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Cited by 233RNA Interference and Gene DeliveryNanoparticle-Based Drug DeliveryAdvancements in Transdermal Drug Delivery

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Long-term storage of lipid-like nanoparticles for mRNA delivery

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Zhao, Pengxuan, Hou, Xucheng, Yan, Jingyue, Du, Shi, Xue, Yonger, Li, Wenqing, Xiang, Guangya, Dong, Yizhou·2020·Bioactive Materials
DOI 10.1016/j.bioactmat.2020.03.001PMC PMC7078456
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Abstract

Lipid-like nanoparticles (LLNs) have been extensively explored for messenger RNA (mRNA) delivery in various biomedical applications. However, the long-term storage of these nanoparticles is still a challenge for their clinical translation. In this study, we investigated a series of conditions for the long-term storage of LLNs with encapsulation of mRNA. We evaluated the stability of LLNs with different concentrations of cryoprotectants (sucrose, trehalose or mannitol) under the conditions of freezing or lyophilization processes. Through in vitro and in vivo mRNA delivery studies, we identified the optimal storage condition, and found that the addition with 5% (w/v) sucrose or trehalose to LLNs could remain their mRNA delivery efficiency for at least three months in the liquid nitrogen storage condition.

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Graphical abstract CC BY-NC-ND · PubMed Central
Fig. 1 The long-term stability of LLNs stored at 4 °C in aqueous condition. (A) Size of LLNs at different time points. (B) Relative luminescence intensity of LLNs. All data are presented as mean ± s.d (n = 3). Statistical significance was analyzed by the two-tailed Student's t-test. **P < 0.01, ***P < 0.001. ns: not significant.
Fig. 1 The long-term stability of LLNs stored at 4 °C in aqueous condition. (A) Size of LLNs at different time points. (B) Relative luminescence intensity of LLNs. All data are presented as mean ± s.d (n = 3). Statistical significance was analyzed by the two-tailed Student's t-test. **P < 0.01, ***P < 0.001. ns: not significant.CC BY-NC-ND · PubMed Central
Fig. 2 The stability of LLNs containing cryoprotectants with different concentrations (w/v) after three freeze–thaw cycles. (A) Size of LLNs. (B) Relative luminescence intensity of LLNs. All data are presented as mean ± s.d (n = 3). Statistical significance was analyzed by the two-tailed Student's t-test. **P < 0.01, ***P < 0.001.
Fig. 2 The stability of LLNs containing cryoprotectants with different concentrations (w/v) after three freeze–thaw cycles. (A) Size of LLNs. (B) Relative luminescence intensity of LLNs. All data are presented as mean ± s.d (n = 3). Statistical significance was analyzed by the two-tailed Student's t-test. **P < 0.01, ***P < 0.001.CC BY-NC-ND · PubMed Central
Fig. 3 The stability of LLNs containing cryoprotectants with different concentrations (w/v) stored in liquid nitrogen. (A) Size of LLNs. (B) Relative luminescence intensity of LLNs. All data are presented as mean ± s.d (n = 3). Statistical significance was analyzed by the two-tailed Student's t-test. **P < 0.01.
Fig. 3 The stability of LLNs containing cryoprotectants with different concentrations (w/v) stored in liquid nitrogen. (A) Size of LLNs. (B) Relative luminescence intensity of LLNs. All data are presented as mean ± s.d (n = 3). Statistical significance was analyzed by the two-tailed Student's t-test. **P < 0.01.CC BY-NC-ND · PubMed Central
Fig. 4 The stability of lyophilized LLNs (A) Size of LLNs. (B) Relative luminescence intensity of LLNs. All data are presented as mean ± s.d (n = 3). Statistical significance was analyzed by the two-tailed Student's t-test. **P < 0.01, ***P < 0.001.
Fig. 4 The stability of lyophilized LLNs (A) Size of LLNs. (B) Relative luminescence intensity of LLNs. All data are presented as mean ± s.d (n = 3). Statistical significance was analyzed by the two-tailed Student's t-test. **P < 0.01, ***P < 0.001.CC BY-NC-ND · PubMed Central
Fig. 5 In vivo mRNA delivery efficiency. Total bioluminescence signal (A, C) and normalized bioluminescence signal with tissue weight (B, D) of different LLNs. (A, B) were LLNs stored for 1 week, (C, D) were LLNs stored for 3 months. All data are presented as mean ± s.d (n = 3). Statistical significance was analyzed by the two-tailed Student's t-test. ***P < 0.001.
Fig. 5 In vivo mRNA delivery efficiency. Total bioluminescence signal (A, C) and normalized bioluminescence signal with tissue weight (B, D) of different LLNs. (A, B) were LLNs stored for 1 week, (C, D) were LLNs stored for 3 months. All data are presented as mean ± s.d (n = 3). Statistical significance was analyzed by the two-tailed Student's t-test. ***P < 0.001.CC BY-NC-ND · PubMed Central

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Metadata

Authors
Zhao, P., Hou, X., Yan, J., Du, S., Xue, Y., Li, W., Xiang, G., & Dong, Y.
Affiliations
  • Huazhong University of Science and Technology (CN)
  • The Ohio State University (US)
Corresponding author
  • Xiang, Guangya
  • Dong, Yizhou
ORCID
Venue
Bioactive Materials
Publisher
Elsevier BV
Published
2020-06
Volume
5
Issue
2
Pages
358-363
ISSN
  • 2097-1192 (ISSN-L)
  • 2452-199X
DOI
10.1016/j.bioactmat.2020.03.001
PMC
PMC7078456
OpenAlex
W3011313923
Cited by
233
References
34
Citations by year
2026: 17 · 2025: 39 · 2024: 48 · 2023: 53 · 2022: 47 · 2021: 24 · 2020: 4
FWCI
10.38
Type
journal article
Language
en
Open access
Licence
Locations
  • Bioactive Materials (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
Ohio State University
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
  • Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmaceutical Science › Advancements in Transdermal Drug Delivery
Keywords
mRNA delivery, long-term storage, cryoprotectants, sucrose, trehalose, mannitol, freeze-drying, freezing, liquid nitrogen storage, nanoparticle stability

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