Dissociation of fluorescently labeled lipids from liposomes in biological environments challenges the interpretation of uptake studies

Münter, Rasmus; Kristensen, Kasper; Pedersbæk, Dennis; Larsen, Jørgen Hannover; Simonsen, Jens B.

doi:10.1039/c8nr07755j

Cited by 74 publications

(73 citation statements)

References 37 publications

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“…We confirmed that DiI was stably incorporated into the lipid-based NPs (Supplementary Information Fig. S1), as consistent with previous studies using similar types of fluorophores 15,18 . We emphasize that all the lipid-based NPs were prepared with equal mol% of fluorophores (0.5% DiI), and that the concentration of the NPs was adjusted such that the amount of fluorophores (based on absorbance) was equal in each sample (Supplementary Information Fig.…”

Section: Preparation and Characterization Of The Npssupporting

confidence: 91%

“…The used gains are listed for each NP. The method is similar to what have been described elsewhere 15,18,58 . The DiI seems relatively stably incorporated.…”

Section: Image Analysismentioning

confidence: 99%

“…Briefly, liposomes (Lip and PEG-Lip) are some of the most commonly used drug carriers. They are spherical vesicles comprised of a lipid bilayer and an aqueous core, while surface grafting of polyethylene glycol (PEG) is frequently used to increase their blood circulation time 2,15,16 . LPX are complexes of nucleic acids that are stabilized by cationic lipids 13 , and are usually used for gene-targeting applications 2 .…”

Section: Introductionmentioning

confidence: 99%

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Head-to-head comparison of the penetration efficiency of lipid-based nanoparticles in a 3D tumor spheroid model

Niora¹,

Pedersbæk²,

Lassen³

et al. 2020

Preprint

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<p>Most tumor-targeted drug delivery systems must overcome a large variety of physiological barriers before reaching the tumor site and diffuse through the tight network of tumor cells. Many studies focus on optimizing the first part, the accumulation of drug carriers at the tumor site, ignoring the penetration efficiency, i.e., a measure of the ability of a drug delivery system to overcome tumor surface adherence and uptake. We used 3D tumor spheroids in combination with light-sheet fluorescence microscopy in a head-to-head comparison of a variety of commonly used lipid-based nanoparticles, including liposomes, PEGylated liposomes, lipoplexes and reconstituted high-density lipoproteins (rHDL). Whilst PEGylation of liposomes only had minor effects on the penetration efficiency, we show that lipoplexes mainly associated to the periphery of tumor spheroids, possibly due to their positive surface charge leading to fusion with the cells at the spheroid surface or aggregation. Surprisingly, the rHDL showed significantly higher penetration efficiency and high accumulation inside the spheroid. While these findings indeed could be relevant when designing novel drug delivery systems based on lipid-based nanoparticles, we stress that the used platform and detailed image analysis is a versatile tool for in vitro studies of the penetration efficiency of nanoparticles in tumors.</p><div><br></div>

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Section: Preparation and Characterization Of The Npssupporting

confidence: 91%

“…The used gains are listed for each NP. The method is similar to what have been described elsewhere 15,18,58 . The DiI seems relatively stably incorporated.…”

Section: Image Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Head-to-head comparison of the penetration efficiency of lipid-based nanoparticles in a 3D tumor spheroid model

Niora¹,

Pedersbæk²,

Lassen³

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…However, several studies have demonstrated that up to 75% of such dyes dissociated from vesicles, when incubated in plasma. 53,54 In addition, these commonly used dyes can spontaneously form EVlike particles. 54 Radiolabeling of exosomes presents an alternative imaging strategy; however, few published reports have investigated the biodistribution of postinserted radiolabeled EVs.…”

Section: Mex Biodistributionmentioning

confidence: 99%

“…53,54 In addition, these commonly used dyes can spontaneously form EVlike particles. 54 Radiolabeling of exosomes presents an alternative imaging strategy; however, few published reports have investigated the biodistribution of postinserted radiolabeled EVs. 55,56 An alternative labeling method of engineering MEX with Cre-recombinase in Lox reporter mice has been reported, but it remains unclear whether such engineering methods modulate the functions of the resulting vesicles.…”

Section: Mex Biodistributionmentioning

confidence: 99%

Preclinical translation of exosomes derived from mesenchymal stem/stromal cells

et al. 2019

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Exosomes are nanovesicles secreted by virtually all cells. Exosomes mediate the horizontal transfer of various macromolecules previously believed to be cell-autonomous in nature, including nonsecretory proteins, various classes of RNA, metabolites, and lipid membrane-associated factors. Exosomes derived from mesenchymal stem/stromal cells (MSCs) appear to be particularly beneficial for enhancing recovery in various models of disease. To date, there have been more than 200 preclinical studies of exosome-based therapies in a number of different animal models. Despite a growing number of studies reporting the therapeutic properties of MSC-derived exosomes, their underlying mechanism of action, pharmacokinetics, and scalable manufacturing remain largely outstanding questions. Here, we review the global trends associated with preclinical development of MSC-derived exosome-based therapies, including immunogenicity, source of exosomes, isolation methods, biodistribution, and disease categories tested to date. Although the in vivo data assessing the therapeutic properties of MSC-exosomes published to date are promising, several outstanding questions remain to be answered that warrant further preclinical investigation.

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In Vivo Quantification of Nanoparticle Association with Immune Cell Subsets in Blood

Ong

Rose

Johnston

2021

Adv Healthcare Materials

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Nanoparticles offer great promise for more effective drug delivery. However, their particulate nature typically results in rapid systemic clearance by immune cells in blood. Currently, to understand these interactions, nanoparticle association is probed ex vivo with whole blood. While ex vivo assays give important information about the relative cell association, they do not consider changes in immune cell homeostasis or the complex mixing behavior that occurs in vivo. To address this, a nanoparticle in vivo immune‐cell association assay is developed to study the in vivo association of unmodified and poly(ethylene glycol) modified liposomes with immune cells, and compared this to the ex vivo association in static whole blood. In vivo, it is observed that neutrophils play a significantly greater role in nanoparticle binding than suggested by ex vivo assays. The increased influence of neutrophils in vivo is largely due to a significant increase in number of circulating neutrophils after intravenous injection. Conversely, the number of circulating monocytes significantly decreased after intravenous injection, leading to significantly less total association of liposomes to monocytes compared to ex vivo. This novel in vivo immune cell binding assay sheds new light on the fate of nanoparticles following intravenous delivery.

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Dissociation of fluorescently labeled lipids from liposomes in biological environments challenges the interpretation of uptake studies

Abstract: Many commonly used fluorescently labeled lipids were found to dissociate from liposomes during incubation in blood plasma.

Cited by 74 publications

References 37 publications

Head-to-head comparison of the penetration efficiency of lipid-based nanoparticles in a 3D tumor spheroid model

Head-to-head comparison of the penetration efficiency of lipid-based nanoparticles in a 3D tumor spheroid model

Preclinical translation of exosomes derived from mesenchymal stem/stromal cells

In Vivo Quantification of Nanoparticle Association with Immune Cell Subsets in Blood

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