Fucoidans are algal polysaccharides that exhibit protective properties against oxidative stress. The aim of this study was to investigate different fucoidans from brown seaweeds for their ability to protect against iron-dependent oxidative stress (ferroptosis), a main hallmark of retinal and brain diseases, including hemorrhage. We investigated five new high-molecular weight fucoidan extracts from Fucus vesiculosus, F. serratus, and F. distichus subsp. evanescens, a previously published Laminaria hyperborean extract, and commercially available extracts from F. vesiculosus and Undaria pinnatifida. We induced oxidative stress by glutathione depletion (erastin) and H2O2 in four retinal and neuronal cell lines as well as primary cortical neurons. Only extracts from F. serratus, F. distichus subsp. evanescens, and Laminaria hyperborea were partially protective against erastin-induced cell death in ARPE-19 and OMM-1 cells, while none of the extracts showed beneficial effects in neuronal cells. Protective fucoidans also attenuated the decrease in protein levels of the antioxidant enzyme GPX4, a key regulator of ferroptosis. This comprehensive analysis demonstrates that the antioxidant abilities of fucoidans may be cell type-specific, besides depending on the algal species and extraction method. Future studies are needed to further characterize the health-benefiting effects of fucoidans and to determine the exact mechanism underlying their antioxidative abilities.
Fouling is a major problem in seaweed aquaculture and one of the main obstacles during the domestication process for new culture species. During first attempts to cultivate Fucus vesiculosus and Fucus serratus in the Kiel fjord (Western Baltic Sea), fouling by the epizoans Electra pilosa, Mytilus sp., and Amphibalanus improvisus rendered the production of marketable biomass impossible. This study tested (1) if regular desiccation by air exposure is effective in decreasing the abundance and size of foulers and (2) if and how regular desiccation affects the growth performance of the cultivated Fucus thalli. For this purpose, thalli of F. vesiculosus and F. serratus were cultivated freely floating in baskets directly deployed in the fjord and desiccated to defined percentages of the wet weight (ww) by air exposure. The treatments comprised controls and desiccations of different intensities (from 90 to 40% of ww) and at different frequencies (1× week −1 , 3× week −1). Growth rates of both Fucus species were not or only slightly reduced by the desiccation treatments. The final harvested biomass of F. vesiculosus under frequent mild desiccations (3× week −1 to 80% of ww) was even higher than the biomass of undesiccated controls. The size of the epizoans E. pilosa and A. improvisus was significantly reduced by the desiccation treatments and the abundance of all epizoan species was drastically reduced by the desiccation regimes. Frequent mild desiccations (F. vesiculosus: 3× week −1 to 80% of ww, F. serratus: 3× week −1 to 90% of ww) proved to be most effective and decreased the epizoan ww share of the total harvest from 13.0 ± 4.8% in the control to 1.8 ± 0.2% for F. vesiculosus and from 19.1 ± 2.7 to 1.0 ± 0.1% for F. serratus. Thus, desiccation seems to be an effective measure for the production of clean Fucus biomass in culture which is necessary for further valorization. A technical solution for the implementation of this procedure in large-scale cultures remains to be developed.
An experimental farm has been installed in the Kiel Fjord, western Baltic Sea, aiming at the development of a sustainable production process for Fucus species ( Fucus vesiculosus , Fucus serratus ). The envisaged cultivation method includes the unattached rearing of thalli in baskets deployed in the sea and their vegetative reproduction. Fertility (i.e., receptacle formation) is expected to be problematic for this approach, because receptacles are terminated in growth and degrade after gamete release. In culture experiments, natural fertility led to only minimal overall growth in F. vesiculosus and even weight loss in F. serratus . Therefore, we tested if long-term unattached cultivation of formerly attached thalli leads to a lowering of fertility by an acclimatization process. However, fertility after 1 and 2 years of unattached cultivation was statistically equal and still comparable to the high fertility of attached populations. Furthermore, we tested if the only known naturally unattached population in the western Baltic Sea near Glücksburg, which remains largely infertile in the wild, keeps its low fertility if put under culture conditions. During an experimental 1-year cultivation, thalli from this population remained almost entirely vegetative (2.0 ± 3.1% fertile apices). Hence, the Glücksburg population is a promising source of aquacultural seedling biomass. Yet, further tests are necessary to check, if the fertility remains low over several years of cultivation. If unattached populations are used as source for commercial cultures, the collection of seedling material should always be accompanied by strong measures to ensure the continued integrity of these valuable habitats.
