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Product Summary
Source
Exosome derived from Orange
Product Overview
Plant exosomes are nanosized (30-150 nm) membrane vesicles that contain biomolecules. Plant-derived exosomes refer to naturally occurring nanoparticles derived from plants that contain bioactive molecules and proteins. These exosomes have been shown to have multiple benefits in a variety of applications, such as skincare, drug delivery, and biomedicine. Plant-derived exosomes have been found to possess antioxidant, anti-inflammatory, and anti-aging properties, making them an attractive option for the development of new and innovative therapies. Plant-derived natural substances are widely used as cosmeceutical materials because they exert beneficial effects on the human skin, such as antiaging, moisturizing, whitening, regeneration, and nutritional supply. Besides, they could delivery therapeutic compounds to target cells, potentially revolutionizing the way in which drugs are administered. Overall, plant-derived exosomes hold great promise for a wide range of applications in the fields of medicine and biotechnology.
PNExo™ is focused on the production and delivery of high quality plant-derived exosomes products. Exosomes are important tools of intercellular communication with a variety of biological functions, including cell regeneration and immune regulation. PNExo™ products undergo a rigorous screening and purification process that guarantees their high purity and activity. Lyophilization is useful for a long-term storage at 4°C, and frozen liquid should be kept at -20°C to -80°C. Ultracentrifugation and precipitation techniques are mainly used in exosome Isolation. It had been reported that both methods yielded extracellular vesicles in the size range of exosomes and included apoproteins, which can be used in downstream analyses. Creative Biostructure PNExo™ exosome products guarantee higher purity and quality to meet our customer research.
Form
Lyophilized powder
Concentration
> 1x10^6 particles
Storage
Lyophilized powder store at 4 °C. Frozen liquid store at -20°C to -80°C. Recommended to avoid repeated freeze-and-thaw cycles.
Reconstitution
Reconstitute lyophilized exosome by adding deionized water for a desired final concentration. Centrifuge before opening to ensure exosomes are at bottom, resuspend exosomes by pipetting and/or vortex, please avoid bubbles. Centrifuge again and mix well for using.
Scientific Background
Health Benefits of Orange: Bioactive Compounds and Biological Functions
Orange (Citrus sinensis) is well-recognized for its rich profile of bioactive compounds that contribute to antioxidant, anti-inflammatory, metabolic, and cellular regulatory effects across various experimental models. These compounds function through diverse biochemical pathways, supporting redox balance, modulating cellular responses, and influencing immune and metabolic processes. Key bioactive classes include:
- Flavonoids (e.g. Hesperidin, Naringenin): Core polyphenols in orange that regulate oxidative stress and inflammation via inhibition of COX-2, VEGF, and MMPs; support apoptosis in cancer cell lines; and modulate lipid metabolism by reducing apoB secretion.
- Anthocyanins (e.g. Cyanidin-3-glucoside): Natural pigments with strong antioxidant and anti-mutagenic properties, shown to neutralize reactive oxygen species (ROS), chelate metal ions, and enhance molecular stability in biological systems.
- Carotenoids (e.g. Beta-Carotene, Lutein): Lipophilic antioxidants that quench singlet oxygen and peroxyl radicals, protecting cellular membranes from oxidative damage.
- Ascorbic Acid (Vitamin C): A key water-soluble antioxidant that lowers LDL oxidation susceptibility, maintains vascular integrity, and neutralizes ROS/RNS to reduce DNA damage and oxidative stress.
- Phenolic Acids (e.g. Hydroxybenzoic Acid, Caffeic Acid): Secondary metabolites with anti-inflammatory and chemopreventive potential, involved in downregulating COX-2 and modulating phase II detoxification enzymes.
- Other Compounds (e.g. Chlorogenic Acid, Quercetin): Additional phytochemicals reported in bitter orange varieties that may synergize with core components to enhance antioxidant defenses.
What Are Orange-Derived Exosomes?
Orange-derived exosomes are naturally occurring nanosized vesicles (approximately 30-150 nm) isolated from Citrus sinensis juice or pulp. Formed through endocytosis and exocytosis in plant cells, these vesicles are enclosed by lipid bilayers and carry proteins, lipids, and nucleic acids.
Their advantages in research include:
- Natural Origin: Harvested directly from oranges, offering a plant-based model for exploring vesicle-mediated biological processes.
- High Biocompatibility: Low immunogenicity and excellent safety profile make them suitable for in vitro and in vivo studies.
- Efficient Barrier Penetration: Capable of crossing biological barriers and entering target cells efficiently via endocytic pathways.
- Targeted Delivery Potential: Naturally enriched with surface molecules like integrins, and modifiable for enhanced targeting to specific cell types.
- Cargo Protection and Controlled Release: Their membrane structure shields encapsulated molecules, supporting stability and controlled release.
- Scalability and Cost Efficiency: Compared to mammalian exosomes, plant-derived exosomes benefit from abundant raw materials and simplified extraction procedures. Orange-derived exosomes offer a cost-effective alternative for scalable research applications without compromising structural integrity or biological activity.
