Rongfeng Yang

Background: Myocardial ischemia–reperfusion injury (MIRI) remains a major clinical challenge in the management of acute myocardial infarction. Ginsenoside Rg1, a bioactive component from Panax ginseng, exhibits cardioprotective properties but suffers from poor bioavailability and limited tissue targeting. Methods: We designed a novel DNA nanocarrier, Rg1@pTDN, by loading Rg1 onto a tetrahedral DNA nanostructure (TDN) modified with the myocardial-targeting peptide CREKA. The physicochemical characteristics of Rg1@pTDN were evaluated by DLS, zeta potential analysis, AFM, and gel electrophoresis. Biodistribution, biosafety, and cellular uptake were assessed in vitro and in vivo. Cardioprotective efficacy was evaluated in a murine MIRI model and a H/R injury cell model. Mechanistic studies focused on oxidative stress and endoplasmic reticulum (ER) stress pathways. Results: Rg1@pTDN exhibited uniform nanoscale structure, high Rg1 loading efficiency, and good colloidal stability. In vivo imaging revealed preferential accumulation in cardiac tissue following intravenous administration. Rg1@pTDN was well tolerated and improved survival, cardiac function, and myocardial histology in MIRI mice. It significantly reduced serum CK-MB concentrations and oxidative stress markers (MDA), while increasing antioxidant enzyme activities (SOD, GSH-Px). In vitro, Rg1@pTDN suppressed ROS accumulation and H/R–induced apoptosis in H9c2 cells. Furthermore, Rg1@pTDN alleviated H/R–induced ER stress, as shown by decreased GRP78 and CHOP mRNA expression and reduced phosphorylated PERK and CHOP protein levels. Conclusion: Rg1@pTDN represents a promising nanotherapeutic strategy for myocardial ischemia–reperfusion injury through targeted delivery and dual inhibition of oxidative and ER stress. This DNA-based platform offers a versatile approach for enhancing the efficacy of natural compounds in cardiovascular disease.

Skin wound healing is a multifaceted biological process involving dynamic interactions among various cells and signaling molecules. Angiogenesis is a key component of this repair process. Dendrobium officinale, a traditional medicinal plant, has shown therapeutic promise, particularly through its bioactive nanovesicles. This study investigates the therapeutic potential of Dendrobium officinale-derived nanovesicles (DDNVs) in regulating angiogenesis, inflammation, and tissue repair, to promote enhanced skin wound healing. A full-thickness mouse skin wound model was used to evaluate the in vivo effects of DDNVs on wound closure, angiogenesis, and collagen remodeling. Histological staining (H&E and Masson’s trichrome) and CD31 immunofluorescence were performed. In vitro, DDNVs were tested on Human umbilical vein endothelial cells(HUVECs) and Human keratinocyte cells (HaCaT) cells to assess cell proliferation, migration, and angiogenesis. Confocal microscopy was used to track cellular uptake. Activation of the Akt/eNOS pathway and expression of key genes related to inflammation and matrix remodeling were evaluated by Western blotting and qPCR. DDNVs significantly accelerated wound healing and promoted angiogenesis in vivo, as evidenced by enhanced CD31 expression and collagen remodeling. In vitro, DDNVs entered cells efficiently and stimulated HUVEC and HaCaT proliferation and migration. This was accompanied by activation of the Akt/eNOS signaling pathway, increased expression of eNOS and VEGFR-2, upregulation of extracellular matrix(ECM) components (Vimentin, Fibronectin, COL1A1), and suppression of inflammatory markers such as ICAM-1 and IL-1β. DDNVs exhibit strong potential to enhance skin wound healing by promoting angiogenesis, improving tissue repair, and modulating inflammation. These findings support the clinical development of DDNVs as a novel, plant-derived nanotherapeutic for chronic wound treatment and skin regeneration.

