Low Molecular Weight Fucoidans Research Articles

Antioxidant potential of low molecular weight fucoidans from Sargassum autumnale against H2O2-induced oxidative stress in vitro and in zebrafish models based on molecular weight changes

In this study, we investigated the potential antioxidant abilities of low-molecular weight fucoidans from enzyme-assisted hydrolysates of Sargassum autumnale, based on molecular weight changes, in vitro and in vivo. The yield and free radical-scavenging activities of enzyme-assisted hydrolysates of S. autumnale were screened. The protamex-assisted hydrolysate of S. autumnale (SAP) presented the highest yield and 2,2-diphenyl-1-picrylhydrazyl (DPPH)-scavenging activity; therefore, it was chosen for fucoidan purification. Three fucoidan fractions were observed in SAP, and their antioxidant activity was assessed. Fucoidan fraction 3 of protamex-assisted hydrolysate of S. autumnale (SAPF3) offered significant protection against H2O2-induced oxidative stress, and was structurally and physically similar to commercial fucoidan. Fucose and low-molecular weight fucoidans were highly concentrated in SAPF3. The results of our study show that SAPF3, a low-molecular weight fucoidan from S. autumnale, possesses strong antioxidant properties and may be an effective alternative to antioxidant agents in the functional food industry.

Depolymerization of fucoidan with endo-fucoidanase changes bioactivity in processes relevant for bone regeneration

Fucoidans are polysaccharides from brown macroalgae, showing multiple bioactivities important for bone regeneration and bone health. However, the use of fucoidans in medical applications remains sparse due to the heterogeneity in their chemical properties and unclear structure-function relationships. Innovations in extraction techniques and post processing steps are needed to produce homogeneous fucoidan molecules with tailorable bioactivities. Here, we applied enzyme-assisted extraction coupled with enzymatic hydrolysis by Fhf1 fucoidanase to generate low (LMW) and medium molecular weight (MMW) fucoidans from Fucus evanescens. In contrast to the anti-angiogenic properties of the high molecular weight fucoidan, LMW and MMW no longer suppressed the production of pro-angiogenic molecules by bone stem cells, nor impaired the formation of prevascular structures in vitro. In contrast to LMW, a pro-inflammatory response of OEC was observed after treatment with high concentrations of MMW. Thus, fucoidanase hydrolysis could be a useful tool to tailor the bioactivity of fucoidans.

Low molecular weight fucoidan prepared by fucoidanase degradation – A promising browning inhibitor

Low molecular weight (MW) fucoidans have been proved with excellent tyrosinase inhibitory activity, which may proposed as a potential anti-browning ingredient for fruit and vegetable fresh-keeping agent. This study was conducted to obtain a low MW fucoidan fraction with high tyrosinase inhibitory capacity from Laminaria japonica crude fucoidan by fucoidanase degradation and to preliminary explore its type of tyrosinase inhibition and the application in browning inhibition of fresh-cut fruit. Firstly, the wild fucoidan-degrading strain, Flavobacteriaceae RC2-3, was mutated by UV irradiation and the fucoidanase production was improved by 40.5%. Fucoidan, which degraded by purified fucoidanase for 2 h, presented favourable tyrosinase inhibitory ability and were then separated by ultrafiltration. The fucoidan fraction with MW below 5 kDa (FF5) demonstrated the best tyrosinase inhibitory effect and shown obvious browning inhibition of fresh-cut apple slices. The inhibitory kinetics revealed that FF5 was a quite effectively competitive inhibitor of tyrosinase. The results of this study suggested that FF5 could be used as a promising browning inhibitor in the preservation of fruits and vegetables.

Structural Characterization and α-Glucosidase Inhibitory and Antioxidant Activities of Fucoidans Extracted from Saccharina japonica.

