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Preparation and stability of lipid nanoparticles excluding 1,2-distearoyl-sn-glycero-3-phosphocholine

  • Hye Yoon Jung

    Hye Yoon Jung is a senior undergraduate student at Seoul National University of Science and Technology. Her research interests are in biomimetic membranes.

         , Tae Hoon Kim

    Tae Hoon Kim is a senior undergraduate student at Seoul National University of Science and Technology. His research interests are in biomimetic membranes.

         , Kunn Hadinoto

    Kunn Hadinoto received his B.S. degree from the University of Washington, USA, in 2000 and his Ph.D. degree from Purdue University, USA, in 2004. He joined NTU’s School of Chemical & Biomedical Engineering as an assistant professor in 2007. Prior to that, he worked as a research fellow at the Agency of Science Technology & Research (A*STAR) Singapore. His research interests are in nanopharmaceuticals and their applications.

         and Jin-Won Park

    Jin-Won Park is a professor at Seoul National University of Science and Technology. He received his B.S. degree from Korea University, Korea, in 1998 and his M.S. and Ph.D. degrees from Purdue University, USA, in 2003 and 2005, respectively. From 2007 to 2010, he was an assistant professor at Gachon University, Korea. Since 2010, he has been a professor at Seoul National University of Science and Technology. His research interests are in biomimetic membranes and their applications.

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Published/Copyright: October 16, 2024
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Tenside Surfactants Detergents
From the journal Tenside Surfactants Detergents Volume 61 Issue 6

Abstract

Since 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), the component to keep structure of mRNA-lipid nanoparticle (LNP), is known to cause adverse effects, the replacement of DSPC with the combination of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) and 1-stearoyl-2-hydroxy-sn-glycero-3-phospho ethanolamine (SHPE) was investigated. Specifically, when DSPC of mRNA-LNP was replaced by an 11:1 ratio of DSPE:SHPE, it was found that the size and permeability of mRNA-LNP were the same as those of mRNA-LNP containing DSPC in terms of stability. This result appears to be due to lipid geometry – the ratio of lipid volume to head group area and lipid length.

Keywords: lipid nanoparticle; DSPC replacement; stability; size; permeability

1 Introduction

1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) is included in mRNA-LNP (mRNA-lipid nanoparticles), which are also used as a Covid-19 vaccine, and the mRNA-LNP structure is maintained. 1 , 2 As mRNA-LNP is known to be effective in the delivery of erythropoietin, a treatment for renal failure, it is expected that mRNA-LNP will be used as a drug delivery platform in the future. 3 However, during cell metabolism, DSPC is degraded to eicosanoid, a proinflammatory mediator, by phospholipase-A2, producing antigens that induce inflammatory cytokines. 4 Therefore, the generated antigen induces complement hyper-activation, which can cause severe systemic reactions, including shock symptoms such as acute respiratory distress, decreased blood pressure, and loss of consciousness. 4 , 5 Furthermore, the use of some cationic lipids for in vivo administration has demonstrated their potential cytotoxicity. 6 To overcome the toxicity of cationic lipids, FDA-approved ionizable cationic lipids such as DLin-MC3-DMA, ALC-0315, and SM-102 have been synthesized by multistep reactions with low chemical yields. 7 , 8 Cholesterol molecules present in lipid bilayers are highly susceptible to oxidation. Cholesterol oxidation products known as oxysterols lead to chronic inflammation. 9

Although unsaturated lipids such as 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) are known to be the most potent and dominant component, the unsaturated lipids are susceptible to oxidants. 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 The susceptibility leads to an important factor in the pathogenesis of neurodegenerative diseases. 17 Therefore, in this research, alternative approaches were investigated as follows. The role of DSPC in stably maintaining the structure of mRNA-LNP was due to the presence of a phosphotidylcholine group. 18 Therefore, we would like to prepare mRNA-LNPs with a combination of other phospholipids – 1,2-distearoyl-sn-glycero-3-phosphoethanolamine and 1-stearoyl-2-hydroxy-sn-glycero-3-phosphoethanolamine – of the same geometric shape as DSPC without producing pro-inflammatory mediators. Both lipids were considered easy to reach the DSPC geometry because the carbon number of the hydrocarbon group was equal to that of DSPC. 19

In this study, we aim to investigate the particle size and stability of LNPs prepared by replacing DSPC with with the combination of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) and 1-stearoyl-2-hydroxy-sn-glycero-3-phospho- ethanolamine (SHPE) using dynamic light scattering (DLS) over 25 days. These properties may provide a platform for the DSPC-free LNPs for vaccines or therapies.

