Unlocking the Power of Plant Exosomes (PDEVs) – The New Natural Way of Delivering Cosmetic Efficacy

Introduction

Plant exosomes are one of the latest innovations in cosmetics and toiletries and the one emerging technology that market analysis companies, such as Mintel, tell us to watch. Exosomes are a special type of tiny extracellular vesicles (EVs) that offer a natural way to substantially improve product performance. While it is very likely that you have heard of synthetic liposomes and natural subcellular organelles such as melanosomes, lysosomes and plant phytosomes, it is less likely that that you are familiar with plant derived exosomes.

Plant exosomes (PDEVs) have significant advantages over exosomes derived from other sources. For example, plant exosomes are accepted for use in personal care, pharma and food, which is not the case for human derived exosomes, which are not allowed in beauty products ^[1]. Plant exosomes are beginning to be produced on a commercial scale and are currently used to optimise and enhance cosmetic efficacy. Exosomes have been shown to penetrate the skin barrier and help reduce fine lines, improve skin texture and hydration, and enhance skin elasticity. They have been shown to reduce inflammation and the damage caused by sunlight. Potentially they could be valuable tools for combating photodamage and premature skin ageing ^[2].

Exosomes: Hiding in plain sight

Although discovered as recently as 1983 ^{[3, 4]}, exosomes are likely to have been part of cell physiology since life began over 3.8 billion years ago. It seems therefore that exosomes are omnipresent and so have always been present in the food we eat and in the juices we drink. Extracellular vesicles found in food items, referred to as “FoodEVs”, are officially recognized and included in food databases such as the Food and Agriculture Organization (FAO), INFOODS for milk, starchy roots and tubers, nuts and seeds, and fruits. And it is because of their efficacy and safety that FoodEVs are becoming the latest innovation in functional foods ^[5].

Nature’s Microscopic Courier Service

Exosomes are extremely small, varying in size from nano to micro scale, with plant-derived exosomes (PDEVs) measuring between 50 to 150 nm. Figure 1 (see below) shows a transmission electron micrograph (TEM) of isolated PDEVs, which have been negatively stained with phosphotungstic acid to show the double membrane surrounding each exosome. It is this double membrane that makes exosomes particularly robust and able to survive in the hostile environments outside cells. Exosomes act as a type of intercellular-taxi service, protecting and carrying complex bioactive substances and molecular messages to and from cells. They are essential for tissues to function and are remarkable because they can deliver high concentrations of actives, transfer proteins, lipids, and nucleic acids and transform cells. Their functions include regulating the immune response and tissue repair ^[6].

Exosomes form when the cell membrane gradually and sequentially, encloses in on itself and creates tiny bubble-like compartments of multivesicular bodies. These subcellular bubbles mingle with other organelles within the cell and pick up a diverse molecular cargo, that will include DNA, RNA, lipids, various metabolites, along with cytosolic and cell-surface proteins. They are then enveloped in a bilipid membrane (derived from its mother-cell) before budding off and being released. As exosomes contain an assortment of different complex molecules, the properties of each type of exosome depends on the source. Factors such as the mother-species, tissue and type of cell and the local micro-conditions that the cell is experiencing when the exosomes are being produced, all have an influence ^{[5, 7]}.

Plant Exosomes: Protective, Precision Delivery in Cosmetics

Exosomes’ extraordinary ability to carry and deliver therapeutic molecules with precision has captured the attention of researchers and is revolutionizing the way pharma thinks about drug delivery. Various techniques for loading substances into exosomes are being further developed such as incubation, electroporation, sonication, extrusion, freeze-thaw cycling, and transfection ^{[8, 9]}. The discoveries from this research are spilling over into cosmetics and plant exosomes are now poised to make an equally high impact on the beauty sector.

Plant-derived exosomes contain lipids, proteins, nucleic acids, and secondary metabolites including some of the natural antioxidants commonly used in cosmetics. When encapsulated within plant exosomes, ascorbic acid, superoxide dismutase, glutathione, catalase, and melatonin etc., retain their native molecular configurations, whether membrane-bound or freely soluble. Encapsulation within plant exosomes therefore preserves their structural integrity and biological efficacy.

