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. 2019 Nov 18;8(11):1457. doi: 10.3390/cells8111457

Figure 1.

Figure 1

Histogel modulates the angiogenic response. (a) Five percent and 10% of two different Histogel preparations were electrophoretically analyzed on agarose gel and stained with Stains-All solution. p.a., pre-autoclaved. Three different standard molecules (2 × 103, 1.1 × 103, 300 kDa) were used as markers of molecular weight (MK). (b) HUVEC spheroids were embedded in collagen or in collagen:5% Histogel 1:1 (HG) gels. Fifty nanograms per milliliter of VEGFA165 were used as the positive control. The formation of radially growing sprouts was evaluated after 24 h of incubation. Data are the mean ± SEM of three independent experiments (** p < 0.001 vs. CTRL; # p < 0.001 vs. VEGF, one-way ANOVA followed by Bonferroni’s test versus the control). (c) Alginate beads containing vehicle, or 100 ng of VEGFA165, or 5% Histogel (v/v 1:1) were implanted on the top of chick embryo chorioallantoic membrane (CAM) at Day 11 of development. After 72 h, newly formed blood vessels converging toward the implant were counted in ovo at 5× magnification using an STEMI SR stereomicroscope equipped with an objective f equal to 100 mm with adapter ring 475,070 (Carl Zeiss). Data are the mean ± SEM (n = 6–8) (*** p < 0.0001 vs. control; # p < 0.0001 vs. VEGF, one-way ANOVA followed by Bonferroni’s test versus the control). (d) Five percent of liquid alginic acid was mixed with 1.0 µg/mL VEGFA165 in the absence or in the presence of v/v 1:1 of 5% Histogel and injected subcutaneously into the flank of C57BL/6 mice. Plugs with vehicle alone were used as negative controls (CTRL). One week after injection, plugs were harvested. CD31 and CD45 mRNA expression levels were measured by RT-qPCR. Data are the mean ± SEM (n = 10) and are expressed as relative expression ratios (ΔΔCt – fold increase) using one vehicle plug as the reference. * p < 0.05; ** p < 0.01; *** p < 0.005; **** p < 0.001, one-way ANOVA followed by Bonferroni’s test versus the control.