Journal: Nature Communications

Article Title: Charge-assisted stabilization of lipid nanoparticles enables inhaled mRNA delivery for mucosal vaccination

doi: 10.1038/s41467-024-53914-x

Figure Lengend Snippet: a Schematic illustrating the preparation and mechanism of CAS-LNP. By incorporating charged lipids into clinical LNP formulation, the increased electrostatic repulsions among CAS-LNPs enhance LNP stability during nebulization. b Synthetic scheme of DSSC-DOPE. c Formulations of SM102-LNP and CAS-LNPs. d Representative size distribution of LNPs measured by DLS. e The mRNA encapsulation efficiency of LNPs. f A representative Cryo-TEM image of 2.5% CAS-LNP. g pKa of SM102-LNP and 2.5% CAS-LNP measured by 2-( p -toluidino)−6-napthalene sulfonic acid (TNS) assay. h Zeta potentials of SM102-LNP and CAS-LNPs. i Percentage of intact LNPs after nebulization in 0.3×PBS. Data are shown as mean ± standard deviation (SD) ( n = 3 technical replicates). Statistical significance was analyzed by one-way analysis of variance (ANOVA) and Tukey’s multiple comparisons test. Source data are provided as a Source Data file.

Article Snippet: To synthesize DSSC-DOPE, equal moles of DSSC (GL Biochem) and PDP-DOPE were dissolved in dimethyl sulfoxide (DMSO) and mixed in a small glass vial charged with a stir bar.

Techniques: Formulation, Encapsulation, Standard Deviation

Journal: Nature Communications

Article Title: Charge-assisted stabilization of lipid nanoparticles enables inhaled mRNA delivery for mucosal vaccination

doi: 10.1038/s41467-024-53914-x

Figure Lengend Snippet: a Chemical structures of DOPA, HA, and HA-DSSC ( b ). c Zeta potentials of SM102-LNP, 2.5% DOPA-LNP, 2.5% HA-LNP, and 2.5% HA-DSSC-LNP ( n = 3 technical replicates). Data are shown as mean ± SD. d Percentage of intact LNPs after nebulization in 0.1 × PBS ( n = 3 technical replicates). Data are shown as mean ± SD. e Representative IVIS images and ( f ) quantitative analysis of luminescence in mice at 6 h post-administration ( n = 3 biologically independent samples). Data are shown as mean ± SEM. Statistical significance was analyzed by one-way ANOVA and Tukey’s multiple comparisons test. g Chemical structures of DSSC, SSSC, DDSC, and DDDC. h Isoelectric points of DSSC, SSSC, DDSC, and DDDC. i Zeta potentials of CAS-LNP, SSSC-LNP, DDSC-LNP, and DDDC-LNP ( n = 3 technical replicates). Data are shown as mean ± SD. j Percentage of intact LNPs after nebulization in 0.1 × PBS ( n = 3 technical replicates). Data are shown as mean ± SD. k Representative IVIS images of lungs and ( l ) quantitative analysis at 24 h post-administration ( n = 5 biologically independent samples). Data are shown as mean ± SEM. m Chemical structures of D E SSC-DOPE and D E SSC E -DOPE. Either one or both carboxyl groups of DSSC were converted to methyl esters. n Zeta potentials of CAS-LNP, D E SSC-LNP, and D E SSC E -LNP ( n = 3 technical replicates). Data are shown as mean ± SD. o Percentage of intact LNPs after nebulization in 0.1 × PBS ( n = 3 technical replicates). Data are shown as mean ± SD. p Representative IVIS images and ( q ) quantitative analysis of luminescence in mice treated with CAS-LNP, D E SSC-LNP, or D E SSC E -LNP at 6 h post-administration ( n = 5 biologically independent samples). Data are shown as mean ± SEM. r Representative IVIS images of major organs and ( s ) quantitative analysis of lungs at 24 h post-administration ( n = 5 biologically independent samples). Data are shown as mean ± SEM. The statistical significance of this figure was analyzed by one-way ANOVA and Tukey’s multiple comparisons test. Source data are provided as a Source Data file.

Article Snippet: To synthesize DSSC-DOPE, equal moles of DSSC (GL Biochem) and PDP-DOPE were dissolved in dimethyl sulfoxide (DMSO) and mixed in a small glass vial charged with a stir bar.

Techniques: Quantitative Luminescence