This is not necessarily the case when these actives are added directly into the chaos of a cosmetic formulation. The exosome’s lipid bilayer membrane and internal core provide protective native microenvironments that shield these molecules. Also, the biolipid membrane enables exosomes to safely and naturally deliver their cargo into cells via membrane fusion. Interestingly, exosomes can also concentrate biomolecules gathered from the parent cell and so they may contain much higher concentrations of actives than cell lysates, juices, extracts and infusions. Table 1 shows the typical antioxidant content of plant exosomes ^[10].

Edible plants are the ideal source of exosomes for cosmetics. There is mounting evidence showing plant exosomes can establish cross kingdom communication through absorption from dietary sources and control processes such as inflammation and tissue regeneration. They can be produced in the amounts the industry needs and doubtless because of our very long association with plant exosomes in food, exosomes derived from fruits and vegetables are safe and well-tolerated by our immune system ^[5]. To be completely sure that there is no risk of plant exosomes carrying traces of agrochemicals, it is very important that the plants used for exosome production are grown organically, i.e., cultivated on land free from pesticides etc., ^[11]. To use plant exosomes at their full potential, they first need to be carefully extracted from the appropriate plant sources, concentrated, standardised and where necessary, dried.

Going Beyond Nature: Extracting and Blending so Supercharging Nature’s Plant Exosomes

South Korean researchers compared the efficacy of plant exosomes and plant extracts in a thorough in vitro study that used the leaves of Cantella asiatica (Cica), Portulaca oleracea (Purslane), Camellia Sinensis (Green tea) and the roots of Panax ginseng (Ginseng) ^[12]. After hot air drying the plants at 45 °C for 24 hours, the tissues were finely ground using a blender. The fine plant powder was then soaked in distilled water at 80 °C for 3 hours and filtered through a 0.45 µm mesh filter.

These pre-processed samples were subject to high pressure processing (200 MPa at 25 °C for 30 seconds) before passing through a juice extractor at 30 rpm. A mesh filter removed large particles and the filtered pressed juice was divided into two parts. One part was subject to ultracentrifugation at 150,000×g, 4 °C for 70 min to remove exosomes, while exosomes were extracted from the other part using a two-phase extraction system (ATPS). In this way, plant extracts were prepared (with exosomes removed) and exosomes were harvested from the same plant sources using an aqueous two-phase system.

When human keratinocytes cells were exposed to these plant extracts and exosomes for 6 hours, gene expression analysis revealed a clear distinction between the cells treated with extracts and those treated with exosomes. The number of differentially expressed genes (DEGs) of the human keratinocytes are shown in figure 2 (see below), which was redrawn from their paper ^[12]. Around 861 genes were upregulated and 648 downregulated in the exosome groups, compared to 398 upregulated and 438 downregulated in the extract groups. This study shows extracts and exosomes differ in the way they influence skin cells and there are differences between how exosomes from different plants affect small subsets of genes. The study also showed that the plant exosomes positively influenced the genes known to impact aging, regeneration, skin barrier, and so could help maintain healthy skin ^{[12, 13]}.

Research has shown that plant exosomes can become supercharged delivery systems when more than one source of plant exosomes such as those from different fruits (i.e. from apples and papaya), or from various vegetable leaves, stems and roots, are combined.

Following research showing Euglena gracilis derived EVs enhanced skin-regenerative wound healing ^[14], a recent in vitro wound healing study showed human fibroblasts treated for 24 hours with PDEVs, increase their wound healing ability by 52% when compared to untreated wounded fibroblasts (see figure 3, below). The wound healing assay demonstrates the migratory capacity of cells in vitro. In the assay, a wound is made in a monolayer of cells and the rate of wound closure is measured at various time points.

In the experiment shown in figure 3, normal human dermal fibroblasts (NHDF) were grown into continuous compact cell monolayers before wounds were made. The wounded cell monolayers were either treated with PDEVs or left untreated (control). The distance between the two cell fronts in both the PDEV treated and untreated wounds were monitored and compared with the distance when the wounds were first made (time zero). After 24 hours, the cells treated with PDEVs had achieved complete wound closure, while the untreated control cells had only approximately 52% wound closure. This wound assay demonstrated how plant exosomes (PDEVs) can positively influence NHDF cell migration, making plant exosomes excellent candidates for anti-aging products formulated to address fine lines and wrinkles.

As we age, fibroblasts tend to become larger and less efficient, leading to a decline in collagen production and a looser, less firm skin texture. When human fibroblasts were treated with the same plant exosome mixture for 24 hours, there was a 20% reduction in area and a 22% reduction in duplication time compared to untreated fibroblasts (see figure 4, below). These results show the potential for plant exosomes to counteract some of the causes of the signs of skin aging.

