A fucoidan fraction purified from <i>Chnoospora minima</i>; a potential inhibitor of LPS-induced inflammatory responses

Fernando, I.P. Shanura,Sanjeewa, K.K. Asanka,Samarakoon, Kalpa W.,Lee, Won Woo,Kim, Hyun-Soo,Kang, Nalae,Ranasinghe, P.,Lee, Hyi-Seung,Jeon, You-Jin Elsevier 2017 International journal of biological macromolecules Vol.104 No.1

<P><B>Abstract</B></P> <P>Fucoidans are an interesting group of bioactive sulfated polysaccharides abundant in brown seaweeds. The current study highlights the enrichment and extraction of fucoidan from <I>Chnoospora minima</I> by means of enzyme-assistant extraction using Celluclast and evaluation of its anti-inflammatory potential through <I>in vitro</I> and <I>in vivo</I> studies. The purified <I>C. minima</I> fucoidan (F2,4) inhibited the nitrous oxide (NO) production (IC<SUB>50</SUB> =27.82±0.88μg/ml) and expression of PGE<SUB>2</SUB> through the subsequent downregulation of iNOS and COX-2 expression in lipopolysaccharide (LPS) stimulated RAW 264.7 macrophages. F2,4 downregulated TNF-α, IL1-β, and IL-6 in RAW 264.7 macrophages in a dose-dependent manner and suppressed NO and ROS production in LPS stimulated zebrafish embryos while exerting a protective effect against the cell damage caused by LPS. Polysaccharide structural characterization was performed using FTIR, HPAE-PAD analysis of the monosaccharide content and NMR spectroscopy. Current findings confirm the potential anti-inflammatory activity of fucoidan purified from <I>C. minima</I> and elaborate its potential application as a functional ingredient in consumer products.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Fucoidan fraction was purified from <I>Chnoospora minima</I> Celluclast assisted extract. </LI> <LI> Polysaccharides were characterized by FTIR and monosaccharide content analysis. </LI> <LI> Fucoidan fraction inhibited the LPS-induced inflammatory responses in vitro and in vivo. </LI> <LI> INOS, COX-2, PGE<SUB>2</SUB>, TNF-α, IL1-β, and IL-6 were involved in the regulative mechanisms. </LI> <LI> <I>C. minima</I> fucoidan is a promising ingredient for anti-inflammatory applications. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Beijing urban particulate matter-induced injury and inflammation in human lung epithelial cells and the protective effects of fucosterol from <i>Sargassum binderi</i> (Sonder ex J. Agardh)

Fernando, I.P. Shanura,Jayawardena, Thilina U.,Kim, Hyun-Soo,Lee, Won Woo,Vaas, A.P.J.P.,De Silva, H.I.C.,Abayaweera, G.S.,Nanayakkara, C.M.,Abeytunga, D.T.U.,Lee, Dae-Sung,Jeon, You-Jin Academic Press 2019 Environmental research Vol.172 No.-

<P><B>Abstract</B></P> <P>Particulate matter (PM) air pollution has gradually become a widespread problem in East Asia. PM may cause unfamiliar inflammatory responses, oxidative stress, and pulmonary tissue damage, and a comprehensive understanding of the underlying mechanisms is required in order to develop effective anti-inflammatory agents. In this study, fine dust collected from Beijing, China (CPM) (size < PM13 with majority < PM2.5) was evaluated for its oxidative stress- and inflammation-inducing effects, which cause cell damage, in A459 human lung epithelial cells. Oxidative stress was marked by an increase in intracellular ROS levels and the production of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), and heme oxygenase-1 (HO-1). Upon induction of oxidative stress, a marked increase was observed in the expression of key inflammatory mediators such as COX-2 and PGE<SUB>2</SUB> and the pro-inflammatory cytokines TNF-α and IL-6 via NF-kB and MAPK pathways. Cellular damage was marked by a reduction in viability, increased lactate dehydrogenase (LDH) release, formation of apoptotic and necrotic bodies, accumulation of sub-G1 phase cells, and DNA damage. Apoptosis was found to be mediated via the activation of caspases through the mitochondria-mediated pathway. Fucosterol, purified from the brown alga <I>Sargassum binderi</I> (Sonder ex J. Agardh) by bio-assay-guided fractionation and purification, exhibited potential therapeutic effects against CPM-induced detrimental effects. Further studies could focus on developing fucosterol, in forms such as steroidal inhalers, against PM-induced pulmonary tissue inflammation.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Fine dust air pollution is a major reason of pulmonary complications in East Asia. </LI> <LI> Dust particles induce oxidative stress and inflammation damaging the lung epithelial cells. </LI> <LI> Fucosterol suppressed the dust induced cell damage by inhibiting oxidative stress and inflammation. </LI> <LI> Fucosterol may have beneficial effects in alleviating adverse respiratory effects of air pollution. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Apoptotic and antiproliferative effects of Stigmast-5-en-3-ol from <i>Dendronephthya gigantea</i> on human leukemia HL-60 and human breast cancer MCF-7 cells

