Fig. 5. LIFT hydrogel co-encapsulation of CaCO₃ protects lactase activity after oral delivery.

a, Lactase activity, as measured by ONPG assay, after 15 min incubation in PBS or SGF at 37 °C. Absorbances were normalized to that of lactase incubated in PBS. Statistical analysis was performed by two-tailed Student’s t-test, n = 3 independent experiments. b, Lactase activity after hydrogel encapsulation with or without CaCO₃ co-encapsulation and incubation in SGF for 1 h. Absorbances were normalized to that of alginate/CaCO₃. Statistical analysis was performed by two-way ANOVA with post-hoc Šidák’s multiple comparisons test, n = 3 independent experiments. c, Lactase activity of various treatments after trypsin treatment. Absorbances were normalized to that of treatment without trypsin. Statistical analysis was performed by one-way ANOVA with post-hoc Tukey’s multiple comparisons test, n = 3 independent experiments. d, Activity of lactase encapsulated in LIFT hydrogels after 1 h in male Sprague–Dawley rats. CaCO₃ was administered separately (LIFT + CaCO₃) or co-encapsulated (LIFT/CaCO₃). Absorbances were normalized by hydrogel mass. Statistical analysis was performed by one-way ANOVA with post-hoc Tukey’s multiple comparisons test, n = 5 rats per treatment. e, Activity of lactase encapsulated in LIFT hydrogels after 2 h in male Sprague–Dawley rats. Absorbances were normalized by hydrogel mass. Statistical analysis was performed by one-way ANOVA with post-hoc Tukey’s multiple comparisons test, n = 4 (LIFT) or 5 rats (LIFT+CaCO₃, LIFT/CaCO₃). f, Activity of lactase encapsulated in LIFT hydrogels after 6 h in female Yorkshire pigs. Hydrogels were retrieved from porcine stomach and randomly sampled. Absorbances were normalized by hydrogel mass and to control hydrogels without CaCO₃. Statistical analysis was performed by two-tailed Student’s t-test, n = 3 independent pig experiments. All data are presented as mean ± standard deviation.

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