Fucoidans, sulfated polysaccharides extracted from brown algae, are marine products with the potential to modulate bone formation and vascularization processes. The bioactivity and safety of fucoidans are highly associated with their chemical structure, which may vary with algae species and extraction method. Thus, in depth evaluation of fucoidan extracts in terms of endotoxin content, cytotoxicity, and their detailed molecular biological impact on the individual cell types in bone is needed. In this study, we characterized fucoidan extracts from three different Fucus species including Fucus vesiculosus (Fv), Fucus serratus (Fs), and Fucus distichus subsp. evanescens (Fe) for their chemical features, endotoxin content, cytotoxicity, and bioactive effects on human outgrowth endothelial cells (OEC) and human mesenchymal stem cells (MSC) as in vitro models for bone function and vascularization. Extracts contained mainly high molecular weight (HMW) fucoidans and were free of endotoxins that may cause inflammation or influence vascularization. OEC tolerated fucoidan concentrations up to 200 µg/mL, and no indication of cytotoxicity was observed. The inflammatory response, however, investigated by real-time PCR (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) and endothelial barrier assessed by impedance measurement differed for the individual extracts. MSC in comparison with endothelial cells were more sensitive to fucoidans and showed partly reduced metabolic activity and proliferation at higher doses of fucoidans. Further results for MSC indicated impaired osteogenic functions in alkaline phosphatase and calcification assays. All tested extracts consistently lowered important molecular mediators involved in angiogenesis, such a VEGF (vascular endothelial growth factor), ANG-1 (angiopoietin 1), and ANG-2 (angiopoietin 2), as indicated by RT-PCR and ELISA. This was associated with antiangiogenic effects at the functional level using selected extracts in co-culture models to mimic bone vascularization processes during bone regeneration or osteosarcoma.
Ingredients of brown seaweed like fucoidans are often described for their beneficial biological effects, that might be interesting for a medical application. In this study, we tested an extract from Dictyosiphon foeniculaceus (DF) to evaluate the effects in glioblastoma and uveal melanoma, looking for a possible anti-cancer treatment. We investigated toxicity, VEGF (vascular endothelial growth factor) secretion and gene expression of tumor and non-tumor cells. SVGA (human fetal astrocytes), the human RPE (retinal pigment epithelium) cell line ARPE-19, the tumor cell line OMM-1 (human uveal melanoma), and two different human primary glioblastoma cultures (116-14 and 118-14) were used. Tests for cell viability were conducted with MTS-Assay (3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium), and the proliferation rate was determined with cell counting. VEGF secretion was assessed with ELISA (enzyme-linked immunosorbent assay). The gene expression of VEGF receptor 1 (VEGFR1), VEGF receptor 2 (VEGFR2) and VEGF-A was determined with real-time qPCR (quantitative polymerase chain reaction). DF lowered the cell viability of OMM-1. Proliferation rates of ARPE-19 and OMM-1 were decreased. The VEGF secretion was inhibited in ARPE-19 and OMM-1, whereas it was increased in SVGA and 116-14. The expression of VEGFR1 was absent and not influenced in OMM-1 and ARPE-19. VEGFR2 expression was lowered in 116-14 after 24 h, whereas VEGF-A was increased in 118-14 after 72 h. The extract lowered cell viability slightly and was anti-proliferative depending on the cell type investigated. VEGF was heterogeneously affected. The results in glioblastoma were not promising, but the anti-tumor properties in OMM-1 could make them interesting for further research concerning cancer diseases in the human eye.
