The effect of Schizophyllan on the differentiation of osteoclasts and osteoblasts.

Schizophyllan (SPG), a ß-glucan from Schizophyllum commune, is recognized for its antioxidant, immunoregulatory, and anticancer activities. In this study, its effects on bone cells, particularly osteoclasts and osteoblasts, were examined. We demonstrated that SPG dose-dependently inhibited osteoclastogenesis and reduced gene expression associated with osteoclast differentiation. SPG also decreased bone resorption and F-actin ring formation. This inhibition could have been due to the downregulation of transcription factors c-Fos and nuclear factor of activated T cells 1 (NFATc1) via the MAPKs (JNK and p38), IκBα, and PGC1ß/PPARγ pathways. In coculture, SPG lowered osteoclastogenic activity in calvaria-derived osteoblasts by reducing macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor-κB ligand (RANKL) expression. In addition, SPG slightly enhanced osteoblast differentiation, as evidenced by increased differentiation marker gene expression and alizarin red staining. It also exhibited antiresorptive effects in a lipopolysaccharide-induced calvarial bone loss model. These results indicated a dual role of SPG in bone cell regulation by suppressing osteoclastogenesis and promoting osteoblast differentiation. Thus, SPG could be a therapeutic agent for bone resorption-related diseases such as osteoporosis, rheumatoid arthritis, and periodontitis.

Schizophyllan from Schizophyllum commune BRM 060008: Potential application as an inhibitor of lipase.

ß-D-glucan has significant implications in regulating lipid metabolism and preventing diseases associated with lipid accumulation. Schizophyllan (SPG) from Schizophyllum commune fungus is a commercially important ß-glucan with applications in the health food industry, pharmacy, and cosmetics. However, SPG was obtained by submerged culture of the wood-rotting and filamentous fungus S. commune BRM 060008, which may have been isolated from the Cerrado Biome of Brazil. In this study, to confirm that the polysaccharide produced by BRM 060008 strain fermentation was indeed (1→3)(1→6)-ß-D-glucan, it was purified and characterized using Fourier transform infrared spectroscopy, thermogravimetric analysis, high-performance size exclusion chromatography, nuclear magnetic resonance, and methylation analysis. The polysaccharide produced was identified as the ß-D-glucan expected with a high molecular weight (1.093 × 106 g/mol) and the thermogravimetric analysis indicated a maximum degradation temperature of ~324 °C and a 60 % residual weight, lower than commercial SPG. The molecular structure and thermal properties of the ß-D-glucan were similar to the commercial sample. Additionally, the in vitro pancreatic lipase inhibitory activity was evaluated, investigating anti-obesity and anti-lipidemic properties. The results showed unprecedented lipase inhibition activity to SPG prepared using the S. commune strain BRM 060008, making it promising for food and pharmaceutical applications.

Schizophyllan promotes osteogenic differentiation of human adipose tissue-derived mesenchymal stem cells in vitro.

BACKGROUND: Bioactive polysaccharides are a promising way for bone disease prevention with high efficiency. Schizophyllan (SPG) is a polysaccharide derived from a species of fungus with anticancer, antitumor, and anti-inflammatory effects. In the present study, for the first time, the cell proliferation, osteogenic markers, mineral deposition, and osteogenic gene expression of human adipose tissue-derived mesenchymal stem cells (hADMSCs) grown on SPG were evaluated by in vitro assays. METHODS AND RESULTS: The cytotoxicity of SPG was measured using the MTT assay and acridine orange staining. Differentiation of hADMSCs was assessed using alkaline phosphatase (ALP) activity test, cellular calcium content assay, and mineralized matrix staining. To this end, Alizarin red S, von Kossa staining, and the expression of bone-specific markers, including ALP, Runx2, and osteonectin, were used by real-time RT-PCR over a 2-week period. According to the results, SPG at 10 µg/ml concentration was determined as the optimal dosage for differentiation studies. The results of osteogenic differentiation tests showed that compared to the control groups in vitro, SPG enhanced the osteogenic markers and mineralization as well as upregulation of the expression of bone specific genes in differentiated hADMSCs during differentiation. CONCLUSIONS: The results revealed that SPG could be applied as effective factor for osteogenic differentiation in the future. The current study provides insights into the hADMSC-based treatment and introduces promising therapeutic material for individuals who suffer from bone defects and injuries.

