Exosome-like Nanoparticles, High in Trans-δ-Viniferin Derivatives, Produced from Grape Cell Cultures: Preparation, Characterization, and Anticancer Properties Yury Shkryl, Zhargalma Tsydeneshieva, Ekaterina Menchinskaya, Tatiana Rusapetova, Olga Grishchenko, Anastasia Mironova, Dmitry Bulgakov, Tatiana Gorpenchenko, Vitaly Kazarin, Galina Tchernoded, Victor Bulgakov, Dmitry Aminin, Yulia Yugay Biomedicines, 2024 Background: Recent interest in plant-derived exosome-like nanoparticles (ENs) has surged due to their therapeutic potential, which includes antioxidant, anti-inflammatory, and anticancer activities. These properties are attributed to their cargo of bioactive metabolites and other endogenous molecules. However, the properties of ENs isolated from plant cell cultures remain less explored. Methods: In this investigation, grape callus-derived ENs (GCENs) were isolated using differential ultracentrifugation techniques. Structural analysis through electron microscopy, nanoparticle tracking analysis, and western blotting confirmed that GCENs qualify as exosome-like nanovesicles. Results: These GCENs contained significant amounts of microRNAs and proteins characteristic of plant-derived ENs, as well as trans-δ-viniferin, a notable stilbenoid known for its health-promoting properties. Functional assays revealed that the GCENs reduced the viability of the triple-negative breast cancer cell line MDA-MB-231 in a dose-dependent manner. Moreover, the GCENs exhibited negligible effects on the viability of normal human embryonic kidney (HEK) 293 cells, indicating selective cytotoxicity. Notably, treatment with these GCENs led to cell cycle arrest in the G1 phase and triggered apoptosis in the MDA-MB-231 cell line. Conclusions: Overall, this study underscores the potential of grape callus-derived nanovectors as natural carriers of stilbenoids and proposes their application as a novel and effective approach in the management of cancer.
Influence of Growth Regulators and Different Spectra of Monochromatic Radiation on the Growth and Biosynthetic Characteristics of Callus Culture of Ipomoea batatas (L.) Lam. Yu. A. Yugay, O. V. Grishchenko, E. A. Vasyutkina, V. P. Grigorchuk, E. N. Chukhlomina, Zh. L. Tsydeneshieva, O. D. Kudinova, Yu. L. Yaroshenko, A. I. Degtyarenko, E. P. Subbotin, V. P. Bulgakov, Yu. N. Kulchin, Yu. N. Shkryl Russian Journal of Plant Physiology, 2023 Abstract The work studied the influence of plant growth regulators as well as the effect of monochromatic radiation of different spectral composition on the growth and accumulation of secondary metabolites in the callus culture of sweet potato (Ipomoea batatas L.). It was found that auxin analogues in low concentrations (0.5 mg/L) significantly stimulated the growth of cell biomass, while the effect of 4-chlorophenoxyacetic acid (4-CPA) was more pronounced (stimulation 16 times) than that of 2,4-dichlorophenoxyacetic acid (2,4-D) (13.5 times stimulation). Both the increase in the concentration of 2,4-D and 4-CPA in the medium and the addition of cytokinin, 6-benzylaminopurine (BAP), led to a significant inhibition of culture growth (up to three times). In contrast to 2,4-D, the addition of 4-CPA led to an eightfold increase in the total content of polyphenolic compounds in cultured cells. Activating effect of 4-CPA on the biosynthetic characteristics of I. batatas cell culture persisted even with the addition of BAP, while an increase in the concentration of 4‑CPA led to the leveling of the activating effect. Monochromatic radiation—white, red (660 and 630 nm), yellow, green, and blue (440 and 460 nm) light—inhibited growth (up to 1.5 times) and the total accumulation of secondary metabolites in I. batatas cells (up to 1.8 times). At the same time, the white, bright blue, and red spectra differentially activated the formation of individual compounds 3,4-dicaffeoylquinic acid and 3-feruloyl-5-caffeoylquinic acid. Thus, the authors have established that both auxin analogues and spectral radiation exhibit different effects on the growth and biosynthetic characteristics of the I. batatas calli.
Isolation and Characterization of Extracellular Vesicles from Arabidopsis thaliana Cell Culture and Investigation of the Specificities of Their Biogenesis Yulia Yugay, Zhargalma Tsydeneshieva, Tatiana Rusapetova, Olga Grischenko, Anastasia Mironova, Dmitry Bulgakov, Vladimir Silant’ev, Galina Tchernoded, Victor Bulgakov, Yury Shkryl Plants, 2023 Over recent years, extracellular vesicles (EVs), commonly termed exosomes, have gained prominence for their potential as natural nanocarriers. It has now been recognized that plants also secrete EVs. Despite this discovery, knowledge about EV biogenesis in plant cell cultures remains limited. In our study, we have isolated and meticulously characterized EVs from the callus culture of the model plant, Arabidopsis thaliana. Our findings indicate that the abundance of EVs in calli was less than that in the plant’s apoplastic fluid. This difference was associated with the transcriptional downregulation of the endosomal sorting complex required for transport (ESCRT) genes in the calli cells. While salicylic acid increased the expression of ESCRT components, it did not enhance EV production. Notably, EVs from calli contained proteins essential for cell wall biogenesis and defense mechanisms, as well as microRNAs consistent with those found in intact plants. This suggests that plant cell cultures could serve as a feasible source of EVs that reflect the characteristics of the parent plant species. However, further research is essential to determine the optimal conditions for efficient EV production in these cultured cells.
Plant Exosomal Vesicles: Perspective Information Nanocarriers in Biomedicine Yury Shkryl, Zhargalma Tsydeneshieva, Anton Degtyarenko, Yulia Yugay, Larissa Balabanova, Tatiana Rusapetova, Victor Bulgakov Applied Sciences Switzerland, 2022 Exosomal nanoparticles (exosomes or nanovesicles) are biogenic membrane vesicles secreted by various cell types and represent a conservative mechanism of intercellular and interspecies communication in pro- and eukaryotic organisms. By transporting specific proteins, nucleic acids, and low molecular weight metabolites, the exosomes are involved in the regulation of developmental processes, activation of the immune system, and the development of a protective response to stress. Recently, the plant nanovesicles, due to an economical and affordable source of their production, have attracted a lot of attention in the biomedical field. Being a natural transport system, the plant exosomes represent a promising platform in biomedicine for the delivery of molecules of both endogenous and exogenous origin. This review presents current data on the biogenesis of plant exosomes and their composition, as well as mechanisms of their loading with various therapeutic compounds, which are determining factors for their possible practical use. We believe that further research in this area will significantly expand the potential of targeted therapy, particularly targeted gene regulation via the small RNAs, due to the use of plant exosomes in clinical practice.
