Anti-inflammatory potential of plant-derived extracellular vesicles from Solanum nigrum L. integrated in gelatine-dopamine hydrogel on RAW 264.7 and MC3T3 cells

Original scientific article

Authors

  • Anggraini Barlian Department of Biotechnology, Bandung Institute of Technology, Bandung, West Java, Indonesia, Department of Biology, Bandung Institute of Technology, Bandung, West Java, Indonesia and Scientific Imaging Center, Institut Teknologi Bandung, Bandung 40132, Indonesia https://orcid.org/0000-0002-0826-3134
  • Tasya Fediarisa Department of Biotechnology, Bandung Institute of Technology, Bandung, West Java, Indonesia and Scientific Imaging Center, Institut Teknologi Bandung, Bandung 40132, Indonesia https://orcid.org/0009-0003-6533-2604
  • Aida Fitri Kamila Department of Biology, Bandung Institute of Technology, Bandung, West Java, Indonesia and Scientific Imaging Center, Institut Teknologi Bandung, Bandung 40132, Indonesia https://orcid.org/0009-0008-5426-2539
  • Noviana Vanawati Department of Biology, Bandung Institute of Technology, Bandung, West Java, Indonesia https://orcid.org/0000-0001-7569-253X
  • Yung-Hsin Cheng Department of Materials Science and Engineering, National Taiwan University of Science and Technology, Taipei, Taiwan https://orcid.org/0000-0003-4630-6999

DOI:

https://doi.org/10.5599/admet.3149

Keywords:

Cell-free therapy, lyophilization, macrophage cells, osteoblast cells, enzyme-linked immunosorbent assay

Abstract

Background and purpose: Plant-derived extracellular vesicles (PDEV) from Solanum nigrum L. fruit show promise as a cell-free regenerative and inflammatory therapy for bone defects due to their anti-inflammatory properties. However, challenges such as storage stability and targeted delivery efficiency remain in PDEV's applications. Strategies such as lyophilization and injectable hydrogel delivery systems offer potential solutions. Experimental approach: In this study, lyophilized PDEVs derived from Solanum nigrum L. berries were incorporated into a thermosensitive injectable gelatine-dopamine (Gel-Dop) hydrogel and evaluated by in vitro for their anti-inflammatory potential using MC3T3 pre-osteoblast cells and RAW 264.7 macrophage cells. Key results: The isolated PDEVs show a spherical morphology, an average size of approximately 132.6 nm, a polydispersity index of 0.197, and a protein concentration of 509 μg mL-1. These PDEVs were efficiently internalized by MC3T3 and RAW 264.7 cells after 12 hours of incubation and showed no cytotoxic effects at concentrations up to 10 μg mL-1. The release profile confirmed that the hydrogel effectively released the PDEVs, which remained non-toxic and were internalized by cells after 12 hours of incubation. Subsequently, treatment of lipopolysaccharide (LPS) stimulated MC3T3 and RAW 264.7 cells with PDEVs led to a reduction in IL-6 protein expression. Conclusion: These findings suggest that lyophilized PDEVs from Solanum nigrum L. berries, when incorporated into Gel-Dop hydrogel, hold promise for future development as an anti-inflammatory agent in bone therapy. This study is the first to characterize and incorporate lyophilized PDEVs from Solanum nigrum L. into thermosensitive injectable Gel-Dop hydrogel and demonstrate their anti-inflammatory potential through the suppression IL-6 expression in LPS-stimulated MC3T3 and RAW 264.7 cells.

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References

[1] B. Clarke. Normal bone anatomy and physiology. Clinical Journal of American Society of Nephrology 3 (2008) S131-S139. https://doi.org/10.2215/cjn.04151206 DOI: https://doi.org/10.2215/CJN.04151206

[2] X. Song, L. Xu, W. Zhang. Biomimetic synthesis and optimization of extracellular vesicles for bone regeneration. Journal of Controlled Release 355 (2023) 18-41. https://doi.org/10.1016/j.jconrel.2023.01.057 DOI: https://doi.org/10.1016/j.jconrel.2023.01.057

[3] R. Quarto, P. Giannoni. Bone Tissue Engineering: Past-Present-Future. Methods in Molecular Biology 1416 (2016) 21-33. https://doi.org/10.1007/978-1-4939-3584-0_2 DOI: https://doi.org/10.1007/978-1-4939-3584-0_2