Angiogenesis, the formation of new blood vessels from existing ones, is an essential process for successful bone regeneration. Further, angiogenesis is a key factor for the development of bone-related disorders like osteosarcoma or arthritis. Fucoidans, sulfated polysaccharides from brown algae, have been shown to affect angiogenesis as well as a series of other physiological processes including inflammation or infection. However, the chemical properties of fucoidan which define the biological activity vary tremendously, making a prediction of the bioactivity or the corresponding therapeutic effect difficult. In this study, we compare the effect of four chemically characterized high molecular weight fucoidan extracts from Fucus distichus subsp. evanescens (FE_crude and fractions F1, F2, F3) on angiogenic and osteogenic processes in bone-related primary mono- and co-culture cell systems. By determining the gene expression and protein levels of the regulatory molecules vascular endothelial growth factor (VEGF), angiopoietin-1 (ANG-1), ANG-2 and stromal-derived factor 1 (SDF-1), we show that the extracted fucoidans negatively influence angiogenic and osteogenic processes in both the mono- and co-culture systems. We demonstrate that purer fucoidan extracts with a high fucose and sulfate content show stronger effects on these processes. Immunocytochemistry of the co-culture system revealed that treatment with FE_F3, containing the highest fucose and sulfate content, impaired the formation of angiogenic tube-like structures, indicating the anti-angiogenic properties of the tested fucoidans. This study highlights how chemical properties of fucoidan influence its bioactivity in a bone-related context and discusses how the observed phenotypes can be explained on a molecular level—knowledge that is indispensable for future therapies based on fucoidans.
We report the experience of the FucoSan InterReg project that had the ambition to generate commercialization opportunities for biotechnology research in a marine environment. Fucoidan, a promising biomarine polysaccharide extracted from seaweed, offers a broad array of potential applications; however, the supporting innovation value chain is still under development. We explore how the use of business modelling tools can contribute to building a shared understanding of commercialization opportunities across a diverse range of research and development actors. We analyze data (interviews, workshops, and surveys) from a German-Danish network of actors involved in the FucoSan InterReg project to identify how the tools contribute to setting up a base to support future activities across a potential innovation value chain. The results point towards the direct and indirect positive effects of engaging in the co-creation of a shared understanding of the functionality and possibilities of promising biomarine products. The findings support the idea that interdisciplinary and multilateral interactions help actors to identify the necessary connections and interdependencies to build a sustainability-driven innovation value chain.
The fucose-containing sulfated polysaccharides (syn. fucoidans) from brown seaweeds exhibit multiple biological activities. The aim of the present study was to evaluate the suitability of crude fucoidans from six different brown algal species for further research and development. For this, fucoidans were extracted from Fucus vesiculosus (F.v.), F. serratus (F.s.), F. evanescens (F.e.), Dictyosiphon foeniculaceus (D.f.), Laminaria digitata (L.d.), and Saccharina latissima (S.l.) and compared concerning their yields, their basic characteristics, degree of sulfation (DS), molecular mass, monosaccharide composition, their contents of polyphenols, laminarin and protein as well as their radical-scavenging capacity, three exemplary bioactivities and their fluorescence intensity (FI) increasing effect on the sulfated glycan sensor Polymer-H. The activities positively correlated with the DS, fucose content and the FI of Polymer H, whereas the antioxidant capacity correlated with the coextracted polyphenols. The best predictive power for the activities showed to have the combination of FI of Polymer-H and DS, which can thus be used for an initial screening of algae extracts. D.f. turned out to be unsuitable as fucoidan source, as the extract had only a low DS (0.14) and fucose content (38.7%) as well as the weakest bioactivities. The fucoidans from S.l. and F.e., which had the fucose content, highest DS, purity and yield, exhibited the best activities and thus proved to be the fucoidans most suitable for further investigations.