2 Experimental procedures

2.1 Materials

1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), 1-stearoyl-2-hydroxy-sn-glycero-3-phosphoethanolamine (SHPE), cholesterol (Chol), and 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG2000) were purchased from Merck (Rahway, NJ, USA) and used without further purification. Cationic lipid, 1,2-di-O-octadecenyl-3-trimethylammonium propane (DOTMA), was purchased from MedChemExpress (Monmouth Junction, NJ, USA). Erythropoietin mRNA (858 nucleotides) was obtained from TriLink Biotechnologies (San Diego, CA, USA).

2.2 LNP preparation

LNPs were prepared by the previously established ethanol injection method. 19 Lipids were dissolved in ethanol at 12.5 mM and mixed in a desired molar ratio. Considering that the ratios of lipid volume to head group area and lipid length were 0.5–1, 1, and less than 0.33333 for DSPC, DSPE, and SHPE, respectively, the ratios of DOTMA:DSPE:SHPE:Chol:DMG-PEG2000 were 61:11:1:24:3, 61:10:2:24:3, and 61:9:3:24:3, respectively. 20 , 21 As a contrast, the solution of DOTMA:DSPC:Chol:DMG-PEG2000 (61:12:24:3) was also prepared. mRNA was dissolved in RNase-free sodium citrate buffer (50 mM, pH 3) to a concentration of 0.1 mg/ml. 22 After the solutions were passed through a 200 nm polyethersulfone filter from Merck (Rahway, NJ, USA), the aqueous and ethanolic solutions were mixed in a volume ratio of 3:1 by the injection method to produce LNPs with a DOTMA to nucleotide phosphate ratio of 6:1. 23 Immediately prior to injection, the lipids were heated above their transition temperature. After a further 10 min of mixing, the extrusion of the collected LNPs through the filter was repeated until the injection flow became little prohibited. The amount of mRNA not incorporated into the LNPs was characterized using RiboGreen® from Thermo Fisher Scientific (Waltham, MA, USA) and subtracted from the input to estimate the amount encapsulated. 24 The loading efficiency of mRNA was (85 ± 5) % and indistinguishable based on the ratios.

2.3 LNP characterization and stability

The size of the LNPs was determined by DLS measurements (nano SAQLA, Otsuka). For this purpose, the sample from the extrusion described above was diluted 40 times in 10 mM PBS buffer of pH 7.4, and stored at 4 °C for 30 days. Additionally, the size was monitored on the 10th, 20th, and 30th day from the date of the preparation. The number distributions were estimated using the refractive index of water (1.33), as the LNPs were dispersed in an aqueous state. For the measurement of the permeability across the LNP layer, a pH-sensitive dye (pyranine) was added to the PBS buffer. Since the pH-sensitive dye was encapsulated inside the mRNA-LNPs, the release from the inside to the outside, the permeability, was estimated by measuring the fluorescence intensity of the dye.

3 Results

3.1 LNP size

The size distribution of the LNPs was determined by dynamic light scattering. All conditions were maintained except DSPC or DSPE & SHPE to investigate the effect of the combination of DSPE and SHPE on LNP. Since the headgroup of SHPE was relatively larger compared to the size of tail group, it was expected that the size of the LNP would become smaller as the ratio of SHPE increased, as shown in Figure 1. The size decreases as the curvature of the particle increases, and the curvature theoretically increases as the ratio of the hydrophobic region to the hydrophilic region exposed to the external aqueous region decreases. The average diameter of each ratio was 75 nm, 84 nm, and 94 nm in order of the SHPE ratio increase. The delivery of mRNA-LNP is known to be restricted by the size of the fenestrae in healthy liver, which is approximately (100–160) nm. 25 The size of the LNPs prepared with DOTMA:DSPE:SHPE:Chol:DMG-PEG2000 (61:11:1:24:3) was almost identical to that of the contrast, LNPs prepared with DOTMA:DSPC:Chol:DMG-PEG2000 (61:12:24:3). The retention of LNP size was dependent on the ratios. Up to day 10, the size remained at all of the ratios. On day 20, it was observed that the change in size was inversely proportional to the ratio of the SHPE. Additionally, the retention up on 30th day was also identical between the LNPs prepared with DOTMA:DSPE:SHPE:Chol:DMG-PEG2000 (61:11:1:24:3) and those prepared with DOTMA:DSPC:Chol:DMG-PEG2000 (61:12:24:3).