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Characterization of nanovesicles isolated from orange juice (ONVs). (A) Size distribution of ONVs measured by nanoparticle tracking analysis. (B) Transmission electron microscopy (TEM) images of ONVs isolated from different types of orange juice (scale bars: 200 nm for a and b, 500 nm for c). (C) Protein profile of ONVs from various juice preparations revealed by silver-stained polyacrylamide gels. (D) Lipid composition of 1 mg ONV pellet, expressed as percentage of total lipids; two independent preparations are shown in different colors. (E) Representative metabolites detected in orange juice and ONVs by proton NMR spectroscopy. (Berger E, et al., 2020)
Potential Applications of Orange-derived Exosomes
Intestinal Health and Inflammation Regulation
Orange exosomes have shown protective effects on gut barrier function. In vitro, they modulate inflammation-related genes (e.g., ICAM1, HMOX-1) and enhance expression of tight junction markers such as CLDN1 and OCLN in colonic epithelial cells. In vivo, oral administration reversed gut epithelial alterations in obese mice, increased villi size, and improved nutrient absorption capacity.
Immune System Interaction
Orange-derived exosomes have been observed to influence mucosal immunity. In a mouse model of IgA nephropathy, oral delivery of dexamethasone-loaded orange vesicles reduced proteinuria and renal inflammation by modulating intestinal lymphocyte function, indicating their potential for gut-immune axis research.
Targeted Delivery in Cancer Research
Leveraging their natural membrane properties, orange exosomes can efficiently penetrate tissues and deliver therapeutic cargo. In ovarian cancer models, they demonstrated enhanced tumor accumulation via transcytosis and supported intracellular drug delivery, resulting in significant in vivo efficacy.
Plant Disease Control and RNA Delivery
In plant systems, orange exosomes serve as a natural vector for delivering functional RNAs. They have been used to transfer endogenous microRNAs to inhibit fungal pathogens like Penicillium italicum, reducing disease progression in citrus fruits. Additionally, they have been engineered to carry exogenous dsRNA for crop protection, suppressing mycotoxin production and enhancing disease resistance.
References
- Berger E, Colosetti P, Jalabert A, et al. Use of nanovesicles from orange juice to reverse diet-induced gut modifications in diet-induced obese mice. Molecular Therapy Methods & Clinical Development. 2020, 18: 880-892.
- Bruno S P, Paolini A, D'Oria V, et al. Extracellular vesicles derived from citrus sinensis modulate inflammatory genes and tight junctions in a human model of intestinal epithelium. Frontiers in Nutrition. 2021, 8: 778998.
- Zhang W, Yuan Y, Li X, et al. Orange-derived and dexamethasone-encapsulated extracellular vesicles reduced proteinuria and alleviated pathological lesions in IgA nephropathy by targeting intestinal lymphocytes. Frontiers in Immunology. 2022, 13: 900963.
- Yin C, Zhu H, Lao Y, et al. MicroRNAs in the exosome-like nanoparticles from orange juice inhibit Citrus blue mold caused by Penicillium italicum. LWT, 2023, 182: 114781.
- Long F, Pan Y, Li J, et al. Orange-derived extracellular vesicles nanodrugs for efficient treatment of ovarian cancer assisted by transcytosis effect. Acta Pharmaceutica Sinica B. 2023, 13(12): 5121-5134.
- Yin C, Lao Y, Xie L, et al. Citrus exosome-modified exogenous dsRNA delivery reduces plant pathogen resistance and mycotoxin production. Pesticide Biochemistry and Physiology. 2024, 205: 106151.
Case Study
Case Study 1: Oral Delivery of Orange-Derived Exosome-Encapsulated Dexamethasone in IgA Nephropathy (Zhang W, 2022)
This study developed orange-derived extracellular vesicles (OEVs) as oral carriers for dexamethasone phosphate (DexP) to modulate intestinal immunity in IgA nephropathy. EVs-DexP showed high encapsulation efficiency (70-80%) and remained stable in gastrointestinal conditions.
In vitro, EVs-DexP suppressed CD4⁺CD69⁺ lymphocyte activation more effectively than DexP alone (50.98% vs. 58.48%, p = 0.0175). In an IgAN mouse model, EVs-DexP reduced urinary ACR from 173.3 to 84.3 μg/mg and alleviated mesangial proliferation and IgA deposition.
Further analysis revealed decreased IgA⁺B220⁺ cells in Peyer's patches (3.98% vs. 8.37%, p = 0.016) and reduced LIGHT⁺CD4⁺ T cells, indicating inhibition of T cell-dependent IgA synthesis. These results highlight OEVs as a promising strategy for targeted steroid delivery with reduced systemic exposure.
