Light sheet fluorescence microscopy versus confocal microscopy: in quest of a suitable tool to assess drug and nanomedicine penetration into multicellular tumor spheroids

Lazzari, Gianpiero; Vinciguerra, Daniele; Balasso, Anna; Nicolas, Valérie; Goudin, Nicolas; Garfa-Traoré, Meriem; Fehér, András; Dinnyés, András; Nicolas, Julien; Couvreur, Patrick; Mura, Simona

doi:10.1016/j.ejpb.2019.06.019

Cited by 64 publications

(63 citation statements)

References 63 publications

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“…Recently, a doxorubicin-loaded polymeric NP diffusion was tracked via confocal scanning microscopy in a complex pancreatic cancer 3D model made of cancer, endothelial, and fibroblast cells. Results showed that confocal scanning microscopy is limited in its ability to provide accurate deep penetration (>100 μm) data on the diffusion of small molecules due to the progressive loss of their fluorescence signal 37 . This problem may be addressed by utilizing different characterization instruments based on the specific light-matter properties needed.…”

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confidence: 99%

Tracking Gold Nanorods’ Interaction with Large 3D Pancreatic-Stromal Tumor Spheroids by Multimodal Imaging: Fluorescence, Photoacoustic, and Photothermal Microscopies

Darrigues

Nima

Nedosekin

et al. 2020

Sci Rep

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pancreatic cancer is one of the most complex types of cancers to detect, diagnose, and treat. However, the field of nanomedicine has strong potential to address such challenges. When evaluating the diffusion and penetration of theranostic nanoparticles, the extracellular matrix (ECM) is of crucial importance because it acts as a barrier to the tumor microenvironment. in the present study, the penetration of functionalized, fluorescent gold nanorods into large (>500 μm) multicellular 3D tissue spheroids was studied using a multimodal imaging approach. the spheroids were generated by coculturing pancreatic cancer cells and pancreatic stellate cells in multiple ratios to mimic variable tumor-stromal compositions and to investigate nanoparticle penetration. fluorescence live imaging, photothermal, and photoacoustic analysis were utilized to examine nanoparticle behavior in the spheroids. Uniquely, the nanorods are intrinsically photoacoustic and photothermal, enabling multiimaging detection even when fluorescence tracking is not possible or ideal. Nanomedicine for cancer treatments-the use of nanosized structures in cancer therapeutics and/or imaginghas been heavily investigated over the last two decades 1. Nano-sized delivery systems are useful theragnostic agents for enhanced tumor penetration, accumulation, and targeting potency. Studies of nanoparticle (NP)-based therapies have mainly focused on targeted cancer cell treatment but also include cancer stem cells 2 , stromal cancer microenvironment 3-5 , and/or cellular immune system 6,7. As a physical barrier that limits NP penetration and distribution, the extracellular matrix (ECM) is a crucial concern when evaluating the effects of nanoscale cancer therapies. Recent work showed that the ECM can be regulated by using gold NPs to interrupt the crosstalk between cancer cells and stellate cells, which reeducates the stellate cells 8,9. However, the development of nanosystems for cancer treatment has been hampered by the limitations of current in vitro models, which are generally based on two-dimensional (2D) cell cultures. These 2D models struggle to provide an accurate representation of the in vivo environment and its components, which include the dynamic tumor microenvironment (TME), cell heterogeneity, and nutrient and pH gradient interaction between cells and the ECM 10. The real TME is composed not only of cancer cells but also of others cells such as fibroblasts, cancer-associated fibroblasts (CAFs), stromal cells, myofibroblasts, endothelial cells, adipocytes, various immune cells, and extra-abundant compromised ECM. These heterocellular components can induce adaptive survival mechanisms of cancer such as treatment resistance, a leading cause of cancer-related mortality and one of the greatest challenges in cancer treatment 11. The cellular responses and cell signaling that take place in the TME often cannot be mimicked in a 2D in vitro model.

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confidence: 99%

Tracking Gold Nanorods’ Interaction with Large 3D Pancreatic-Stromal Tumor Spheroids by Multimodal Imaging: Fluorescence, Photoacoustic, and Photothermal Microscopies

Darrigues

Nima

Nedosekin

et al. 2020

Sci Rep

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“…Then the MTSs were incubated with DFMs@NR, nDFMs@NR, or SCMs@NR (20 mg mL À1 , NR) for 3 h, followed by incubation in the presence of 20 mM GSH for 24 h (see the ESI † for details), 22,43 aer which they were washed in phosphate buffer saline buffer and detected by light sheet uorescence microscopy (LSFM). 67 As compared in Fig. 3, with the nDFMs@NR, each equatorial section of A549 MTS showed low uorescence signals, revealing limited penetration.…”

Section: Long-distance Travel Of Seed Scms From Dfmsmentioning

confidence: 79%

Dandelion flower-like micelles

Yao

Zhu

et al. 2020

Chem. Sci.

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“… 23 − 25 Even though this simple system is grown from monocultures, it still preserves the essential features present in cancer tumors in vivo, i.e., rapid proliferation at the surface and slow metabolism, or necrosis, in the center of spheroids. 26 The majority of studies of NP distributions in tumor spheroids use tumor spheroid fixation, 27 − 35 often in combination with histological sectioning. 7 , 36 − 39 Nevertheless, there are also reports of NP penetration in live tumor spheroids using confocal-scanning microscopy.…”

Section: Introductionmentioning

confidence: 99%

“… 57 With this setup, we combine deep imaging of NPs with single-cell resolution of tumor spheroids of hundreds of micrometers. 27 , 58 …”

Section: Introductionmentioning

confidence: 99%

Head-to-Head Comparison of the Penetration Efficiency of Lipid-Based Nanoparticles into Tumor Spheroids

et al. 2020

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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 three-dimensional (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 are mainly associated with 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 the detailed image analysis are a versatile tool for in vitro studies of the penetration efficiency of nanoparticles in tumors.

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Light sheet fluorescence microscopy versus confocal microscopy: in quest of a suitable tool to assess drug and nanomedicine penetration into multicellular tumor spheroids

Cited by 64 publications

References 63 publications

Tracking Gold Nanorods’ Interaction with Large 3D Pancreatic-Stromal Tumor Spheroids by Multimodal Imaging: Fluorescence, Photoacoustic, and Photothermal Microscopies

Tracking Gold Nanorods’ Interaction with Large 3D Pancreatic-Stromal Tumor Spheroids by Multimodal Imaging: Fluorescence, Photoacoustic, and Photothermal Microscopies

Dandelion flower-like micelles

Head-to-Head Comparison of the Penetration Efficiency of Lipid-Based Nanoparticles into Tumor Spheroids

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