Figure 1: Size distribution of mRNA-LNP in DOTMA:DSPE:SHPE:Chol:DMG-PEG2000 at different mixing ratios: A) 64:11:1:24:3, B) 64:10:2:24:3, and C) 64:9:3:24:3.
Figure 1:

Size distribution of mRNA-LNP in DOTMA:DSPE:SHPE:Chol:DMG-PEG2000 at different mixing ratios: A) 64:11:1:24:3, B) 64:10:2:24:3, and C) 64:9:3:24:3.

3.2 LNP stability

The permeability, the flux of protons across the LNP layer, was monitored using pyranine, whose fluorescence intensity decreased with decreasing pH. 26 , 27 The aqueous solution inside the LNP had a pH of 3.0, while the outside had a pH of 7.4. When the proton was released from the LNPs, the intensity of the pyranine decreased. Therefore, the fluorescence intensity was measured over time. The intensity was maintained longer as the SHPE ratio decreased, as shown in Figure 2. This result was consistent with that of the size according to the SHPE ratio. Again, the intensity was identical between the LNPs prepared with DOTMA:DSPE:SHPE:Chol:DMG-PEG2000 (61:11:1:24:3) and those prepared with DOTMA:DSPC:Chol:DMG-PEG2000 (61:12:24:3).

Figure 2: Permeability through the layer from inside to outside of mRNA LNPs for the investigated ratios of DOTMA:DSPE:SHPE:Chol:DMG-PEG2000.
Figure 2:

Permeability through the layer from inside to outside of mRNA LNPs for the investigated ratios of DOTMA:DSPE:SHPE:Chol:DMG-PEG2000.

Both the size and the permeability results have been interpreted in terms of lipid geometry. The ratios of lipid volume to the product of its headgroup area and length are 0.3, close to less than 1, and 1 for lyso-PE, PC with two fatty-acid tails, and PE with two fatty-acid tails, respectively. 23 In reality, lyso-PE is more favorable for the formation of micelles than structures with an aqueous interior. Therefore, more lyso-PE leads to the instability of the LNP. However, only PE with two fatty-acid tails is not suitable to form LNP, either. Considering the lipid geometry, PC with two fatty-acid tails is the best molecule to keep the structure stable, similar to the spherical lipid bilayer. This is the reason why DSPC has so far been used for the production of mRNA-LNP. 28 Considering the ratios further, one SHPE per every 11 DSPE corresponded to 0.94 as the area fraction of the hydrophobic inner region compared to the hydrophilic outer region. In addition, the fractions for 61:10:2:24:3, and 61:9:3:24:3 were 0.88 and 0.83, respectively. Both experimental results described above seemed to support the fractional calculation. Other lipids can be considered instead of DSPE and SHPE. However, when saturation and unsaturation coexist, phase separation occurs in the lipid layer. 29 Also, the use of lipids with different carbon numbers may lead to mismatches between the hydrophobic tails of each lipid. Therefore, the selection of different lipids may lead to instability in the structural interactions.

4 Conclusions

In this study, the mRNA-LNP was prepared without DSPC which plays a role in maintaining the structure of conventional mRNA-LNP. DSPC was replaced by a combination of DSPE and SHPE, specifically DOTMA:DSPC:Chol:DMG-PEG2000 (61:12:24:3) with DOTMA:DSPE:SHPE:Chol:DMG-PEG2000 (61:11:1:24:3). Only the DSPE and SHPE ratios have been changed, 11:1, 10:2, and 9:3. At the ratio of 11:1, the size and the permeability of mRNA-LNPs were identical to those of DSPC-included mRNA-LNPs. The present study can provide a platform to reduce the adverse effect, that DSPC may cause.