Fernando, I.P. Shanura,Sanjeewa, K.K. Asanka,Ann, Yong-Seok,Ko, Chang-ik,Lee, Seung-Hong,Lee, Won Woo,Jeon, You-Jin Elsevier 2018 Toxicology in vitro Vol.52 No.-

<P><B>Abstract</B></P> <P>The genus <I>Dendronephthya</I> encompasses marine soft corals that produce a wide spectrum of biofunctional terpenoids. Anticancer properties of these metabolites are widely exploited as potential chemotherapeutic agents. The present study reports the purification and isolation of a potential antiproliferative constituent, stigmast-5-en-3-ol from the 70% ethanol extract of the soft coral <I>Dendronephthya gigantea</I>. Among several other 3β-hydroxy-Δ5-steroidal congeners, stigmast-5-en-3-ol indicated prominent antiproliferative effects on HL-60 (leukemia) and MCF-7 (breast cancer) cell lines with IC<SUB>50</SUB> values of 37.82 and 45.17 μg/ml respectively. Stigmast-5-en-3-ol increased apoptotic body formation, accumulation of sub G1 apoptotic cells, and DNA damage in HL-60 and MCF-7 cells. It increased the expression of Bax, caspases, and PARP cleavage while decreasing Bcl-xL levels in both cancer cell lines indicating that the effects are arbitrated via the mitochondria-mediated apoptotic pathway. Steroidal derivatives were identified by GC MS/MS and the identity of stigmast-5-en-3-ol was confirmed by NMR spectra. The present study suggests that stigmast-5-en-3-ol could be a promising candidate for anticancer drug research.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Stigmast-5-en-3-ol was isolated from a sterol rich fraction of <I>Dendronephthya gigantea</I>. </LI> <LI> Stigmast-5-en-3-ol decreased HL-60 and MCF-7 cancer cell proliferation. </LI> <LI> Stigmast-5-en-3-ol induced apoptosis via mitochondria-mediated pathway. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Identification of sterols from the soft coral <i>Dendronephthya gigantea</i> and their anti-inflammatory potential

Fernando, I.P.Shanura,Sanjeewa, K.K.Asanka,Kim, Hyun-Soo,Kim, Seo-Young,Lee, Seung-Hong,Lee, Won Woo,Jeon, You-Jin Elsevier 2017 Environmental toxicology and pharmacology Vol.55 No.-

<P><B>Abstract</B></P> <P>Exploration of anti-inflammatory phytochemicals has received tremendous attention worldwide owing to the rapid increase in inflammatory diseases. Current study reveals the identification of eight 3β-hydroxy-Δ<SUP>5</SUP>-steroidal congeners from a nonpolar column fraction of the ethanol solubles from the soft coral <I>Dendronephthya gigantea</I> collected from Jeju Island South Korea, using GC–MS/MS analysis. The sterol-rich fraction (DGEH21) showed a significant anti-inflammatory activity as exhibited by the inhibition of NO production (IC<SUB>50</SUB> 4.33±0.50μg/mL) and PGE<SUB>2</SUB> production in LPS-stimulated RAW 264.7 macrophages. It also suppressed the expression levels of proinflammatory cytokines, TNF-α, IL-1β, and IL-6 in a dose-dependent manner. Furthermore, DGEH21 effectively downregulated the expression levels of iNOS, and COX-2 and reduced NO and ROS production as well as cell death in LPS-stimulated <I>in-vivo</I> zebrafish embryo model. However, DGEH21 at relatively high concentrations indicated cytotoxicity in both RAW cells and zebrafish embryos with RAW cell viability being nearly 80% after treatment with 25μg/mL DGEH21. This study highlights the synergistic anti-inflammatory activity of several steroids found in <I>D. gigantea</I>. Their actions may be useful in the development of anti-inflammatory cosmeceuticals, pharmaceutical agents, and other consumer products.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Eight 3β-hydroxy-Δ5-steroidal congeners were identified from the <I>Dendronephthya gigantea</I>. </LI> <LI> Synergistically the sterols indicated a profound anti-inflammatory potential in LPS-induced RAW cells. </LI> <LI> Sterols downregulated inflammatory mediators in LPS-induced RAW cells. </LI> <LI> Indicated anti-inflammatory effects in in vivo LPS-induced zebra fish model. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Reduction of heavy metal (Pb²⁺) biosorption in zebrafish model using alginic acid purified from <i>Ecklonia cava</i> and two of its synthetic derivatives

