PNExo™ Exosome-Lemon(PNE-FL08)

Benefits of Lemon: Bioactive Components and Functional Properties

Lemon (Citrus limon) is widely valued for its diverse range of bioactive compounds that contribute to its antioxidant, anti-inflammatory, antimicrobial, and metabolic regulatory effects in experimental models. These components act through multiple biochemical pathways, supporting cellular resilience, modulating oxidative stress, and influencing microbial dynamics. Below are the major classes of bioactives found in lemon and their associated functions based on published scientific studies:

• Flavonoids (e.g. Hesperidin, Limocitrin): Key phenolic compounds in lemon that reduce oxidative stress and inflammatory markers via modulation of COX-2, TNF-α, and IL-6, support vascular tone, and inhibit tyrosinase activity to help regulate pigmentation and skin homeostasis.
• Phenolic Acids (e.g. Ferulic Acid, p-Hydroxybenzoic Acid): Strong radical scavengers that work synergistically with vitamin C to enhance photoprotection, reduce UV-induced skin damage, and prevent lipid peroxidation in cell membranes.
• Monoterpenes (e.g. D-Limonene, β-Pinene, γ-Terpinene): Dominant components of lemon essential oil, shown to exert antimicrobial, anti-inflammatory, and skin-penetration-enhancing effects, as well as modulate tumor cell viability in vitro via apoptosis-related signaling.
• Limonoids (e.g. Limonin): Citrus-specific triterpenoids with reported neuroprotective and metabolic regulatory effects, including modulation of fat accumulation, insulin sensitivity, and neuronal apoptosis, though requiring dose attention due to potential mitochondrial stress.
• Ascorbic Acid (Vitamin C): A potent antioxidant that suppresses NF-κB-driven inflammation, supports immune defense mechanisms, and contributes to collagen biosynthesis and barrier function.
• Organic Acids (e.g. Citric Acid): Naturally occurring acids that support mineral absorption by adjusting gut pH, and aid in the dissolution of urinary calcium crystals, thus promoting renal and digestive health.

What are Lemon-Derived Exosomes?

Lemon exosome-like nanoparticles (LELNs) are naturally occurring nanosized (approximately 100 nm) extracellular vesicles derived from Citrus limon. These advanced vesicles are generated by plant cells through a coordinated process of endocytosis, vesicle fusion, and exocytosis, and they encapsulate a variety of biomolecules, including proteins, lipids, and nucleic acids.

The appeal of lemon-derived exosomes in research stems from several key advantages:

• Natural Origin: Sourced directly from lemons, they represent a natural platform for exploring biological interactions.
• Biocompatibility: Their plant origin often suggests favorable compatibility with biological systems, making them promising for various research applications.
• Bioactive Cargo Delivery: These exosomes naturally carry a diverse array of bioactive molecules, allowing for their exploration as delivery vehicles for specific compounds.
• Inherent Properties: Research indicates that plant-derived exosomes, including those from lemons, may possess inherent antioxidant, anti-inflammatory, and anti-aging properties, contributing to their broad research potential.

Physical and morphological analysis of nanovesicles derived from Citrus limon L. (A) Citrus lemon before and after juice extraction. (B) Transmission electron microscopy (TEM) image showing nanovesicles at 50,000× magnification. Scale bar = 100 nm. (C) Nanoparticle tracking analysis (NTA) showing size distribution and concentration profile of the nanovesicles. (Takakura H, et al., 2022)

Potential Applications of Lemon-Derived Exosomes

Antioxidant and Cellular Protection

Lemon exosome-like nanoparticles (LELNs) activate key cytoprotective pathways, including AhR and Nrf2, enhancing the expression of antioxidant enzymes and reducing intracellular ROS accumulation. These effects contribute to the maintenance of redox homeostasis and cellular integrity under oxidative stress, particularly in liver cells and inflamed tissues. Their ability to stabilize redox balance supports their use as natural antioxidants in experimental oxidative damage models.

Anti-inflammatory and Immunomodulatory Effects

Studies have shown that lemon-derived exosomes suppress pro-inflammatory signaling pathways such as ERK and NF-κB, while promoting anti-inflammatory responses through macrophage polarization. In animal models, they have been observed to alleviate tissue inflammation, reduce cytokine production, and aid in rebalancing immune responses, highlighting their potential as natural immunomodulators.

Microbiome and Gut Health Support

LELNs enhance the survival and function of beneficial gut bacteria by improving their resistance to bile salts and promoting stress adaptation. Additionally, these vesicles can modulate the gut microbial environment, indirectly inhibiting pathogenic species by supporting probiotic growth and metabolic function. Such findings underscore their potential as prebiotic enhancers in microbiota-related research.

