title: Identification of a Pro-Angiogenic Potential and Cellular Uptake Mechanism of a LMW Highly Sulfated Fraction of Fucoidan from Ascophyllum nodosum.
abstract: Herein we investigate the structure/function relationships of fucoidans fromAscophyllum nodosumto analyze their pro-angiogenic effect and cellular uptake in native and glycosaminoglycan-free (GAG-free) human endothelial cells (HUVECs). Fucoidans are marine sulfated polysaccharides, which act as glycosaminoglycans mimetics. We hypothesized that the size and sulfation rate of fucoidans influence their ability to induce pro-angiogenic processes independently of GAGs. We collected two fractions of fucoidans, Low and Medium Molecular Weight Fucoidan (LMWF and MMWF, respectively) by size exclusion chromatography and characterized their composition (sulfate, fucose and uronic acid) by colorimetric measurement and Raman and FT-IR spectroscopy. The high affinities of fractionated fucoidans to heparin binding proteins were confirmed by Surface Plasmon Resonance. We evidenced that LMWF has a higher pro-angiogenic (2D-angiogenesis on Matrigel) and pro-migratory (Boyden chamber) potential on HUVECs, compared to MMWF. Interestingly, in a GAG-free HUVECs model, LMWF kept a pro-angiogenic potential. Finally, to evaluate the association of LMWF-induced biological effects and its cellular uptake, we analyzed by confocal microscopy the GAGs involvement in the internalization of a fluorescent LMWF. The fluorescent LMWF was mainly internalized through HUVEC clathrin-dependent endocytosis in which GAGs were partially involved. In conclusion, a better characterization of the relationships between the fucoidan structure and its pro-angiogenic potential in GAG-free endothelial cells was required to identify an adapted fucoidan to enhance vascular repair in ischemia.
authors.affiliation: Inserm U1148, LVTS, Université Paris 13, Sorbonne Paris Cité, Paris 75018, France. nicolas.marinval@inserm.fr.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Marinval
authors.affiliation: Inserm U1148, LVTS, Université Paris 13, Sorbonne Paris Cité, Paris 75018, France. pierre.saboural@univ-paris13.fr.
authors.forename: Pierre
authors.initials: P
authors.lastname: Saboural
authors.affiliation: Inserm U1148, LVTS, Université Paris 13, Sorbonne Paris Cité, Paris 75018, France. haddad.oualid@univ-paris13.fr.
authors.forename: Oualid
authors.initials: O
authors.lastname: Haddad
authors.affiliation: Inserm U1148, LVTS, Université Paris 13, Sorbonne Paris Cité, Paris 75018, France. murielle.maire@univ-paris13.fr.
authors.forename: Murielle
authors.initials: M
authors.lastname: Maire
authors.affiliation: Inserm U1148, LVTS, Université Paris 13, Sorbonne Paris Cité, Paris 75018, France. bassand.k@gmail.com.
authors.forename: Kevin
authors.initials: K
authors.lastname: Bassand
authors.affiliation: Laboratoire CSPBAT, CNRS UMR 7244, UFR SMBH, Université Paris 13, Sorbonne Paris Cité, Bobigny F-93017, France. frederic.geinguenaud@univ-paris13.fr.
authors.forename: Frederic
authors.initials: F
authors.lastname: Geinguenaud
authors.affiliation: Laboratoire CSPBAT, CNRS UMR 7244, UFR SMBH, Université Paris 13, Sorbonne Paris Cité, Bobigny F-93017, France. nadia.djaker@univ-paris13.fr.
authors.forename: Nadia
authors.initials: N
authors.lastname: Djaker
authors.affiliation: Laboratoire CSPBAT, CNRS UMR 7244, UFR SMBH, Université Paris 13, Sorbonne Paris Cité, Bobigny F-93017, France. khadijabenakrout@hotmail.fr.
authors.forename: Khadija
authors.initials: K
authors.lastname: Ben Akrout
authors.affiliation: Laboratoire CSPBAT, CNRS UMR 7244, UFR SMBH, Université Paris 13, Sorbonne Paris Cité, Bobigny F-93017, France. marc.lamydelachapelle@univ-paris13.fr.
