Submit to BMT
Cite this article
5
Download
36
Views
Journal Browser
Volume | Year
Issue
Search
Forthcoming Issue
Current Issue
View All
News and Announcements
View All
ORIGINAL RESEARCH

A green biomineralization strategy for efficient encapsulation and long-term preservation of extracellular vesicles

Sisi Jing1# ,  Zheyu Wu1# ,  Yanwen Xu2# ,  Tianbai He1 ,  Shuangquan Gou1 ,  Yongfeng Lu1 ,  Meixi Lin1 ,  Anyu Wang1 ,  Yichen Dai1 ,  Haipeng Hu1 ,  Xiaolin Cui1 ,  Zhefan Stephen Chen3,4 ,  Xiangyan Shi5 ,  Jinhua Qiu5 ,  Qiyu Guo6 ,  Yanxia Zhou7* ,  Chen-zhong Li1,8* ,  Cheng Jiang1,8*
Show Less
1 Department of Biomedical Engineering, School of Medicine, The Chinese University of Hong Kong, Shenzhen, Guangdong, China
2 Translational Medicine Institute, Shenzhen Second People’s Hospital, The First Affiliated Hospital of Shenzhen University Health Science Center, Shenzhen, Guangdong, China
3 School of Life Sciences, Faculty of Science, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China
4 Gerald Choa Neuroscience Institute, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China
5 Department of Biology, Shenzhen MSU-BIT University, Shenzhen, Guangdong, China
6 Department of Neurology, Huizhou First Hospital, Huizhou, Guangdong, China
7 Department of Neurology, Shenzhen Second Peopleʼs Hospital, The First Affiliated Hospital of Shenzhen University Health Science Center, Shenzhen, Guangdong, China
8 Guangdong Basic Research Center of Excellence for Aggregate Science, School of Science and Engineering, The Chinese University of Hong Kong, Shenzhen, Guangdong, China
BMT, 193 https://doi.org/10.12336/bmt.25.00193
Submitted: 23 September 2025 | Revised: 17 January 2026 | Accepted: 21 January 2026 | Published: 6 April 2026
© 2026 by the Author(s). Licensee Biomaterials Translational, USA. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 (CC BY-NC-SA 4.0) (https://creativecommons.org/licenses/by-nc-sa/4.0/deed.en)
Download PDF
Cite
XML
Abstract

Extracellular vesicles (EVs) are pivotal in mediating intercellular communication, facilitating signal transduction, and enabling biomarker discovery. However, the challenge of maintaining EV bioactivity and the high costs of storage and transportation significantly limit their further application. Hence, this study reports a promising green biomineralization strategy for the long-term preservation of EVs. Specifically, zeolitic imidazolate framework-8 (ZIF-8) is employed to encapsulate EVs through biomineralization self-assembly (EVs@ZIF-8), aiming to improve EV stability and preserve bioactivity at 4℃. The preservation effect of ZIF-8 encapsulation is systematically investigated, with comparisons made to conventional −80°C cryopreservation and lyophilization. The results demonstrate that EVs@ZIF-8 retained bioactivity comparable to both EVs stored at −80°C for >1 month and lyophilized EVs, with significantly greater stability than EVs stored at 4°C. Moreover, EVs@ZIF-8 showed significantly higher protein content after multiple freeze–thaw cycles than pristine EVs. In vitro cellular experiments further reveal that EVs@ZIF-8 exhibited higher cellular uptake efficiency and more consistent bioactivity than lyophilized samples. These findings suggest that ZIF-8 can effectively preserve the stability and bioactivity of EVs, address the challenges associated with traditional preservation methods, and provide new possibilities for the clinical applications of EVs.

Keywords
Extracellular vesicles
Zeolitic imidazolate framework-8
Biomineralization
Long-term storage
Green preservation
Funding
This study was supported by the General Program of National Natural Science Foundation of China (82471500), the University Development Fund (UDF01002573), the Shenzhen Science and Technology Program (2023SC0005), the general program of GuangDong Basic and Applied Basic Research Foundation (2024A515012060, 2022A1515110206), the Pengcheng Peacock Research Fund (2024TC0150, KQ002573), the Shenzhen Peacock Team (KQTD20240729102029016), the Shenzhen International Science and Technology Cooperation Project (GJHZ20220913144002005), the Major Special Projects of Shenzhen Science and Technology Program (KJZD20230923115228056), the Guangdong Basic Research Center of Excellence for Aggregate Science and major research plan of Key Area Research and Development Program of Guangdong Province (2024B0101040001), and the Research Fund for International Senior Scientists of NSFC (W2531016).
Conflict of interest
The authors declare no conflict of interest.
References
  1. Théry C, Zitvogel L, Amigorena S. Exosomes: Composition, biogenesis and function. Nat Rev Immunol. 2002;2(8):569-579. doi: 10.1038/nri855