Fernando, I.P. Shanura,Sanjeewa, K.K. Asanka,Kim, Seo-Young,Lee, Jung-Suck,Jeon, You-Jin Elsevier 2018 International journal of biological macromolecules Vol.106 No.-

<P><B>Abstract</B></P> <P>Heavy metal contamination has become a major problem that causes severe environmental and health issues due to their biosorption, bioaccumulation, and toxicity. This study was designed to evaluate heavy metal chelating abilities of alginic acid (AA) extracted from the brown seaweed <I>Ecklonia cava</I> and two of its derivatives prepared by the partial oxidation of the 2° OH groups (OAA) and partial carboxylation of the monomeric units (CAA) upon reducing the heavy metal biosorption in zebrafish (<I>Danio rerio</I>) modal. Metal ions were quantified using ICP-OES and biopolymers were characterized by FTIR and XRD analysis. All investigated biopolymers indicated potential ability for chelating Pb<SUP>2+</SUP>, Cu<SUP>2+</SUP>, Cd<SUP>2+</SUP>, As<SUP>3+</SUP>, and Ag<SUP>+</SUP>. The sorption capacities were in the order of CAA>OAA>AA. All biopolymers indicated a comparatively higher chelation towards Pb<SUP>2+</SUP>. AA, OAA, and CAA could effectively reduce Pb<SUP>2+</SUP> induced toxicity and Pb<SUP>2+</SUP> stress-induced ROS production in zebrafish embryos. Besides, they could reduce the biosorption of Pb<SUP>2+</SUP> in adult zebrafish which could lead to bioaccumulation. Since alginic acid purified from <I>E. cava</I> and its derivatives could be utilized as seaweed derived biopolymers to purify heavy metals contaminated water and as a dietary supplement to reduce heavy metal biosorption in organisms.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Heavy metal contamination of food and water has become a major issue in certain regions. </LI> <LI> We purified alginic acid from <I>Ecklonia cava</I> and performed chemical modifications. </LI> <LI> Partial oxidation and carboxylation increased heavy metal chelation ability of alginic acid. </LI> <LI> Highest sorption was towards Pb<SUP>2+</SUP> followed by Cu<SUP>2+</SUP>, Ag<SUP>+</SUP>, Cd<SUP>2+</SUP>, and As<SUP>3+</SUP>. </LI> <LI> These derivatives could reduce Pb<SUP>2+</SUP> uptake from zebrafish alimentary canal. </LI> </UL> </P>

Anti-inflammatory potential of alginic acid from <i>Sargassum horneri</i> against urban aerosol-induced inflammatory responses in keratinocytes and macrophages

Fernando, I.P. Shanura,Jayawardena, Thilina U.,Sanjeewa, K.K. Asanka,Wang, Lei,Jeon, You-Jin,Lee, Won Woo Elsevier 2018 Ecotoxicology and environmental safety Vol.160 No.-