The bioactive sulfated polysaccharide from brown algae, fucoidan, can be used for a wide array of applications. As with other natural products, there are seasonal variabilities as well as variability within the investigated species, across regions, and from using different extraction procedures. In this study, the use of hot demineralized water and two variations of hot acidified water (10 mM sulfuric acid and 100 mM hydrochloric acid) as extraction solvents for microwave extraction of fucoidan from three different brown algae of the Fucus genus (F. vesiculosus, F. serratus, and F. evanescens) were investigated. The effect on yield of fucoidan from the different solvents at temperatures 80 °C, 100 °C, 120 °C was tested. The Fucus used in this study were harvested in the Baltic Sea in the Kiel Fjord, Germany, during Summer and Autumn of 2017. Air dried F. vesiculosus from Brittany in France was also analyzed and used for optimization of the extraction method and as a reference sample. The extraction procedure was adapted and modified from the method provided by Fletcher et al. (2017). The extracts were purified by performing dialysis. The results showed that fucoidan yield is maximized by extracting with 10 mM sulfuric acid for all species investigated. A large seasonal variance between species was observed, and large differences in yield were also dependent on species. These results suggest that to maximize fucoidan yield, one should tailor the extraction method to the specific algae species used, however, microwave assisted extraction (MAE) with 10 mM sulfuric acid proves a good general extraction method.
The marine origin polysaccharide fucoidan combines multiple biological activities. As demonstrated by various studies in vitro and in vivo, fucoidans show anti-viral, anti-tumor, anti-oxidant, anti-inflammatory and anti-coagulant properties, although the detailed molecular action remains to be elucidated. The aim of the present study is to assess the impact of crude fucoidan extracts, on the formation of vascular structures in co-culture models relevant for bone vascularization during bone repair and for vascularization processes in osteosarcoma. The co-cultures consisted of bone marrow derived mesenchymal stem cells, respectively the osteosarcoma cell line MG63, and human blood derived outgrowth endothelial cells (OEC). The concentration dependent effects on the metabolic activity on endothelial cells and osteoblast cells were first assessed using monocultures of OEC, MSC and MG63 suggesting a concentration of 100 µg/mL as a suitable concentration for further experiments. In co-cultures fucoidan significantly reduced angiogenesis in MSC/OEC but also in MG63/OEC co-cultures suggesting a potential application of fucoidan to lower the vascularization in bone tumors such as osteosarcoma. This was associated with a decrease in VEGF (vascular endothelial growth factor) and SDF-1 (stromal derived factor-1) on the protein level, both related to the control of angiogenesis and furthermore discussed as crucial factors in osteosarcoma progression and metastasis. In terms of bone formation, fucoidan slightly lowered on the calcification process in MSC monocultures and MSC/OEC co-cultures. In summary, these data suggest the suitability of lower fucoidan doses to limit angiogenesis for instance in osteosarcoma.
Fucoidans represent an intriguing class of fucose-containing sulfated polysaccharides. The biological activities of these polysaccharides are related to their compositional and structural parameters, whereby their degree of sulfation, as well as molecular weight (MW) distribution and chain conformation play important roles. Modern NMR and mass spectrometry techniques allow elucidating details of the glycan structure, but not the structure of the whole molecules in their native state. Accordingly, the knowledge about the latter of the fucoidans is currently still limited. Contrary to traditional MW determination by SEC with column calibration, SEC with triple detection provides not only the absolute Mw, but can also give information on additional molecule characteristics. In the present study, we used this method to compare six fucoidans extracted from Fucus vesiculosus (FV), F. serratus (FS), F. evanescens (FE), Dictyosiphon foeniculaceus (DF), Laminaria digitata (LD), and Saccharina latissima (SL) concerning their molar mass (Mw, Mn, Mp, dispersity) and size (rms radius, Rh) characteristics and distribution as well as their chain conformation solution. The tested fucoidans displayed considerable structural diversity including large differences in their MW profiles and showed to be heterogeneously composed. Fuc-FV and Fuc-SL showed the broadest MW distributions, those from Fuc-FE and Fuc-DF the narrowest ones. Most of the fucoidan fractions (except for Fuc-DF) turned out to exist as expanded flexible chains in PBS solution. The conformation data suggest branched structures with partly long side chains. The knowledge obtained by this study is useful for further fractionation and structural characterization as well as the interpretation of the bioactivity differences between the various fucoidans.