 

  1. Simons M, Raposo G. Exosomes--vesicular carriers for intercellular communication. Curr Opin Cell Biol. 2009;21(4):575-581. doi: 10.1016/j.ceb.2009.03.007

 

  1. Gusachenko ON, Zenkova MA, Vlassov VV. Nucleic acids in exosomes: Disease markers and intercellular communication molecules. Biochemistry (Mosc). 2013;78:1-7. doi: 10.1134/s000629791301001x

 

  1. Rezaie J, Feghhi M, Etemadi T. A review on exosomes application in clinical trials: Perspective, questions, and challenges. Cell Commun Signal. 2022;20(1):145. doi: 10.1186/s12964-022-00959-4

 

  1. Fan Z, Jiang C, Wang Y, et al. Engineered extracellular vesicles as intelligent nanosystems for next-generation nanomedicine. Nanoscale Horiz. 2022;7(7):682-714. doi: 10.1039/d2nh00070a

 

  1. Zheng J, Jiang X, Bai S, et al. Clinically accurate diagnosis of Alzheimer’s disease via single-molecule bioelectronic label-free profiling of multiple blood extracellular vesicle biomarkers. Adv Mater. 2025;37:e2505262. doi: 10.1002/adma.202505262

 

  1. Jiang C, Hopfner F, Katsikoudi A, et al. Serum neuronal exosomes predict and differentiate Parkinson’s disease from atypical parkinsonism. J Neurol Neurosurg Psychiatry. 2020;91(7):720-729. doi: 10.1136/jnnp-2019-322588

 

  1. Yan S, Zhang W, Li X, et al. Single extracellular vesicle detection assay identifies membrane-associated α-synuclein as an early-stage biomarker in Parkinson’s disease. Cell Rep Med. 2025;6(3):101999. doi: 10.1016/j.xcrm.2025.101999

 

  1. Yan S, Jiang C, Janzen A, et al. Neuronally derived extracellular vesicle α-synuclein as a serum biomarker for individuals at risk of developing Parkinson disease. JAMA Neurol. 2024;81(1):59-68.doi: 10.1001/jamaneurol.2023.4398

 

  1. Jiang C, Hopfner F, Berg D, et al. Validation of α-synuclein in L1CAM-immunocaptured exosomes as a biomarker for the stratification of parkinsonian syndromes. Mov Disord. 2021;36(11):2663-2669. doi: 10.1002/mds.28591

 

  1. Yu J, Zhou R, Liu S, et al. Electrochemical biosensors for the detection of exosomal microRNA biomarkers for early diagnosis of neurodegenerative diseases. Anal Chem. 2025;97(10):5355-5371. doi: 10.1021/acs.analchem.4c02619

 

  1. Zheng J, Zhou R, Wang B, et al. Electrochemical detection of extracellular vesicles for early diagnosis: A focus on disease biomarker analysis. Extracell Vesicles Circ Nucleic Acids. 2024;5(2):165-179. doi: 10.20517/evcna.2023.72

 

  1. Li Y, Zhang H, Jiang Y, Yang J, Cai D, Bai X. The application of extracellular vesicles in orthopedic disease. Interdiscip Med. 2024;2(3):e20230055. doi: 10.1002/inmd.20230055

 

  1. Mehdizadeh S, Mamaghani M, Hassanikia S, Pilehvar Y, Ertas YN. Exosome-powered neuropharmaceutics: Unlocking the blood-brain barrier for next-gen therapies. J Nanobiotechnol. 2025;23(1):329. doi: 10.1186/s12951-025-03352-8

 

  1. Liang Y, Iqbal Z, Lu J, et al. Cell-derived nanovesicle-mediated drug delivery to the brain: Principles and strategies for vesicle engineering. Mol Ther. 2023;31(5):1207-1224. doi: 10.1016/j.ymthe.2022.10.008

 

