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. 2022 Mar 18;13:1479. doi: 10.1038/s41467-022-29121-x

Fig. 4. In vitro and in vivo physicochemical properties, residence stability, and drug release behavior of thermosensitive hydrogels formed from F127-g-Gelatin.

Fig. 4

a Concentration-dependent sol-gel transition properties of F127-g-Gelatin. b Representative SEM image of F127-g-Gelatin hydrogel (n = 3). c Concentration-dependent storage (G') and loss (G'') modulus of F127-g-Gelatin at 37 °C (n = 3 for 4.5 wt.%, n = 4 for 5.5, 6.0, 6.5, and 7.0 wt.%, and n = 5 for 4.0 and 5.0 wt.%.). d, e Cumulative release of (d) NO2− + GSNO (n = 4) and (e) Alexa FluorTM 647 labeled aCTLA-4 (n = 3) from 4.5 wt% F127-g-Gelatin into PBS with or without MMP9. f, g In vitro residence stability of (f) GSNO (n = 4) and (g) Alexa FluorTM 647 labeled aCTLA-4 (n = 3) containing 4.5 wt% F127-g-Gelatin in PBS with or without MMP9. h DLS size distribution of aCTLA-4 and supernatants released from F127-g-Gelatin hydrogel (F127-g-Gelatin) or F127-g-Gelatin hydrogel containing aCTLA-4 (aCTLA-4/F127-g-Gelatin) (n = 12). The left and right insets represent the average size (n = 12) and zeta potentials (n = 6 for aCTLA-4, and n = 12 for the other) of materials, respectively. i Representative TEM image of F127-g-Gelatin micelles in situ released from F127-g-Gelatin thermosensitive hydrogel (n = 3). j FRET analysis with aCTLA-4-TRITC and F127-g-Gelatin-FITC at FITC excitation and TRITC emission (n = 4). k Competitive assay to verify the activity of aCTLA-4 released from F127-g-Gelatin hydrogel (n = 3 for None+None, and n = 4 for the other). l ALT/AST activities of blood taken from mice 2 d after subcutaneous administration of 4.5 wt.% F127-g-Gelatin hydrogel (n = 5). m In vivo residence stability of 4.5 wt.% F127-g-Gelatin hydrogel quantified by time-resolved volume of hydrogel remaining at the injection site (n = 4). n In vivo quantification of aCTLA-4-AF647 remaining at the injection site by using IVIS® resulting from formulation in the 4.5 wt.% F127-g-Gelatin hydrogel (n = 4). Data are presented as mean ± SD for c–h and j, k, and mean ± SEM for l–n. *****p < 0.0001, ****p < 0.001, ***p < 0.01, **p < 0.05, and *p < 0.1. Exact p-values for d–h, k, m, and n are reported in the source file. d–g, m, n Two-way ANOVA using Tukey post-hoc statistical hypothesis. h, k One-way ANOVA using Tukey post-hoc statistical hypothesis. l Two-tailed Student t-test. Source data are available in a Source Data file.