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Protective Effect of Mesenchymal Stem Cell-Derived Extracellular Vesicles on Inner Ear Sensorineural Cells Affected
Maria Perde-Schrepler1,2, Ioana Brie2, Mihai Cenariu3
1Department of Otoryhinolaryngology, Iuliu Hatieganu University of Medicine and Pharmacy, 400012 Cluj-Napoca, Romania.
Abstract:
Background and Objectives: Extracellular vesicles (EVs) derived from mesenchymal stem cells have gained much attention as potential therapeutic agents in many diseases, including hearing disorders such as sensorineural hearing loss (SNHL). EVs inherit similar therapeutic effects, including the stimulation of tissue regeneration from the parental cells. The aim of our study was to isolate EVs produced by MSCs and use them to treat inner ear cells in culture to evaluate their protective potential against the damaging effect of an ototoxic drug. Materials and Methods: We isolated MSC-derived EVs by precipitation and characterized them by number, size, and morphology using nanoparticle tracking analysis and TEM, evaluated the protein concentration by BCA assay and the presence of EV markers CD9, CD63, and CD81 by the Dot Blot immunoblotting method. HEI-OC1 inner ear cell line was treated with EVs either alone or followed by Cisplatin. We assessed the uptake of EVs in HEI-OC1 cells by fluorescence microscopy after PKH26 labeling, ROS production by the DCFDA (dichlorfluorescein diacetate) assay, cellular viability by Alamar Blue assay, and apoptosis with the Annexin V/Propidium Iodide method. Results: The isolated EVs had mean dimensions of 184.4 nms and the concentration of the EV suspension was 180 × 106 particles/mL. TEM analysis showed intact vesicular structures with lipid-bilayer membranes having similar sizes with those measured by NTA. The PKH26-labeled EVs were observed in the HEI-OC1 cells after 24 h incubation, the amount increasing with the concentration. EVs reduced ROS production and increased the number of viable cells both alone and as pretreatment before Cisplatin, dose-dependently. Cells in early apoptosis were inhibited by EVs, while those in late apoptosis were enhanced, both with and without Cisplatin. Conclusions: EVs secreted by MSC protected HEI-OC1 cells against Cisplatin toxicity, reduced ROS production, and stimulated cell viability and the elimination of damaged cells by apoptosis, protecting the HEI-OC1 cells against Cisplatin-induced damage.
Insights
Mesenchymal stem cell-derived extracellular vesicles (EVs) protect inner ear cells from drug-induced damage. These EVs reduce oxidative stress and promote cell survival, offering potential therapeutic benefits for sensorineural hearing loss.
Area of Science:
- Regenerative Medicine
- Ototoxicity Research
- Cell Biology
Background:
- Mesenchymal stem cell-derived extracellular vesicles (EVs) show therapeutic potential for diseases, including sensorineural hearing loss (SNHL).
- EVs can stimulate tissue regeneration, mirroring the effects of their parent cells.
- Investigating EV protective effects against ototoxic drug damage in inner ear cells is crucial.
Purpose of the Study:
- To isolate and characterize extracellular vesicles (EVs) from mesenchymal stem cells (MSCs).
- To evaluate the protective capacity of MSC-derived EVs against cisplatin-induced damage in cultured inner ear cells (HEI-OC1).
- To assess the impact of EVs on cell viability, reactive oxygen species (ROS) production, and apoptosis.
Main Methods:
- Isolation and characterization of MSC-derived EVs using nanoparticle tracking analysis, TEM, BCA assay, and Dot Blot.
- Treatment of HEI-OC1 cells with EVs, alone or followed by cisplatin exposure.
- Assessment of EV uptake, ROS levels, cell viability (Alamar Blue), and apoptosis (Annexin V/PI staining).
Main Results:
- Isolated EVs exhibited characteristic morphology and size (mean 184.4 nm).
- EVs were successfully internalized by HEI-OC1 cells.
- EVs significantly reduced ROS production, enhanced cell viability, and modulated apoptosis in cisplatin-treated cells.
Conclusions:
- Mesenchymal stem cell-derived extracellular vesicles (EVs) demonstrate significant protective effects against cisplatin-induced ototoxicity in vitro.
- EVs mitigate cellular damage by reducing oxidative stress and promoting cell survival.
- These findings support the potential of MSC-derived EVs as a therapeutic strategy for sensorineural hearing loss.