Two sulfated fucoidan fractions (Lj3 and Lj5) were extracted from Saccharina japonica and then subjected to acid hydrolysis to obtain Lj3h and Lj5h. Lj3h and Lj5h were characterized using IR, methylation analysis, and mass spectrometry. It was found that Lj3h and Lj5h were homogeneous low molecular weight fucoidans. Specifically, Lj3h was composed of the main chain of 1,3-linked α-L-fucopyranose residues with sulfate at C-2 and/or C-4 and three different monosaccharides (galactose, glucose, mannose) branched at C-2 and/or C-4 of fucose residue. Lj5h contained backbones of alternating galactopyranose residues and fucopyranose residues attached via a 1→3 linkage (galactofucan) and 1→6 linked galactan. The sulfation pattern was mainly located at C2/C4 fucose or galactose residues and more branches occupied at C-4 of fucose residue and C-2, C-3 or/and C-6 of galactose residue. In vitro assay indicated that, among the four fucoidans tested, only Lj5 showed potent α-glucosidase inhibitory activity with IC50 of 153.27±22.89 μg/mL, and the two parent fucoidans, Lj3 and Lj5, showed better antioxidant activity than their derivatives. These findings highlight the structure and bioactivity diversity of Saccharina japonica-derived fucoidans.

Antibacterial and Antioxidant Capacities and Attenuation of Lipid Accumulation in 3T3-L1 Adipocytes by Low-Molecular-Weight Fucoidans Prepared from Compressional-Puffing-Pretreated Sargassum Crassifolium.

In this study, we extracted fucoidan from compressional-puffing-pretreated Sargassum crassifolium by hot water. The crude extract of fucoidan (SC) was degraded by various degradation reagents and four low-molecular-weight (LMW) fucoidans, namely SCO (degradation by hydrogen peroxide), SCA (degradation by ascorbic acid), SCOA (degradation by hydrogen peroxide + ascorbic acid), and SCH (degradation by hydrogen chloride) were obtained. The degradation reagents studied could effectively degrade fucoidan into LMW fucoidans, as revealed by intrinsic viscosity, agarose gel electrophoresis, and molecular weight analyses. These LMW fucoidans had higher uronic acid content and sulfate content than those of SC. It was found that SCOA exhibited antibacterial activity. All LMW fucoidans showed antioxidant activities as revealed by DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt), and FRAP (ferric reducing antioxidant power) methods. Biological experiments showed that SC and SCOA had relatively high activity for the reversal of H2O2-induced cell death in 3T3-L1 adipocytes, and SCOA showed the highest effect on attenuation of lipid accumulation in 3T3-L1 adipocytes. Therefore, for the LMW fucoidans tested, SCOA showed antibacterial activity and had a high fucose content, high sulfate content, high activity for the reversal of H2O2-induced cell death, and a marked effect on attenuation of lipid accumulation. It can thus be recommended as a natural and safe antibacterial and anti-adipogenic agent for food, cosmetic, and nutraceutical applications.

Melanogenesis inhibitory effect of low molecular weight fucoidan from Undaria pinnatifida

In this study, fucoidans with different molecular weight that were isolated from the brown alga Undaria pinnatifida (Phaeophyceae, Laminariales) were investigated for their ability to inhibit melanogenesis and scavenge superoxide and hydroxyl radicals. Fucoidan samples with low molecular weights of 89, 35, 17, and 6 kDa were prepared by radiation-degradation of a 378 kDa fucoidan isolated from U. pinnatifida. The inhibitory activity of fucoidan against melanin biosynthesis in B16BL6 melanoma cells was enhanced for low molecular weight samples. To investigate the increase in melanogenesis inhibition exhibited by the low molecular weight fucoidan, tyrosinase inhibition activity and radical scavenging activities were measured. There was an increase in the tyrosinase inhibition activity for low molecular weight samples. Additionally, the radical scavenging activity was increased for lower molecular weight fucoidans. These results suggest that low molecular weight fucoidans from seaweeds may have beneficial biological properties.

A mechanism of low molecular weight fucoidans degraded by enzymatic and acidic hydrolysis for the prevention of UVB damage

Fucoidans have been long used as a food supplement due to their diverse pharmacological effects on human health. Low molecular weight fucoidan (LF) is a common form of fucoidans that has been shown to have enhanced biological activity. In the present study, fucoidans were extracted from the brown alga Sargassum hemiphyllum and enzyme-hydrolyzed into low and high molecular weight fucoidans (HF). The skin protective effects of LF, HF, and other fucoidans derivatives against ultraviolet B (UVB) damage were determined by measuring the expression levels of matrix metalloproteinase genes encoding collagenases (MMP-1, MMP-2, MMP-8, MMP-9, MMP-13), transforming growth factor β receptor II (TGFβRII), and type I procollagen. The results show that LF protects against UVB damage to the skin by inhibiting UVB-induced transcription factor activator protein-1 (AP-1)-stimulated transcription of MMP genes encoding collagenases (MMP-1, MMP-8, and MMP-13) and increasing TGFβRII mRNA levels to prevent the loss of TGFβRII that occurs during UVB-induced collagen degradation. Moreover, this study reveals that the biological properties of fucoidans are highly dependent on the fucose content, sulfate content, and molecular weight.

