 nm; + 63 mV 385 nm; + 62 mV 459 nm; +72 mV 475 nm; +71 mV 685 nm; +74 mV |
Dihydroethidium (DHE) probe (72 h incubation) | Mouse bone marrow-derived hematopoietic stem cells | 250–1,000 μg/mL | ROS production was not significantly altered following exposure to chitosan NPs | – | Omar Zaki et al., 2015 |
| Chitosan NPs | 75–85% 50–190 kDa |
173 nm + 23 mV |
Dichlorofluorescin diacetate (DCFH-DA) probe (24 h incubation) | HEK-293 cells | 100 μg/mL | Chitosan NPs had no effect on ROS production | Bulk chitosan was also tested and had no effect in ROS production | Arora et al., 2016 |
| PLA NPs | Poly(D,L-lactide) (PDLLA) 1,01,782 g/mol and 0.68 dL/g | 188 nm −24 mV (water) 78 nm −0.4 mV (DMEMb) |
2′,7′-Dichlorofluorescin diacetate (DCFH-DA) probe (24 h incubation) | RAW 264.7 cells | 4.3, 17, 34, 340 μg/mL | PLA NPs with 78 nm in DMEM caused a significant increase in ROS production for the highest concentration tested (340 μg/mL) | The increase in ROS production was related to cytotoxicity. The sample and concentration that induced ROS production decreased cell viability to values close to 70%. All the other concentrations were close to 100% | Da Silva et al., 2019 |
| PLA NPs | Poly(D,L-lactide) (PDLLA) 1,01,782 g/mol and 0.68 dL/g | 109 nm −7 mV (water) 154 nm −0.7 mV (DMEM) |
2′,7′-Dichlorofluorescin diacetate (DCFH-DA) probe (24 h incubation) | RAW 264.7 cells | 8.6, 34, 69, 690 μg/mL | No ROS production observed | – | Da Silva et al., 2019 |
| PLA NPs | na | 176 nm −58 mV In cell culture: 212 nm −24 mV |
2′,7′-Dichlorofluorescin diacetate (DCFH-DA) probe (72 h incubation) | Schneider's Drosophila melanogaster line 2 (S2) cells | 0.5–500 μg/mL | ROS production was only observed at the highest tested concentration (500 μg/mL) indicating a concentration dependent effect | – | Legaz et al., 2016 |
| PLGA NPs | Resomer® RG503H, acid terminated, 50:50, Mw 24,000–38,000 | 80 nm −25 mV |
2′,7′-Dichlorofluorescin diacetate (DCFH-DA) probe (3 h incubation) | 16HBE14o-, L5178Y, and TK6 cells | 40 μg/mL | No increase in ROS production in 16HBE14o-, L5178Y, and TK6 cells, in comparison to the control | The L5178Y mouse lymphoma and TK6 human B-lymphoblastoid cells, are routinely used in in vitro regulatory genotoxic assays. The human bronchial epithelial cells 16HBE14o-, a cell line is suitable for toxicity studies of inhaled NPs as it is highly similar to the primary bronchial epithelium | Platel et al., 2016 |
| hexadecyltrimethylammonium bromide (CTAB) stabilized PLGA NPs | Resomer® RG503H, acid terminated, 50:50, Mw 24,000–38,000 and PEG 2,000 | 82 nm +15 mV |
2′,7′-Dichlorofluorescin diacetate (DCFH-DA) probe (3 h incubation) | 16HBE14o-, L5178Y, and TK6 cells | 40 μg/mL | Significant increase in ROS production in 16HBE14o-, L5178Y, and TK6 cells, in comparison to the control | The L5178Y mouse lymphoma and TK6 human B-lymphoblastoid cells, are routinely used in in vitro regulatory genotoxic assays. The human bronchial epithelial cells 16HBE14o-, a cell line is suitable for toxicity studies of inhaled NPs as it is highly similar to the primary bronchial epithelium | Platel 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 |
2′,7′-Dichlorofluorescin diacetate (DCFH-DA) probe (24 h incubation) | MDAMB-231 cells | na | Increased proxidant effect of CH-OA-B-PLGA was two times higher than plain OA | 100 mg/kg is the double of the OA effective dose | Sharma et al., 2017 |
| Poly-lactic-co-glycolic acid–polyethylene oxide (PLGA–PEO) NPs | (Purchased from Advancell) | 140 nm −43 mV (in cell culture medium) |
Hydroethidine probe (24–48 h incubation) | 16HBE14o- and A549 cells | 37.5 and 75 μg/cm2 | Weak production of intracellular ROS at the highest concentrations used, only in the A549 cell line | – | Guadagnini et al., 2013b |
| PLGA NPs | 75:25 Resomer® RG756 | 170 nm −45 mV (200 nm in cell culture medium) |
2′,7′-Dichlorofluorescin diacetate (DCFH-DA) probe (5 min−48 h incubation) | THP-1 cell culture | 0.1 or 1 mg/mL | No Induction of ROS production at 0.1 mg/mL At 1 mg/mL, a transient increase in ROS production was verified at 5 min |
