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Comparative Study
. 2021 Mar 17;19(3):156.
doi: 10.3390/md19030156.

Poly- and Oligosaccharide Ulva sp. Fractions from Enzyme-Assisted Extraction Modulate the Metabolism of Extracellular Matrix in Human Skin Fibroblasts: Potential in Anti-Aging Dermo-Cosmetic Applications

Mathilde Fournière  1   2 , Gilles Bedoux  1 , Nicolas Lebonvallet  3 , Raphaël Leschiera  3 , Claudie Le Goff-Pain  2 , Nathalie Bourgougnon  1 , Thomas Latire  1   2
Affiliations
Comparative Study

Poly- and Oligosaccharide Ulva sp. Fractions from Enzyme-Assisted Extraction Modulate the Metabolism of Extracellular Matrix in Human Skin Fibroblasts: Potential in Anti-Aging Dermo-Cosmetic Applications

Mathilde Fournière et al. Mar Drugs. .

Abstract

Ulva sp. is known to be a source of bioactive compounds such as ulvans, but their biological activity on human dermal fibroblast extracellular matrix (ECM) is poorly reported. In this work, the regulation of ECM has been investigated for the first time at both proteomic and transcriptomic levels in normal human skin dermal fibroblasts, after 48 h of incubation with poly- and oligosaccharide fractions from Ulva sp. obtained after enzyme-assisted extraction and depolymerization. Cell proliferation enhancement (up to +68%) without exhibiting any cytotoxic effect on fibroblasts was demonstrated at 50 and 1000 µg/mL by both fractions. At the proteomic level, polysaccharide fractions at 1000 µg/mL enhanced the most the synthesis of glycosaminoglycans (GAGs, up to +57%), total collagen, especially types I (up to +217%) and III, as well as the synthesis and activity of MMP-1 (Matrix Metalloproteinase-1, up to +309%). In contrast, oligosaccharide fractions had no effect on GAGs synthesis but exhibited similarities for collagens and MMP-1 regulation. At the transcriptomic level, the decrease of COL1A1 and COL1A2 expression, and increase of COL3A1 and MMP-1 expression, confirmed the modulation of ECM metabolism by both fractions. Our research emphasizes that poly- and oligosaccharide Ulva sp. fractions exhibit interesting biological activities and supports their potential use in the area of skin renewal for anti-aging dermo-cosmetic applications.

Keywords: Ulva sp.; collagen; extracellular matrix; human dermal fibroblast; matrix metalloproteinase; seaweed.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Effect of poly- and oligosaccharide fractions from Ulva sp. on the proliferation of fibroblasts evaluated by WST-1 assay. Normal human dermal fibroblasts (NHDF) were cultured for 48 h with two concentrations (50 and 1000 µg/mL) of fractions (n = 9). Significant differences between fractions and control are indicated by * p < 0.05 and *** p < 0.001.
Figure 2
Figure 2
Effect of poly- and oligosaccharide fractions from Ulva sp. on the cytotoxicity of fibroblasts evaluated by LDH (lactate dehydrogenase) assay. NHDF were cultured for 48 h with two concentrations (50 and 1000 µg/mL) of fractions (n = 3). No statistically significant differences were shown in comparison with the control.
Figure 3
Figure 3
Effect of poly- and oligosaccharidic fractions from Ulva sp. on fibroblast glycosaminoglycans (GAGs) synthesis evaluated by Alcian blue staining assay. NHDF were cultured for 48 h with two concentrations (50 and 1000 µg/mL) of fractions (n = 4). Significant differences between fractions and control are indicated by * p < 0.05 and ** p < 0.01.
Figure 4
Figure 4
Effect of poly- and oligosaccharide fractions from Ulva sp. on total collagen synthesis evaluated by Red sirius assay. NHDF were cultured for 48 h with two concentrations (50 and 1000 µg/mL) of fractions (n = 6 and n = 7, respectively). Significant differences between fractions and control are indicated by * p < 0.05 and ** p < 0.01.
Figure 5
Figure 5
Effect of poly- and oligosaccharidic fractions from Ulva sp. on fibroblast (a) type I collagen synthesis (n = 6) and (b) MMP-1 (Matrix Metalloproteinase-1) synthesis (n = 4) determined by ELISA assays. NHDF were cultured for 48 h with two concentrations (50 and 1000 µg/mL) of fractions. Significant differences between fractions and control are indicated by * p < 0.05.
Figure 6
Figure 6
Effect of poly- and oligosaccharide Ulva sp. fractions at 1000 µg/mL on (c) type I mature collagen synthesis (n = 5), (d) MMP-1 production (n = 6), (g) type III mature collagen synthesis (n = 4) and (h) tissue inhibitor of metalloproteinase--1 (TIMP-1) production (n = 5) in fibroblasts evaluated by Western blot. Representative blots of Western blot of (a) type I collagen synthesis, (b) MMP-1 production, (e) type III collagen synthesis, and (f) TIMP-1 production are shown. Significant differences between fractions and control are indicated by * p < 0.05.
Figure 7
Figure 7
Effect of poly- and oligosaccharide Ulva sp. fractions at 1000 µg/mL on (b) MMP-1 activity of NHDF (n = 4) determined by zymography assay. Representative gel of zymogram (a) MMP-1 activity is shown. No statistically significant differences were shown in comparison with the control.
Figure 8
Figure 8
Effect of poly- and oligosaccharide fractions from Ulva sp. on (a) COL1A1, (b) COL1A2, (c) MMP-1, (d) COL3A1, and (e) TIMP-1 mRNA expression levels in fibroblasts. NHDF (n = 4) were cultured for 48 h with the fractions at two concentrations (50 and 1000 µg/mL) for the assessment of genes. Significant differences between fractions and control are indicated by * p < 0.05 and ** p < 0.01.
Figure 9
Figure 9
Detailed processes of poly- and oligosaccharide fractions production from Ulva sp.
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