  1. Görgens A, Corso G, Hagey DW, et al. Identification of storage conditions stabilizing extracellular vesicles preparations. J Extracell Vesicles. 2022;11(6):e12238. doi: 10.1002/jev2.12238

 

  1. Wu JY, Li YJ, Hu XB, Huang S, Xiang DX. Preservation of small extracellular vesicles for functional analysis and therapeutic applications: A comparative evaluation of storage conditions. Drug Deliv. 2021;28(1):162-170. doi: 10.1080/10717544.2020.1869866

 

  1. Maroto R, Zhao Y, Jamaluddin M, et al. Effects of storage temperature on airway exosome integrity for diagnostic and functional analyses. J Extracell Vesicles. 2017;6(1):1359478. doi: 10.1080/20013078.2017.1359478

 

  1. Zhang Y, Bi J, Huang J, Tang Y, Du S, Li P. Exosome: A review of its classification, isolation techniques, storage, diagnostic and targeted therapy applications. Int J Nanomedicine. 2020;15:6917-6934. doi: 10.2147/ijn.s264498

 

  1. Huang A, Tong L, Kou X, et al. Structural and functional insights into the biomineralized zeolite imidazole frameworks. ACS Nano. 2023;17(23):24130-24140. doi: 10.1021/acsnano.3c09118

 

  1. Wang Q, Astruc D. State of the art and prospects in metal-organic framework (MOF)-based and MOF-derived nanocatalysis. Chem Rev. 2019;120(2):1438-1511. doi: 10.1021/acs.chemrev.9b00223

 

  1. Venna SR, Jasinski JB, Carreon MA. Structural evolution of zeolitic imidazolate framework-8. J Am Chem Soc. 2010;132(51):18030-18033. doi: 10.1021/ja109268m

 

  1. Kaur H, Mohanta GC, Gupta V, Kukkar D, Tyagi S. Synthesis and characterization of ZIF-8 nanoparticles for controlled release of 6-mercaptopurine drug. J Drug Deliv Sci Technol. 2017;41:106-112. doi: 10.1016/j.jddst.2017.07.004

 

  1. Park KS, Ni Z, Côté AP, et al. Exceptional chemical and thermal stability of zeolitic imidazolate frameworks. Proc Natl Acad Sci U S A. 2006;103(27):10186-10191. doi: 10.1073/pnas.0602439103

 

  1. Lee YR, Jang MS, Cho HY, Kwon HJ, Kim S, Ahn WS. ZIF-8: A comparison of synthesis methods. Chem Eng J. 2015;271:276-280. doi: 10.1016/j.cej.2015.02.094

 

  1. Lian X, Fang Y, Joseph E, et al. Enzyme-MOF (metal-organic framework) composites. Chem Soc Rev. 2017;46(11):3386-3401. doi: 10.1039/c7cs00058h

 

  1. Peng F, Xiang Y, Wang H, Hu Y, Zhou R, Hu Y. Biomimetic assembly of spore@ZIF‐8 microspheres for vaccination. Small. 2022;18(38):e2204011. doi: 10.1002/smll.202204011

 

  1. Ji Z, Zhang H, Liu H, Yaghi OM, Yang P. Cytoprotective metal-organic frameworks for anaerobic bacteria. Proc Natl Acad Sci U S A. 2018;115(42):10582-10587. doi: 10.1073/pnas.1808829115

 

  1. Feng Y, Wang H, Zhang S, et al. Antibodies@MOFs: An in vitro protective coating for preparation and storage of biopharmaceuticals. Adv Mater. 2019;31(2):e1805148. doi: 10.1002/adma.201805148

 

  1. Liang K, Ricco R, Doherty CM, et al. Biomimetic mineralization of metal-organic frameworks as protective coatings for biomacromolecules. Nat Commun. 2015;6(1):7240. doi: 10.1038/ncomms8240

 

  1. Jia J, Kong D, Liu Y, Zhang H, Liang X, Li Q. A biomimetic mineralization strategy for the long‐term preservation of exosomes through non‐destructive encapsulation within zeolite imidazolate frameworks‐8. Small. 2025;21(18):e2412264. doi: 10.1002/smll.202412264

 

  1. Luzuriaga MA, Welch RP, Dharmarwardana M, et al. Enhanced stability and controlled delivery of MOF-encapsulated vaccines and their immunogenic response in vivo. ACS Appl Mater Interfaces. 2019;11(10):9740-9746. doi: 10.1021/acsami.8b20504

