Comparative Characterization of Extracellular Vesicles Isolated from Aloe vera Leaf Peel and Gel Using Ultracentrifugation and PEG Precipitation

Document Type : Original Article

Authors
1 Department of biology, Faculty of science, University of Jiroft, Jiroft.Iran
2 Department of Biology, Faculty of Science, University of Jiroft, Jiroft, Iran
3 Department of Biology, Faculty of Science, University of Jiroft, Jiroft, Iran.
Abstract
This study aimed to evaluate different isolation methods, including ultracentrifugation (UC) and polyethylene glycol (PEG)-based precipitation of extracellular vesicles (EVs) from Aloe vera and investigate their yield, purity, and physicochemical properties. Plant-derived extracellular vesicles (PDEVs) have attracted increasing attention as natural nanocarriers for biomedical, nutraceutical, and food-related applications. In this study, EVs were isolated from Aloe vera leaf peel and gel using UC and PEG methods. The EVs were characterized by nanoparticle tracking analysis (NTA), NanoDrop protein quantification, and zeta potential measurements. Aloe vera gel-derived EVs using the UC method showed higher particle concentrations, more consistent size distributions, and lower protein contamination compared with peel and gel PEG-derived samples. The PEG isolation markedly increased apparent protein concentration, especially in the gel, indicating co-precipitation of non-vesicular plant components and polymer-associated artifacts. Zeta potential analysis further revealed significant surface charge variation on PEG-derived vesicles, both in peel and gel, particularly after high-speed centrifugation. In conclusion, these results demonstrate that UC provides superior purity and physicochemical stability of Aloe vera EVs, while PEG precipitation inflates protein yield and alters vesicle surface properties. The present study highlights the importance of the isolation method, which affects its potential applications.