Corresponding author: Jin-Won Park, Department of Chemical and Biomolecular Engineering, College of Energy and Biotechnology, Seoul National University of Science and Technology, Seoul, Republic of Korea; and SeoulTech-KIRAMS Graduate School of Medical Sciences, Seoul National University of Science and Technology, Seoul, Republic of Korea E-mail:

About the authors

Hye Yoon Jung

Hye Yoon Jung is a senior undergraduate student at Seoul National University of Science and Technology. Her research interests are in biomimetic membranes.

Tae Hoon Kim

Tae Hoon Kim is a senior undergraduate student at Seoul National University of Science and Technology. His research interests are in biomimetic membranes.

Kunn Hadinoto

Kunn Hadinoto received his B.S. degree from the University of Washington, USA, in 2000 and his Ph.D. degree from Purdue University, USA, in 2004. He joined NTU’s School of Chemical & Biomedical Engineering as an assistant professor in 2007. Prior to that, he worked as a research fellow at the Agency of Science Technology & Research (A*STAR) Singapore. His research interests are in nanopharmaceuticals and their applications.

Jin-Won Park

Jin-Won Park is a professor at Seoul National University of Science and Technology. He received his B.S. degree from Korea University, Korea, in 1998 and his M.S. and Ph.D. degrees from Purdue University, USA, in 2003 and 2005, respectively. From 2007 to 2010, he was an assistant professor at Gachon University, Korea. Since 2010, he has been a professor at Seoul National University of Science and Technology. His research interests are in biomimetic membranes and their applications.

Acknowledgments

This study was supported by the Research Program funded by the SeoulTech (Seoul National University of Science and Technology).

  1. Research ethics: Not applicable.

  2. Informed consent: Not applicable.

  3. Author contributions: Hye Yoon Jung conceptualized the study, carried out all experiments, analyzed all data, and wrote the first draft of the manuscript. Tae Hoon Kim carried out all experiments and analzed data. Kunn Hadinoto analyzed all data and reviewed the manuscript. Jin-Won Park conceptualized the study, supervised the study, reviewed, and edited the manuscript. All authors contributed to and approved the final draft of the manuscript.

  4. Use of Large Language Models, AI and Machine Learning Tools: None declared.

  5. Conflict of interests: The authors state no conflict of interest.

  6. Research funding: None declared.

  7. Data availability: Not applicable.

References

1. Baden, L. R.; El Sahly, H. M.; Essink, B.; Kotloff, K.; Frey, S.; Novak, R.; Diemert, D.; Spector, S. A.; Rouphael, N.; Creech, C. B.; McGettigan, J.; Khetan, S.; Segall, N.; Solis, J.; Brosz, A.; Fierro, C.; Schwartz, H.; Neuzil, K.; Corey, L.; Gilbert, P.; Janes, H.; Follmann, D.; Marovich, M.; Mascola, J.; Polakowski, L.; Ledgerwood, J.; Graham, B. S.; Bennett, H.; Pajon, R.; Knightly, C.; Leav, B.; Deng, W.; Zhou, H.; Han, S.; Ivarsson, M.; Miller, J.; Zaks, T. Efficacy and Safety of themRNA-1273 SARS-CoV-2 Vaccine. N. Engl. J. Med. 2021, 384, 403–416. https://doi.org/10.1056/nejmoa2035389.Search in Google Scholar PubMed PubMed Central

2. Polack, F.; Thomas, S. J.; Kitchin, N.; Absalon, J.; Gurtman, A.; Lockhart, S.; Perez, J. L.; Marc, G. P.; Moreira, E. D.; Zerbini, C.; Bailey, R.; Swanson, K. A.; Roychoudhury, S.; Koury, K.; Li, P.; Kalina, W. V.; Cooper, D.; Frenck, R. W.; Hammitt, L. L.; Türeci, Ö.; Nell, H.; Schaefer, A.; Ünal, S.; Tresnan, D. B.; Mather, S.; Dormitzer, P. R.; Şahin, U.; Jansen, K. U.; Gruber, W. C. Safety and Efficacy of the BNT162b2 mRNA Covid-19 Vaccine. N. Engl. J. Med. 2020, 383, 2603–2615. https://doi.org/10.1056/nejmoa2034577.Search in Google Scholar

