Figure 1
Staphylococcus epidermidis treated in duplicate with BE (biotransformed transgenic soybean extract), NBE (non-biotransformed soybean extract), GEN (genistein), and LNZ (linezolid). Blue wells represent bacterial growth inhibition and pink wells indicate bacterial growth. The numbers in the left panel indicate the concentration of the compounds in each well. The negative control (NC) was a natural bacterial growth, shown in pink color. The positive control (PC) was a sterilized culture medium, with no growth, shown in blue color.
Figure 2
Fluorescence microscopy of primary human dermal fibroblasts adult (HDFa) labeled with anti-procollagen type I antibody (green) and DAPI (blue) after 24 h exposure to treatments. A, Cells with culture medium only (NC, negative control); B, Cells treated with biotransformed soy extract (BE, 1.33 μg/mL); C, Cells treated with non-biotransformed soy extract (NBE, 1.33 μg/mL); D, Cells treated with commercial soy extract (CE, 0.023 μg/mL); E, Cells treated with genistein + daidzein (G, 1.90 and D, 2.27 ng/mL); F, Cells treated with β-estradiol (50 ng/mL); G, Cells treated with PHTPP (1 μM); H, Cells treated with PHTPP (1 μM) + BE (1.33 μg/mL). Magnification 40×, scale bar 50 μm. I, Quantification of collagen-I expression in HDFa. Data are reported as means±SE of three independent experiments. *P<0.05 vs NC, (ANOVA and Tukey post hoc test).
Figure 3
Collagen-I expression in human dermal fibroblasts adult (HDFa) cells after 24 h of treatment. A, Expression of type 1 collagen protein. B, Expression of ß-tubulin protein. C, Quantification of collagen expression (fold of control). Data are reported as means±SE of three independent experiments. *P<0.05 vs NC (ANOVA and Tukey post-test). Negative control (NC, medium only); biotransformed soy extract (BE, 1.33 µg/mL); non-biotransformed soy extract (NBE, 1.33 µg/mL); commercial soy extract (CE, 0.023 μg/mL); PHTPP (1 μM); PHTPP (1 μM) + BE (1.33 μg/mL); β-estradiol (50 ng/mL); genistein + daidzen (G+D, 1.90 + 2.27 ng/mL).
Figure 4
Macroscopic appearance of cream formulations with or without the incorporation of extracts, A, Placebo (formulation only); B, Formulation incorporated with non-biotransformed soybean extract; C, Formulation incorporated with biotransformed soybean extract; D, Formulation incorporated with commercial soybean extract.
Figure 5
pH values of cream formulations incorporated or not (placebo) with the biotransformed soy extract (BE). Data are reported as means±SE (ANOVA); n=6 measurements, performed on newly prepared formulations (Cycle 0) and at the end of each cycle.
Figure 6
Water loss of cream formulations incorporated or not (placebo) with the biotransformed soy extract (BE). Data are reported as means±SE; n=6 measurements, performed on newly prepared formulations (Cycle 0) and at the end of each cycle. ****P=0.006 (ANOVA and Tukey post hoc test). nd: not detected.
Figure 7
Retention (A) and permeation (B) in the Franz cells of isoflavones daidzein and genistein after the pig's ear skin treatment with cream formulations incorporated or not (placebo) with the biotransformed soybean extract (BE). Data are reported as means±SE of six independent experiments. *P=0.05 vs placebo (ANOVA and t-test). nd: not detected.
Figure 8
Processing of human skin fragments. The skin fragment (A and B) was separated from the adipose tissue (C), and the explants were cut (2 cm2) and cultivated on stainless steel grids inside 90×15-mm plates in contact with the culture medium (D), kept at 37°C and 5% CO2, so that it was possible to add the formulation under study (E and F).
Figure 9
Fragments of human skin at 0 days (T0), 15 days (T15), and 30 days (T30) of cultivation. Images are shown under the same conditions of analysis, and show skin fragments without cream application (CONTROL), with cream application (PLACEBO), with application of biotransformed soy extract incorporated into the cream (BE), and with application of non-biotransformed soy extract incorporated into the cream (NBE). Scale bar 2.5 cm.
Figure 10
Ferric reducing ability of plasma (FRAP) in human skin extracts. Results are reported as means±SE of three independent experiments, performed in triplicate with human skin extracts treated for 0, 15, and 30 days with biotransformed soy extract (BE), non-biotransformed soy extract (NBE), and placebo, in addition to skin without any application as a negative control. *P<0.05 compared to control (ANOVA and Tukey post hoc test).
Figure 11
Quantification of total protein in human skin extracts. Results are reported as means±SE of three independent experiments, performed in triplicate with human skin extracts treated for 0, 15, and 30 days with biotransformed soy extract (BE), non-biotransformed soy extract (NBE), and placebo, in addition to skin without any application as a negative control (ANOVA and Tukey post hoc test).
Figure 12
Micrographs in polarized light (A.2, B.2, C.2) and without polarized light (A.1, B.1, C.1) of human skin samples after 30 days of treatments, showing in orange type I collagen (picrosirius red). A, human skin explant; B, human skin explant treated with placebo; C, human skin explant treated with biotransformed soybean extract (BE) cream formulation. Magnification 40×, scale bar 50 μm. D, Quantification of collagen-I. Data are reported as means±SE of three independent experiments. *P<0.05 vs negative control (ANOVA and Tukey post hoc test).
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