<P><B>Abstract</B></P> <P>The airborne particulate pollutants originating in the deserts of Mongolia and China which becomes contaminated with industrial effluents and traffic emissions while moving with the wind currents towards East Asia has recently become a serious environmental and health issue in the region. They cause asthma, collateral lung tissue damage, oxidative stress, allergic reactions, and inflammation. The current study was undertaken to evaluate the protective effects of alginate extracted from the invasive alga <I>Sargassum horneri</I> (SHA) against fine dust collected from Beijing, China (Chinese fine dust; CFD). It was found that CFD induces inflammation in HaCaT keratinocytes and inhibits macrophage activation. All of the particulate matter (PM) comprising CFD was < PM13 majority being < PM2.5 which is defined for mineral elements and polycyclic aromatic hydrocarbons. SHA attenuated PGE<SUB>2</SUB> levels in CFD-induced HaCaT keratinocytes. The IC<SUB>50</SUB> for SHA was 36.63 ± 4.11 µg mL<SUP>-l</SUP>. SHA also reduced the levels of COX-2, IL-6, and TNF-α, and inhibited certain key molecular mediators of the NF-κB and MAPK pathways in keratinocytes. SHA substantially reduced the levels of CFD-derived metal ions like Pb<SUP>2+</SUP> and Ca<SUP>2+</SUP> in keratinocytes attributable to its metal ion chelating properties. CFD-induced HaCaT keratinocyte culture media increased inflammatory responses in RAW 264.7 macrophages. These cells presented with increased levels of NO, iNOS, COX-2, PGE<SUB>2</SUB>, and pro-inflammatory cytokines. It was found that the aforementioned effects could be reversed in RAW 264.7 macrophages when keratinocytes were treated with SHA. Therefore, SHA could be used against fine dust-induced inflammation in keratinocytes.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Beijing, China urban fine dust increase keratinocytes and macrophages inflammation. </LI> <LI> Fine dust contains metal ions and polycyclic aromatic hydrocarbons. </LI> <LI> Alginate from <I>Sargassum horneri</I> ameliorates the inflammatory effects. </LI> <LI> Alginate chelate metal ions and reduce inflammatory mediators and cytokines. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Isolation and purification of fucoidan fraction in <i>Turbinaria ornata</i> from the Maldives; Inflammation inhibitory potential under LPS stimulated conditions in in-vitro and in-vivo models

Jayawardena, Thilina U.,Fernando, I.P. Shanura,Lee, Won Woo,Sanjeewa, K.K. Asanka,Kim, Hyun-Soo,Lee, Dae-Sung,Jeon, You-Jin Elsevier 2019 International journal of biological macromolecules Vol.131 No.-

<P><B>Abstract</B></P> <P>Fucoidan, referred to as fucose containing sulfated polysaccharides (FCSP), is a polymer from brown algae cell wall that is reported to exhibit potential anti-inflammatory activity. In the present study, the fucoidans are extracted from <I>Turbinaria ornata</I> (<I>TO</I>) from the Maldives. The method involves enzyme assisted extraction and is modified in order to improve the effectiveness and purity of final product. Purified fucoidan fraction was identified as F10, and its chemical properties were verified via FTIR, <SUP>1</SUP>H NMR and monosaccharide analysis. Selected inflammatory mediators were studied to evaluate the anti-inflammatory potential using RAW 264.7 macrophages. F10 successfully inhibited NO production (IC<SUB>50</SUB> = 30.83 ± 1.02 μg mL<SUP>−1</SUP>). F10 dose-dependently down-regulated iNOS, COX-2, and pro-inflammatory cytokines including PGE<SUB>2</SUB> levels. The in vivo experiments were assisted by zebrafish embryo model. This exhibited reduction in ROS, NO expression levels. To our knowledge, this is the first report to illustrate potential anti-inflammatory activity of FCSPs' extracted from the brown algae <I>T. ornata</I>. Concisely, the results suggest that fucoidan purified from <I>T. ornata</I> increases the macrophage cellular and zebrafish embryo resistance against LPS-induced inflammation. Based on the observations, the fucoidans are promising candidates to be used in the pharmaceutical and cosmeceutical sectors.</P>

FTIR characterization and antioxidant activity of water soluble crude polysaccharides of Sri Lankan marine algae

I.P. Shanura Fernando,K. K. Asanka Sanjeewa,Kalpa W. Samarakoon,이원우,김현수,김은아,U. K. D. S. S. Gunasekara,D. T. U. Abeytunga,Chandrika Nanayakkara,E. D. de Silva,이희승,전유진 한국조류학회I 2017 ALGAE Vol.32 No.1

Polysaccharides of marine algae exhibit different structural characteristics and interesting biological functions. In this study, crude polysaccharides (CP) of eleven Sri Lankan marine algae obtained through hot water extraction and ethanol precipitation were investigated for DPPH, alkyl, and hydroxyl radical scavenging activities using electron spin resonance spectrometry and for intracellular reactive oxygen species scavenging activity in the Chang liver cell line. Characterization of CPs was done by Fourier transform infrared (FTIR) spectroscopy and by analysis of the monosaccharide composition. Time-dependent density functional theory quantum-chemical calculations at the RB3LYP/6-31G(d,p) level for constructed dimeric units of the corresponding polysaccharides were used to resolve the FTIR spectra. CPs from Chnoospora minima showed the highest DPPH and alkyl radical scavenging activities and higher intracellular reactive oxygen species scavenging effects for both AAPH and H2O2 induced ROS production in “Chang” cells. The major polysaccharide constituent in C. minima CP was identified as fucoidan and it displayed a higher sulfate content. The degree of sulfation of these polysaccharides suggests a positive correlation with the observed antioxidant properties.