3. Yang, T.; Li, C.; Wang, X.; Zhao, D.; Zhang, M.; Cao, H.; Liang, Z.; Xiao, H.; Liang, X.-J.; Weng, Y.; Huanga, Y. Efficient Hepatic Delivery and Protein Expression Enabled by Optimized mRNA and Ionizable Lipid Nanoparticle. Bioact. Mater. 2020, 5, 1053–1061. https://doi.org/10.1016%2Fj.bioactmat.2020.07.003.10.1016/j.bioactmat.2020.07.003Search in Google Scholar PubMed PubMed Central

4. Kiaie, S. H.; Zolbanin, N. M.; Ahmadi, A.; Bagherifar, R.; Valizadeh, H.; Kashanchi, F.; Jafari, R. Recent Advances in mRNA-LNP Therapeutics: Immunological and Pharmacological Aspects. J. Nanobiotechnol. 2022, 20, 276. https://doi.org/10.1186/s12951-022-01478-7.Search in Google Scholar PubMed PubMed Central

5. Allen-Redpath, K.; Aldrovandi, M.; Lauder, S. N.; Gketsopoulou, A.; Tyrrell, V. J.; Slatter, D. A.; Andrews, R.; Watkins, W. J.; Atkinson, G.; McNeill, E.; Gilfedder, A.; Protty, M.; Burston, J.; Johnson, S. R. C.; Rodrigues, P. R. S.; Jones, D. O.; Lee, R.; Handa, A.; Channon, K.; Obaji, S.; Alvarez-Jarreta, J.; Krönke, G.; Ackermann, J.; Jenkins, P. V.; Collins, P. W.; O’Donnell, V. B. Phospholipid Membranes Drive Abdominal Aortic Aneurysm Development through Stimulating Coagulation Factor Activity. Proc. Natl. Acad. Sci. U.S.A. 2019, 16, 8038–8047. https://doi.org/10.1073/pnas.1814409116.Search in Google Scholar PubMed PubMed Central

6. Kedmi, R.; Ben-Arie, N.; Peer, D. The Systemic Toxicity of Positively Charged Lipid Nanoparticles and the Role of Toll-like Receptor 4 in Immune Activation. Biomaterials 2010, 31, 6867–6875. https://doi.org/10.1016/j.biomaterials.2010.05.027.Search in Google Scholar PubMed

7. Jayaraman, M.; Ansell, S. M.; Mui, B. L.; Tam, Y. K.; Chen, J.; Du, X.; Butler, D.; Eltepu, L.; Matsuda, S.; Narayanannair, J. K.; Rajeev, K. G.; Hafez, I. M.; Akinc, A.; Maier, M. A.; Tracy, M. A.; Cullis, P. R.; Madden, T. D.; Manoharan, M.; Hope, M. J. Maximizing the Potency of siRNA Lipid Nanoparticles for Hepatic Gene Silencing in V, Maximizing the Potency of siRNA Lipid Nanoparticles for Hepatic Gene Silencing In Vivo. Angew. Chem. 2012, 124, 8657–8661. https://doi.org/10.1002/ange.201203263.Search in Google Scholar

8. Sabnis, S.; Kumarasinghe, E. S.; Salerno, T.; Mihai, C.; Ketova, T.; Senn, J. J.; Lynn, A.; Bulychev, A.; McFadyen, I.; Chan, J.; Almarsson, Ö.; Stanton, M. G.; Benenato, K. E. A Novel Amino Lipid Series for mRNA Delivery: Improved Endosomal Escape and Sustained Pharmacology and Safety in Non-human Primates. Mol. Ther. 2018, 26, 1509–1519. https://doi.org/10.1016/j.ymthe.2018.03.010.Search in Google Scholar PubMed PubMed Central

9. Kulig, W.; Cwiklik, L.; Jurkiewicz, P.; Rog, T.; Vattulainen, I. Cholesterol Oxidation Products and Their Biological Importance. Chem. Phys. Lipids 2016, 199, 144–160. https://doi.org/10.1016/j.chemphyslip.2016.03.001.Search in Google Scholar PubMed

10. Kulkarni, J. A.; Myhre, J. L.; Chen, S.; Tam, Y. Y. C.; Danescu, A.; Richman, J. M.; Cullis, P. R. Design of Lipid Nanoparticles for In Vitro and In Vivo Delivery of Plasmid DNA. Nanomed.-Nanotechnol. Biol. Med. 2017, 13, 1377–1387. https://doi.org/10.1016/j.nano.2016.12.014.Search in Google Scholar PubMed