Vimentin is an important intermediate filament protein that provides structural support and anchorage for organelles within the cell. Fibroblasts release vimentin into the extracellular space after injury, where it directs cell migration to the damage-site and so is critical for regulating healing. It is exciting to see that fibroblasts treated with the mixture of plant exosomes, increased the release of Vimentin into the extracellular space by 31% (see figure 5, below).

Type I collagen is the extracellular protein essential for maintaining the structure of skin. It is important for the strength of the extracellular matrix. Type I collagen also promotes elastin synthesis and speeds up wound healing. The progressive decrease in collagen production that takes place as we age results in a loss of skin tone and an increase in fine lines and wrinkles. Fibroblasts synthesise more type I collagen than other proteins. These calls are therefore used for in vitro studies on antiaging actives. When NHDF cells, isolated from the dermal layer of healthy donors, were treated in vitro for 24 hours with plant exosomes mix, fluorescence microscopy revealed a 60% increase in collagen type 1 expression (see figure 6, below).

While the exact composition of the plant exosome mixture in these studies remains proprietary, this does not diminish the significance of the results, which serve to highlight the promising antiaging effects of blends of plant-derived exosomes could have on skin.

Clinical Performance of Plant Exosomes

In an in vivo placebo controlled clinical study designed to demonstrate the effectiveness of plant exosomes on facial skin, a panel of 40 volunteers (aged between 40 and 50 years old) applied daily, either the basic serum formula without plant exosomes or the serum formula with plant exosomes. They applied 1mL of the serum daily for the first 20 days, and then 1mL twice daily for the remaining 25 days. The serum’s INCI was as follows, INCI: Aqua, Malus domestica fruit extract, Citrus reticulata fruit extract, Carica papaya fruit extract, Sorbitol, Propanediol, Benzyl alcohol, Polyglyceryl-4 caprate, Glucomannan, Silybum marianum ethyl ester, Tetrasodium glutamate diacetate, Potassium sorbate.

At the start of the trial, (T0), then at 20 days, (T20) and 45 days, (T45), blinded evaluations of skin elasticity (Cutometer^®) and of skin roughness (Antera 3D camara) were determined. Figure 7 below shows the results for facial skin elasticity. It can be clearly seen that, when compared with skin treated with the placebo, at day 20, the serum containing the plant exosomes had caused a dramatic (17.5%) improvement, which continued to increase to 22.5% after 45 days.

Unlike the placebo, which had no effect, the serum containing the plant exosomes produced a similarly dramatic improvement in skin roughness (see figure 8, below).

These findings demonstrate that a specific blend of plant exosomes, formulated into a basic serum, has remarkable efficacy against two key signs skin aging. This study highlights the potential to develop plant exosome blends tailored to target many other cosmetic benefits. By carefully combining exosomes derived from various botanical sources, it will be possible to create a near infinite range of multifunctional, synergistic exosome blends suitable to satisfy the needs of the beauty industry.

Sustainable and Ethical Sourcing

Not only are plant exosomes highly effective in cosmetics and a natural alternative to many conventional cosmetic actives, but because they must be organic to be free from agrochemicals, they are an attractive choice, for environmentally conscious brands and consumers. Plant exosomes can be prepared with zero waste and so align with the principles of circular economy. This is important for the growing number of consumers who are trying to reduce their environmental impact. Plant exosomes are suitable actives for ecofriendly, ethical, vegetarian, or vegan beauty products and can be tailored to meet the needs of the most demanding consumers.

Closing Thoughts: Benefits for Beauty

Plant exosomes are far more than a natural, more stable and effective replacement for other delivery vesicles such as liposomes. Their unique properties allow for fresh approaches to achieving a variety of cosmetic benefits. By strategically blending exosomes isolated from various organic plant sources, each with its own distinct profile of bioactive constituents, researchers can create a near infinite number of new natural cosmetic actives and discover synergistic combinations that unlock efficacies that otherwise could not be achieved. As microscopic couriers driving cell communication, plant exosome blends are perfect ingredients for the latest trends such as for Beauty-AI, topical skin Biohacking products and Sophisticated Simplicity, where consumers demand the highest natural efficacy. Without a doubt, plant exosomes are poised to revolutionize the beauty industry.