Fucoidan from Fucus vesiculosus Fails to Improve Outcomes Following Intracerebral Hemorrhage in Mice.

Intracerebral hemorrhage (ICH) is the most fatal stroke subtype, with no effective therapies. Hematoma expansion and inflammation play major roles in the pathophysiology of ICH, contributing to primary and secondary brain injury, respectively. Fucoidan, a polysaccharide from the brown seaweed Fucus vesiculosus, has been reported to activate a platelet receptor that may function in limiting bleeding, and to exhibit anti-inflammatory effects. As such, the aim of the present study was to examine the effects of fucoidan on hemorrhaging and neurological outcomes after ICH. Male CD-1 mice were subjected to experimental ICH by infusion of bacterial collagenase. Animals were randomly divided into the following groups: sham, ICH + vehicle, ICH + 25 mg/kg fucoidan, ICH + 75 mg/kg fucoidan, and ICH + 100 mg/kg fucoidan. Brain water content, neurobehavioral outcomes, and hemoglobin content were evaluated at 24 h post ICH. Our findings show that fucoidan failed to attenuate the ICH-induced increase in BWC. The neurological deficits that result from ICH also did not differ in the treatment groups at all three doses. Finally, we found that fucoidan had no effect on the hemoglobin content after ICH. We postulate that fucoidan treatment did not improve the measured outcomes after ICH because we used crude fucoidan, which has a high molecular weight, in our study. High-molecular-weight fucoidans are reported to have less therapeutic potential than low molecular weight fucoidans. They have been shown to exhibit anti-coagulant and pro-apoptotic properties, which seem to outweigh their anti-inflammatory and potential procoagulant abilities. We propose that using a low-molecular-weight fucoidan, or fractionating the crude polysaccharide, may be effective in treating ICH. Future studies are needed to confirm this.

Isolation and characterization of fucoidans from five brown algae and evaluation of their antioxidant activity

In this study, we evaluated the chemical property and antioxidant activity of fucoidans isolated from brown algae, Laminaria japonica (LJF), Lessonia nigrescens (LNF), Lessonia trabeculata (LTF), Ascophyllum mackaii (AMF), and Ecklonia maxima (EMF). LJF was less in sulfate content (14.16%) and more in galactose and mannose content (1.08 and 0.68) than the documented early. EMF contained 20%-30% of sulfate and fucose, 0.97 in molar ratio which was lower than that of sulfate to other four fucoidans (1.21-1.41). AMF (162 kDa) and EMF (150 kDa) were the first two largest in molecular weight, which were followed by LJP (126 kDa), LNF (113 kDa) and LTF (105 kDa). The fucoidans isolated these algae showed a wide range of antioxidant activity in vitro. It was found that the reducing power of the isolated fucoidans was positively correlated with their sulfate content and molecular weight. In addition, LNF and LTF at low concentrations exhibited high superoxide and hydroxyl radical scavenging activity. This demonstrated that low molecular weight fucoidans may perform a high antioxidant activity.

Degradation of fucoidans from Sargassum fulvellum and their biological activities

A fucoidan extracted from Sargassum fulvellum, was degraded by ultrasound (US) or electron beam (EB) irradiation in the presence of hydrogen peroxide aqueous solution. From the energetic point of view, the most effective method for the fucoidan degradation was found to be EB radiation method in H2O2 with a yield of scission Gs 3.310×10−8mol/J at the reaction condition of 1.5% hydrogen peroxide and irradiation 2.5kGy. The degradation took place by the formation of a reactive hydroxyl radical due to the dissociation of H2O2 in the presence of US or EB. The low molecular weight fucoidans (LMWFs) prevented P-selectin binding to Sialyl Lewis X with an IC50 (inhibitory concentration 50) of 20nM as compared to 400nM for heparin and 25,000nM for dextran sulfate. The LMWFs showed no hemolytic activity at concentrations up to 950μg/ml.

Fucoidan in a 3D scaffold interacts with vascular endothelial growth factor and promotes neovascularization in mice.

