PNExo™ Exosome-Grape(PNE-FG84)

Key Bioactive Compounds and Benefits of Grapes

Grapes (Vitis vinifera) are a natural reservoir of diverse polyphenolic compounds, which are primarily found in the skin, seeds, and pulp. These bioactive constituents perform a broad range of biological functions, including antioxidative, anti-inflammatory, and cell-regulatory activities. Their effects on human health at the molecular level are being increasingly investigated in scientific research. The following are the main categories of active ingredients present in grapes, along with their corresponding functional properties:

• Anthocyanins (e.g., cyanidin-3-glucoside): These pigments are responsible for the red to purple color of grapes and have been shown to promote apoptosis in abnormal cells by activating caspase-3 and inducing DNA fragmentation. They also downregulate VEGF pathways, thereby suppressing angiogenesis.
• Flavanols (e.g., catechin, epicatechin, and proanthocyanidins): Known for their strong radical-scavenging capacity, proanthocyanidins also inhibit lipid peroxidation, contributing to membrane integrity. They can arrest the cell cycle in G1 phase and inhibit tumor cell proliferation.
• Resveratrol: A stilbene compound concentrated in grape skin, resveratrol supports endothelial nitric oxide release, enhances vascular function, and modulates SIRT1 signaling involved in cellular homeostasis and aging.
• Flavonols (e.g., quercetin, kaempferol): These compounds are known for downregulating inflammatory mediators such as TNF-α and MCP-1, and for blocking NF-κB signaling, thereby reducing inflammatory cascade reactions.
• Phenolic Acids (e.g., hydroxycinnamic acid, hydroxybenzoic acid): These molecules contribute to antioxidant activity and may support the suppression of oxidative stress-induced cellular damage.

What Are Grape-Derived Exosomes?

Grape-derived exosomes, also known as grape exosome-like nanoparticles (GENs), are nanoscale vesicles naturally secreted by grape cells. These vesicles, typically ranging from 30 to 150 nanometers, are enclosed by a lipid bilayer and act as biological couriers, carrying plant-based biomolecules such as proteins, lipids, RNAs (mRNA, miRNA, lncRNA), and small antioxidants like procyanidins and resveratrol. Increasingly studied as a type of plant-derived extracellular vesicle, GENs show great promise in molecular delivery, intercellular communication, and stability under biological conditions.

Key Advantages of Grape-Derived Exosomes

• Natural Molecular Cargo: GENs contain diverse plant biomolecules including regulatory RNAs and polyphenols, which may influence gene expression and oxidative balance when taken up by mammalian cells.
• Cholesterol-Free Composition: Unlike exosomes from animal sources, GENs contain minimal to no cholesterol, making them structurally unique and potentially less immunogenic in certain research applications.
• Efficient Cellular Uptake: GENs are readily absorbed by various mammalian cells, such as macrophages, lymphocytes, and epithelial cells, primarily through energy-dependent endocytosis.
• High Stability: These vesicles maintain structural integrity under physiological temperatures and can withstand long-term frozen storage and repeated freeze-thaw cycles without degradation.
• Antioxidant Encapsulation: Their bilayer membrane structure protects and delivers active antioxidant compounds, enhancing their bioavailability and potentially amplifying their cellular effects.
• Plant-Based and Biocompatible: As naturally sourced from grapes without synthetic additives or solvents, GENs offer a cleaner, safer alternative for plant nanovesicle research.

Morphological and molecular characterization of grape-derived exosome-like nanoparticles (ENs). (A, B) Scanning electron microscopy (SEM) images and nanoparticle tracking analysis show GCENs (from calluses) and GENs (from juice) have sizes ranging from ~90 to 250 nm with mildly irregular spherical morphology. (C) Western blot analysis confirms the presence of exosomal markers HSP70 and TET8 in both EN samples. (Shkryl Y, et al, 2024)

Potential Applications of Grape Exosome-like Nanoparticles

Intestinal Health and Stem Cell Regulation

Grape exosome-like nanoparticles have been shown to influence intestinal homeostasis. In in vivo models, they promoted the expression of key intestinal stem cell markers such as Lgr5, SOX2, Nanog, and KLF4, supporting epithelial renewal and mucosal barrier function. These findings suggest their potential utility in studies related to gastrointestinal regeneration and inflammation.

Anti-Inflammatory and miRNA Modulation

Grape exosomes are enriched in plant-derived miRNAs, including members of the miR169 family, which have shown sequence similarity with mammalian miRNAs. Some of these molecules have been implicated in downregulating pro-inflammatory genes such as IL-6 and IL-1β, making them relevant in research on immune modulation and inflammation-related signaling.

