Quantifying nanotherapeutic penetration using a hydrogel-based microsystem as a new 3D <i>in vitro</i> platform

Goodarzi, Saba; Prunet, Audrey; Rossetti, Fabien; Bort, Guillaume; Tillement, Olivier; Porcel, Erika; Lacombe, S.; Wu, Ting-Di; Guerquin‐Kern, Jean‐Luc; Delanoë-Ayari, Hélène; Lux, François; Rivière, Charlotte

doi:10.1039/d1lc00192b

Cited by 17 publications

(22 citation statements)

References 66 publications

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“… 27 Therefore, a satisfactory penetration depth and uniform NP diffusion are usually required to maximize the therapeutic effects. We used a 3D MCTS model that simulates the complex situation of the tumor site 28 to evaluate the ability of the IGP@P NPs to penetrate into the tumor with the assistance of LIFU irradiation. In the absence of LIFU irradiation, we found that the red fluorescent DiI-labeled IGP@P NPs were mainly distributed on the surface of the 3D MCTSs, and only a small amount of the NPs penetrated into the model.…”

Section: Resultsmentioning

confidence: 99%

Theranostic Nanoplatform with Sequential SDT and ADV Effects in Response to Well-Programmed LIFU Irradiation for Cervical Cancer

Zhou¹,

Hou²,

Liu³

et al. 2021

IJN

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Background: Some patients with cervical cancer have the need to preserve fertility; therefore, a minimally invasive treatment option that can effectively inactivate tumors in these patients is necessary. Methods: In this paper, we designed and prepared nanoparticles (NPs) carrying IR780 and perfluorohexane (PFH) and characterized their properties. We focused on the promotion of programmed low-intensity focused ultrasound (LIFU) irradiation on the penetration and treatment of cervical cancer. First we used penetration-enhancing LIFU irradiation to promote the penetration of the NPs into 3D multicellular tumor spheroids (MCTSs) and tumors in tumor-bearing nude mice. Then we used re-therapeutic LIFU irradiation to achieve antitumor effects in vitro and in vivo. Photoacoustic (PA) and magnetic resonance (MR) imaging were used to monitor and evaluate the targeting and therapeutic effects of these NPs on tumor tissues. Results:The NPs prepared in this paper exhibited high affinity for HeLa cells, and can selectively achieve mitochondrial localization in the cell due to IR780 assistance. The penetration-enhancing LIFU irradiation have the ability to promote the penetration of the NPs into cervical cancer models in vivo and in vitro. Under LIFU irradiation, the cytotoxic reactive oxygen species (ROS) produced by IR780 during the first half of the re-therapeutic LIFU irradiation and the physical acoustic droplet vaporization (ADV) effect after PFH phase transition during the second half of the re-therapeutic LIFU irradiation can achieve synergistic minimally invasive treatment of tumors, which can be visualized and evaluated by PA and MR imaging in vivo. Conclusion:Well-programmed LIFU irradiation can promote NP penetration into deep tumor tissue and achieve antitumor effects simultaneously. Linking ROS + ADV effects can induce cell coagulation necrosis and lead to a comprehensive, long-term impact on tumor tissue, providing a conceptual theranostic nanoplatform for cervical cancer.

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Section: Resultsmentioning

confidence: 99%

Theranostic Nanoplatform with Sequential SDT and ADV Effects in Response to Well-Programmed LIFU Irradiation for Cervical Cancer

Zhou¹,

Hou²,

Liu³

et al. 2021

IJN

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“…While these particles have attracted great interest, the complexity of their design and release profiles has impeded their translation into clinical use [ 15 , 30 ]. Methodologies that can improve our understanding of how the biophysical properties of the nanoparticles impact their cellular uptake have potential to enrich existing methods for nanoparticle evaluation, to collectively advance nanoparticle research [ 5 , 31 , 32 , 33 , 34 , 35 ].…”

Section: Discussionmentioning

confidence: 99%

Application of Rapid Fluorescence Lifetime Imaging Microscopy (RapidFLIM) to Examine Dynamics of Nanoparticle Uptake in Live Cells

