<i>In Vivo</i> Real-Time Imaging of Extracellular Vesicles in Liver Regeneration <i>via</i> Aggregation-Induced Emission Luminogens

Cao, Hongmei; Yue, Zhiwei; Hong, Gao; Chen, Chao; Cui, Kaige; Zhang, Kaiyue; Cheng, Yuanqiu; Shao, Guoqiang; Kong, Deling; Li, Zongjin; Ding, Dan; Wang, Yuebing

doi:10.1021/acsnano.8b09776

Cited by 77 publications

(54 citation statements)

References 52 publications

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“…PKH26 is a membrane dye that intercalates the aliphatic portion of exposed lipid bilayers, and has been widely used to label extracellular vesicles (Cao et al, 2019). Given that MDVs were also membrane structures, we labeled MDVs using a PKH26 Red Fluorescent Cell Linker kit according to the manufacturer's protocol.…”

Section: Fluorescent Labeling and Mitotracker Stainingmentioning

confidence: 99%

Mitochondrial-Derived Vesicles Protect Cardiomyocytes Against Hypoxic Damage

Zhao

et al. 2020

Front. Cell Dev. Biol.

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Myocardial ischemia is a condition with insufficient oxygen supporting the heart tissues, which may result from myocardial infarction or trauma-induced hemorrhagic shock. In order to develop better preventive and therapeutic strategies for myocardial ischemic damage, it is important that we understand the mechanisms underlying this type of injury. Mitochondrial-derived vesicles (MDVs) have been proposed as a novel player in maintaining mitochondrial quality control. This study aimed to investigate the role and possible mechanisms of MDVs in ischemia/hypoxia-induced myocardial apoptosis. H9C2 cardiomyocytes were used for the cellular experiments. A 40% fixed blood volume hemorrhagic shock rat model was used to construct the acute general ischemic models. MDVs were detected using immunofluorescence staining with PDH and TOM20. Exogenous MDVs were reconstituted in vitro from isolated mitochondria under different hypoxic conditions. The results demonstrate that MDV production was negatively correlated with cardiomyocyte apoptosis under hypoxic conditions; exogenous MDVs inhibited hypoxia-induced cardiomyocyte apoptosis; and MDV-mediated protection against hypoxia-induced cardiomyocyte apoptosis was accomplished via Bcl-2 interactions in the mitochondrial pathway. This study provides evidence that MDVs protect cardiomyocytes against hypoxic damage by inhibiting mitochondrial apoptosis. Our study used a novel approach that expands our understanding of MDVs and highlights that MDVs may be part of the endogenous response to hypoxia designed to mitigate damage. Strategies that stimulate cardiomyocytes to produce cargo-specific MDVs, including Bcl-2 containing MDVs, could theoretically be helpful in treating ischemic/hypoxic myocardial injury.

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Section: Fluorescent Labeling and Mitotracker Stainingmentioning

confidence: 99%

Mitochondrial-Derived Vesicles Protect Cardiomyocytes Against Hypoxic Damage

Zhao

et al. 2020

Front. Cell Dev. Biol.

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“…[ 44 ] Besides PKH26 (red) 3,30‐octadecyloxacarbacyanate perchlorate (DiO, cell membrane green fluorescent dye) and DAPI were used for cytomembrane and nucleus staining. [ 45–47 ] E‐EVs were greatly internalized by BMSCs after incubation for 2 h, while L‐EVs were rarely engulfed ( Figure A,B). Then, TEM scanning was performed to further reveal the intracellular localization of E‐EVs.…”

Section: Resultsmentioning

confidence: 99%

Autologous Versatile Vesicles‐Incorporated Biomimetic Extracellular Matrix Induces Biomineralization

Wei

Shi

Zhang

et al. 2020

Adv Funct Materials

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Restoring extracellular matrix (ECM) with supportive and osteoinductive abilities is of great significance for bone tissue regeneration. Current approaches involving cell-based scaffolds or nanoparticle-modified biomimic-ECM have been met with additional biosafety concerns. Herein, the natural biomineralization process is first analyzed and is found that mesenchymal stem cells-derived extracellular vesicles (EVs) from early and late stages of osteoinduction play different roles during the mineralization process. The functional EVs hierarchically with blood-derived autohydrogel (AH) are then incorporated to form an osteoinductive biomimetic extracellular matrix (BECM). The alkaline phosphatase-rich EVs are released from the outer layers to induce osteoblast differentiation during early stages. Thereafter, as the degradation of AH occurred, calcium/phosphorus (Ca/P)rich EVs are liberated to promote the nucleation of extracellular mineral crystals. Additionally, BECM contains considerable collagen fibrils that provide additional nucleation sites for crystallites deposition, thus reaching self-mineralization in situ. In conclusion, this research provides a promising, versatile mineralization-instructive platform to tackle the challenges faced in bone-tissue engineering. Scheme 1. Illustration of the BECM induced osteogenesis and biomineralization. In the early stage of mineralization, BECM secreted ALP-rich E-EVs. The E-EVs moved to intracellular mitochondria and provided ALP for phosphate metabolism, contributing to the formation of Ca/P-rich vesicles. Later, with the degradation of BECM, Ca/P-rich L-EVs were released gradually. The L-EVs aligned to extracellular collagen and promoted the nucleation of extracellular mineral crystals. Besides, L-EVs inside BECM aggregate around the internally collagen fibrils and initiated in situ mineralization. Finally, with the complete degradation of BECM, the internal mineralized components were released to the extracellular matrix, promoting the deposition formation together to realize an ideal self-mineralization process for bone tissue regeneration.www.afm-journal.de www.advancedsciencenews.com (3 of 15)

