Specific labelling of phagosome-derived vesicles in macrophages with a membrane dye delivered with microfabricated microparticles

Cheng, Wen‐Hao; Kim, Sundol; Zivkovic, Sandra; Chung, Hoyong; Ren, Yi; Guan, Jingjiao

doi:10.1016/j.actbio.2022.01.028

Cited by 4 publications

(12 citation statements)

References 51 publications

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“…To fluorescently label the microparticles, red fluorescent sulfate-modified polystyrene nanoparticles with a diameter of 100 nm were dispersed in the microparticles. A microcontact-printing method that we previously developed was modified to fabricate the microparticles. , Specifically, a thin film of PNIPAM containing the nanoparticles was spin-coated onto a PDMS stamp carrying an array of 5 μm-radius pillars using an aqueous solution of PNIPAM and the nanoparticles. The resultant film on the pillars was then transferred onto a glass coverslip via a conformal contact, generating an array of circular microparticles over a centimeter-wide printing area.…”

Section: Resultsmentioning

confidence: 99%

“…To further confirm that phagosomes were ruptured, we labeled the microparticle-containing phagosomes with LysoView 488 dye, which can accumulate in acidic phagosomes and emit green fluorescence . After dissolving microparticles that were not phagocytosed with 22 °C PBS, we found that some phagocytosed microparticles, indicated by strong red fluorescence, exhibited the strong green fluorescence of LysoView 488.…”

Section: Resultsmentioning

confidence: 99%

“…Poly(N-isopropylacrylamide-co-fluorescein-O-acrylate) (PNIPAM-fluorescein hereafter) was synthesized and characterized as described previously. 14 2.3. Cell Culture.…”

Section: Synthesis and Characterization Of Poly(n-isopropylacrylamide...mentioning

confidence: 99%

“…PDMS stamps were prepared as we previously did. 14 The stamps carried 5 μm-radius circular pillars in a hexagonal lattice with a pillar-to-pillar distance of 30 μm. The height of the pillars was ∼6 μm.…”

Section: Synthesis and Characterization Of Poly(n-isopropylacrylamide...mentioning

confidence: 99%

“…The osmotic pressure generated by PNIPAM has been extensively studied both experimentally and theoretically, − allowing us to use a well-established polymer physics theory to calculate the osmotic pressure generated by a PNIPAM microparticle in a phagosome. Third, it utilizes a microfabrication technique to produce the PNIPAM microparticles with monodispersed geometry and composition. , Use of such microparticles enables a reliable interpretation of experimental results.…”

Section: Introductionmentioning

confidence: 99%

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Osmotically Rupturing Phagosomes in Macrophages Using PNIPAM Microparticles

Cheng

Fukuda

Kim

et al. 2023

ACS Appl. Mater. Interfaces

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The rupture of macrophage phagosomes has been implicated in various human diseases and plays a critical role in immunity. However, the mechanisms underlying this process are complex and not yet fully understood. This study describes the development of a robust engineering method for rupturing phagosomes based on a welldefined mechanism. The method utilizes microfabricated microparticles composed of uncrosslinked linear poly(N-isopropylacrylamide) (PNIPAM) as phagocytic objects. These microparticles are internalized into phagosomes at 37 °C. By exposing the cells to a cold shock at 0 °C, the vast majority of the microparticle-containing phagosomes rupture. The percentage of phagosomal rupture decreases with the increase of the cold-shock temperature. The osmotic pressure in the phagosomes and the tension in the phagosomal membrane are calculated using the Flory−Huggins theory and the Young−Laplace equation. The modeling results indicate that the osmotic pressure generated by dissolved microparticles is probably responsible for phagosomal rupture, are consistent with the experimentally observed dependence of phagosomal rupture on the cold-shock temperature, and suggest the existence of a cellular mechanism for resisting phagosomal rupture. Moreover, the effects of various factors including hypotonic shock, chloroquine, tetrandrine, colchicine, and L-leucyl-L-leucine O-methyl ester (LLOMe) on phagosomal rupture have been studied with this method. The results further support that the osmotic pressure generated by the dissolved microparticles causes phagosomal rupture and demonstrated usefulness of this method for studying phagosomal rupture. This method can be further developed, ultimately leading to a deeper understanding of phagosomal rupture.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

“…Poly(N-isopropylacrylamide-co-fluorescein-O-acrylate) (PNIPAM-fluorescein hereafter) was synthesized and characterized as described previously. 14 2.3. Cell Culture.…”

Section: Synthesis and Characterization Of Poly(n-isopropylacrylamide...mentioning

confidence: 99%

Section: Synthesis and Characterization Of Poly(n-isopropylacrylamide...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Osmotically Rupturing Phagosomes in Macrophages Using PNIPAM Microparticles

Cheng

Fukuda

Kim

et al. 2023

ACS Appl. Mater. Interfaces

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Features of Anisotropic Drug Delivery Systems

Kudryavtseva,

Sukhorukov

2024

Advanced Materials

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Natural materials are anisotropic. Delivery systems occurring in nature, such as viruses, blood cells, pollen, and many others, do have anisotropy, while delivery systems made artificially are mostly isotropic. There is apparent complexity in engineering anisotropic particles or capsules with micron and submicron sizes. Nevertheless, some promising examples of how to fabricate particles with anisotropic shapes or having anisotropic chemical and/or physical properties have been developed. Anisotropy of particles, once they face biological systems, influences their behaviour. Internalisation by the cells, flow in the bloodstream, biodistribution over organs and tissues, directed release, and toxicity of particles regardless of the same chemistry are all reported to be factors of anisotropy of delivery systems. Here, we review the current methods to introduce anisotropy to particles or capsules, including loading with various therapeutic cargo, variable physical properties primarily by anisotropic magnetic properties, controlling directional motion, and making Janus particles. The advantages of combining different anisotropy in one entity for delivery and common problems and limitations for fabrication are under discussion.This article is protected by copyright. All rights reserved

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