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. 2020 Mar 18;5(2):358-363.
doi: 10.1016/j.bioactmat.2020.03.001. eCollection 2020 Jun.

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

Pengxuan Zhao  1   2 , Xucheng Hou  1 , Jingyue Yan  1 , Shi Du  1 , Yonger Xue  1 , Wenqing Li  1 , Guangya Xiang  2 , Yizhou Dong  1   3   4   5   6   7
Affiliations

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

Pengxuan Zhao et al. Bioact Mater. .

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.

Keywords: Lipid-like nanoparticles; Long-term storage; mRNA delivery.

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Conflict of interest statement

The authors disclose no conflicts.

Figures

Image 1
Graphical abstract
Fig. 1
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. 2
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. 3
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. 4
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. 5
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.
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