Targeted Cellular Regulation

Lemon nanovesicles exhibit selective activity against cancer cells by triggering apoptosis pathways, particularly via TRAIL signaling. They have been shown to inhibit proliferation in various tumor cell lines and reduce tumor burden in vivo. Furthermore, their structural compatibility enables functionalization for drug encapsulation and targeted delivery, expanding their utility in therapeutic delivery system design.

Regenerative and Wound Healing Research

In tissue engineering studies, LELNs have been incorporated into biomaterials such as hydrogels to promote wound healing, especially in chronic or diabetic contexts. By modulating immune cell behavior and stimulating tissue repair mechanisms, they demonstrate promising potential for regenerative medicine applications.

References

1. Raimondo S, Naselli F, Fontana S, et al. Citrus limon-derived nanovesicles inhibit cancer cell proliferation and suppress CML xenograft growth by inducing TRAIL-mediated cell death. Oncotarget. 2015, 6(23): 19514.
2. Lei C, Mu J, Teng Y, et al. Lemon exosome-like nanoparticles-manipulated probiotics protect mice from C. diff infection. Iscience. 2020, 23(10).
3. Lei C, Teng Y, He L, et al. Lemon exosome-like nanoparticles enhance stress survival of gut bacteria by RNase P-mediated specific tRNA decay. Iscience. 2021, 24(6).
4. Takakura H, Nakao T, Narita T, et al. Citrus limon L.-derived nanovesicles show an inhibitory effect on cell growth in p53-inactivated colorectal cancer cells via the macropinocytosis pathway. Biomedicines. 2022, 10(6): 1352.
5. Raimondo S, Urzì O, Meraviglia S, et al. Anti-inflammatory properties of lemon-derived extracellular vesicles are achieved through the inhibition of ERK/NF-κB signalling pathways. Journal of Cellular and Molecular Medicine. 2022, 26(15): 4195-4209.
6. Urzì O, Cafora M, Ganji N R, et al. Lemon-derived nanovesicles achieve antioxidant and anti-inflammatory effects activating the AhR/Nrf2 signaling pathway. Iscience. 2023, 26(7).
7. Tinnirello V, Zizzo M G, Conigliaro A, et al. Industrial-produced lemon nanovesicles ameliorate experimental colitis-associated damages in rats via the activation of anti-inflammatory and antioxidant responses and microbiota modification. Biomedicine & Pharmacotherapy. 2024, 174: 116514.
8. Gasparro R, Gambino G, Duca G, et al. Protective effects of lemon nanovesicles: evidence of the Nrf2/HO-1 pathway contribution from in vitro hepatocytes and in vivo high-fat diet-fed rats. Biomedicine & Pharmacotherapy. 2024, 180: 117532.
9. Jamshidi Z, Dehghani S, Nameghi M A, et al. Engineering extracellular vesicles derived from lemons for delivering chemotherapeutic drug employing periostin targeting. Journal of Drug Delivery Science and Technology. 2024, 99: 106011.
10. Jin E, Yang Y, Cong S, et al. Lemon-derived nanoparticle-functionalized hydrogels regulate macrophage reprogramming to promote diabetic wound healing. Journal of Nanobiotechnology. 2025, 23(1): 68.

Case Study 1: Antioxidant and Anti-inflammatory Properties of Lemon-derived Nanovesicles in Skin and Inflammation Models (Urzì O, 2023)

A study investigated lemon-derived nanovesicles (LNVs) for their antioxidant and anti-inflammatory properties. In human dermal fibroblasts, LNVs (10 - 25 µg/mL) were internalized without cytotoxicity and significantly reduced ROS levels induced by H₂O₂ and UVB exposure. This effect was linked to the activation of the AhR/Nrf2 signaling pathway and upregulation of antioxidant enzymes. LNVs also enhanced extracellular matrix gene expression (COL1A1, HAS2) and promoted wound closure under oxidative stress. In zebrafish models, LNV pre-treatment reduced neutrophil recruitment and ROS production following both sterile injury and LPS-induced inflammation, confirming systemic antioxidant and immunomodulatory effects.

Figure 1. Protective effects of LNVs against oxidative damage and UVB-induced stress in HDFα cells. (A) Quantification of ROS levels after H₂O₂ (left) and UVB (right) stimulation in cells pre-treated with LNVs (10 and 25 µg/mL) for 24 h. (B) Wound healing assay in HDFα cells pre-treated with LNVs (10 and 25 µg/mL). Cells were scratched, exposed to UVB for 25 s, and imaged at 0, 3, and 6 h. Percentage of wound closure was quantified.