THP-1 monocytes differentiation into macrophages was performed using 12-o-tetradecanoylphorbol-13-acetate (PMA) | Grabowski et al., 2015 |
| PVA stabilized PLGA NPs | 75:25 Resomer® RG756 and PVA (87–89% hydrolyzed, 30–70 kDa) | Ratio PVA:PLGA 11.5:100 230 nm −1 mV (210 nm in cell culture medium) |
2′,7′-Dichlorofluorescin diacetate (DCFH-DA) probe (5 min−48 h incubation) | THP-1 cell culture | 0.1 or 1 mg/mL | No Induction of ROS production at 0.1 mg/mL At 1 mg/mL, a transient increase in ROS production was verified at 5 min |
THP-1 monocytes differentiation into macrophages was performed using 12-o-tetradecanoylphorbol-13-acetate (PMA) | Grabowski et al., 2015 |
| Chitosan stabilized PLGA NPs | 75:25 Resomer® RG756 and Protasan® UP CL113, 75–90% deacetylation, 50–150 kDa | Ratio chi:PVA:PLGA 15.3:30.4:100 230 nm + 40 mV (270 nm in cell culture medium) |
2′,7′-Dichlorofluorescin diacetate (DCFH-DA) probe (5 min−48 h incubation) | THP-1 cell culture | 0.1 or 1 mg/mL | No Induction of ROS production at 0.1 mg/mL At 1 mg/mL, a transient increase in ROS production was verified at 5 min |
THP-1 monocytes differentiation into macrophages was performed using 12-o-tetradecanoylphorbol-13-acetate (PMA) | Grabowski et al., 2015 |
| Pluronic stabilized PLGA NPs | 75:25 Resomer® RG756 and Pluronic F68 | Ratio F68:PLGA 15.5:100 230 nm −30 mV (315 nm in cell culture medium) |
2′,7′-Dichlorofluorescin diacetate (DCFH-DA) probe (5 min−48 h incubation) | THP-1 cell culture | 0.1 or 1 mg/mL | No Induction of ROS production at 0.1 and 1 mg/mL | THP-1 monocytes differentiation into macrophages was performed using 12-o-tetradecanoylphorbol-13-acetate (PMA) | Grabowski et al., 2015 |
| PLGA NPs | 50:50c (intrinsic viscosity 0.60 g/dl) 65:35c (intrinsic viscosity 0.64 g/dl) 75:25c (intrinsic viscosity 0.72 g/dl) 85:15c (intrinsic viscosity 0.62 g/dl) |
210 nm −14 mV 211 nm −8.70 mV 218 nm −12.7 mV 243 nm −12.7 mV |
2′,7′-Dichlorofluorescin diacetate (DCFH-DA) probe (24 h incubation) | RAW 264.7 cells | 10, 30, 100, and 300 μg/mL | No effect on ROS production up to 100 μg/ml concentration; 300 μg/ml showed 1.5- to 2-fold stimulation of ROS production A further increase in NPs concentration to 1,000 μg/ ml interfered with ROS assay due to fluorescence quenching |
No significant differences were found in these assays between these NPs | Singh and Ramarao, 2013 |
| PLA NPs | DL-PLA (MW 10,000) | 256 nm −17.1 mV |
2′,7′-Dichlorofluorescin diacetate (DCFH-DA) probe (24 h incubation) | RAW 264.7 cells | 10, 30, 100, and 300 μg/mL | No effect on ROS production up to 100 μg/ml concentration; 300 μg/ml showed 1.5- to 2-fold stimulation of ROS production A further increase in NPs concentration to 1,000 μg/ ml interfered with ROS assay due to fluorescence quenching |
– | Singh and Ramarao, 2013 |
| PCL NPs | PCL (intrinsic viscosity 1.07 g/dl) | 268 nm −9.10 mV |
2′,7′-Dichlorofluorescin diacetate (DCFH-DA) probe (24 h incubation) | RAW 264.7 cells | 10, 30, 100, and 300 μg/mL | No effect on ROS production up to 100 μg/ml concentration; 300 μg/ml showed 1.5- to 2-fold stimulation of ROS production A further increase in NPs concentration to 1,000 μg/ ml interfered with ROS assay due to fluorescence quenching |
– | Singh and Ramarao, 2013 |
| Poly(lactide-co-caprolactone) (PLCL) NPs | PLCL 25:75 (intrinsic viscosity 0.71 g/dl) PLCL 80:20 (intrinsic viscosity 0.77 g/dl |
261 nm −15.3 mV 261 nm −15.4 mV |
2′,7′-Dichlorofluorescin diacetate (DCFH-DA) probe (24 h incubation) | RAW 264.7 cells | 10, 30, 100, and 300 μg/mL | No effect on ROS production up to 100 μg/ml concentration; 300 μg/ml showed 1.5- to 2-fold stimulation of ROS production A further increase in NPs concentration to 1,000 μg/ ml interfered with ROS assay due to fluorescence quenching |
– | Singh and Ramarao, 2013 |
a
DD, deacetylation degree.
b
DMEM, Dulbecco's Modified Eagle Medium.
c
PLGA lactic to glycolic acid.