Bystander effect of metal byproducts released from electroporated cells after electroporation in vitro

Bioelectrochemistry, Год журнала: 2025, Номер 164, С. 108940 - 108940

Опубликована: Фев. 13, 2025

Electrodes dissolution during electroporation releases metal ions into the medium, altering microenvironment of electroporated cells and allowing to penetrate cell membrane. During membrane repair, homeostasis restoration or activation death pathways, eliminate excess metals from cytoplasm This study assessed effects post-electroporation byproducts on untreated (non-electroporated) in vitro. CHO HCT116 were with three pulse protocols (unipolar: 100 μs, 5 ms; bipolar: 2 μs) using either aluminum stainless-steel electrodes. After electroporation, transferred fresh growth medium incubated for 4 h. Incubation period allowed recovery leading accumulation incubation medium. Stainless-steel electrodes ms protocol caused a considerable increase iron, chromium nickel compared other protocols. Metal toxicity non-electroporated cells, disrupting cycle function inducing death. The observed results combined cellular functions mechanisms use protect themselves overload.

Язык: Английский

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Expulsion of iron-rich ferritin via CD63-mediated exosome drives ferroptosis resistance in ovarian cancer cells

Frontiers in Cell and Developmental Biology, Год журнала: 2025, Номер 13

Опубликована: Март 10, 2025

Introduction Ferroptosis is a promising new target for ovarian cancer (OVCA) treatment. However, some OVCA cell types resist the induction of ferroptosis by limiting intracellular accumulation labile iron pool (LIP). Methods HEY, COV318 and PEO4 were treated with erastin assessed viability using PI flow cytometry assays. Erastin-affected metabolism was analysed FerroOrange assay, Western Blot (WB) analysis ferritin heavy chain (FtH), transferrin receptor (CD71), ferroportin (FPN). Mitochondrial reactive oxygen species (mitROS) lipid peroxidation quantified via MitoSOX BODIPY-C11 assays, respectively. Exosomes (EVs) collected from culture media through ultracentrifugation then enumerated analyzed Nanoparticale Tracking Analysis (NTA) transmission electron microscopy (TEM). CD63 protein expression in EVs measured WB CD9 as loading control. Loss-of-function assays FtH performed siRNA-mediated transient transfection. Results We demonstrate that treatment (8 µM, 8 h) accompanied release iron-rich EV pathway cells, thus failing to exert cytotoxic effects. Mechanistically, causes upregulation CD63, tetraspanin involved forming multivesicular bodies (MVBs) EVs, increase MBVs microscopy. Consistent these findings, isolation followed nanoparticle tracking revealed significant EVs/cell erastin-treated cells. Notably, harvested cells contained FtH, major iron-storage protein. Inhibition biogenesis GW4869 prevented restored LIP accumulation, peroxidation, sensitivity Discussion Overall, our results indicate can utilize CD63+ secrete mechanism evade erastin-induced ferroptosis. These findings suggest combining inhibitors could offer strategy overcoming resistance OVCA.

Язык: Английский

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Portable Cell Tracking Velocimetry for Quantification of Intracellular Fe Concentration of Blood Cells

Micromachines, Год журнала: 2025, Номер 16(2), С. 126 - 126

Опубликована: Янв. 23, 2025

Hematological analysis is crucial for diagnosing and monitoring blood-related disorders. Nevertheless, conventional hematology analyzers remain confined to laboratory settings due their high cost, substantial space requirements, maintenance needs. Herein, we present a portable cell tracking velocimetry (CTV) device the precise measurement of magnetic susceptibility biological entities at single-cell level, focusing on red blood cells (RBCs) in this work. The system integrates microfluidic channel positioned between permanent magnets that generate well-defined field gradient (191.82 TA/mm2). When are injected into chamber, particular response recorded used estimate properties quantify intracellular hemoglobin (Hb) concentration. We successfully track over 400 RBCs per condition using imaging trajectory analysis, enabling detailed characterizations physical properties. A comparison mean corpuscular measurements revealed strong correlation our CTV standard ultraviolet-visible (UV-Vis) spectrophotometry (23.1 ± 5.8 pg vs. 22.4 3.9 pg, p > 0.05), validating accuracy measurements. system's resolution reveals population distributions unobtainable through bulk methods. Thus, technology provides rapid, label-free approach characterization, offering new possibilities point-of-care hematological field-based research applications.

Язык: Английский

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Injectable ECM-mimetic dynamic hydrogels abolish ferroptosis-induced post-discectomy herniation through delivering nucleus pulposus progenitor cell-derived exosomes

Nature Communications, Год журнала: 2025, Номер 16(1)

Опубликована: Апрель 1, 2025

Discectomy-induced ferroptosis of nucleus pulposus cells (NPCs) contributes to postoperative lumbar disc herniation (LDH) recurrence and intervertebral degeneration (IDD). We discover that progenitor (NPPCs) could imprint resistance into NPCs through exosome-dependent intercellular transmission miR-221-3p. Based on these findings, we first develop synthetically-tailored NPPC-derived exosomes with enhanced miR-221-3p expression NPC uptake capacity, which are integrated an injectable hydrogel based extracellular matrix (ECM) analogues. The ECM-mimetic (HACS) serves as a biomimetic filler for the post-operative care herniated discs, be facilely injected discectomy-established (NP) cavity localized treatment. HACS-mediated in-situ exosome release in NP enables marked inhibition not only prevents LDH but also reverses IDD symptoms, leading robust restoration structure functions. In summary, this study offers promising approach treating herniation.

Язык: Английский

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Extracellular Vesicles in Viral Liver Diseases

Viruses, Год журнала: 2024, Номер 16(11), С. 1785 - 1785

Опубликована: Ноя. 17, 2024

Extracellular vesicles (EVs) are bilayer released by cells in the microenvironment of liver including parenchymal and non-parenchymal cells. They third important mechanism communications between cells, besides secretion cytokines chemokines direct cell-to-cell contact. The aim this review is to discuss role EVs viral disease, as there increasing evidence that transportation proteins, all types RNA, particles complete virions implicated pathogenesis both cirrhosis viral-related hepatocellular carcinoma. biogenesis discussed their diseases presented. Their use diagnostic prognostic biomarkers also analyzed. Most importantly, significance possible novel treatment strategies for fibrosis carcinoma presented, although available data based on experimental clinical trials have not been reported.

Язык: Английский

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Dental pulp stem cells derived exosomes inhibit ferroptosis via regulating the Nrf2-keap1/GPX4 signaling pathway to ameliorate chronic kidney disease injury

Tissue and Cell, Год журнала: 2024, Номер 93, С. 102670 - 102670

Опубликована: Дек. 7, 2024

Язык: Английский

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