Antioxidant and Anti-Aging Investigations

Due to their natural content of resveratrol, myricetin, and procyanidins, grape-derived exosomes have demonstrated strong antioxidant properties. Studies suggest their role in scavenging free radicals and mitigating oxidative stress, with applications in models of skin photoaging and epithelial damage. Their vesicular structure enhances compound stability and delivery to target cells.

Targeted Biodistribution and Uptake

Fluorescence-tracking experiments have revealed that grape-derived exosomes can localize to the small intestine, colon, brain, lungs, and liver, depending on the route of administration. This suggests their promise as naturally derived nanocarriers for research on targeted delivery across biological barriers.

Exploratory Cancer Research

Preliminary findings have reported that GENs derived from grape cell cultures, especially those rich in trans-δ-viniferin derivatives, may exhibit cytotoxic effects on tumor cell lines (in vitro). Additionally, studies on breast cancer models have investigated GENs as biocompatible delivery vehicles for drug molecules. These studies pave the way for further exploration of their role in oncology research.

References

1. Ju S, Mu J, Dokland T, et al. Grape exosome-like nanoparticles induce intestinal stem cells and protect mice from DSS-induced colitis. Molecular Therapy. 2013, 21(7): 1345-1357.
2. Record M. Exosome-like nanoparticles from food: Protective nanoshuttles for bioactive cargo. Molecular Therapy. 2013, 21(7): 1294-1296.
3. Pérez-Bermúdez P, Blesa J, Soriano J M, et al. Extracellular vesicles in food: Experimental evidence of their secretion in grape fruits. European Journal of Pharmaceutical Sciences. 2017, 98: 40-50.
4. Ghiasi M R, Rahimi E, Amirkhani Z, et al. Leucine-rich repeat-containing G-protein coupled receptor 5 gene overexpression of the rat small intestinal progenitor cells in response to orally administered grape exosome-like nanovesicles. Advanced Biomedical Research. 2018, 7(1): 125.
5. Teng Y, He J, Zhong Q, et al. Grape exosome-like nanoparticles: A potential therapeutic strategy for vascular calcification. Frontiers in Pharmacology. 2022, 13: 1025768.
6. Farheen J, Iqbal M Z, Lu Y, et al. Vitis vinifera Kyoho-derived exosome-like nanoparticles-based drug delivery and therapeutic modalities for breast cancer therapy. Journal of Drug Delivery Science and Technology. 2024, 92: 105332.
7. Shkryl Y, Tsydeneshieva Z, Menchinskaya E, et al. Exosome-like nanoparticles, high in trans-δ-Viniferin derivatives, produced from grape cell cultures: Preparation, characterization, and anticancer properties. Biomedicines. 2024, 12(9): 2142.
8. Wang M, Chen J, Chen W, et al. Grape-Derived Exosome-Like Nanovesicles Effectively Ameliorate Skin Photoaging by Protecting Epithelial Cells. Journal of Food Science. 2025, 90(6): e70309.

Case Study 1: Anticancer Activity of Grape Callus-Derived Nanovesicles (Shkryl Y, 2024)

Researchers explored grape callus-derived exosome-like nanoparticles (GCENs) for their effects on cancer cells. These vesicles, averaging 79 nm in size, were rich in trans-δ-viniferin glycosides, with the most abundant reaching 480 µg/g dry weight. In vitro tests showed GCENs were taken up by MDA-MB-231 breast cancer cells within 4-24 hours and triggered apoptosis and G1-phase cell cycle arrest in a dose-dependent manner. At 1.2 × 10⁹ particles/mL, early apoptotic cells increased from 8% to 37%. Importantly, GCENs had minimal impact on normal HEK-293 cells, indicating selective cytotoxicity. Proteomic and miRNA profiling confirmed these vesicles carry antioxidant and stress-related proteins, along with plant-specific miRNAs. The findings support GCENs as a natural nanocarrier with potential in cancer research.

Figure 1. Cellular Uptake and Cytotoxicity of GCENs in Human Cell Lines. (A) Confocal fluorescence images of PKH26-labeled GCENs internalized by MDA-MB-231 cells after 4 and 24 hours of incubation. Red fluorescence (PKH26) indicates GCEN localization; blue fluorescence (DAPI) marks nuclei. (B) Light microscopy images showing MDA-MB-231 cell morphology after treatment with varying GCEN concentrations. (C) Cytotoxic effects of GCENs on MDA-MB-231 and HEK-293 cells analyzed by MTT assay. Scale bar = 20 µm.