[4] H.M. Torres, K.M. Arnold, M. Oviedo, J.J. Westendorf, S.R. Weaver. Inflammatory processes affecting bone health and repair. Current Osteoporosos Reports 21 (2023) 842-853. https://doi.org/10.1007/s11914-023-00824-4 DOI: https://doi.org/10.1007/s11914-023-00824-4

[5] R. Hardy, M.S. Cooper. Bone loss in inflammatory disorders. Journal of Endocrinology 201 (2009) 309-320. https://doi.org/10.1677/joe-08-0568 DOI: https://doi.org/10.1677/JOE-08-0568

[6] B. Langdahl, S. Ferrari, D.W. Dempster. Bone modeling and remodeling: potential as therapeutic targets for the treatment of osteoporosis. Therapeutic Advances in Musculoskeletal Disease 8 (2016) 225-235. https://doi.org/10.1177/1759720x16670154 DOI: https://doi.org/10.1177/1759720X16670154

[7] E.F. Eriksen. Cellular mechanisms of bone remodeling. Reviews in Endocrine and Metabolic Disorders 11 (2010) 219-227. https://doi.org/10.1007/s11154-010-9153-1 DOI: https://doi.org/10.1007/s11154-010-9153-1

[8] M.M. Weivoda, E.W. Bradley. Macrophages and bone remodeling. Journal of Bone and Mineral Research 38 (2023) 359-369. https://doi.org/10.1002/jbmr.4773 DOI: https://doi.org/10.1002/jbmr.4773

[9] R.H. Straub, M. Cutolo, R. Pacifici. Evolutionary medicine and bone loss in chronic inflammatory diseases--A theory of inflammation-related osteopenia. Seminars in Arthritis and Rheumatism 45 (2015) 220-228. https://doi.org/10.1016/j.semarthrit.2015.04.014 DOI: https://doi.org/10.1016/j.semarthrit.2015.04.014

[10] M.A. Terkawi, G. Matsumae, T. Shimizu, D. Takahashi, K. Kadoya, and N. Iwasaki. Interplay between inflammation and pathological bone resorption: insights into recent mechanisms and pathways in related diseases for future perspectives. International Journal of Molecular Sciences 23 (2022). https://doi.org/10.3390/ijms23031786 DOI: https://doi.org/10.3390/ijms23031786

[11] F. Huang, P. Wong, J. Li, Z. Lv, L. Xu, G. Zhu, M. He, Y. Luo. Osteoimmunology: the correlation between osteoclasts and the Th17/Treg balance in osteoporosis. Journal of Cellular and Molecular Medicine 26 (2022) 3591-3597. https://doi.org/10.1111/jcmm.17399 DOI: https://doi.org/10.1111/jcmm.17399

[12] P. Zhou, T. Zheng, B. Zhao. Cytokine-mediated immunomodulation of osteoclastogenesis. Bone 164 (2022) 116540. https://doi.org/10.1016/j.bone.2022.116540 DOI: https://doi.org/10.1016/j.bone.2022.116540

[13] G. Fernandez de Grado, L. Keller, Y. Idoux-Gillet, Q. Wagner, A.M. Musset, N. Benkirane-Jessel, F. Bornert, and D. Offner. Bone substitutes: a review of their characteristics, clinical use, and perspectives for large bone defects management. Journal of Tissue Engineering (2018). https://doi.org/10.1177/2041731418776819 DOI: https://doi.org/10.1177/2041731418776819

[14] H.J. Haugen, S.P. Lyngstadaas, F. Rossi, G. Perale. Bone grafts: which is the ideal biomaterial? Journal of Clinical Periodontology (2019) 92-102. https://doi.org/10.1111/jcpe.13058 DOI: https://doi.org/10.1111/jcpe.13058

[15] A.A. Ivanov, A.V. Kuznetsova, O.P. Popova, T.I. Danilova, O.O. Yanushevich. Modern approaches to acellular therapy in bone and dental regeneration. International Journal of Molecular Science 22 (2021). https://doi.org/10.3390/ijms222413454 DOI: https://doi.org/10.3390/ijms222413454

