Exosomes Isolated from Herbages

Recently, exosomes of plant origin have attracted considerable attention, particularly from herbages known for their medicinal activities. These plant exosomes have opened new avenues of research and application in therapy, agriculture, environmental remediation, cosmetics, and personal care.

Creative Biostructure's PNExo™ exosome products, isolated from various herbages, represent a breakthrough solution for research and industrial applications. All products are of high purity and quality. In addition to our product list, we provide GMP exosome production and CDMO services for your unique project.

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Background

Exosomes are small membrane-bound vesicles secreted by cells that play a crucial role in cell-cell communication. These membrane-bound particles contain lipids, proteins, and RNA. They were first identified in mammals, such as exosomes isolated from stem cell lines, exosomes isolated from cancer cell lines, and exosomes isolated from body fluids. However, recent studies have pointed to the presence of exosomes isolated from plant. Plant exosomes are rich in bioactive molecules, which are relevant to plant stress responses, inter-plant communication, and interactions with microbes.

Herbage-derived exosomes often carry beneficial compounds such as antioxidants, anti-inflammatory agents, and compounds that can enhance the immune response. For example, exosomes from Angelica sinensis (Dong Quai) are known for their antioxidant properties, while exosomes derived from Panax quinquefolius support immune system function and promote longevity.

A summary of the contents in various common exosomes isolated from herbages is shown below:

Biogenesis of Exosomes in Plants

The biogenesis of exosomes in herbages begins with forming multivesicular bodies (MVBs) in the cytoplasm. These MVBs are membrane-bound compartments that contain internal vesicles. The MVBs eventually fuse with the plasma membrane, releasing their internal vesicles into the extracellular space as exosomes.

In herbages, the exosomal cargo comprises a wide array of molecules, such as lipids, proteins, RNAs (including microRNAs, mRNAs, and long non-coding RNAs), and metabolites. These cargo molecules reflect the plant's metabolic state and environmental response. For example, stress conditions like drought, heavy metal exposure, or pathogen attack can trigger changes in the composition of exosomal cargo, which may play a role in signaling neighboring cells or other organisms.

Figure. 1: Plant-derived extracellular vesicles. A. Representative figure showing the overview structure and generalized composition of plant-derived extracellular vesicles (P-EVs). P-EVs contain various biomolecules in their lumen and surface. B. Schematic representation of the biological composition of P-EVs and their function in pathogen defense. (Nemati M et al., 2022)

Isolation Techniques for Plant Exosomes

The isolation of exosomes from herbages involves several steps, with different methods employed based on the specific properties of the plant material and the desired exosome yield. The following are common techniques used to isolate and purify plant-derived exosomes:

• Ultracentrifugation is the most common exosome for isolating exosomes from plant tissues. The process involves subjecting the liquid medium (typically a plant extract) to high-speed centrifugation to pellet the exosomes.
• Size-Exclusion Chromatography (SEC) is another effective method to separate exosomes from other particles based on size. It uses a column filled with porous material that allows smaller molecules to pass through more easily, separating them from larger vesicles. SEC is advantageous for purifying exosomes with high yields and lower contamination levels than ultracentrifugation.
• Density Gradient Centrifugation (DGC) isolates exosomes based on their density. Layering solutions with increasing sucrose concentrations creates a sucrose gradient. The sample containing exosomes is placed at the top and centrifuged. During centrifugation, the exosomes move to a specific point in the gradient where their density matches the sucrose solution. This allows them to be separated from other cellular components, ensuring high purity.
• Polymer Precipitation remove larger particles from plant extracts, followed by exosome precipitation using reagents like polyethylene glycol (PEG). This technique is relatively simple and cost-effective but may result in lower purity of the exosomal preparation.
• Microfluidics (MF) uses small channels and controlled fluid flow to isolate exosomes. By applying forces such as hydrodynamic or electrokinetic forces, microfluidic devices can separate exosomes based on size, charge, or surface markers. This technique is precise, fast, and scalable, offering a more efficient and controlled way to isolate exosomes than traditional methods.
• Tangential Flow Filtration (TFF) isolates exosomes by passing a sample through a membrane filter with a specific pore size. Unlike traditional filtration, TFF operates tangentially to the filter, which prevents clogging and allows continuous filtration. This method efficiently separates exosomes from larger particles and other impurities, providing high-purity isolates.

Figure. 2: Schematic diagram of the GELNs isolation and purification methods. Fresh ginger was first processed using a breaker or blender to obtain ginger juice, and the juice was subjected to differential centrifugation to obtain crude GELNs, followed by further purification to obtain pure GELNs. Currently reported methods for GELNs purification include sucrose gradient ultracentrifugation, membrane filtration, and PEG-based precipitation. The obtained GELNs showed a 50–200 nm phospholipid bilayer structure containing lipids, protein, small RNA, and active ingredients. GELNs, ginger-derived exosome-like nanoparticles. PEG, polyethylene glycol. (Zhu H and He W, 2023)

Applications of Exosomes Isolated from Herbages

Therapeutic Applications

Herbages-derived exosomes, including microRNAs, proteins, and lipids, carry bioactive molecules that have therapeutic potential. These exosomes are promising for developing new drugs, especially for diseases with limited traditional treatments. As natural delivery vehicles, herbage exosomes can target specific cells or tissues. They deliver therapeutic agents directly to the intended areas, reducing the risk of side effects.

