Figure 3
From: Vitamin C–squalene bioconjugate promotes epidermal thickening and collagen production in human skin
Effects of ex-vivo topical treatment of human skin explants (donor 1) for 10 days with three formulations containing 3 wt% Vit C, upon collagen type I expression in whole skin and GAGs staining at dermal–epidermal junction (DEJ). (A) Collagen type I was labeled by a rabbit anti-human collagen I antibody with biotin/streptavidin amplification and FITC fluorescence revelation (green) and cell nuclei stained with propidium iodide (red). Pictures were realized with a tri CCD DXC 390P camera (Sony) and stored with Leica IM1000 software, version 1.10 (www.leica-microsystems.com). (B) Staining of fixed skin sections with Schiff reagent revealed the neutral GAGs as purple pink near DEJ (arrows), at the bottom of upper epidermis layer. (C) One-way ANOVA with Tukey’s post-test was used to compare collagen I signal (n = 8 images; mean of 3 measures per image), relative to untreated control (not significant). (D) GAGs coloration measured as percent of DEJ surface (n = 8 to 9 images; mean of 3 measures per image) from each explant was compared in formulations and untreated control by one-way ANOVA with Tukey’s post-test (*p < 0.05). Graphs were done with GraphPad Prism version 5.0.0 for Windows (GraphPad Software, San Diego, California USA, www.graphpad.com). Scale bar = 50 µm panel A, and 100 μm panel C.