Preparation and characterization of carboxymethylated Aureobasidium pullulans ß-(1 → 3, 1 → 6)-glucan and its in vitro antioxidant activity.

Aureobasidium pullulans ß-(1 â†’ 3, 1 â†’ 6)-glucan (APG) has a high degree of ß-(1 â†’ 6)-glucosyl branching and a regular triple helical structure similar to that of schizophyllan. In this study, APG was carboxymethylated to different degrees of substitution (DS = 0.51, 1.0, and 2.0, denoted CMAPG 1-3, respectively) using a heterogeneous reaction. With increasing DS, the triple-helix structure drastically decreased and converted to a random coil structure in CMAPG 3. Further, aqueous solutions of CMAPG changed from pseudoplastic fluids to perfect Newtonian liquids with increasing DS, indicating that the intra- and intermolecular hydrogen bonds had been cleaved by the substituents to form a random coil structure. In addition, APG and CMAPG solutions exhibited scavenging ability against hydroxyl, organic, and sulfate radicals. It was also found that the carboxymethylation of APG drastically enhanced the organic radical scavenging ability. On the basis of the relationship between the DS and radical scavenging ability of the CMAPG samples, we believe hydroxyl and organic radicals were preferably scavenged by the donation of hydrogen atoms from the glucose rings and the methylene moieties of the carboxymethyl groups, respectively. Considering the obtained results, CMAPG and APG are expected to have applications in pharmaceuticals, functional foods, and cosmetics as antioxidant polysaccharides.

Unlocking the benefits of split gill mushroom: Chemical analysis and prebiotic properties of schizophyllan extract.

The edible split gill mushroom is considered both a nutritive and therapeutic superfood, as well as rich in schizophyllan and protein. Prebiotic properties and other biological effects distinguish the schizophyllan (ß-glucan). Thus, this research investigates the identity of the mushroom strain, the nutritional composition of this mushroom, and the schizophyllan extract for further analysis, including its prebiotic activity and so on. The experimental results revealed that this mushroom was identified as Schizophyllum commune, comprising more excellent carbohydrates, protein, crude fiber, lower fat, and no heavy metal detection. Moreover, this extract consisted of pharmaceutical hydrophobin (14.0-18.5 kDa), lectin protein (21-35 kDa), bioactive purpurin or red pigment, including the prebiotic ß-glucan stimulating the proliferation of probiotic bacteria isolated from yogurt. Therefore, both S. commune and the schizophyllan extract can be used as a prebiotic candidate, functional food, and nutraceutical product.

Structural analysis of polysaccharide/antisense DNA complexes during cytoplasmic target mRNA hybridization.

We previously demonstrated antisense oligonucleotides (AS-ODNs) delivery system based on the complex formed with poly (dA) and schizophyllan, a type of ß-1,3-glucan. This complex enables efficient intracellular delivery of AS-ODNs. In this communication, we investigated the cytoplasmic translocation of the complex itself and its mechanism of action on mRNA. As a result, we found that the complex moved into the cytoplasm while keeping its structure, and AS-ODN hybridized with the target mRNA. This result encourages pharmaceutical applications of the complex.

Efficient production and skincare activity evaluation of schizophyllan, a ß-glucan derived from Schizophyllum commune NTU-1.

