Direct Observation of Endocytosis Dynamics of Anti-ErbB Modified Single Nanocargoes

Ge, Feng; Du, Yi; He, Yan

doi:10.1021/acsnano.2c00184

Cited by 13 publications

(13 citation statements)

References 48 publications

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“…All three forms of dynamics were observed regardless of whether the particle ultimately desorbed from the cell or whether it was internalized. Previous studies have also reported both confined motion, 31,40,45,46 as well as particles that explore the membrane prior to particle internalization. 30,31,45,46,70 Once within the cell, particles also displayed various dynamics: Many particles disappeared immediately after internalization (Figure 5a).…”

Section: T H I S C O N T E N T Imentioning

confidence: 99%

Rapid Internalization of Nanoparticles by Human Cells at the Single Particle Level

Richards,

Burgers,

Vlijm

et al. 2023

ACS Nano

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Nanoparticle uptake by cells has been studied for applications both in nanomedicine and in nanosafety. While the majority of studies have focused on the biological mechanisms underlying particle internalization, less attention has been given to questions of a more quantitative nature, such as how many nanoparticles enter cells and how rapidly they do so. To address this, we exposed human embryonic kidney cells to 40–200 nm carboxylated polystyrene nanoparticles and the particles were observed by live-cell confocal and super-resolution stimulated emission depletion fluorescence microscopy. How long a particle remained at the cell membrane after adsorbing onto it was monitored, distinguishing whether the particle ultimately desorbed again or was internalized by the cell. We found that the majority of particles desorb, but interestingly, most of the particles that are internalized do so within seconds, independently of particle size. As this is faster than typical endocytic mechanisms, we interpret this observation as the particles entering via an endocytic event that is already taking place (as opposed to directly triggering their own uptake) or possibly via an as yet uncharacterized endocytic route. Aside from the rapidly internalizing particles, a minority of particles remain at the membrane for tens of seconds to minutes before desorbing or being internalized. We also followed particles after cell internalization, observing particles that appeared to exit the cell, sometimes as rapidly as within tens of seconds. Overall, our results provide quantitative information about nanoparticle cell internalization times and early trafficking.

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Section: T H I S C O N T E N T Imentioning

confidence: 99%

Rapid Internalization of Nanoparticles by Human Cells at the Single Particle Level

Richards,

Burgers,

Vlijm

et al. 2023

ACS Nano

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“…Unfortunately, a full understanding of the complicated interaction mechanisms, particularly the relation between interactions at different scales and the association between molecular diffusion and biological function, is still lacking. [36][37][38][39][40] The permeabilization of substances into cells is also controlled by lipids, which are the basic structural constituents of cell membranes. Instead of simple random motion, membrane lipids exhibit complex diffusion behaviors [41][42][43] that are governed by various interaction mechanisms, such as crowding and trapping, both between lipid molecules and between lipids and proteins.…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Transition between Different Diffusion Modes of Individual Lipids during the Membrane‐Specific Action of As‐CATH4 Peptides

Cheng

et al. 2023

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The cell membrane permeabilization ability of immune defense antimicrobial peptides (AMPs) is widely applied in biomedicine. Although the mechanisms of peptide–membrane interactions have been widely investigated, analyses at the molecular level are still lacking. Herein, the membrane‐specific action of a native AMP, As‐CATH4, is investigated using a single‐lipid tracking method in combination with live cell and model membrane assays conducted at different scales. The peptide–membrane interaction process is characterized by analyzing single‐lipid diffusion behaviors. As‐CATH4 exhibits potent antimicrobial activity through bacterial membrane permeabilization, with moderate cytotoxicity against mammalian cells. In‐plane diffusion analyses of individual lipids show that the lipid molecules exhibit non‐Gaussian and heterogeneous diffusion behaviors in both pristine and peptide‐treated membranes, which can be decomposed into two Gaussian subgroups corresponding to normal‐ and slow‐diffusive lipids. Assessment of the temporal evolution of these two diffusion modes of lipids reveal that the peptide action states of As‐CATH4 include surface binding, transmembrane defect formation, and dynamic equilibrium. The action mechanisms of As‐CATH4 at varying concentrations and against different membranes are distinguished. This work resolves the simultaneous mixed diffusion mechanisms of single lipids in biomimetic cell membranes, especially during dynamic membrane permeabilization by AMPs.

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“…We probe the particle transport with fine spatiotemporal details within different bacterial turbulences by using single-particle tracking (SPT) of plasmonic spherical gold nanoparticles (AuNPs, with a diameter of 90 nm) under dark-field microscopy. Previously, by using single plasmonic nanoparticles as tracers, we have illustrated the spatiotemporal heterogeneity of some important complex processes such as polymer gelation, liquid–liquid phase separation, and endocytosis of nanocargoes . With deteriorating habitat, the superdiffusion of the nanotracers, whose size is similar to that of the nanocargoes or vesicles secreted by bacteria, is pushed to the limit of ballistic diffusion thanks to the collective swarming behavior.…”

Section: Introductionmentioning

confidence: 99%

“…Previously, by using single plasmonic nanoparticles as tracers, we have illustrated the spatiotemporal heterogeneity of some important complex processes such as polymer gelation, 30 liquid−liquid phase separation, 31 and endocytosis of nanocargoes. 32 With deteriorating habitat, the superdiffusion of the nanotracers, whose size is similar to that of the nanocargoes or vesicles secreted by bacteria, is pushed to the limit of ballistic diffusion thanks to the collective swarming behavior. Surprisingly, we found that rather than lengthening of all individuals, a very low percentage of lengthened members within the population is sufficient to enhance the mobility of the crowd group, implying a possible adaptive strategy to resist the antibiotic.…”

Section: Introductionmentioning

confidence: 99%

Direct Observation of Nanotracer Transport in Swarming Bacteria during Antibiotic Adaptation

et al. 2023

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Microorganisms inevitably encounter environmental variations and thus need to develop necessary strategies to adjust the colonies for survival. Here, we use cooperating Serratia marcescens bacteria to reveal how the whole population responds to a gradually deteriorating habitat. When subjected to antibiotics with increasing doses, the swarming bacteria transform weak homogeneous turbulent flows to nematic jet flows with defects and vortices on a large scale, by which bacteria exploit these coherent flows to transfer material and/or information. We elucidate a complete view of detailed spatiotemporal transport behavior in such microscale active turbulence with single-nanoparticle tracking. The nanoparticles in these active flows are brought into the state with the up limit of superdiffusion by the bacterial collective response to the stronger antibiotic stimulation. Strikingly, we found that, under the strengthening stimulation from antibiotics, bacteria with only a small fraction of their community get elongated and facilitate the drastic turbulence transition and an enhanced superdiffusion. These findings imply a possible collective response mechanism against environmental variations.

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Direct Observation of Endocytosis Dynamics of Anti-ErbB Modified Single Nanocargoes

Cited by 13 publications

References 48 publications

Rapid Internalization of Nanoparticles by Human Cells at the Single Particle Level

Rapid Internalization of Nanoparticles by Human Cells at the Single Particle Level

Transition between Different Diffusion Modes of Individual Lipids during the Membrane‐Specific Action of As‐CATH4 Peptides

Direct Observation of Nanotracer Transport in Swarming Bacteria during Antibiotic Adaptation

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