The aim of this study was to functionalize 3D porous cross-linked scaffolds with natural non-animal sulfated polysaccharide fucoidans in order to allow a delivery of vascular endothelial growth factor (VEGF) and potentiate its angiogenic activity. Microporous (20μm) and macroporous (200μm) scaffolds were functionalized with low, medium, or high molecular weight fucoidans (named LMWF, MMWF, and HMWF, respectively). In vitro, addition of fucoidans promoted endothelial progenitor cells proliferation in both micro- and macroporous scaffolds. While control scaffolds without fucoidans loaded with VEGF165 (100ng) showed a fast burst release in PBS during the first 24h, MMWF significantly reduced the VEGF165 release (p < 0.001). Surface plasmon resonance experiments confirmed a direct interaction between MMWF and VEGF165, characterized by an affinity K D (K d/K a) of 1 × 10(-9)M. In a subcutaneous angiogenesis model in mice, fucoidan functionalized scaffolds showed a more intense vascularization response than control groups. Expression of isolectin-B4 and α-smooth muscle actin, as well as confinement of erythrocytes, demonstrated the neoformed blood vessels functionality. There was a significant difference in neovessel area and neovessel density between MMWF scaffolds or VEGF165 scaffolds and MMWF+VEGF165 scaffolds (p < 0.001 for all cases). Here, we demonstrate that fucoidan sequesters VEGF165 and delivers biological cues promoting angiogenesis. In conclusion, this study shows that hydrogels functionalized with fucoidan can direct the formation of mature vasculature through a local release of VEGF165 and can be a useful tool in ischemic tissues to guide therapeutic angiogenesis.

A comparative study of antithrombotic and antiplatelet activities of different fucoidans from Laminaria japonica

IntroductionFucoidans extracted from brown algae represent an intriguing group of natural fucose-enriched sulfated polysaccharide, with excellent anticoagulant, antimetastatic, antiangiogenic and anti-inflammatory activities. In the present study, we compared antithrombotic activities of four fucoidan fractions with different molecular weight and sulfated ester content from Laminaria japonica in an electrical induced arterial thrombosis and their potential mechanism underlying such activity. Results and ConclusionsIn vivo middle molecular weight (MMW) fucoidan fractions with molecular weight about 28000 and 35000 exhibited better antithrombotic activity in electrical induced arterial thrombosis than low molecular weight (LMW) fucoidan LF1 and LF2 (Mw 7600 and 3900). Inhibition of arterial thrombosis occurred at dose of 0.1-0.25mg/kg for MMW fucoidans, accompanied with moderate anticoagulant activity and significant decrease of whole blood viscosity and hematocrit. The antithrombotic effects of MMW Fucoidans might be related with promotion of TFPI content and decrease of TXB2 content, without affecting platelet aggregation and 6-keto-PGF1α content in vivo. In contrast, LMW fucoidans showed a correlation among anticoagulant, antiplatelet and antithrombotic effects in vivo. Antithrombotic action of LF1 and LF2 required high dose of 2.5-10mg/kg, concomitantly with anticoagulant activity and specific inhibition of platelet aggregation in vivo. Their antithrombotic effect might be related to their promotion of TFPI and 6-keto-PGF1α, down regulation of TXB2, without affecting hemorheology. These findings suggested that fucoidan fractions with different molecular weight acted on the antithrombotic action by different mechanism. By comparison, highly sulfated fucoidan LF2 with molecular weight of 3900 seemed to be a more suitable choice of antithrombotic drug for its antithrombotic activity accompanied with specific inhibitory activity on platelet aggregation, low anticoagulant activity and low hemorrhagic risk in vivo.

Relationship between oversulfation and conformation of low and high molecular weight fucoidans and evaluation of their in vitro anticancer activity.

Low and high molecular weight fucoidans (F5-30K and F>30K) were chemically modified through the addition of sulfate groups, and the effect of oversulfation on the in vitro anticancer activity was investigated. After the addition of sulfate groups, a considerable increase of 35.5 to 56.8% was observed in the sulfate content of the F5-30K fraction, while the sulfate content of the F>30K fraction increased to a lesser extent (from 31.7 to 41.2%). Significant differences in anticancer activity were observed between the oversulfated F5–30K and F>30K fractions, with activities of 37.3–68.0% and 20.6–35.8%, respectively. This variation in the anticancer activity of oversulfated fucoidan derivatives was likely due to differences in their sulfate content. The results suggest that the molecular conformation of these molecules is closely related to the extent of sulfation in the fucan backbones and that the sulfates are preferably substituted when the fucoidan polymers are in a loose molecular conformation.