Schizophyllan (SPG), a ß-glucan produced by the fungus Schizophyllum commune, possesses a ß-(1 â†’ 3)-linked backbone with single ß-(1 â†’ 6)-linked glucose side chains at approximately every third residue. In this study, we screened SPG-producing strains of S. commune from different provinces in China. A candidate strain (NTU-1) with a high SPG yield was chosen, and the fermentation conditions were optimized. The optimal carbon and nitrogen sources were sucrose (40 g/L) and yeast extract (20 g/L), respectively. The optimal conditions for pH and temperature were 5.0 and 28 °C, respectively. Inclusion of 0.2 mg/L of 2,4-Dichlorophenoxyacetic acid in the medium further increased the SPG concentration. In a 5-L bioreactor, the fermentation cycle was reduced from the initial seven days to five days, and the concentration of SPG obtained was 21.3 g/L, which is the highest reported to date. In addition, we evaluated the bioactivity of the SPG prepared using strain NTU-1. The results showed that SPG had certain characteristics of anti-oxidation, anti-photoaging, and inhibition of melanin production, making it a promising reagent for skin care.

Production and Anti-Inflammatory Performance of PVA Hydrogels Loaded with Curcumin Encapsulated in Octenyl Succinic Anhydride Modified Schizophyllan as Wound Dressings.

Amphiphilic polysaccharides can be used as wall materials and applied to encapsulate hydrophobic active chemicals; moreover, there is significant demand for novel medical high-molecular-weight materials with various functions. In order to prepare amphiphilic schizophyllan (SPG), octenyl succinic anhydride (OSA) was chosen to synthesize OSA-modified schizophyllan (OSSPG) using an esterified reaction. The modification of OSSPG was demonstrated through FT-IR and thermal analysis. Moreover, it was found that OSSPG has a better capacity for loading curcumin, and the loading amount was 20 µg/mg, which was 2.6 times higher than that of SPG. In addition, a hydrogel made up of PVA, borax, and C-OSSPG (OSSPG loaded with curcumin) was prepared by means of the one-pot method, based on the biological effects of curcumin and the immune-activating properties of SPG. The mechanical properties and biological activity of the hydrogel were investigated. The experimental results show that the dynamic cross-linking of PVA and borax provided the C-OSSPG/BP hydrogel dressing with exceptional self-healing properties, and it was discovered that the C-OSSPG content increased the hydrogel's swelling and moisturizing properties. In fibroblast cell tests, the cells treated with hydrogel had survival rates of 80% or above. Furthermore, a hydrogel containing C-OSSPG could effectively promote cell migration. Due to the excellent anti-inflammatory properties of curcumin, the hydrogel also significantly reduces the generation of inflammatory factors, such as TNF-α and IL-6, and thus has a potential application as a wound dressing medicinal material.

Enhanced Therapeutic Efficacy of Immunostimulatory CpG-ODN by Silencing SOCS-1 with Polysaccharide/miR-155 Complexes.

For the induction of antigen-specific immune responses, adjuvants as well as antigens are essential. CpG-ODN is a potent agonist of toll-like receptor 9 (TLR9) and is known as an adjuvant to induce cellular immune responses. We previously developed a therapeutic oligonucleotide delivery system based on the formation of a complex between schizophyllan (SPG), a kind of ß-1,3-glucan, and poly(dA), which actively delivered CpG-ODN to antigen-presenting cells (APCs) in the draining lymph nodes and induced antigen-specific immune responses. However, unfortunately, the signaling pathway of TLR9 is negatively regulated by an intracellular protein called suppressor of cytokine signaling-1 (SOCS-1), which suppresses the adjuvant effect of CpG-ODN. To solve this, we focused on microRNA-155 (miR-155), which regulates innate and autoimmune processes by targeting SOCS-1. In this study, we proposed a strategy of combining miR-155 and CpG-ODN, each complexed with SPG (denoted as SPG/miR-155 and SPG/CpG, respectively), to induce a more potent immune response. As a result, we showed that the efficient delivery of miR-155 to APCs by a complex form could induce much more potent cellular immune responses than SPG/CpG alone. Furthermore, the mice treated with the combination of SPG/miR-155 and SPG/CpG showed a long delay in tumor growth occurrence and improved survival after tumor inoculation. These results indicate the possibility of therapeutic strategies for cancer.

Co-production of schizophyllan and cellulolytic enzymes from bagasse by Schizophyllum commune.

