Sea Bass Primary Cultures versus RTgill-W1 Cell Line: Influence of Cell Model on the Sensitivity to Nanoparticles
Abstract
:1. Introduction
2. Materials and Methods
2.1. Preparation of Nanoparticle Stock Dispersions
2.2. Characterization
2.3. Protein Corona Quantification
2.4. Cell Cultures
2.4.1. Sea Bass Dicentrarchus labrax Gill Primary Cells
2.4.2. Rainbow Trout Oncorhynchus mykiss Gill Cell Line
2.5. Scanning Electron Microscopy (SEM)
2.6. Cytotoxicity Screening
2.7. Data Treatment
2.7.1. Toxicity Assays: Endpoints and Decision Trees
2.7.2. Integrative Biological Response Index
3. Results
3.1. Nanoparticle Characterization
3.2. Cell Culture Morphology and Characterization
3.3. Toxicity Assays
3.3.1. Decision Trees and Individual Biomarkers
3.3.2. Exposure to TiO2 Nanoparticles
3.3.3. Exposure to Polystyrene (PS) Nanoparticles
3.3.4. Exposure to Ag Nanoparticles
3.3.5. Integrated Biological Response (IBR) Index
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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Material | Coating | Media | Z-Ave (nm) | St.Dev | Z-Pot (mV) | St.Dev |
---|---|---|---|---|---|---|
TiO2 | ||||||
NH2 | H2O | 73.86 | 1.78 | 58.6 | 5.41 | |
NH2 | L15 | 137.33 | 10.21 | −16.53 | 2.35 | |
NH2 | SW | 1567.3 | 60.8 | −11.9 | 1.529 | |
COOH | H2O | 24.82 | 5.37 | −29.2 | 3.62 | |
COOH | L15 | 178.7 | 16.3 | −14.9 | 1.627 | |
COOH | SW | 2103.3 | 45.6 | −9.6 | 1.816 | |
Polystyrene | ||||||
NH2 | H2O | 141.7 | 2.04 | 54.6 | 1.92 | |
NH2 | L15 | 918.0 | 61.6 | −9.3 | 1.519 | |
NH2 | SW | 5542.5 | 928.6 | −8.1 | 1.627 | |
COOH | H2O | 61.02 | 1.3 | −60.04 | 0.987 | |
COOH | L15 | 7640.7 | 609.6 | −12.5 | 2.003 | |
COOH | SW | 9643.3 | 1682.1 | −10.9 | 0.123 | |
Ag | ||||||
NH2 | H2O | 93.33 | 2.04 | 18.4 | 0.1 | |
NH2 | L15 | 5561.3 | 1576.1 | −13.9 | 0.354 | |
NH2 | SW | 1.90 × 107 | 5.985 | −12 | 0.168 | |
COOH | H2O | 89.61 | 0.285 | −61.1 | 1.4 | |
COOH | L15 | 7.15 × 106 | 6.18 × 106 | −5.6 | 1.106 | |
COOH | SW | 8417.0 | 666.3 | −4.7 | 0.026 |
Scheme 1. | RTgill W1 | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|
TiO2 | TiO2 | ||||||||||
Assay | Factor 1 | Sig f1 | Factor 2 | Sig f2 | Factor 3 | Sig f3 | Assay | Factor 1 | Sig f1 | Factor 2 | Sig f2 |
ROS | Time | 0.003 | Coating (48 h) | 0.046 | ROS | ||||||
NRU | ExpMedia | <0.001 | Time (L15) | <0.001 | Weathering (48 h) | 0.011 | NRU | Time | 0.027 | ||
WST | Time | 0.001 | Protein (48 h) | 0.01 | WST | Concentration | 0.004 | ||||
AlaB | Weathering | <0.001 | Time (Y) | 0.003 | ExpMedia (48 h) | 0.008 | AlaB | Weathering | <0.001 | Coating (Y) | 0.032 |
PS | PS | ||||||||||
Assay | Factor 1 | Sig f1 | Factor 2 | Sig f2 | Factor 3 | Sig f3 | Assay | Factor 1 | Sig f1 | Factor 2 | Sig f2 |
ROS | ExpMedia | <0.001 | Weathering (SW) | <0.001 | ROS | Time | <0.001 | ||||
Time (L15) | <0.001 | ||||||||||
NRU | Coating | 0.007 | ExpMedia (a) | 0.01 | Time (a, L15) | 0.023 | NRU | Conc | 0.003 | ||
WST | n.a | WST | Weathering | 0.012 | |||||||
AlaB | Coating | <0.001 | AlaB | ||||||||
Ag | Ag | ||||||||||
Assay | Factor 1 | Sig f1 | Factor 2 | Sig f2 | Factor 3 | Sig f3 | Assay | Factor 1 | Sig f1 | Factor 2 | Sig f2 |
ROS | Time | 0.001 | Weathering/ ExpMedia | 0.044/0.009 | ExpMedia/Weathering | <0.001/0.015) | ROS | Time | <0.001 | Conc | 0.032 |
NRU | Weathering | <0.001 | Time (non w) | 0.011 | ExpMedia (non w) | NRU | Time | <0.001 | |||
WST | Time | 0.001 | WST | Conc | 0.001 | ||||||
AlaB | Time | 0.002 | Weathering (24 h) | <0.001 | AlaB | Conc | 0.002 | Time (conc<10) | 0.002 | ||
Coating (48 h) | <0.001 |
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Jimeno-Romero, A.; Gwinner, F.; Müller, M.; Mariussen, E.; Soto, M.; Kohl, Y. Sea Bass Primary Cultures versus RTgill-W1 Cell Line: Influence of Cell Model on the Sensitivity to Nanoparticles. Nanomaterials 2021, 11, 3136. https://doi.org/10.3390/nano11113136
Jimeno-Romero A, Gwinner F, Müller M, Mariussen E, Soto M, Kohl Y. Sea Bass Primary Cultures versus RTgill-W1 Cell Line: Influence of Cell Model on the Sensitivity to Nanoparticles. Nanomaterials. 2021; 11(11):3136. https://doi.org/10.3390/nano11113136
Chicago/Turabian StyleJimeno-Romero, Alba, Frederik Gwinner, Michelle Müller, Espen Mariussen, Manu Soto, and Yvonne Kohl. 2021. "Sea Bass Primary Cultures versus RTgill-W1 Cell Line: Influence of Cell Model on the Sensitivity to Nanoparticles" Nanomaterials 11, no. 11: 3136. https://doi.org/10.3390/nano11113136