 

  1. Mao Y, Wang L, Xu Z, et al. Developing a selection framework for zinc ion-based biomaterial design: Guided by the biosafety assessment of ZIF-8 and ZnO. ACS Biomater Sci Eng. 2024;10(5):2967-2982. doi: 10.1021/acsbiomaterials.3c01693

 

  1. Zhuang J, Kuo CH, Chou LY, Liu DY, Weerapana E, Tsung CK. Optimized metal-organic-framework nanospheres for drug delivery: Evaluation of small-molecule encapsulation. ACS Nano. 2014;8(3):2812-2819. doi: 10.1021/nn406590q

 

  1. Wang Y, Jia M, Zheng X, et al. Microvesicle-camouflaged biomimetic nanoparticles encapsulating a metal-organic framework for targeted rheumatoid arthritis therapy. J Nanobiotechnology. 2022;20(1):253. doi: 10.1186/s12951-022-01447-0

 

  1. Qin B, Zhang Q, Hu X, et al. How does temperature play a role in the storage of extracellular vesicles? J Cell Physiol. 2020;235(11):7663-7680. doi: 10.1002/jcp.29700

 

  1. Wu Y, Deng W, Klinke DJ 2nd. Exosomes: Improved methods to characterize their morphology, RNA content, and surface protein biomarkers. Analyst. 2015;140(19):6631-6642. doi: 10.1039/c5an00688k

 

  1. Lee M, Ban JJ, Im W, Kim M. Influence of storage condition on exosome recovery. Biotechnol Bioprocess Eng. 2016;21:299-304. doi: 10.1007/s12257-015-0781-x

 

  1. Jennings I, Kitchen DP, Woods TAL, Kitchen S, Preston FE, Walker ID. Stability of coagulation proteins in lyophilized plasma. Clin Lab Haematol. 2015;37(4):495-502. doi: 10.1111/ijlh.12318

 

  1. Gil L, Olaciregui M, Luño V, Malo C, González N, Martínez F. Current status of freeze-drying technology to preserve domestic animals sperm. Reprod Domest Anim. 2014;49:72-81. doi: 10.1111/rda.12396

 

  1. Charoenviriyakul C, Takahashi Y, Nishikawa M, Takakura Y. Preservation of exosomes at room temperature using lyophilization. Int J Pharm. 2018;553(1-2):1-7. doi: 10.1016/j.ijpharm.2018.10.032

 

  1. Chen C, Han D, Cai C, Tang X. An overview of liposome lyophilization and its future potential. J Control Release. 2010;142(3):299-311. doi: 10.1016/j.jconrel.2009.10.024

 

  1. Zhou G, Zhou Q, Li R, et al. Synthetically engineered bacterial extracellular vesicles and IL-4-encapsulated hydrogels sequentially promote osteoporotic fracture repair. ACS Nano. 2025;19(16):16064-16083. doi: 10.1021/acsnano.5c03106

 

  1. Liu H, Song P, Zhang H, et al. Synthetic biology-based bacterial extracellular vesicles displaying BMP-2 and CXCR4 to ameliorate osteoporosis. J Extracell Vesicles. 2024;13(4):e12429. doi: 10.1002/jev2.12429

 

  1. Ji N, Wang F, Wang M, Zhang W, Liu H, Su J. Engineered bacterial extracellular vesicles for central nervous system diseases. J Control Release. 2023;364:46-60. doi: 10.1016/j.jconrel.2023.10.027

 

  1. Liu H, Zhang H, Han Y, et al. Bone-targeted engineered bacterial extracellular vesicles delivering miRNA to treat osteoporosis. Compos Part B Eng. 2023;267:111047. doi: 10.1016/j.compositesb.2023.111047

 

  1. Liu H, Zhang Q, Wang S, Weng W, Jing Y, Su J. Bacterial extracellular vesicles as bioactive nanocarriers for drug delivery: Advances and perspectives. Bioact Mater. 2022;14:169-181. doi: 10.1016/j.bioactmat.2021.12.006

 

  1. Liu H, Geng Z, Su J. Engineered mammalian and bacterial extracellular vesicles as promising nanocarriers for targeted therapy. Extracell Vesicles Circ Nucl Acids. 2022;3(2):63-86. doi: 10.20517/evcna.2022.04
Share
Back to top
We use cookies on our website to ensure you get the best experience.
Read more about our cookies here
Accept