Ahmed-Cox

Macmillan

Pandžić

et al. 2022

Cells

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A key challenge in nanomedicine stems from the continued need for a systematic understanding of the delivery of nanoparticles in live cells. Complexities in delivery are often influenced by the biophysical characteristics of nanoparticles, where even subtle changes to nanoparticle designs can alter cellular uptake, transport and activity. Close examination of these processes, especially with imaging, offers important insights that can aid in future nanoparticle design or translation. Rapid fluorescence lifetime imaging microscopy (RapidFLIM) is a potentially valuable technology for examining intracellular mechanisms of nanoparticle delivery by directly correlating visual data with changes in the biological environment. To date, applications for this technology in nanoparticle research have not been explored. A PicoQuant RapidFLIM system was used together with commercial silica nanoparticles to follow particle uptake in glioblastoma cells. Importantly, RapidFLIM imaging showed significantly improved image acquisition speeds over traditional FLIM, which enabled the tracking of nanoparticle uptake into subcellular compartments. We determined mean lifetime changes and used this to delineate significant changes in nanoparticle lifetimes (>0.39 ns), which showed clustering of these tracks proximal to both extracellular and nuclear membrane boundaries. These findings demonstrate the ability of RapidFLIM to track, localize and quantify changes in single nanoparticle fluorescence lifetimes and highlight RapidFLIM as a valuable tool for multiparameter visualization and analysis of nanoparticle molecular dynamics in live cells.

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“…Manufacturing process has already been detailed in Goodarzi et al 23,24 . Briefly, 200µm diameter micro-wells were molded with 2% agarose solution over PDMS counter-moulds.…”

Section: Microfabricated Devicementioning

confidence: 99%

“…21 The proportion of CSCs within a given cell population derived from either tumor samples or in vitro cell cultures can therefore be evaluated by functional tests that are based on their capacity to survive as single cell and/or reconstitute a population of cancer cells in vitro 22 . Therefore, in order to provide time-resolved characterization of CSCs populations at individual cell level, we have generated data sets from LFBM time-lapses of patient-derived brain CSCs in suspension, cultured in micro-wells of a microfabricated chip recapitulating in this way classical stem cell culture conditions (Figure 1) 23,24 . This system can generate high-throughput images of micro-wells ( circa 15 micro-wells per second) and requires fast and accurate analysis of micro-wells content to assess cell fate.…”

Section: Introductionmentioning

confidence: 99%

Machine learning-based detection of label-free cancer stem-like cell fate

Chambost

Berabez

Cochet‐Escartin

et al. 2022

Preprint

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Most imaging methods rely on labelling biological samples in order to provide specific and easily detectable features. However, label-free imaging is a non-invasive and non-toxic alternative that requires accurate image analysis algorithm based on cell morphology. Such analysis has to deal with a high image variability while fewer features are extractable, so far, fast analysis of label-free brightfield microscopy (LFBM) images remains a challenging task. With the development of microfabricated devices during the last decades, high throughput image generation makes it possible to use machine learning-based algorithms in order to analyse LFBM images. Fast algorithms are also crucially needed to analyze high throughput experiments. In this paper, we provide a data-driven study in order to assess the complexity of LFBM time-lapses monitoring isolated cancer stem-like cells (CSCs) fate in non-adherent conditions. We combined for the first time individual cell fate and cell state temporality analysis in a unique algorithm. Several image analysis algorithms of increasing processing capacities were tested: a classical computer vision algorithm (CCVA), a shallow learning-based algorithm (SLBA) and a deep learning-based algorithm (DLBA). We show that our optimized DLBA has by far the best accuracy compared to CCVA and SLBA, is at least as accurate as other state-of-the-art DLBAs while being faster. With this study, we demonstrate that optimizing our DLBA accordingly to the image analysis problem can overall provide better results than pretrained models. Such a fast and accurate DLBA is therefore compatible with the generation of high throughput data and opens the route for on-the-fly analysis of CSC fate from LFBM time-lapses.

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Quantifying nanotherapeutic penetration using a hydrogel-based microsystem as a new 3D in vitro platform

Abstract: The huge gap between 2D in vitro assays used for drug screening, and the in vivo 3D-physiological environment hampered reliable predictions for the route and accumulation of nanotherapeutics in vivo....

Cited by 17 publications

References 66 publications

Theranostic Nanoplatform with Sequential SDT and ADV Effects in Response to Well-Programmed LIFU Irradiation for Cervical Cancer

Theranostic Nanoplatform with Sequential SDT and ADV Effects in Response to Well-Programmed LIFU Irradiation for Cervical Cancer

Application of Rapid Fluorescence Lifetime Imaging Microscopy (RapidFLIM) to Examine Dynamics of Nanoparticle Uptake in Live Cells

Machine learning-based detection of label-free cancer stem-like cell fate

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