Corinne Sinquin

Further data on the structure of brown seaweed fucans: relationships with anticoagulant activity.

The composition, molecular weight (MW), anticoagulant activity and nuclear magnetic resonance spectra of various low-molecular-weight fucans (LMWFs) obtained by partial hydrolysis or radical depolymerization of a crude fucoidan extracted from the brown seaweed Ascophyllum nodosum are compared. Fucose units were found mainly sulfated at O-2, to a lesser extent at O-3, and only slightly at O-4, contrary to previously published results for fucoidans from other brown seaweeds, and fucose 2, 3-O-disulfate residues were observed for the first time. As the sulfation pattern excluded an alpha-(1-->2)-linked fucose backbone and a high proportion of alpha-(1-->4) linkages was found, it would appear that the concept of fucoidan structure needs to be revised. Anticoagulant activity is apparently related not only to MW and sulfation content, as previously determined, but also (and more precisely) to 2-O-sulfation and 2,3-O-disulfation levels.

Relationship between Sulfate Groups and Biological Activities of Fucans

The anticoagulant and antiproliferative effects of low molecular weight fucans with different sulfate content were examined. The anticoagulant activity was determined by activated partial thromboplastin time and the antiproliferative one was achieved in vitro on CCL39 fibroblast cell line. The results showed that inhibitory effects of fucans on both coagulation and cell proliferation are dependent on their sulfation degree. Decreased sulfation diminishes the two activities not in the same manner: some low molecular weight fucan fractions with no anticoagulant activity retain their ability to inhibit cell growth.

Marine Polysaccharides: A Source of Bioactive Molecules for Cell Therapy and Tissue Engineering

The therapeutic potential of natural bioactive compounds such as polysaccharides, especially glycosaminoglycans, is now well documented, and this activity combined with natural biodiversity will allow the development of a new generation of therapeutics. Advances in our understanding of the biosynthesis, structure and function of complex glycans from mammalian origin have shown the crucial role of this class of molecules to modulate disease processes and the importance of a deeper knowledge of structure-activity relationships. Marine environment offers a tremendous biodiversity and original polysaccharides have been discovered presenting a great chemical diversity that is largely species specific. The study of the biological properties of the polysaccharides from marine eukaryotes and marine prokaryotes revealed that the polysaccharides from the marine environment could provide a valid alternative to traditional polysaccharides such as glycosaminoglycans. Marine polysaccharides present a real potential for natural product drug discovery and for the delivery of new marine derived products for therapeutic applications.

Relationships between chemical characteristics and anticomplementary activity of fucans.

We have shown previously that a low-molecular-weight fucan extracted from the brown seaweed Ascophylum nodosum strongly inhibited human complement activation in vitro and its mechanism of action was largely elucidated. We further investigated the influence of molecular weight and chemical composition of fucan on its anticomplementary activity. The capacity of 12 fragments of fucan (ranging from a molecular weight of 4100 to 214,000) to prevent complement-mediated haemolysis of sheep erythrocytes (classical pathway) and of rabbit erythrocytes (alternative pathway) increased with increasing molecular weight, and reached a plateau for 40,000 and 13,500, respectively. The most potent fucan fractions were 40-fold more active than heparin in inhibiting the classical pathway. They were, however, as active as heparin in inhibiting the alternative pathway. In addition, we have developed a haemolytic test based on the CH50 protocol, which allows discrimination between activators and inhibitors of complement proteins. Although the mannose content within the different fucan fragments did not vary, the galactose and glucuronic acid contents increased with increasing activity, suggesting that these residues should be essential for full anticomplementary activity. Meanwhile, sulphate groups appeared to be necessary, but were clearly not a sufficient requirement for anticomplementary activity of fucans. Taken together, these data illustrate the prospects for the use of fucans as potential anti-inflammatory agents.

Antiviral activities of sulfated polysaccharides isolated from Sphaerococcus coronopifolius (Rhodophytha, Gigartinales) and Boergeseniella thuyoides (Rhodophyta, Ceramiales).