The present study aimed to prepare a kind of controlled-releasing insulin-like growth factor 1 (IGF-1)/spider silk protein nanofibrous membrane using a electrostatic spinning method and evaluated its effect on the cell viability of endothelial progenitor cells (EPCs). Recombinant spidroin named as GMCDRSSP-IgF-1 was electro-spun into nanofibrous membrane which can be degraded by protease and be capable of sustained-release of IGF-1. The membrane can be degraded after being treated with thrombin. The release assay results showed that IGF-1 concentration could be maintained at 20 ng/ml for a long time with treatment of Tobacco Etch Virus (TEV) protease. The viability of EPCs on GMCDRSSP-IgF-1 nanofibrous membrane was significantly increased with the presence of TEV protease. The controlled and sustained release of IGF-1 from the nanofibrous membrane could promote the adhesion and viability of EPCs. In summary, the nanofibrous membrane that exhibits controlled degradation and sustained release of IGF-1 was prepared with electrostatic spinning from genetically modified recombinant spider silk protein. The nanofibrous membrane exhibited good blood compatibility and cytocompatibility. With the presence of TEV protease, the sustained-release of IGF-1 significantly promoted the adhesion and viability of EPCs. The new nanofibrous membrane can be potentially used as a scaffold for EPCs culture in vitro and future in vivo studies.

Accumulating evidence has shown that endothelial progenitor cell‐derived exosomes (EPC‐Exos) can ameliorate myocardial fibrosis. The purpose of the present study was to investigate the effects of EPC‐Exos‐derived microRNAs (miRNAs) on myocardial infarction (MI). A miRNA‐Seq dataset of miRNAs differentially expressed between EPCs and exosomes was collected. Quantitative real‐time polymerase chain reaction (qRT‐PCR) was used to validate the miRNA expression indicated by miRNA‐Seq. Immunofluorescence, cell proliferation, and angiogenesis assays were employed to investigate the effects of miRNAs on cardiac fibroblasts (CFs) in vitro. Interactions between miRNAs and their respective targets were examined via immunoblotting, qRT‐PCR, and luciferase reporter assays. An MI rat model was constructed, and various staining and immunohistochemical assays were performed to explore the mechanisms underlying the miRNA‐mediated effects on MI. miR‐363‐3p and miR‐218‐5p were enriched in EPC‐Exos, and miR‐218‐5p and miR‐363‐3p mimic or inhibitor enhanced or suppressed CF proliferation and angiogenesis, respectively. miR‐218‐5p and miR‐363‐3p regulated p53 and junction‐mediating and regulatory protein (JMY) by binding to the promoter region of p53 and the 3 ^′ untranslated region of JMY. Additionally, treatment of CFs with Exo‐miR‐218‐5p or Exo‐miR‐363‐3p upregulated p53 and downregulated JMY expression, promoted mesenchymal‐endothelial transition, and inhibited myocardial fibrosis. Administration of exosomes containing miR‐218‐5p mimic or miR‐363‐3p mimic ameliorated left coronary artery ligation‐induced MI and restored myocardial tissue integrity in the MI model rats. In summary, these results show that the protective ability of EPC‐Exos against MI was mediated by the shuttled miR‐218‐5p or miR‐363‐3p via targeting of the p53/JMY signaling pathway.

Hyperlipidemia is an important factor in the induction of cardiovascular diseases. However, the molecular mechanisms underlying the vascular injury involved in hyperlipidemia remains unclear. This study aimed to investigate the Notch pathway of endothelial progenitor cells (EPCs) in reendothelialization after vascular injury and to explore the involvement of Notch pathway in the senescence of EPCs. Our results demonstrated that high-fat diet (HFD) treatment inhibited reendothelialization after vascular injury in the mice model. In vitro studies showed that 7-ketocholesterol (7-keto) stimulation induced senescence in the isolated EPCs from mice. In addition, 7-keto markedly upregulated the protein expression of Notch1 and Delta-like ligand 4 and induced the transport of notch intracellular domain (NICD) to the nucleus. Mechanistically, treatment with NICD inhibitor reduced the senescence of the EPCs stimulated by cholesterol. In summary, our results showed that HFD treatment caused the disruption of reendothelialization after vascular injury in the mouse model. In vitro studies indicated that 7-keto-induced senescence of EPCs was at least via the activation of the Notch1 pathway. Mechanistic data suggested that 7-keto may activate the Notch1 pathway by regulating the generation and transport of NICD to the nucleus. Future investigations are warranted to confirm the role of Notch1 in the dysfunction of EPCs during obesity.