[16] A. Sha, Y. Luo, W. Xiao, J. He, X. Chen, Z. Xiong, L. Peng, L. Zou, B. Liu, Q. Li. Plant-Derived Exosome-like Nanoparticles: A Comprehensive Overview of Their Composition, Biogenesis, Isolation, and Biological Applications. International Journal of Molecular Science 25 (2024) 12092. https://doi.org/10.3390/ijms252212092 DOI: https://doi.org/10.3390/ijms252212092

[17] J. Kim, S. Li, S. Zhang, J. Wang. Plant-derived exosome-like nanoparticles and their therapeutic activities. Asian Journal of Pharmaceutical Sciences 17 (2022) 53-69. https://doi.org/10.1016/j.ajps.2021.05.006 DOI: https://doi.org/10.1016/j.ajps.2021.05.006

[18] X. Chen, X. Dai, Y. Liu, Y. Yang, L. Yuan, X. He, G. Gong. Solanum nigrum Linn.: an insight into current research on traditional uses, phytochemistry, and pharmacology. Frontier in Pharmacology 13 (2022) 918071. https://doi.org/10.3389/fphar.2022.918071 DOI: https://doi.org/10.3389/fphar.2022.918071

[19] N. Emmanuela, D.R. Muhammad, Iriawati, C.H. Wijaya, Y.M.D. Ratnadewi, H. Takemori, I.D. Ana, R. Yuniati, W. Handayani, T.D.K. Wungu, Y. Tabata, and A. Barlian. Isolation of plant-derived exosome-like nanoparticles (PDEVs) from Solanum nigrum L. berries and their effect on interleukin-6 expression as a potential anti-inflammatory agent. PLoS One 19 (2024) e0296259. https://doi.org/10.1371/journal.pone.0296259 DOI: https://doi.org/10.1371/journal.pone.0296259

[20] T. Burnouf, V. Agrahari, V. Agrahari. Extracellular vesicles as nanomedicine: hopes and hurdles in clinical translation. International Journal of Nanomedicine (2019) 8847-8859. https://doi.org/10.2147/ijn.s225453 DOI: https://doi.org/10.2147/IJN.S225453

[21] M. Guarro, F. Suñer, M. Lecina, S. Borrós, C. Fornaguera. Efficient extracellular vesicles freeze-dry method for direct formulations preparation and use. Colloids and Surfaces B: Biointerfaces 218 (2022) 112745. https://doi.org/10.1016/j.colsurfb.2022.112745 DOI: https://doi.org/10.1016/j.colsurfb.2022.112745

[22] L. Liu, W. Liu, Z. Han, Y. Shan, Y. Xie, J. Wang, H. Qi, and Q. Xu. Extracellular vesicles-in-hydrogel (EViH) targeting pathophysiology for tissue repair. Bioactive Materials 44 (2025) 283-318. https://doi.org/10.1016/j.bioactmat.2024.10.017 DOI: https://doi.org/10.1016/j.bioactmat.2024.10.017

[23] H.T. A Alamir, G.L. Ismaeel, A.T. Jalil, W.H. Hadi, I.K. Jasim, A.F. Almulla, Z.A. Radhea. Advanced injectable hydrogels for bone tissue regeneration. Biophysical Reviews 15 (2023) 223-237. https://doi.org/10.1007/s12551-023-01053-w DOI: https://doi.org/10.1007/s12551-023-01053-w

[24] M.A. Rider, S.N. Hurwitz, D.G. Meckes Jr. ExtraPEG: a polyethylene glycol-based method for enrichment of extracellular vesicles. Scientific Reports 6 (2016) 23978. https://doi.org/10.1038/srep23978 DOI: https://doi.org/10.1038/srep23978

[25] S. Bosch, L. de Beaurepaire, M. Allard, M. Mosser, C. Heichette, D. Chrétien, D. Jegou, and J.M. Bach. Trehalose prevents aggregation of exosomes and cryodamage. Scientific Reports 6 (2016) 36162. https://doi.org/10.1038/srep36162 DOI: https://doi.org/10.1038/srep36162

[26] V. Kuete. Physical, hematological, and histopathological signs of toxicity induced by African medicinal plants. In: Toxicological Survey of African Medicinal Plants, Elsevier, Dschang, Cameroon, 2014, p. 635-657. https://doi.org/10.1016/B978-0-12-800018-2.00022-4 DOI: https://doi.org/10.1016/B978-0-12-800018-2.00022-4