REFERENCES

1. Annex II, Ref. 416, of the EU Cosmetics Regulation (EC) No. 1223/2009. https://ec.europa.eu/growth/tools-databases/cosing/
2. Thakur, A., Shah, D., Rai, D., Parra, D. C., Pathikonda, S., Kurilova, S., Cili, A. (2023). Therapeutic Values of Exosomes in Cosmetics, Skin Care, Tissue Regeneration, and Dermatological Diseases. Cosmetics. 10(2):65. https://doi.org/10.3390/cosmetics10020065
3. Harding, nd Stahl, P. (1983). Transferrin recycling in reticulocytes: pH and iron are important determinants of ligand binding and processing. Biochemical and Biophysical Research Communications. 113 (2): 650–8. https://doi.org/10.1016/0006-291X(83)91776-X
4. Pan, B. and Johnstone, R. M. (1983). Fate of the transferrin receptor during maturation of sheep reticulocytes in vitro: selective externalization of the receptor. Cell. 33 (3): 967–78. https://doi.org/10.1016/0092-8674(83)90040-5
5. Logozzi, M., Di Raimo, R., Mizzoni, D., and Fais, S. (2022). The Potentiality of Plant-Derived Nanovesicles in Human Health-A Comparison with Human Exosomes and Artificial Nanoparticles. Int J Mol Sci. 23(9):4919. https://doi.org/10.3390/ijms23094919
6. De Robertis, M., Sarra, A., D’Oria, V., Mura, F., Bordi, F., Postorino, P., and Fratantonio, D. (2020). Blueberry-derived exosome-like nanoparticles counter the response to TNF-Α-Induced change on gene expression in EA. hy926 cells. Biomolecules. 10(5):742. https://doi.org/10.3390/biom10050742
7. Kalluri, R., and LeBleu, V. S. (2020). The biology, function, and biomedical applications of exosomes. Science. 367(6478): eaau6977. https://doi.org/10.1126/science.aau6977
8. Subha, D. K., Harshnii, K. G., Madhikiruba, M., Nandhini, M., and Tamilselvi, K. S. (2023). Plant derived exosome- like Nanovesicles: an updated overview. Plant Nano Biology, (3Α.1016/j.plana.2022.100022.
9. Zeng, H., Guo, S., Ren, X., Wu, Z., Liu, S., and Yao, X. (2023). Current Strategies for Exosome Cargo Loading and Targeting Delivery. Cells. 12(10):1416. https://doi.org/10.3390/cells12101416
10. Logozzi, M., Di Raimo, R., Mizzoni, D., and Fais, S. (2021). Nanovesicles from Organic Agriculture-Derived Fruits and Vegetables: Characterization and Functional Antioxidant Content. Int J Mol Sci. 22(15):8170. https://doi.org/10.3390/ijms22158170
11. Orefice, N. S., Di Raimo, R., Mizzoni, D., Logozzi, M., and Fais, S. (2023). Purposing plant-derived exosomes-like nanovesicles for drug delivery: patents and literature review. Expert Opinion on Therapeutic Patents, 33(2), 89–100. https://doi.org/10.1080/13543776.2023.2195093
12. Cho, J. H., Hong, Y. D., Kim, D., Park, S. J., Kim, J. S., Kim, H-M., Yoon, E. J., and Cho, J-S. (2022). Confirmation of plant-derived exosomes as bioactive substances for skin application through comparative analysis of keratinocyte transcriptome. Appl Biol Chem. 65, 8. https://doi.org/10.1186/s13765-022-00676-z
13. Kırbas¸, O. K., Bozkurt, B. T., Asutay, A. B., Mat, B., Ozdemir, B., Öztürkog˘lu, D., Ölmez, H., I˙s¸lek, Z., S¸ahin, F., Tas¸lı, P. N. (2019). Optimized Isolation of Extracellular Vesicles From Various Organic Sources Using Aqueous Two-Phase System. Sci Rep. 16;9(1):19159. https://doi.org/10.1038/s41598-019-55477-0
14. Ko, Y., Baek, H., Hwang, J-H., Kim, Y., Lim, K.-M., Kim, J., and Kim, J. W. (2023). Nonanimal Euglena gracilis-Derived Extracellular Vesicles Enhance Skin-Regenerative Wound Healing. Adv Mater Interfaces 10(4) 2202255. https://doi.org/10.1002/admi.202202255

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