Water-soluble sulfated polysaccharides isolated from two red algae Sphaerococcus coronopifolius (Gigartinales, Sphaerococcaceae) and Boergeseniella thuyoides (Ceramiales, Rhodomelaceae) collected on the coast of Morocco inhibited in vitro replication of the Human Immunodeficiency Virus (HIV) at 12.5 μg/mL. In addition, polysaccharides were capable of inhibiting the in vitro replication of Herpes simplex virus type 1 (HSV-1) on Vero cells values of EC50 of 4.1 and 17.2 μg/mL, respectively. The adsorption step of HSV-1 to the host cell seems to be the specific target for polysaccharide action. While for HIV-1, these results suggest a direct inhibitory effect on HIV-1 replication by controlling the appearance of the new generations of virus and potential virucidal effect. The polysaccharides from S. coronopifolius (PSC) and B. thuyoides (PBT) were composed of galactose, 3,6-anhydrogalactose, uronics acids, sulfate in ratios of 33.1, 11.0, 7.7 and 24.0% (w/w) and 25.4, 16.0, 3.2, 7.6% (w/w), respectively.

Exopolysaccharides produced by marine bacteria and their applications as glycosaminoglycan-like molecules

Although polysaccharides are ubiquitous and the most abundant renewable bio-components, their studies, covered by the glycochemistry and glycobiology fields, remain a challenge due to their high molecular diversity and complexity. Polysaccharides are industrially used in food products; human therapeutics fall into a more recent research field and pharmaceutical industry is looking for more and more molecules with enhanced activities. Glycosaminoglycans (GAGs) found in animal tissues play a critical role in cellular physiological and pathological processes as they bind many cellular components. Therefore, they present a great potential for the design and preparation of therapeutic drugs. On the other hand, microorganisms producing exopolysaccharides (EPS) are renewable resources meeting well the actual industrial demand. In particular, the diversity of marine microorganisms is still largely unexplored offering great opportunities to discover high value products such as new molecules and biocatalysts. EPS-producing bacteria from the marine environment will be reviewed with a focus on marine-derived EPS from bacteria isolated from deep-sea hydrothermal vents. Information on chemical and structural features, putative pathways of biosynthesis, novel strategies for chemical and enzymatic modifications and potentialities in the biomedical field will be provided. An integrated approach should be used to increase the basic knowledge on these compounds and their applications; new clean environmentally friendly processes for the production of carbohydrate bioactive compounds should also be proposed for a sustainable industry.

Pharmacological Modulation of Human Mesenchymal Stem Cell Chondrogenesis by a Chemically Oversulfated Polysaccharide of Marine Origin: Potential Application to Cartilage Regenerative Medicine

Mesenchymal stem cells (MSCs) are considered as an attractive source of cells for cartilage engineering due to their availability and capacity for expansion and multipotency. Differentiation of MSC into chondrocytes is crucial to successful cartilage regeneration and can be induced by various biological agents, including polysaccharides that participate in many biological processes through interactions with growth factors. Here, we hypothesize that growth factor‐induced differentiation of MSC can be increased by chemically oversulfated marine polysaccharides. To test our hypothesis, human adipose tissue‐derived MSCs (hATSCs) were cultured in pellets with transforming growth factor (TGF)‐β1‐supplemented chondrogenic medium containing either the polysaccharide GY785 DR or its oversulfated isoform GY785 DRS. Chondrogenesis was monitored by the measurement of pellet volume, quantification of DNA, collagens, glycosaminoglycans (GAGs), and immunohistological staining. Our data revealed an increase in pellet volume, total collagens, and GAG production with GY785 DRS and chondrogenic medium. The enhanced chondrogenic differentiation of hATSC was further demonstrated by the increased expression of several chondrogenic markers by real‐time reverse transcription‐polymerase chain reaction. In addition, surface plasmon resonance analyses revealed that TGF‐β1 bound GY785 DRS with higher affinity compared to GY785 DR. In association with TGF‐β1, GY785 DRS was found to upregulate the phosphorylation of extracellular signal‐regulated kinase 1/2, indicating that oversulfated polysaccharide affects the mitogen activated protein kinase signaling activity. These results demonstrate the upregulation of TGF‐β1‐dependent stem cell chondrogenesis by a chemically oversulfated marine polysaccharide. This polysaccharide of marine origin is easily producible and therefore could be considered a promising additive to drive efficient and reliable MSC chondrogenesis for cartilage tissue engineering. STEM CELLS 2012;30:471–480