The differential effect of high and low molecular weight fucoidans on the severity of collagen-induced arthritis in mice.

Fucoidans have been extensively studied for their various biological activities but the exact role of fucoidans on the inflammatory processes associated with arthritic disease has not been studied. The effect of the treatment of high, medium and low molecular weight fucoidans (HMWF, MMWF and LMWF, respectively) on the progression of collagen-induced arthritis (CIA) was tested. A daily oral administration of HMWF enhanced the severity of arthritis, inflammatory responses in the joint cartilage and the levels of collagen-specific antibodies, while LMWF reduced the severity of arthritis and the levels of Th1-dependent collagen-specific IgG(2a). Further in vitro analyses, using macrophage cell lines, revealed that the HMWF induced the expression of various inflammatory mediators, and enhanced the cellular migration of macrophages. These stimulatory effects of fucoidan decreased in fucoidans with lower molecular weights and LMWF did not exhibit any pro-inflammatory effects. Interestingly, the oral administration of HMWF enhanced the production of IFN-gamma, one of the Th1 cytokines, in collagen-stimulated spleen cells that had been isolated from CIA mice, while LMWF had the opposite effect. These results indicate that HMWF enhances arthritis through enhancing the inflammatory activation of macrophages while LMWF reduces arthritis through the suppression of Th1-mediated Immune reactions.

Modulation of protease nexin-1 activity by polysaccharides

Protease nexin-1 (PN-1) is a non-circulating pericellular serpin expressed by vascular cells. PN-1 inhibits different proteases but when associated with glycosaminoglycans, its activity is mainly directed towards thrombin. Fucoidans are sulphated polysaccharides which can interact with several serpins and have antithrombotic and anticoagulant properties in vivo with a lower hemorrhagic risk than heparin. The purpose of this study was to compare the effects of low (LMW) or high molecular weight (HMW) fucoidans to those of standard heparin and LMW heparin on PN-1 properties. Using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) and affinity coelectrophoresis, we observed that polysaccharides bound to thrombin, PN-1 and the thrombin/PN-1 complex. Progress curve kinetics showed that LMW and HMW fucoidans accelerate thrombin inhibition by PN-1 (111 and 402 fold, respectively) whereas the acceleration by LMW heparin and standard heparin was only of 36- and of 307-fold, respectively. Moreover, the formation of PN-1/(125)I-thrombin complex was increased in the presence of heparin, HMW and LMW fucoidans, but barely by LMW heparin. The dose response followed a bell shape curve, again suggesting the formation of ternary complexes between thrombin, PN-1 and polysaccharides. We also investigated the ability of polysaccharides to remove PN-1 bound to the cell membrane of smooth muscle cells in culture. PN-1 was detached by fucoidans and heparins and was still able to inhibit thrombin. In conclusion, fucoidans reduce cell-associated PN-1 and thrombin/PN-1 complexes and increase the antithrombin activity of PN-1. The capacity of PN-1 to regulate the pericellular activity of thrombin amongst other proteases reinforces the therapeutical interest of fucoidans.

Effects of middle molecular weight fucoidans on in vitro and ex vivo angiogenesis of endothelial cells

Native fucoidan, sulfated L-fucose polymer isolated from marine brown algae, has diverse biological activities including anticoagulant, anti-angiogenic and anti-tumor effects. Its anti-angiogenic effect is of interest and the mechanisms have been studied. However, recently, proangiogenic effect of low molecular weight (LMW) fucoidans has been demonstrated. To clarify the opposite effects of fucoidan and LMW fucoidans, it is necessary to address the relationship between the molecular size and the effect on angiogenesis. In this study, the effects of middle molecular weight (MMW) fucoidans (15-30 kDa) on human umbilical vein endothelial cell functions were examined. Fucoidan (30 kDa) had similar effects to native fucoidan; inhibition of HUVEC tube formation and angiogenesis in an ex vivo model, although their effects were weaker than native fucoidan. On the other hand, 15-20 kDa fucoidan enhanced HUVEC migration, but did not inhibit HUVEC tube formation. Thus, 15-20 kDa fucoidan would have proangiogenic effect on angiogenesis. These results elucidate that 20-30 kDa would be a critical point to characterize the role of fucoidans on angiogenesis.