For example, exosomes from Turmeric contain curcumin, a compound known for its anti-inflammatory and anticancer effects. These exosomes could be used in targeted cancer therapies, offering a novel approach that reduces systemic side effects. Similarly, exosomes from Yarrow are recognized for their wound-healing properties. These could be incorporated into new skincare products or wound dressings, helping to speed up tissue regeneration.

Agricultural Biotechnology

Exosomes isolated from herbage hold significant potential in agriculture. These exosomes can serve as natural biostimulants, improving plant growth, enhancing stress resistance, and promoting overall plant health. They transfer RNA molecules or proteins involved in stress responses, helping crops cope with environmental challenges such as drought, high salinity, and pest attacks.

Exosomes from plants like Sedum lineare and Typhae pollen can be used to develop sustainable agricultural solutions. These exosomes help plants adapt better to changing environmental conditions. By enhancing crop resilience without chemical fertilizers or pesticides, herbage exosomes offer an eco-friendly alternative to traditional agricultural practices.

Environmental Remediation

Herbage exosomes may also contribute to environmental remediation, particularly in detoxifying polluted environments. They carry molecules that can bind to heavy metals and other toxins. This enables herbage exosomes to be used in phytoremediation, where plants absorb or break down pollutants from contaminated soils or water.

For example, exosomes from White Birch and Viola yedoensis may contain molecules that help break down environmental pollutants. These exosomes could be applied to innovative techniques to clean up polluted sites or treat wastewater. This approach offers an environmentally friendly solution to environmental remediation.

Cosmetic and Personal Care

The cosmetic industry is becoming more interested in herbage-derived exosomes because of their potential benefits for skin rejuvenation, anti-aging, and wound healing. Exosomes from herbages like Trichosanthes kirilowii are especially valuable. They are packed with bioactive molecules, including flavonoids. These flavonoids have been shown to have antioxidant and anti-inflammatory effects, making them great for improving skin health.

Case Studies

Case Study 1: Salvia miltiorrhiza-Derived Exosome-like Nanoparticles as Angiogenesis Promoters for Myocardial Ischemia-Reperfusion Injury

Exosomes have been found to promote angiogenesis in myocardial ischemia-reperfusion (MI/R) injury. However, mammalian cell-based exosomes are not used in a broad clinical context because of their low yield. Therefore, there is an alternative option—exosome-like nanoparticles extracted from Salvia miltiorrhiza, a traditional Chinese medicine with angiogenic properties. These nanoparticles are high-yielding, cost-effective, and stable with ideal particle size and Zeta potential. In this study, Salvia miltiorrhiza-derived exosome-like nanoparticles have been showed to improv the viability, proliferation, and migration of Human Umbilical Vein Endothelial Cells (HUVECs). These nanoparticles significantly improved cardiac function in MI/R mouse models, as reflected by higher ejection fraction (EF%) and fractional shortening (FS%). Immunohistochemical staining showed a higher density of microvessels in the peri-infarct zone, implying that Salvia miltiorrhiza exosome-like nanoparticles are a potential treatment for cardiac healing and angiogenesis after MI/R MI/R damage.

Figure 3. Functional analysis of DDN in vivo. (A) Cardiac function of C57BL/6 mice 21 days after MI/R (n = 3); (B) angiogenesis of C57BL/6 mice in each group (n = 3). ** p < 0.01, *** p < 0.001, ns means no significant difference, scale bar = 50 µm (Zhang S, et al., 2024)

Case Study 2: Anticancer Effects of Centella Asiatica-Derived Nanovesicles in HepG2 Cells

The researchers tested Centella asiatica-derived exosome-like nanovesicles (ADNVs) as potential anticancer agents to address both cost and toxicity associated with nanoparticle-based cancer treatments. They demonstrated in vitro studies that ADNVs suppress HepG2 proliferation, cell cycle arrest at G1, and apoptosis. Analysis revealed that ADNVs significantly increase reactive oxygen species (ROS) levels and cause mitochondrial damage. More interestingly, gene expression studies showed that ADNVs regulated apoptotic genes such as Bax and CASP3 and downregulated Bcl-2. Metabolomics analysis revealed that cellular metabolites, especially amino acid metabolism and lipid biosynthesis, were dramatically altered after ADNVs. These data suggest that ADNVs affect essential metabolic pathways, such as L-arginine and L-proline. Overall, ADNVs exhibit potent anticancer effects, offering a promising, low-cost, and effective alternative for cancer treatment by targeting metabolic and apoptotic pathways in HepG2 cells.