Figure 2. Antioxidant and immunomodulatory effects of prophylactic LNV treatment in a zebrafish inflammation model. (A) Schematic diagram of Pa-LPS-induced inflammatory stimulus in 48 hpf embryos. (B) Representative images at 4 hours post-injection (hpi) showing neutrophil (mpx⁺) and ROS (DHE⁺) localization in whole embryo and trunk region with or without 25 µg/mL LNV pre-treatment. White asterisks indicate mpx⁺DHE⁺ cells. (C - E) Quantification of DHE⁺ cells (C), mpx⁺ cells (D), and dual-positive mpx⁺DHE⁺ cells (E) in the region of interest.

Case Study 2: Lemon-Derived Nanoparticles Enhance Diabetic Wound Healing via Macrophage Modulation (Jin E, 2025)

A study demonstrated that lemon-derived nanoparticle-functionalized hydrogels (LNV-gel) can significantly accelerate diabetic wound healing by modulating macrophage polarization. In vitro, LNVs promoted M2 macrophage phenotype markers (CD206, Arg-1) while reducing M1-associated genes (iNOS, TNF-α). In a streptozotocin-induced diabetic mouse model, wounds treated with LNV-gel showed a 91.8% closure rate by day 14, significantly outperforming both blank hydrogels and untreated controls.

Histological analysis revealed enhanced collagen deposition and angiogenesis in LNV-gel groups, supported by elevated VEGF and TGF-β expression. The study highlights LNVs as bioactive nanocarriers capable of reprogramming immune responses and supporting tissue regeneration, offering promising potential in chronic wound management and regenerative biomaterials research.

Figure 1. Immunomodulatory effects of GelMA/DAS hydrogel loaded with lemon-derived exosomes (Exo) on RAW264.7 macrophages. (A) Schematic illustration showing macrophage phenotype modulation by GelMA/DAS/Exo hydrogel. (B - C) RT-qPCR results displayed as a bar graph and heatmap showing decreased iNOS and TNF-α (M1 markers) and increased Arg-1 and IL-10 (M2 markers). (D) Immunofluorescence (IF) images show increased Arg-1⁺ and decreased iNOS⁺ cells after hydrogel treatment. (E - F) Quantification of fluorescence intensities of Arg-1 and iNOS. (G - H) Western blot analysis and quantification of Arg-1 and iNOS protein expression. (I) Flow cytometry shows reduction in CD86⁺ (M1) and increase in CD206⁺ (M2) macrophages. These results suggest that GelMA/DAS/Exo promotes M1 to M2 polarization via NF-κB pathway, offering therapeutic potential in diabetic wound healing.

Figure 2. Evaluation of wound-healing efficacy of GelMA/DAS/Exo hydrogel in a diabetic full-thickness skin defect rat model. (A) Schematic showing the diabetic skin injury model and grouping strategy (PBS, Lemon Exosome, GelMA/DAS, GelMA/DAS/Exo). (B) Visual observation of wound healing over 14 days shows enhanced closure in the GelMA/DAS/Exo and Lemon Exosome groups compared to controls. (C) H&E staining of skin sections at day 14 shows improved epithelial regeneration and reduced inflammation in the GelMA/DAS/Exo group. (D) Masson’s trichrome staining highlights enhanced collagen deposition in the hydrogel-treated wounds, indicating effective tissue remodeling. Scale bars: 2 mm (2×) and 400 µm (10×).

References

1. Urzì O, Cafora M, Ganji N R, et al. Lemon-derived nanovesicles achieve antioxidant and anti-inflammatory effects activating the AhR/Nrf2 signaling pathway. Iscience. 2023, 26(7).
2. Jin E, Yang Y, Cong S, et al. Lemon-derived nanoparticle-functionalized hydrogels regulate macrophage reprogramming to promote diabetic wound healing. Journal of Nanobiotechnology. 2025, 23(1): 68.

• Can lemon extracellular vesicles be used for skin-related research?

  Yes. Lemon extracellular vesicles have demonstrated strong potential in skin models, including reducing oxidative stress, modulating inflammation, and supporting collagen-related gene expression, making them valuable in dermatological and cosmetic research.

• Can lemon exosomes be engineered for targeted delivery?

  Yes. Like other plant-derived vesicles, lemon exosomes can be functionalized or loaded with specific molecules, offering promise as natural delivery systems in targeted drug or gene delivery research.

• How are lemon exosomes isolated and purified?

  Our lemon-derived exosomes are extracted using standardized ultracentrifugation and filtration methods, followed by rigorous quality control to ensure purity, structural integrity, and consistent nanoparticle size.

• How are lemon exosomes typically stored and handled for research use?

  Our lemon-derived exosomes are supplied as lyophilized powder for long-term storage at 4°C. Once reconstituted, they should be stored at -20°C to -80°C and protected from repeated freeze-thaw cycles to maintain activity.

• Is PNExo™ Exosome-Lemon suitable for DIY use?

  No. This product is designed exclusively for research and industrial applications. It is not approved for use in homemade formulations.

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