Influence of Liposomes’ and Lipoplexes’ Physicochemical Characteristics on Their Uptake Rate and Mechanisms by the Placenta
Abstract
:1. Introduction
2. Results and Discussion
2.1. Liposomes and Lipoplexes Characterization
2.2. Cytotoxicity Assay of Liposomes and Lipoplexes
2.3. Assessment of Charge Effect on Liposomes’ Internalization by Placental Cells
2.3.1. Qualitative Evaluation on Human Villous Placental Explants
2.3.2. Quantitative Assessment of Liposomes’ Uptake by Placental Cells Using HPLC and FACS
HPLC Dosage of Fluorescent Liposomes Inside Human Villous Placental Explants
FACS Evaluation of Liposomes’ Internalization in BeWo Cells
2.4. Assessment of Uptake Mechanisms of Liposomes and Lipoplexes by Trophoblasts
3. Materials and Methods
3.1. Materials
3.2. Liposomes and Lipoplexes Preparation
3.3. In Vitro Evaluation of Liposomes and Lipoplexes Uptake by Trophoblasts
3.3.1. Cell Culture
3.3.2. MTT Cytotoxicity Assay
3.3.3. Formulations’ Uptake Evaluation Using Fluorescence Activated Cell Sorting (FACS)
3.4. Ex Vivo Evaluation of Liposomes and Lipoplexes Uptake by Villous Placental Explants
3.4.1. Ethical Statement
3.4.2. Incubation of Liposomes and Lipoplexes with Villous Placental Explants
3.4.3. Qualitative Analysis of Liposomes Uptake by Confocal Microscopy
3.4.4. Quantitative Analysis of Liposomes Uptake by HPLC Method
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Formulation | Composition | % of DSPE-PEG2000 | Z-Average (nm ± SD) | PdI | Zeta Potential (mV ± SD) |
---|---|---|---|---|---|
Cationic liposomes | DMAPAP/DOPE/DSPE-PEG2000/DOPE-NBD | 5 | 108.5 ± 0.9 | 0.07 | 31 ± 0.7 |
1 | 106.3 ± 2.2 | 0.08 | 36.1 ± 1.1 | ||
Neutral liposomes | DOPC/Chol/DSPE-PEG2000/DOPE-NBD | 5 | 130.3 ± 3.7 | 0.07 | 3.2 ± 7.2 |
1 | 124.9 ± 1.1 | 0.11 | −10.3 ± 0.3 | ||
Lipoplexes | DMAPAP/DOPE/DSPE-PEG2000/DOPE-NBD + non-coding siRNA + sodium alginate | 5 | 199.0 ± 13.3 | 0.19 | 10.7 ± 5.5 |
1 | 189.6 ± 17.4 | 0.23 | 6.7 ± 1.5 |
Liposomes Incubated | Sample Name | Weight of Villi (mg) | Total Lipid Concentration (µmol/L) | DOPE-NBD Concentration (µg/L) | DOPE-NBD Concentration Extracted (µg/L) | Mass of DOPE-NBD Extract (µg) | Amount of DOPE-NBD in 1 g of Villi (µg) |
---|---|---|---|---|---|---|---|
N5% | C3 | 18 | 150 | 136 | <1 | nd | nd |
C7 | 25 | 300 | 272 | <1 | nd | nd | |
C11 | 12 | 500 | 454 | 2.0 | 0.001 | 0.040 | |
C5% | B3 | 12 | 150 | 136 | 28.1 | 0.014 | 1.172 |
B7 | 9 | 300 | 272 | 69.8 | 0.035 | 3.875 | |
B11 | 17 | 500 | 454 | 145.7 | 0.073 | 4.285 |
Type of Liposomes | Composition | % of DSPE-PEG2000 | % of DOPE-NBD |
---|---|---|---|
Cationic liposomes | DMAPAP/DOPE/DSPE-PEG2000/DOPE-NBD | 5 | 1 |
5 | 0 | ||
1 | 1 | ||
1 | 0 | ||
Neutral liposomes | DOPC/Chol/DSPE-PEG2000/DOPE-NBD | 5 | 1 |
5 | 0 | ||
1 | 1 | ||
1 | 0 | ||
Lipoplexes | DMAPAP/DOPE/DSPE-PEG2000/DOPE-NBD + non-coding siRNA + sodium alginate | 5 | 1 |
5 | 0 | ||
1 | 1 | ||
1 | 0 |
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Fliedel, L.; Alhareth, K.; Seguin, J.; El-Khashab, M.; Chissey, A.; Mignet, N.; Fournier, T.; Andrieux, K. Influence of Liposomes’ and Lipoplexes’ Physicochemical Characteristics on Their Uptake Rate and Mechanisms by the Placenta. Int. J. Mol. Sci. 2022, 23, 6299. https://doi.org/10.3390/ijms23116299
Fliedel L, Alhareth K, Seguin J, El-Khashab M, Chissey A, Mignet N, Fournier T, Andrieux K. Influence of Liposomes’ and Lipoplexes’ Physicochemical Characteristics on Their Uptake Rate and Mechanisms by the Placenta. International Journal of Molecular Sciences. 2022; 23(11):6299. https://doi.org/10.3390/ijms23116299
Chicago/Turabian StyleFliedel, Louise, Khair Alhareth, Johanne Seguin, Marwa El-Khashab, Audrey Chissey, Nathalie Mignet, Thierry Fournier, and Karine Andrieux. 2022. "Influence of Liposomes’ and Lipoplexes’ Physicochemical Characteristics on Their Uptake Rate and Mechanisms by the Placenta" International Journal of Molecular Sciences 23, no. 11: 6299. https://doi.org/10.3390/ijms23116299