Figure 2. GCEN-Induced Cell Cycle Arrest in MDA-MB-231 Cells. (A) Flow cytometry plots showing the distribution of MDA-MB-231 cells in different phases of the cell cycle after 24 h of treatment with grape callus-derived exosome-like nanoparticles. (B) Quantitative analysis of the percentage of cells in G1 (green), S (yellow), and G2/M (blue) phases. Untreated cells served as the control group.

Case Study 2: Grape Exosome-like Nanoparticles Promote Intestinal Repair and Reduce Colitis Symptoms (Ju S, 2013)

This study evaluated the biological effects of grape exosome-like nanoparticles (GELNs) in a mouse model of colitis. Oral administration of GELNs led to significant uptake in the small intestine and colon, where they were internalized by intestinal stem cells (ISCs). GELNs upregulated key ISC markers, including Lgr5, Ascl2, and OLFM4, indicating enhanced epithelial regeneration. In mice with DSS-induced colitis, GELN-treated groups showed reduced weight loss, colon shortening, and histological damage compared to controls. Inflammatory cytokines such as TNF-α and IL-6 were also significantly lower. Importantly, miRNA analysis revealed that GELNs carry small RNAs capable of modulating inflammatory signaling in host cells. These findings highlight the potential of plant-derived vesicles as natural carriers for gut-targeted bioactive molecules.

Figure 1. Uptake of GELNs by intestinal stem cells and epithelial cells. (a) In vivo tracking of DiR-labeled GELNs in C57BL/6 mice over 48 hours, with quantitative analysis of signal intensity. (b) Confocal imaging and quantification of PKH26+/Lgr5-EGFP+ intestinal stem cells in Lgr5-EGFP-IRES-CreERT2 mice after oral administration of PKH26-labeled GELNs. (c) Confocal imaging of PKH26-labeled GELN uptake by CT26 cells stained for EPCAM. (d) Inhibition of GELN uptake by CT26 cells with specific inhibitors, analyzed by flow cytometry. (e) Confocal microscopy and quantification of colon tissue from Lgr5-EGFP-IRES-CreERT2 mice treated with cytochalasin D and PKH26-labeled GELNs, showing reduced internalization of GELNs by EGFP+ stem cells.

Figure 2. Oral administration of GELNs alleviates dextran sulfate sodium (DSS)-induced colitis in mice. (a) Representative image showing improved body condition in GELN-treated mice on day 7. (b) Survival analysis comparing GELN and PBS groups. (c) Intestinal length measurements and (d) villus height in H&E-stained small intestine sections. (e) Confocal images showing co-localization of EGFP+ and Ki67+ cells in intestinal crypts; arrowheads indicate proliferating stem cells. (f) qPCR analysis of crypt-specific gene expression. (g) Confocal analysis reveals nuclear translocation of β-catenin and increased phosphorylation of GSK-3β (Ser9) in intestinal crypts post-GELN treatment.

References

1. Shkryl Y, Tsydeneshieva Z, Menchinskaya E, et al. Exosome-like nanoparticles, high in trans-δ-Viniferin derivatives, produced from grape cell cultures: Preparation, characterization, and anticancer properties. Biomedicines. 2024, 12(9): 2142.
2. Ju S, Mu J, Dokland T, et al. Grape exosome-like nanoparticles induce intestinal stem cells and protect mice from DSS-induced colitis. Molecular Therapy. 2013, 21(7): 1345-1357.

• What is the source of the grape-derived exosomes?

  These exosomes are isolated from high-quality Vitis vinifera grape tissues using standardized ultracentrifugation and filtration protocols.

• Can I customize the concentration or formulation for my study?

  Custom concentrations, bulk quantities, and additional formulation support are available upon request. Please contact our technical team for tailored solutions.

• How are grape exosomes characterized for purity and size?

  Each batch undergoes rigorous quality control using techniques such as nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), and Western blotting for exosome markers.

• Are there any known contaminants or solvent residues in the preparation?

  No. The exosomes are prepared without organic solvents, and purification steps ensure they are free from detectable contaminants or cytotoxic residues.

• Do grape exosomes show batch-to-batch consistency?

  Yes, all products are produced under standardized conditions with strict QC criteria to ensure consistency in size, concentration, and cargo profiles across batches.

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