Keywords

Subjects


Abdal Dayem, A., Kwak, Y., Jeun, H. and Cho, S.G., 2025. Recent Insights into Organoid-Derived Extracellular Vesicles and Their Biomedical Applications. Journal of Personalized Medicine, 15(10), p.492.
Abid, A., Javed, M., Zafar, S., Hamdani, S.A.Z., Shah, S.H.B.U., Abid, J. and Ahmad, A.M.R., 2025. The green healer; an updated review on the phytochemical profile and therapeutic potential of Aloe vera. Frontiers in Nutrition, 12, p.1689700.
Battistelli, M. and Falcieri, E., 2021. Apoptotic bodies: particular extracellular vesicles involved in intercellular communication. Advances in Medical Biochemistry, Genomics, Physiology, and Pathology, pp.473-486.
Choi, S.H., Eom, J.Y., Kim, H.J., Seo, W., Kwun, H.J., Kim, D.K., Kim, J. and Cho, Y.E., 2023. Aloe-derived nanovesicles attenuate inflammation and enhance tight junction proteins for acute colitis treatment. Biomaterials Science, 11(16), pp.5490-5501.
Fawzy, M.P., Hassan, H.A., Amin, M.U., Preis, E., Bakowsky, U. and Fahmy, S.A., 2025. Deploying nucleic acids-loaded plant-derived exosomes as green nano gadget in cancer gene therapy. Materials Advances.
Feng, J., Xiu, Q., Huang, Y., Troyer, Z., Li, B. and Zheng, L., 2023. Plant‐derived vesicle‐like nanoparticles as promising biotherapeutic tools: present and future. Advanced Materials, 35(24), p.2207826.
Gharavi, A.T., Niknejad, A., Irian, S., Rahimi, A. and Salimi, M., 2024. Polyethylene Glycol-Mediated Exosome Isolation: A Method for Exosomal RNA Analysis. Iranian Biomedical Journal, 28(2-3), p.132.
Huang, D., Chen, J., Zhao, M., Shen, H., Jin, Q., Xiao, D., Peng, Z., Chen, T., Zhang, Y., Rao, D. and Liu, M., 2025. Plant-derived extracellular vesicles: composition, function and clinical potential. Journal of Translational Medicine, 23(1), pp.1-17.
Huang, Q., Wang, J., Ning, H., Liu, W. and Han, X., 2025. Exosome isolation based on polyethylene glycol (PEG): a review. Molecular and Cellular Biochemistry, 480(5), pp.2847-2861.
Kalarikkal, S.P., Prasad, D., Kasiappan, R., Chaudhari, S.R. and Sundaram, G.M., 2020. A cost-effective polyethylene glycol-based method for the isolation of functional edible nanoparticles from ginger rhizomes. Scientific reports, 10(1), p.4456.
Kim, M. and Park, J.H., 2022. Isolation of aloe saponaria-derived extracellular vesicles and investigation of their potential for chronic wound healing. Pharmaceutics, 14(9), p.1905.
Kim, M.K., Choi, Y.C., Cho, S.H., Choi, J.S. and Cho, Y.W., 2021. The antioxidant effect of small extracellular vesicles derived from aloe vera peels for wound healing. Tissue engineering and regenerative medicine, 18(4), pp.561-571
Langellotto, M.D., Rassu, G., Serri, C., Demartis, S., Giunchedi, P. and Gavini, E., 2025. Plant-derived extracellular vesicles: a synergetic combination of a drug delivery system and a source of natural bioactive compounds. Drug delivery and translational research, 15(3), pp.831-845.
Lo, K.J., Wang, M.H., Ho, C.T. and Pan, M.H., 2024. Plant-derived extracellular vesicles: a new revolutionization of modern healthy diets and biomedical applications. Journal of agricultural and food chemistry, 72(6), pp.2853-2878.
Ludwig, A.K., De Miroschedji, K., Doeppner, T.R., Börger, V., Ruesing, J., Rebmann, V., Durst, S., Jansen, S., Bremer, M., Behrmann, E. and Singer, B.B., 2018. Precipitation with polyethylene glycol followed by washing and pelleting by ultracentrifugation enriches extracellular vesicles from tissue culture supernatants in small and large scales. Journal of extracellular vesicles, 7(1), p.1528109.
MartínezGreene, J.A., Hernández‐Ortega, K., Quiroz‐Baez, R., Resendis‐Antonio, O., Pichardo‐Casas, I., Sinclair, D.A., Budnik, B., Hidalgo‐Miranda, A., Uribe‐Querol, E., Ramos‐Godínez, M.D.P. and Martínez‐Martínez, E., 2021. Quantitative proteomic analysis of extracellular vesicle subgroups isolated by an optimized method combining polymer‐based precipitation and size exclusion chromatography. Journal of Extracellular Vesicles, 10(6), p.e12087.
Matei, C.E., Visan, A.I. and Cristescu, R., 2025. Aloe Vera Polysaccharides as Therapeutic Agents: Benefits Versus Side Effects in Biomedical Applications. Polysaccharides, 6(2), p.36.
Mensah, E.O., Adadi, P., Asase, R.V., Kelvin, O., Mozhdehi, F.J., Amoah, I. and Agyei, D., 2025. Aloe vera and its byproducts as sources of valuable bioactive compounds: Extraction, biological activities, and applications in various food industries. PharmaNutrition, p.100436.
Midekessa, G., Godakumara, K., Ord, J., Viil, J., Lattekivi, F., Dissanayake, K., Kopanchuk, S., Rinken, A., Andronowska, A., Bhattacharjee, S. and Rinken, T., 2020. Zeta potential of extracellular vesicles: toward understanding the attributes that determine colloidal stability. ACS omega, 5(27), pp.16701-16710.
Mitra, A., Singh, M., Banga, A., Pandey, J., Tripathi, S.S. and Singh, D., 2023. Bioactive compounds and therapeutic properties of Aloe vera—A review. Plant Science Today, 10, pp.1-7.
Mu, N., Li, J., Zeng, L., You, J., Li, R., Qin, A., Liu, X., Yan, F. and Zhou, Z., 2023. Plant-derived exosome-like nanovesicles: current progress and prospects. International Journal of Nanomedicine, pp.4987-5009.
Naini, M.A., Zargari-Samadnejad, A., Mehrvarz, S., Tanideh, R., Ghorbani, M., Dehghanian, A., Hasanzarrini, M., Banaee, F., Koohi-Hosseinabadi, O., Tanideh, N. and Iraji, A., 2021. Anti‐inflammatory, antioxidant, and healing‐promoting effects of Aloe vera extract in the experimental colitis in rats. Evidence‐Based Complementary and Alternative Medicine, 2021(1), p.9945244.
Rahmatinejad, F., Kharat, Z., Jalili, H., Renani, M.K. and Mobasheri, H., 2024. Comparison of morphology, protein concentration, and size distribution of bone marrow and Wharton's jelly-derived mesenchymal stem cells exosomes isolated by ultracentrifugation and Polymer-based precipitation techniques. Tissue and Cell, 88, p.102427.
Ramírez, O., Pomareda, F., Olivares, B., Huang, Y.L., Zavala, G., Carrasco-Rojas, J., Álvarez, S., Leiva-Sabadini, C., Hidalgo, V., Romo, P. and Sánchez, M., 2024. Aloe vera peel-derived nanovesicles display anti-inflammatory properties and prevent myofibroblast differentiation. Phytomedicine, 122, p.155108.
Rawat, S., Arora, S., Dhondale, M.R., Khadilkar, M., Kumar, S. and Agrawal, A.K., 2025. Stability Dynamics of Plant-Based Extracellular Vesicles Drug Delivery. Journal of Xenobiotics, 15(2), p.55.
Rogers, N.M., Hicks, E., Kan, C., Martin, E., Gao, L., Limso, C., Hendren, C.O., Kuehn, M. and Wiesner, M.R., 2023. Characterizing the transport and surface affinity of extracellular vesicles isolated from yeast and bacteria in well-characterized porous media. Environmental Science & Technology, 57(35), pp.13182-13192.
Ruf, A., Oberkofler, L., Robatzek, S. and Weiberg, A., 2022. Spotlight on plant RNA-containing extracellular vesicles. Current Opinion in Plant Biology, 69, p.102272.
Shen, J., Wei, T., Li, M., Jiang, Y., Zhang, J., Qi, Y., Chen, C., Li, X., Huang, P. and Qu, J., 2025. Aloe vera-derived extracellular vesicle-like particles suppress pancreatic carcinoma progression through triggering pyroptosis via ROS-GSDMD/E signaling pathway. Chinese Medicine, 20(1), p.101.
Solaberrieta, I., Jiménez, A. and Garrigós, M.C., 2022. Valorization of Aloe vera skin by-products to obtain bioactive compounds by microwave-assisted extraction: antioxidant activity and chemical composition. Antioxidants, 11(6), p.1058.
Yang, M., Liu, X., Luo, Q., Xu, L. and Chen, F., 2020. An efficient method to isolate lemon derived extracellular vesicles for gastric cancer therapy. Journal of nanobiotechnology, 18(1), p.100.
Zhu, Y., Zhao, J., Ding, H., Qiu, M., Xue, L., Ge, D., Wen, G., Ren, H., Li, P. and Wang, J., 2024. Applications of plant‐derived extracellular vesicles in medicine. MedComm, 5(10), p.e741.
Volume 1, Issue 1
Winter 2026
Pages 70-79

  • Receive Date 22 December 2025
  • Revise Date 02 February 2026
  • Accept Date 07 February 2026
  • First Publish Date 07 February 2026
  • Publish Date 01 February 2026