[27] F. Re, E. Gabusi, C. Manferdini, D. Russo, G. Lisignoli. Bone regeneration improves with mesenchymal stem cell-derived extracellular vesicles (EVs) combined with scaffolds: a systematic review. Biology 10 (2021) 579. https://doi.org/10.3390/biology10070579 DOI: https://doi.org/10.3390/biology10070579

[28] Y. Ju, Y. Hu, P. Yang, X. Xie, B. Fang. Extracellular vesicle-loaded hydrogels for tissue repair and regeneration. Materials Today Bio 18 (2023) 100522. https://doi.org/10.1016/j.mtbio.2022.100522 DOI: https://doi.org/10.1016/j.mtbio.2022.100522

[29] R. Schrieber, H. Gareis. Gelatine Handbook: Theory and Industrial Practice, John Wiley & Sons, 2007. https://doi.org/10.1002/9783527610969.fmatter DOI: https://doi.org/10.1002/9783527610969

[30] B. Sarker, R. Singh, R. Silva, J.A. Roether, J. Kaschta, R. Detsch, D. W Schubert., I. Cicha, A. R Boccaccini. Evaluation of fibroblasts adhesion and proliferation on alginate-gelatin crosslinked hydrogel. PLoS One 9 (2014). https://doi.org/10.1371/journal.pone.0107952 DOI: https://doi.org/10.1371/journal.pone.0107952

[31] W. Ma, H. Chen, S. Cheng, C. Wu, L. Wang, M. Du. Gelatin hydrogel reinforced with mussel-inspired polydopamine-functionalized nanohydroxyapatite for bone regeneration. International Journal of Biological of Macromolecules 240 (2023) 124287. https://doi.org/10.1016/j.ijbiomac.2023.124287 DOI: https://doi.org/10.1016/j.ijbiomac.2023.124287

[32] R. Cui, F. Chen, Y. Zhao, W. Huang, C. Liu. A novel injectable starch-based tissue adhesive for hemostasis. Journal od Materials Chemistry B 8 (2020) 8282-8293. https://doi.org/10.1039/D0TB01562H DOI: https://doi.org/10.1039/D0TB01562H

[33] S. Chen, A.F.U.H. Saeed, Q. Liu, Q. Jiang, H. Xu, G.G. Xiao, L. Rao, and Y. Duo. Macrophages in immunoregulation and therapeutics. Signal Transduction and Targeted Therapy 8 (2023) 207. https://doi.org/10.1038/s41392-023-01452-1 DOI: https://doi.org/10.1038/s41392-023-01452-1

[34] C. Yunna, H. Mengru, W. Lei, C. Weidong. Macrophage M1/M2 polarization. European Journal of Pharmacology 877 (2020) 173090. https://doi.org/10.1016/j.ejphar.2020.173090 DOI: https://doi.org/10.1016/j.ejphar.2020.173090

[35] D. Jacho, P. Babaniamansour, R. Osorio, M. Toledano, A. Rabino, R. Garcia-Mata, E. Yildirim-Ayan. Deciphering the Cell-Specific Effect of Osteoblast-Macrophage Crosstalk in Periodontitis. Tissue Engineering Part A 29 (2023) 579-593. https://doi.org/10.1089/ten.tea.2023.0104 DOI: https://doi.org/10.1089/ten.tea.2023.0104

[36] M.G. Tu, Y.W. Chen, M.Y. Shie. Macrophage-mediated osteogenesis activation in co-culture with osteoblast on calcium silicate cement. Journal of Materials Science: Materials in Medicine 26 (2015) 276. https://doi.org/10.1007/s10856-015-5607-z DOI: https://doi.org/10.1007/s10856-015-5607-z

[37] G. Vallés, E. Gil-Garay, L. Munuera, N. Vilaboa. Modulation of the cross-talk between macrophages and osteoblasts by titanium-based particles. Biomaterials 29 (2008) 2326-2335. https://doi.org/10.1016/j.biomaterials.2008.02.011 DOI: https://doi.org/10.1016/j.biomaterials.2008.02.011

