Preparation of trehalose biomimetic nanovesicles and its protective effect on hypoxic-induced myocardial cell injury in vitro

LYU Yuan-yuan; ZHAO Zhi-han; GU Zheng-hui; ZHONG Guang-hui

doi:10.3969/j.issn.1672-5301.2025.11.013

Abstract：Objective In order to explore new therapeutic strategies for myocardial ischemic injury,this study was conducted to construct trehalose biomimetic nanovesicles(Tre NVs),observe their physicochemical properties,and explore their protective effects on hypoxic-induced myocardial cell injury in vitro.Methods A microfilter was used to continuously extrude the bone marrow mesenchymal stem cells to extract the nanovesicles(NVs).Trehalose(Tre)was actively loaded into NVs via sonication to generate Tre NVs.The particle size,zeta potential,morphology and cytotoxicity of Tre NVs were systematically characterized.The cardioprotective efficacy of Tre NVs under hypoxic stress was assessed by flow cytometry and fluorescent staining.Result Tre NVs were successfully prepared.Tre NVs exhibited a morphology similar to typical extracellular vesicles,forming tea tray shaped vesicles with a double-layer membrane structure.The particle size of Tre NVs was concentrated at(133.9±1.27)nm,and the potential was(-47.1±2.23)mV.The efficiency of Tre loaded on vesicles was determined to be(40.42±0.85)%by high performance liquid chromatography(HPLC)analysis.In addition,the CCK-8 experiment detected that the increase in Tre NVs concentration did not significantly affect the activity of co cultured cardiomyocytes,and the survival rate of cardiomyocytes remained at around 85%.Through laser confocal microscopy,it was observed that Tre NVs labeled with DiI red fluorescent dye could be taken up by H9C2 cells and dispersed around the nucleus of the myocardial cells.Under simulated myocardial ischemia conditions in vitro,the myocardial cells could still effectively uptake Tre NVs,and the apoptosis of the myocardial cells was significantly reduced by flow cytometry after treatment with Tre NVs.By using the DCFH-DA reactive oxygen species detection kit to detect ROS generation,it could be observed that after Tre NVs administration,the ROS generation levels in the hypoxia induced cardiomyocytes were significantly reduced.Tre NVs could significantly attenuate the high reactive oxygen species environment after myocardial ischemia injury and played a protective role on the hypoxic-induced myocardial cell injury models.Conclusion The biomimetic Tre NVs delivery platform efficiently encapsulates trehalose,markedly enhancing its bioavailability.Tre NVs ameliorate the oxidative microenvironment following myocardial ischemia and effectively protect cardiomyocytes from hypoxia-provoked injury.