Effect of low‐molecular‐weight fucoidan on experimental arterial thrombosis in the rabbit and rat

was used to perform an extra-corporeal arteriovenous shunt between the carotid artery and the jugular vein ; mean times for formation of an occlusive thrombus in the carotid artery were determined in control and treated animals. Both fucoidan and unfractionated heparin, injected intravenously 5 min before thrombosis induction, inhibited thrombus formation significantly. LMW fucoidan and heparin increased mean time-to-occlusion about three-fold in the same concentration range (2 mg/kg fucoidan 43.7 + 5.5 min and 1 mg/kg heparin 55.4 + 2.6 versus control 15.6 + 1.6 min ; 5 animals per group ; p < 0.001). The anti-thrombotic effect of fucoidan was also evaluated in this model after subcutaneous injection 2 h before thrombosis induction. With a dose of 7.5 mg/kg, time-to-occlusion was prolonged significantly (22.8 + 3.4 min in the treated group

Structural data on a bacterial exopolysaccharide produced by a deep-sea Alteromonas macleodii strain

Some marine bacteria collected around deep-sea hydrothermal vents are able to produce, in laboratory conditions, complex and innovative exopolysaccharides. In a previous study, the mesophilic strain Alteromonas macleodii subsp. fijiensis biovar deepsane was collected on the East Pacific Rise at 2600 m depth. It was isolated from a polychaete annelid Alvinella pompejana and is able to synthesise and excrete the exopolysaccharide deepsane. Biological activities have been screened and some protective properties have been established. Deepsane is commercially available in cosmetics under the name of Abyssine(®) for soothing and reducing irritation of sensitive skin against chemical, mechanical and UVB aggression. This study presents structural data for this original and complex bacterial exopolysaccharide and highlights some structural similarities with other known EPS produced by marine Alteromonas strains.

An in vitro study of two GAG-like marine polysaccharides incorporated into injectable hydrogels for bone and cartilage tissue engineering

Natural polysaccharides are attractive compounds with which to build scaffolds for bone and cartilage tissue engineering. Here we tested two non-standard ones, HE800 and GY785, for the two-dimensional (2-D) and three-dimensional (3-D) culture of osteoblasts (MC3T3-E1) and chondrocytes (C28/I2). These two glycosaminoglycan-like marine exopolysaccharides were incorporated into an injectable silylated hydroxypropylmethylcellulose-based hydrogel (Si-HPMC) that has already shown its suitability for bone and cartilage tissue engineering. Results showed that, similarly to hyaluronic acid (HA) (the control), HE800 and GY785 significantly improved the mechanical properties of the Si-HPMC hydrogel and induced the attachment of MC3T3-E1 and C28/I2 cells when these were cultured on top of the scaffolds. Si-HPMC hydrogel containing 0.67% HE800 exhibited the highest compressive modulus (11kPa) and allowed the best cell dispersion, especially of MC3T3-E1 cells. However, these cells did not survive when cultured in 3-D within hydrogels containing HE800, in contrast to C28/I2 cells. The latter proliferated in the microenvironment or concentrically depending on the nature of the hydrogel. Among all the constructs tested the Si-HPMC hydrogels containing 0.34% HE800 or 0.67% GY785 or 0.67% HA presented the most interesting features for cartilage tissue engineering applications, since they offered the highest compressive modulus (9.5-11kPa) while supporting the proliferation of chondrocytes.