Plant-derived nanovesicles from Ginkgo biloba seeds mitigate LPS-induced endothelial dysfunction and promote vascular homeostasis

Maneea Moubarak; Emese Szilágyi-Tolnai; Ani Barbulova; Immacolata Fiume; Ildikó Kovács-Forgács; Judit Homoki; Georgina Pesti-Asbóth; Endre Szilágyi; Ramila Mammadova; Matic Kisovec; Marjetka Podobnik; Dávid Papp; Gitta Schlosser; Judit Remenyik; Gabriella Pocsfalvi

doi:10.3389/fbioe.2025.1715489

Plant-derived nanovesicles from Ginkgo biloba seeds mitigate LPS-induced endothelial dysfunction and promote vascular homeostasis

Front Bioeng Biotechnol. 2026 Feb 13:13:1715489. doi: 10.3389/fbioe.2025.1715489. eCollection 2025.

Authors

Affiliations

¹ Extracellular Vesicles and Mass Spectrometry Laboratory, Institute of Biosciences and BioResources, National Research Council of Italy, Naples, Italy.
² Complex Systems and Microbiome-Innovations Centre, Faculty of Agricultural and Food Sciences and Environmental Management, University of Debrecen, Debrecen, Hungary.
³ Department of Molecular Biology and Nanobiotechnology, National Institute of Chemistry, Ljubljana, Slovenia.
⁴ MTA-ELTE Lendület Ion Mobility Mass Spectrometry Research Group, Institute of Chemistry, ELTE Eötvös Loránd University, Budapest, Hungary.

^# Contributed equally.

Abstract

Background: Endothelial dysfunction is increasingly recognized as an early and central event in the onset of cardiovascular and neurodegenerative diseases. Ginkgo biloba extracts are known for their vascular-protective properties, including enhancement of endothelial function, antioxidant and anti-inflammatory activity, nitric oxide preservation, and modulation of platelet aggregation. Plant-derived nanovesicles (PDNVs) are emerging as versatile bioactive carriers with demonstrated anti-inflammatory, anticancer, antimicrobial, regenerative, and microbiota-modulating effects. However, their vascular-protective potential remains underexplored. This study aimed to assess the effects of PDNVs isolated from Ginkgo biloba seeds on endothelial responses under inflammatory stress.

Methods: PDNVs were isolated from Ginkgo biloba seed homogenate using differential ultracentrifugation followed by density gradient ultracentrifugation with linear and non-linear iodixanol gradients. Nanoparticle tracking analysis (NTA) and cryo-transmission electron microscopy (cryo-TEM) characterized vesicle size, concentration, and morphology. Untargeted mass spectrometry profiled the protein content of distinct PDNV fractions. Functional assays were conducted on human umbilical vein endothelial cells (HUVECs) exposed to lipopolysaccharide (LPS)-induced inflammatory stress.

Results: Ginkgo PDNV isolates comprise a heterogeneous population of nanometer sized particles, including vesicles with single and double layers. Proteomics revealed seed storage proteins (legumin and ginnacin) and membrane-associated ATPases, HSP90, catalase, phosphoenolpyruvate carboxylase (PEPC), and eEF1A. PDNVs were non-toxic up to 50 μg/mL; at 100 μg/mL, they enhanced mitochondrial activity but triggered early apoptosis and necrosis. PDNVs did not increase ROS production, even in the presence of H₂O₂. At 1 μg/mL, they significantly suppressed LPS-induced expression of IL-1β, TNF-α, IL-6, and IL-8 (mRNA and protein; p ≤ 0.05 to p ≤ 0.001). PDNVs preserved endothelial integrity by downregulating VCAM-1 and upregulating occludin, maintained eNOS expression (p ≤ 0.01), and attenuated COX-1, COX-2, and prostacyclin synthase (PGIS) induction. Thrombotic markers (TXB₂, vWF, and PAI-1) remained unaffected.

Conclusion: Ginkgo seed-derived PDNVs exhibit vascular-protective and anti-inflammatory properties, supporting their potential as safe, multifunctional agents for endothelial modulation. Further studies are warranted to explore their therapeutic applications in vascular biology.

Keywords: Ginkgo biloba; endothelial dysfunction; endothelial inflammation; plant-derived nanovesicles; proteomics; vascular protection.