11. Szlasa, W.; Zendran, I.; Zalesinska, A.; Tarek, M.; Kulbacka, J. Lipid Composition of the Cancer Cell Membrane. J. Bioenerg. Biomembr. 2020, 52 (2), 321–342. https://doi.org/10.1007/s10863-020-09846-4.Search in Google Scholar PubMed PubMed Central

12. Zhi, D.; Zhang, S.; Wang, B.; Zhao, Y.; Yang, B.; Yu, S. Transfection Efficiency of Cationic Lipids with Different Hydrophobic Domains in Gene Delivery. Bioconjugate Chem. 2010, 21 (4), 563–577. https://doi.org/10.1021/bc900393r.Search in Google Scholar PubMed

13. Qiu, M.; Glass, Z.; Chen, J.; Haas, M.; Jin, X.; Zhao, X.; Rui, X.; Ye, Z.; Li, Y.; Zhang, F.; Xu, Q. Lipid Nanoparticle-Mediated Codelivery of Cas9 mRNA and Single-Guide RNA Achieves Liver-specific In Vivo Genome Editing of Angptl3. Proc. Natl. Acad. Sci. 2021, 118 (10). https://doi.org/10.1073/pnas.2020401118.Search in Google Scholar PubMed PubMed Central

14. Tam, A.; Kulkarni, J.; An, K.; Li, L.; Dorscheid, D. R.; Singhera, G. K.; Bernatchez, P.; Reid, G. S. D.; Chan, K. Y. T.; Witzigmann, D.; Cullis, P.; Lim, C. Lipid Nanoparticle Formulations for Optimal RNA-Based Topical Delivery to Murine Airways. Eur. J. Pharm. Sci. 2022, 176, 106234. https://doi.org/10.1016/j.ejps.2022.106234.Search in Google Scholar PubMed

15. van Meer, G.; Voelker, D. R.; Feigenson, G. W. Membrane Lipids: where They Are and How They Behave. Nat. Rev. Mol. Cell Biol. 2008, 9 (2), 112–124. https://doi.org/10.1038/nrm2330.Search in Google Scholar PubMed PubMed Central

16. Martin, D. D.; Robbins, M. E.; Spector, A. A.; Wen, B. C.; Hussey, D. H. The Fatty Acid Composition of Human Gliomas Differs from that Found in Nonmalignant Brain Tissue. Lipids 1996, 31, 1283–1288. https://doi.org/10.1007/bf02587914.Search in Google Scholar

17. Fu, Y.; He, Y.; Phan, K.; Bhatia, S.; Pickford, R.; Wu, P.; Dzamko, N.; Halliday, G. M.; Kim, W. S. Increased Unsaturated Lipids Underlie Lipid Peroxidation in Synucleinopathy brain. Acta Neuropathol. Commun. 2022, 10, 165. https://doi.org/10.1186/s40478-022-01469-7.Search in Google Scholar PubMed PubMed Central

18. Gennis, R. B. Biomembranes: Molecular Structure and Function; Springer: New York, 1989.10.1007/978-1-4757-2065-5Search in Google Scholar

19. Degors, I. M. S.; Wang, C.; Rehman, Z. U.; Zuhorn, I. S. Carriers Break Barriers in Drug Delivery: Endocytosis and Endosomal Escape of Gene Delivery Vectors. Acc. Chem. Res. 2019, 52, 1750–1760. https://doi.org/10.1021/acs.accounts.9b00177.D.Search in Google Scholar

20. Park, Y.; Park, J.-W. Preparation, Characterization, and Stability of Lipid Nanoparticles Including Unsaturated Lipids. Tenside Surfact. Det. 2023, 60, 594–598. https://doi.org/10.1515/tsd-2023-2544.Search in Google Scholar

21. Park, J.-W. Phase Effect of Mixed-Phospholipid Layer on Phospholipase D Reaction-Induced-Vesicle Rupture. Colloids Surf. B: Biointerfaces 2012, 97, 207–210. https://doi.org/10.1016/j.colsurfb.2012.04.034.Search in Google Scholar PubMed