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Figure 1. Evaluation of the stability, safety, and biodistribution of orally delivered extracellular vesicles (EVs). (A) Physicochemical stability of EVs in solutions of varying pH (pH 2, pH 7, pH 8) and artificial gastrointestinal fluids. Size and zeta potential were assessed after 2-hour incubation. (B-D) Assessment of hepatic and renal function in mice following oral (po.) and intravenous (iv.) administration of EVs; data indicate no observable toxicity. (E) Biodistribution analysis of Dil-labeled EVs in mice after oral gavage using fluorescence reflectance imaging; fluorescence was mainly detected in the ileocecal region at 2-4 hours post-administration. (F, G) Colocalization analysis showing Dil-labeled EVs (red) and immune cells in the jejunum (F) and ileum (G): CD4⁺ T cells, CD11b⁺ follicular dendritic cells, and CD11c⁺ macrophages (green). Nuclei were counterstained with DAPI (blue). Scale bars: 10 μm. -3.jpg)
Figure 2. Improved therapeutic efficacy of EVs-DexP in IgAN mice. Twenty-four mice were divided into four groups: control, IgAN model, IgAN treated with DexP, and IgAN treated with EVs-DexP (n = 6 per group). (A, B) Urinary albumin/creatinine ratio (ACR) was measured before modeling and after 12 weeks. (C, D) Liver aminotransferase and serum creatinine levels were assessed at 12 weeks. (E) Confocal immunofluorescence imaging of renal tissue showing IgA (FITC, green) deposition and nuclear DAPI staining (blue); scale bar = 75 μm. (F) PAS staining of glomerular structure; EVs-DexP group showed attenuated mesangial proliferation compared to IgAN group; scale bar = 20 μm. Case Study 2: Transcytosis-Enhanced Delivery of Orange-Derived Nanovesicles in Ovarian Cancer Models (Long F, 2023)
This study explored orange-derived extracellular vesicles (OEVs) as nanocarriers to improve drug penetration and delivery in ovarian cancer. Researchers developed a hybrid nanodrug (DN@OEV) by attaching doxorubicin-loaded, cRGD-modified nanoparticles to OEVs. In SKOV3 ovarian cancer cells, DN@OEVs showed a transcytosis rate of 15.8%, over 10 times higher than that of nanoparticles alone. This was linked to receptor-mediated endocytosis and preferential trafficking through Rab5/Rab11-regulated recycling endosomes, avoiding lysosomal degradation.
In multicellular tumor spheroids, DN@OEVs penetrated more deeply than controls, and Exo1 inhibition confirmed the dependence on active transcytosis. In vivo imaging in orthotopic tumor-bearing mice demonstrated that DN@OEVs accumulated more efficiently in tumor tissues compared to free drug and unmodified carriers. After 16 days of treatment, DN@OEVs reduced tumor volume to 50 mm³ versus 1190 mm³ in the PBS group. No significant weight loss or pro-inflammatory cytokine surge was observed, indicating good biosafety. These results support the potential of OEV-based systems for enhancing drug delivery in solid tumor models.
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Figure 1. Evaluation of the biodistribution and ovarian tumor-targeting efficacy of DN@OEV in vivo. (A) In vivo fluorescence imaging of SKOV3-Luc orthotopic ovarian tumor-bearing nude mice after intraperitoneal injection of free Cy7, Cy7-DN, or Cy7-DN@OEV at various time points. At 96 h post-injection, mice were sacrificed, and major organs and tumors were harvested for ex vivo Cy7 and luciferase signal imaging. (B) Quantitative analysis of fluorescence intensity in organs and tumor tissues from ex vivo imaging. (C) Confocal microscopy images of tumor sections following PBS, DOX, DN, or DN@OEV treatment. Scale bar: 50 μm. (D) Schematic representation of transcytosis verification using Zombie and live mice bearing orthotopic tumors. Zombie mice were fixed with PFA prior to DN@OEV administration. (E) Comparison of DOX, DN, and DN@OEV accumulation in tumor tissues between Zombie and live mice. -5.jpg)
Figure 2. Evaluation of the antitumor effects of DN@OEV in orthotopic SKOV3 ovarian cancer-bearing nude mice. (A) In vivo bioluminescence imaging (IVIS) of mice treated with PBS (control), DOX, DN, or DN@OEV (2.5 mg/kg DOX equivalent), performed every 4 days following intraperitoneal injection. Imaging started on Day 0, which was 3 days after SKOV3 cell implantation. (B) Experimental timeline for drug administration and tumor monitoring. (C) Quantification of luminescent signal intensities and monitoring of body weight in each group. References
- Zhang W, Yuan Y, Li X, et al. Orange-derived and dexamethasone-encapsulated extracellular vesicles reduced proteinuria and alleviated pathological lesions in IgA nephropathy by targeting intestinal lymphocytes. Frontiers in Immunology. 2022, 13: 900963.
- Long F, Pan Y, Li J, et al. Orange-derived extracellular vesicles nanodrugs for efficient treatment of ovarian cancer assisted by transcytosis effect. Acta Pharmaceutica Sinica B. 2023, 13(12): 5121-5134.