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“…Recently, using a mouse acute liver injury (ALI) model, Cao et al developed a novel noninvasive, real‐time, and precise strategy for tracking MSC‐EVs and their therapeutic effects. The in vivo EV tracker in that work (DPA‐SCP, Figure A) was a typical AIEgen with a positive charge, which facilitated attachment to the EV membrane with a negative charge (Figure B), and showed much better labeling efficiency compared with commercial EV trackers (PKH26 and DiI), as revealed by the NTA analysis (AIE‐EVs: ∼86.9%; PKH26‐EVs: 50.7%; DiI‐EVs: 39.7%) and flow cytometry.…”

Section: Monitoring Biological Processesmentioning

confidence: 99%

Section: Monitoring Biological Processesmentioning

confidence: 99%

Promising Applications of AIEgens in Animal Models

Situ

Zhao

et al. 2019

Small Methods

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organisms cannot be achieved from these static findings. Therefore, gaining in-depth insights into biological and physiological functions at the in vivo level is of great significance, and various imaging techniques have been explored for this purpose. [1] In addition to the clinically available magnetic resonance imaging (MRI), ultrasonic imaging, computed tomography (CT), positron emission tomography (PET), and single-photon emission computed tomography, fluorescence imaging (another distinct type of optical imaging [1c] from bioluminescence imaging [2] ) has attracted increasing attention, owing to its advantages of high temporal-spatial resolution and sensitivity, noninvasive, and real-time applications, and the ability to provide dynamic information for basic research and (pre-)clinical settings at cellular and subcellular levels. [3] Fluorescent probes or contrast agents are essential for fluorescence imaging. Compared with fluorescent proteins, [4] inorganic nanoparticles (NPs), [5] such as quantum dots, [6] upconversion NPs, [7] and metallic NPs, [8] and organic dyes [9] (e.g., U.S. Food and Drug Administration (FDA)-approved 5-ALA, indocyanine green (ICG), and methylene blue), organic NPs [10] have obvious advantages, such as easy preparation and good biocompatibility. However, many organic NPs show weak fluorescence because they are made with conventional organic dyes, which suffer from the aggregation-caused quenching (ACQ) effect, i.e., their fluorescence is partially or completely quenched in the aggregated state.The advent of the aggregation-induced emission (AIE) concept offers a new strategy to deal with ACQ. [11] AIE refers to the interesting photophysical phenomenon that luminogens are weakly fluorescent or nonfluorescent in solution but show greatly enhanced emission in the aggregated state, primarily because of restricted intramolecular motion. [12] The past 19 years have witnessed spectacular advances of AIEgens, [13] due to their advantages of structural variety, easy functionalization, strong luminescence, large Stokes shift, high photobleaching-resistance, and so on. In particular, various AIE bioprobes have been designed: [14] (i) water-soluble AIEgens, which are comprised of a hydrophobic AIE core and hydrophilic units (e.g., ionic groups, peptides, carbohydrates, DNA, aptamers, antibodies, and poly(ethylene glycol) (PEG)); (ii) bare AIEgen dots (nanoaggregates); iii) AIEgen/biopolymer dots, which can be obtained by loading AIEgens onto biopolymer matrixes covalently (e.g., chitosan, dextran, and starch) or noncovalently (e.g., bovine serum albumin, human serum albumin (HSA), and fetal bovine serum (FBS)); (iv)In vivo investigations using small animal models are of great significance for the comprehensive understanding of biological and physiological functions-seeing is believing. Fluorescence imaging can provide real-time and dynamic information of living systems. During the past few years, aggregation-induced emission luminogens (AIEgens) are widely and rapidly developed for biolog...

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In Vivo Real-Time Imaging of Extracellular Vesicles in Liver Regeneration via Aggregation-Induced Emission Luminogens

Cited by 77 publications

References 52 publications

Mitochondrial-Derived Vesicles Protect Cardiomyocytes Against Hypoxic Damage

Mitochondrial-Derived Vesicles Protect Cardiomyocytes Against Hypoxic Damage

Autologous Versatile Vesicles‐Incorporated Biomimetic Extracellular Matrix Induces Biomineralization

Promising Applications of AIEgens in Animal Models

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