22. Maugeri, M.; Nawaz, M.; Papadimitriou, A.; Angerfors, A.; Camponeschi, A.; Na, M.; Hölttä, M.; Skantze, P.; Johansson, S.; Sundqvist, M.; Lindquist, J.; Kjellman, T.; Mårtensson, I.-L.; Jin, T.; Sunnerhagen, P.; Östman, S.; Lindfors, L.; Valadi, H. Linkage between Endosomal Escape of LNP-mRNA and Loading into EVs for Transport to Other Cells. Nat. Comm. 2019, 10, 4333. https://doi.org/10.1038/s41467-019-12275-6.Search in Google Scholar PubMed PubMed Central

23. Schoenmaker, L.; Witzigmann, D.; Kulkarni, J. A.; Verbeke, R.; Kersten, G.; Jiskoot, W.; Crommelin, D. J. A. mRNA-lipid Nanoparticle COVID-19 Vaccines: Structure and Stability. Int. J. Pharmaceutics 2021, 601, 120586. https://doi.org/10.1016/j.ijpharm.2021.120586.Search in Google Scholar PubMed PubMed Central

24. Jones, L. J.; Yue, S. T.; Cheung, C.-Y.; Singer, V. L. RNA Quantitation by Fluorescence-Based Solution Assay: RiboGreen Reagent Characterization. Anal. Biochem. 1998, 265, 368–374. https://doi.org/10.1006/abio.1998.2914.Search in Google Scholar PubMed

25. Wisse, E.; Jacobs, F.; Topal, B.; Frederik, P.; De Geest, B. The Size of Endothelial Fenestrae in Human Liver Sinusoids: Implications for Hepatocyte-Directed Gene Transfer. Gene Ther. 2008, 15, 1193–1199. https://doi.org/10.1038/gt.2008.60.Search in Google Scholar PubMed

26. Kano, K.; Fendler, J. H. Pyranine as a Sensitive pH Probe for Liposome Interiors and Surfaces. pH Gradients across Phospholipid Vesicles. Biochim. Biophys. Acta Biomembr. 1978, 509, 289–299. https://doi.org/10.1016/0005-2736(78)90048-2.Search in Google Scholar PubMed

27. Clement, N. R.; Gould, J. M. Pyranine (8-Hydroxy-1,3,6-Pyrenetrisulfonate) as a Probe of Internal Aqueous Hydrogen Ion Concentration in Phospholipid Vesicles. Biochemistry 1981, 20, 1534–1538. https://doi.org/10.1021/bi00509a019.Search in Google Scholar PubMed

28. Cevc, G.; Marsh, D. Phospholipid Bilayers: Physical Principles and Methods; John Wiley and Sons: New York, 1987.Search in Google Scholar

29. Park, J.-W.; Ahn, D. J. Temperature Effect on Nanometer-Scale Physical Properties of Mixed Phospholipid Monolayers. Colloids Surf. B: Biointerfaces 2008, 62, 157–161. https://doi.org/10.1016/j.colsurfb.2007.09.020.Search in Google Scholar PubMed

Received: 2024-07-03
Accepted: 2024-09-25
Published Online: 2024-10-16
Published in Print: 2024-11-26

© 2024 the author(s), published by De Gruyter, Berlin/Boston

This work is licensed under the Creative Commons Attribution 4.0 International License.

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https://doi.org/10.1515/tsd-2024-2610
Keywords for this article
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  9. Triethanolamine-modified poloxamer temperature-sensitive hair care gel for intelligent controlled release of sodium thioglycolat
  10. Drug Delivery Systems
  11. Formulation of Felodipine lipid nanoparticle-loaded oral fast-dissolving films
  12. Applications
  13. The effect of addition of sodium hexadecyl sulfate on the performance properties of lauramidopropyl betaine
  14. The role of presence of wormlike micelles of ionic surfactants on the sterilization efficiency of sodium hypochlorite
  15. Synthesis
  16. Synthesis and performance study of a new surfactant with corrosion inhibition function
  17. Synthesis and surfactant properties of sulfonate Gemini surfactants
  18. Short Communication
  19. Preparation and stability of lipid nanoparticles excluding 1,2-distearoyl-sn-glycero-3-phosphocholine
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