A keratinocyte and integrated fibroblast culture model for studying particulate matter-induced skin lesions and therapeutic intervention of fucosterol

Fernando, I.P. Shanura,Jayawardena, Thilina U.,Kim, Hyun-Soo,Vaas, A.P.J.P.,De Silva, H.I.C.,Nanayakkara, C.M.,Abeytunga, D.T.U.,Lee, WonWoo,Ahn, Ginnae,Lee, Dae-Sung,Yeo, In-Kyu,Jeon, You-Jin Elsevier 2019 Life sciences Vol.233 No.-

<P><B>Abstract</B></P> <P>Increased levels of particulate matter (PM) air pollutants in East Asia have resulted in detrimental health impacts increasing morbidity and mortality. Epidemiological studies suggest a possible relation between the cutaneous exposure of PM and increased oxidative stress and inflammation which lead to skin lesions. The present study utilizes an integrated cell culture model of keratinocytes and fibroblasts to mimic viable skin layers and investigate the possible effects of PM exposure after penetration through corneocytes. The skin perfection is upheld by homeostatic functionality of epidermal cells and the integrity of connective tissues. Exposure to xenobiotics could alter the skin cell homeostasis aggravating premature skin aging. Stimulation of HaCaT keratinocytes by PM collected from Beijing, China (CPM) increased the intracellular ROS levels triggering a cascade of events aggravating inflammatory responses and connective tissue degradation. In HDF fibroblasts, treatment with preconditioned keratinocyte culture media augmented inflammatory responses, cellular differentiation, and connective tissue degradation. Above events were marked by the increased intracellular ROS, inflammatory mediators, pro-inflammatory cytokines, matrix metalloproteinases (MMP)-1 and -2 levels, collagenase, and elastase activity. Fucosterol treatment of keratinocytes dose-dependently attenuated the detrimental effects both in keratinocytes and fibroblasts restoring the conditions near to physiological levels. Further evaluations could be advanced on developing fucosterol, in forms such as rejuvenating cosmeceuticals which could attenuate detrimental responses of CPM exposure.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Inhibition of inflammatory responses elicited by urban fine dust particles in keratinocytes and macrophages by diphlorethohydroxycarmalol isolated from a brown alga Ishige okamurae

I. P. Shanura Fernando,김현수,K. K. Asanka Sanjeewa,오재영,전유진,이원우 한국조류학회I 2017 ALGAE Vol.32 No.3

Fine dust (FD) particles have become a major contributor to air pollution causing detrimental effects on the respiratory system and skin. Although some studies have investigated the effects of FD on the respiratory system, their possible effects on the skin remain under-explored. We investigated the FD mediated inflammatory responses in keratinocytes, present in the outer layers of skin tissues and the transfer of inflammatory potential to macrophages. We further evaluated the anti-inflammatory effects of the polyphenolic derivative, diphlorethohydroxycarmalol (DPHC) isolated from Ishige okamurae against FD-induced inflammation. Size distribution of FD particles was analyzed by scanning electron microscopy. FD particles induced the production of cyclooxygenase-2, prostaglandin E2 (PGE2), interleukin (IL)-1β, and IL-6 in HaCaT keratinocytes and the expression of nitric oxide (NO), inducible nitric oxide synthases (iNOS), PGE2, tumor necrosis factor-α expression in RAW 264.7 macrophages. Further, we evaluated the inflammatory potential of the culture medium of inflammation-induced HaCaT cells in RAW 264.7 macrophages and observed a marked increase in the expression of NO, iNOS, PGE2, and proinflammatory cytokines. DPHC treatment markedly attenuated the inflammatory responses, indicating its effectiveness in suppressing a broad range of inflammatory responses. It also showed anti-inflammatory potential in in-vivo experiments using FD-stimulated zebrafish embryos by decreasing NO and reactive oxygen species production, while eventing cell death caused by inflammation.