Figure 4. Centella Asiatica–derived nanovesicles (ADNVs) were internalized and inhibited the proliferation and apoptosis of HepG2 cells. (A) HepG2 cells were incubated with Dio-labeled ADNVs for 24 h. (B) HepG2 cells were incubated with ADNVs (0, 10, 20, 40, 80, or 120 μg/mL) for 24 and 48 h. Cell viability was determined using the CCK-8 assay. (C) Cells were treated with 20, 40, or 80 μg/mL ADNVs for 48 h. Annexin V-FITC/PI double staining and flow cytometry were used to determine the apoptosis rate. (D) Effect of ADNVs on HepG2 cell apoptosis compared with the control group. (E) Flow cytometry analysis of HepG2 cells treated with 0, 20, 40, and 80 μg/mL ADNVs. (F) DNA content in each cell cycle phase (G1, S, G2) after ADNVs treatment. Comparison with the control group. N = 3, *p <0.05, **p <0.01, ***p <0.001. (Huang J, et al., 2024)

Product Advantages

• High Purity and Quality: Through advanced isolation techniques such as ultracentrifugation, PEG precipitation, and Tangential Flow Filtration (TFF), Creative Biostructure ensures that the exosomes meet the highest purity standards. This guarantees the effectiveness of the exosomes in research and production applications.
• GMP Production and CDMO Services: The company offers GMP-certified production of exosomes and provides Contract Development and Manufacturing Organization (CDMO) services. This ensures customers can rely on Creative Biostructure for large-scale, high-quality exosome production.
• Selectable Form: Customers can request exosomes in either lyophilized powder form for easy storage or frozen liquid to maintain freshness. This flexibility allows for convenience in handling and long-term storage.
• Eco-friendly and Sustainable: Herb-derived exosomes are a sustainable alternative to traditional chemical-based solutions in agriculture and environmental remediation. By harnessing the power of nature, Creative Biostructure's products provide a green solution for modern-day challenges.
• Versatility: The exosomes are sourced from various herbages, offering a diverse range of bioactive compounds with different therapeutic, agricultural, and environmental applications.

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Frequently Asked Questions

• What are herbages-derived exosomes, and do they differ from other types of exosomes?

  Herbages-derived exosomes are extracellular vesicles isolated from medicinal or beneficial herbages. They are rich in bioactive compounds like proteins, lipids, and nucleic acids, which contribute to their unique therapeutic properties. Unlike animal-derived exosomes, they are plant-based, eco-friendly, and free of zoonotic contamination.

• Are exosomes isolated from herbage safe and non-toxic?

  Herbage-derived exosomes are generally considered safe and non-toxic due to their natural origin. They are biocompatible and biodegradable, making them suitable for pharmaceutical, nutraceutical, and cosmetic applications. However, quality control and testing are essential to ensure safety.

• How are exosomes isolated and purified from herbage?

  Creative Biostructure uses several advanced techniques to isolate plant-derived exosomes. These include ultracentrifugation, PEG precipitation, and Tangential Flow Filtration (TFF) technology. These methods ensure that the exosomes are highly pure and functional, suitable for various research and production applications.

• What is the difference between lyophilized powder and frozen liquid exosomes?

  Lyophilized powder exosomes are dehydrated and come in powder form, which makes them easier to store for long periods, whereas frozen liquid exosomes are convenient for immediate use but require careful storage and handling.

• Do you provide exosome isolation services?

  We offer comprehensive exosome isolation services, including GMP production and CDMO solutions, tailored to meet your research and production requirements efficiently and reliably.

At Creative Biostructure, we provide PNExo™ herbages exosome products and expert exosome isolation services. Our exosomes offer a natural, sustainable solution to addressing global challenges. Contact us to discover the ideal solution for your needs!

References

1. Huang J, Cao X, Wu W, Han L, Wang F. Investigating the proliferative inhibition of HepG2 cells by exosome-like nanovesicles derived from Centella Asiatica extract through metabolomics. Biomedicine & Pharmacotherapy. 2024;176:116855.
2. Nemati M, Singh B, Mir RA, et al. Plant-derived extracellular vesicles: a novel nanomedicine approach with advantages and challenges. Cell Commun Signal. 2022;20(1):69.
3. Zhang S, Xia J, Zhu Y, Dong M, Wang J. Establishing salvia miltiorrhiza-derived exosome-like nanoparticles and elucidating their role in angiogenesis. Molecules. 2024;29(7):1599.
4. Zhao B, Lin H, Jiang X, et al. Exosome-like nanoparticles derived from fruits, vegetables, and herbs: innovative strategies of therapeutic and drug delivery. Theranostics. 2024;14(12):4598-4621.
5. Zhu H, He W. Ginger: a representative material of herb-derived exosome-like nanoparticles. Front Nutr. 2023;10.