[38] F. Chellat, Y. Merhi, A. Moreau, L. Yahia. Therapeutic potential of nanoparticulate systems for macrophage targeting. Biomaterials 26 (2005) 7260-7275. https://doi.org/10.1016/j.biomaterials.2005.05.044 DOI: https://doi.org/10.1016/j.biomaterials.2005.05.044

[39] Y. Shkryl, Z. Tsydeneshieva, A. Degtyarenko, Y. Yugay, L. Balabanova, T. Rusapetova, and V. Bulgakov. Plant exosomal vesicles: perspective information nanocarriers in biomedicine. Applied Sciences 12 (2022) 8262. https://doi.org/10.3390/app12168262 DOI: https://doi.org/10.3390/app12168262

[40] M.A. Zhukovsky, A. Filograna, A. Luini, D. Corda, C. Valente. Phosphatidic acid in membrane rearrangements. FEBS Lett 593 (2019) 2428-2441. https://doi.org/10.1002/1873-3468.13563 DOI: https://doi.org/10.1002/1873-3468.13563

[41] J. Xu, L. Yu, F. Liu, L. Wan, Z. Deng. The effect of cytokines on osteoblasts and osteoclasts in bone remodeling in osteoporosis: a review. Frontiers in Immunology 14 (2023) 1222129. https://doi.org/10.3389/fimmu.2023.1222129 DOI: https://doi.org/10.3389/fimmu.2023.1222129

[42] N.A. Sims. Influences of the IL-6 cytokine family on bone structure and function. Cytokine 146 (2021) 155655. https://doi.org/10.1016/j.cyto.2021.155655 DOI: https://doi.org/10.1016/j.cyto.2021.155655

[43] M. Foti. Introduction to cytokines as tissue regulators in health and disease. In: Cytokine Effector Functions in Tissues, Elsevier, Milan, Italy, 2017, p. 3-30. https://dx.doi.org/10.1016/B978-0-12-804214-4.00019-1 DOI: https://doi.org/10.1016/B978-0-12-804214-4.00019-1

[44] P.J. Murray, J.E. Allen, S.K. Biswas, E.A. Fisher, D.W. Gilroy, S. Goerdt, S. Gordon, J.A. Hamilton, L.B. Ivashkiv, T. Lawrence, M. Locati, A. Mantovani, F.O. Martinez, J.L. Mege, D.M. Mosser, G. Natoli, J.P. Saeij, J.L. Schultze, K.A. Shirey, A. Sica, J. Suttles, I. Udalova, J.A. van Ginderachter, S.N. Vogel, T.A. Wynn. Macrophage activation and polarization: nomenclature and experimental guidelines. Immunity 41 (2014) 14-20. https://doi.org/10.1016/j.immuni.2014.06.008 DOI: https://doi.org/10.1016/j.immuni.2014.06.008

[45] J.L. Schultze, S.V. Schmidt. Molecular features of macrophage activation. Seminars in Immunology 27 (2015) 416-423. https://doi.org/10.1016/j.smim.2016.03.009 DOI: https://doi.org/10.1016/j.smim.2016.03.009

[46] S. Zhu, M. Huang, G. Feng, Y. Miao, H. Wu, M. Zeng, Y.M. Lo. Gelatin versus its two major degradation products, prolyl-hydroxyproline and glycine, as supportive therapy in experimental colitis in mice. Food Science and Nutrition 6 (2018) 1023-1031. https://doi.org/10.1002/fsn3.639 DOI: https://doi.org/10.1002/fsn3.639

[47] W. Lu, Z. Ding, F. Liu, W. Shan, C. Cheng, J. Xu, W. He, W. Huang, J. Ma, Z. Yin. Dopamine delays articular cartilage degradation in osteoarthritis by negative regulation of the NF-κB and JAK2/STAT3 signaling pathways. Biomedicine and Pharmacotherapy 119 (2019) 109419. https://doi.org/10.1016/j.biopha.2019.109419 DOI: https://doi.org/10.1016/j.biopha.2019.109419

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06-03-2026

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Pharmaceutics

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Anti-inflammatory potential of plant-derived extracellular vesicles from Solanum nigrum L. integrated in gelatine-dopamine hydrogel on RAW 264.7 and MC3T3 cells: Original scientific article. (2026). ADMET and DMPK, 14, Article 3149. https://doi.org/10.5599/admet.3149

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