Fucoidans extracted from brown algae exert manifold biological activities paving the way for the development of numerous applications including treatments outside tumor therapy such as age-related macular degeneration or tissue engineering. In this study, we investigated the antiproliferative effects of fucoidans extracted from six different algae (Fucus vesiculosus, F. serratus, F. distichus subsp. evanescens, Dictyosiphon foeniculaceus, Laminaria digitata, Saccharina latissima) as well as three reference compounds (Sigma fucoidan, heparin, enoxaparin) on tumor (HL-60, Raji, HeLa, OMM-1, A-375, HCT-116, Hep G2) and non-tumor (ARPE-19, HaCaT) cell lines. All fucoidans were extracted according to a standardized procedure and tested in a commercially available MTS assay. Cell viability was measured after 24 h incubation with test compounds (1-100 µg/mL). Apart from few exceptions, fucoidans and heparins did not impair cell viability. In contrast, fucoidans significantly increased cell viability of suspension cell lines, but not of adherent cells. Fucoidans slightly increased viability of tumor cells and had no impact on the viability of non-tumor cells. The cell viability of HeLa and ARPE-19 cells negatively correlated with protein content and total phenolic content (TPC) of fucoidans, respectively. In summary, none of the tested fucoidans turned out to be anti-proliferative, rendering them interesting for future studies and applications.
Fucoidan extracts may have beneficial effects in age-related macular degeneration(AMD). Over-the-counter fucoidan preparations are generally undefined, crude extracts. In thisstudy, we investigated the effect of a crude fucoidan extract from Fucus distichus subspeciesevanescens (Fe) on the retinal pigment epithelium (RPE). Fe extract was investigated for chemicalcomposition and molar mass. It was tested in primary RPE and RPE cell line ARPE19. Oxidativestress was induced with tert-butyl hydroperoxide, cell viability evaluated with MTT assay, VEGFsecretion assessed in ELISA. Phagocytosis was evaluated in a fluorescence microscopic assay.Wound healing ability was tested in a scratch assay. Additionally, the inhibition of elastase andcomplement system by Fe extract was studied. The Fe extract contained about 61.9% fucose andhigh amounts of uronic acids (26.2%). The sulfate content was not as high as expected (6.9%). It wasnot toxic and not protective against oxidative stress. However, Fe extract was able to reduce VEGFsecretion in ARPE19. Phagocytosis was also reduced. Concerning wound healing, a delay could beobserved in higher concentrations. While some beneficial effects could be found, it seems tointerfere with RPE function, which may reduce its beneficial effects in AMD treatment.
Background: Fucoidans are interesting for potential usage in ophthalmology, and especially age-related macular degeneration. However, fucoidans from different species may vary in their effects. Here, we compare fucoidans from five algal species in terms of oxidative stress protection and vascular endothelial growth factor (VEGF) interference in ocular cells. Methods: Brown algae (Fucus vesiculosus, Fucus distichus subsp. evanescens, Fucus serratus, Laminaria digitata, Saccharina latissima) were harvested and fucoidans isolated by hot-water extraction. Fucoidans were tested in several concentrations (1, 10, 50, and 100 µg/mL). Effects were measured on a uveal melanoma cell line (OMM-1) (oxidative stress), retinal pigment epithelium (RPE) cell line ARPE19 (oxidative stress and VEGF), and primary RPE cells (VEGF). Oxidative stress was induced by H2O2 or tert-Butyl hydroperoxide (TBHP). Cell viability was investigated with methyl thiazolyl tetrazolium (MTT or MTS) assay, and VEGF secretion with ELISA. Affinity to VEGF was determined by a competitive binding assay. Results: All fucoidans protected OMM-1 from oxidative stress. However, in ARPE19, only fucoidan from Saccharina latissima was protective. The affinity to VEGF of all fucoidans was stronger than that of heparin, and all reduced VEGF secretion in ARPE19. In primary RPE, only the fucoidan from Saccharina latissima was effective. Conclusion: Among the fucoidans from five different species, Saccharina latissima displayed the most promising results concerning oxidative stress protection and reduction of VEGF secretion.