Schizophyllum commune is a mushroom-forming fungus well-known for its ability to degrade lignocellulosic materials and production of schizophyllan, a high added-value product for cosmeceutical, pharmaceutical, and biomaterial industries. Conventionally, schizophyllan is produced by submerged fermentation using glucose as a carbon source. In this work, we demonstrate that alkaline pretreated bagasse can be used by Schizophyllum commune as an alternative carbon source for the production of schizophyllan. The influence of different factors was investigated including cultivation time, biomass loading, and culturing media component and a co-product correlation model was proposed. In this lab-scale study, a yield of 4.4 g/L of schizophyllan containing 89% glucose was achieved. In addition to schizophyllan, the cellulolytic enzymes co-produced during this process were isolated and characterized and could find applications in a range of industrial processes. This demonstrates the potential of using agricultural waste as a cheaper alternative feedstock for this biorefinery process.

Association with Imidazole in the Cooperative Order-Disorder Transition in Aqueous Solution of Schizophyllan.

Schizophyllan, a triple helical polysaccharide, exhibits cooperative order-disorder transition (CODT) in aqueous solutions. The transition transforms the ordered structure (triple helix I) formed between the branched side chains and solvent molecules into the disordered structure (triple helix II) without dissociation of the triple helix. The CODT behaviors in H2O-imidazole mixtures containing HCl with different molar ratios of imidazole/HCl were investigated by adiabatic calorimetry and differential scanning calorimetry on two schizophyllan solutions with different molar masses. The transition temperature (Tr) and the transition enthalpy (ΔHr) significantly depended on both of the mole fractions of imidazole and imidazole/HCl. The composition dependences of Tr and ΔHr in H2O-imidazole mixtures were analyzed with linear cooperative transition theory for the solvent-stabilizing effect in the mixture with active compounds. Theoretical analyses confirmed that both imidazole and imidazolium ions in the solutions competitively interact with the side chain of the triple helix.

Increasing-Aeration Strategy: a Practical Approach to Enhance the Schizophyllan Production and Improve the Operational Conditions of Schizophyllum commune Cultivation in the Stirred Tank and Bubble Column Bioreactors.

In the present study, the effect of employing the increasing- aeration strategy (IAS) in the oxygen-limited situation and proportionate to increasing oxygen demand of the fungus Schizophyllum commune (S. commune) has been investigated in both stirred tank (STB) and bubble column (BCB) bioreactors. The purpose was to enhance schizophyllan (SPG) production by preventing oxygen starvation, improve mixing conditions of pseudoplastic culture, and intensify shear stress on fungus pellets to release SPG. At first, a constant-aeration rate of 0.08 vvm was implemented in both bioreactors to evaluate the new strategy compared to the previously studied methods. In the second set of experiments with IAS, along with the increasing oxygen demand of culture, the inlet airflow was increased gradually, while the dissolved oxygen (DO) was maintained higher than zero and below 1%. Using IAS in STB significantly raised productivity by about 100% in 96 h from 0.035 to 0.073 g/L.h. Also, employing this strategy in BCB led to a 30% increase in the maximum SPG production from 3.2 to 4.2 g/L. IAS can effectively help handle the operation of S. commune cultivation on a large scale by improving mixing conditions, mass transfer, and shear stress in both bioreactor types. This method had a significant impact on STB cultivation and its productivity so that it can be a practical approach to SPG's industrial production.

Oligonucleotide delivery to antigen presenting cells by using schizophyllan.

Schizophyllan (SPG), a member of the ß-glucan family, can form novel complexes with homo-polynucleotides such as poly(dA) through hydrogen bonding between two main chain glucoses and the one nucleotide base. Dectin-1, one of the major receptors for ß-glucans, is known to be expressed on antigen presenting cells (APCs) such as macrophages and dendritic cells. This suggests that the above-mentioned complexes could deliver bound functional oligonucleotides (ODNs) including antisense (AS)-ODNs, small interfering RNA, and CpG-ODNs to the APCs. Analysis using a quartz crystal microbalance revealed that a complex consisting of SPG and dA60 with a phosphorothioate backbone was recognized by recombinant Dectin-1 protein. Treatment with this complex containing an AS-ODN for tumor necrosis factor alpha protected mice against lipopolysaccharide-induced hepatitis at a very low AS-ODN dose. Moreover, immunization with CpG-ODN/SPG complex and antigenic proteins induced potent antigen specific immune responses. The present review also represents peptide delivery by conjugation with dA60 and the preparation of a nanogel using DNA-DNA hybridization. These findings indicate that the delivery of a specific ODN using ß-glucans could be used for treating various diseases caused by APCs and for activating antigen specific immune responses.