authors.forename: Marc
authors.initials: M
authors.lastname: Lamy de la Chapelle
authors.affiliation: Inserm U1148, LVTS, Université Paris 13, Sorbonne Paris Cité, Paris 75018, France. robert.romain@gmail.com.
authors.forename: Romain
authors.initials: R
authors.lastname: Robert
authors.affiliation: Inserm U1148, LVTS, Université Paris 13, Sorbonne Paris Cité, Paris 75018, France. olivier.oudar@univ-paris13.fr.
authors.forename: Olivier
authors.initials: O
authors.lastname: Oudar
authors.affiliation: Inserm U1148, LVTS, Université Paris 13, Sorbonne Paris Cité, Paris 75018, France. erwan.guyot@aphp.fr.|Laboratoire de Biochimie, Hôpital Jean Verdier, Assistance Publique-Hôpitaux de Paris, Bondy 93140, France. erwan.guyot@aphp.fr.
authors.forename: Erwan
authors.initials: E
authors.lastname: Guyot
authors.affiliation: Inserm U1148, LVTS, Université Paris 13, Sorbonne Paris Cité, Paris 75018, France. christelle.laguillier@aphp.fr.|Laboratoire de Biochimie, Hôpital Jean Verdier, Assistance Publique-Hôpitaux de Paris, Bondy 93140, France. christelle.laguillier@aphp.fr.
authors.forename: Christelle
authors.initials: C
authors.lastname: Laguillier-Morizot
authors.affiliation: Inserm U1148, LVTS, Université Paris 13, Sorbonne Paris Cité, Paris 75018, France. angela.sutton@aphp.fr.|Laboratoire de Biochimie, Hôpital Jean Verdier, Assistance Publique-Hôpitaux de Paris, Bondy 93140, France. angela.sutton@aphp.fr.
authors.forename: Angela
authors.initials: A
authors.lastname: Sutton
authors.affiliation: Inserm U1148, LVTS, Université Paris 13, Sorbonne Paris Cité, Paris 75018, France. cedric.chauvierre@inserm.fr.
authors.forename: Cedric
authors.initials: C
authors.lastname: Chauvierre
authors.affiliation: Inserm U1148, LVTS, Université Paris 13, Sorbonne Paris Cité, Paris 75018, France. frederic.chaubet@univ-paris13.fr.
authors.forename: Frederic
authors.initials: F
authors.lastname: Chaubet
authors.affiliation: Inserm U1148, LVTS, Université Paris 13, Sorbonne Paris Cité, Paris 75018, France. nathalie.charnaux@aphp.fr.|Laboratoire de Biochimie, Hôpital Jean Verdier, Assistance Publique-Hôpitaux de Paris, Bondy 93140, France. nathalie.charnaux@aphp.fr.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Charnaux
authors.affiliation: Inserm U1148, LVTS, Université Paris 13, Sorbonne Paris Cité, Paris 75018, France. hania.hlawaty@inserm.fr.
authors.forename: Hanna
authors.initials: H
authors.lastname: Hlawaty
mesh_terms: D043925:Angiogenesis Inducing Agents / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D044646:Ascophyllum / Q000737:chemistry*; D051242:Caveolin 1 / Q000737:chemistry; D002465:Cell Movement / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002850:Chromatography, Gel; D002966:Clathrin / Q000737:chemistry; D004705:Endocytosis / Q000187:drug effects; D006025:Glycosaminoglycans / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008970:Molecular Weight; D018919:Neovascularization, Physiologic / Q000187:drug effects; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D013329:Structure-Activity Relationship
chemical_list: D043925:Angiogenesis Inducing Agents; D051242:Caveolin 1; D002966:Clathrin; D006025:Glycosaminoglycans; D011134:Polysaccharides; C007789:fucoidan
keywords: angiogenesis; endocytosis; fucoidan; glycocalyx; glycosaminoglycans