A novel ß-glucan-oligonucleotide complex selectively delivers siRNA to APCs via Dectin-1.

Delivering therapeutic nucleic acids to targeted cells and organs has been a challenge for decades. A novel technology to deliver oligonucleotide therapeutics to immune cells is here described. In this approach, a macromolecular complex of oligonucleotides and the ß-1,3-glucan schizophyllan (SPG) is selectively delivered to cells expressing a lectin receptor, Dectin-1, via SPG-Dectin-1 interaction. Detailed investigation of Dectin-1-expressing cells revealed that Dectin-1 is expressed in all subsets of monocytes as well as dendritic cell (DC) populations, including conventional DCs (cDCs) and plasmacytoid DCs (pDCs), in humans. The expression patterns in mice and humans are comparable, except for the expression in pDCs. The results indicate that Dectin-1 is expressed on cells capable of professional antigen presentation, except for B cells. We chose CD40 as a target gene for small interfering RNA (siRNA) as CD40 expression in antigen-presenting cells (APCs), particularly in DCs, plays critical roles in regulating immune responses. Dose-dependent cellular uptake of siCD40-SPG complexes was confirmed in cells expressing Dectin-1. Gene silencing activity was confirmed in vitro by the reduction of CD40 mRNA and by the site-specific cleavage of CD40 mRNA as determined by the 5' RNA ligase-mediated rapid amplification of cDNA ends (5'RLM-RACE) technique. In vivo activity of siCD40-SPG complexes was demonstrated as the reduced CD40 protein expression in monocytes and DCs in mice. Furthermore, the in vivo activity of siCD40-SPG targeting human CD40 was confirmed in cynomolgus monkeys by the 5'RLM-RACE technique. In conclusion, we have demonstrated the receptor-ligand binding-mediated delivery of siRNA targeting immune-regulating monocytes and DCs via the interaction of SPG and its receptor, Dectin-1. As monocytes and DCs play central roles in inducing and controlling immune responses, Dectin-1-targeted delivery of nucleic acids should provide a useful tool for developing drugs to treat a wide range of diseases, including autoimmune diseases, allergy, and cancer, as well as transplantation.

Efficacy of SPG-ODN 1826 Nanovehicles in Inducing M1 Phenotype through TLR-9 Activation in Murine Alveolar J774A.1 Cells: Plausible Nano-Immunotherapy for Lung Carcinoma.

Alveolar macrophages are the first line of defense against intruding pathogens and play a critical role in cancer immunology. The Toll-like receptor (TLR) family mediates an important role in recognizing and mounting an immune response against intruding microbes. TLR-9 is a member of the intracellular TLR family, which recognizes unmethylated CG motifs from the prokaryotic genome. Upon its activation, TLR-9 triggers downstream of the MyD-88-dependent transcriptional activation of NF-κB, and subsequently results in abundant inflammatory cytokines expression that induces a profound inflammatory milieu. The present exploratory investigation aimed at elucidating the potency of schizophyllan for entrapping ODN 1826 (SPG-ODN 1826)-mediated stimulation of TLR-9 in provoking an inflammatory-type response in murine alveolar macrophages. Schizophyllan (SPG), a representative of the ß-glucan family, was used in the present study as a nanovehicle for endosomal trafficking of CpG ODN 1826. TEM analysis of SPG-ODN 1826 nanovehicles revealed that the prepared nanovehicles are spherical and have an average size of about 100 nm. Interestingly, SPG-ODN 1826 nanovehicles were competent in delivering their therapeutic payload within endosomes of murine alveolar macrophage (J774A.1) cells. Exposure of these nanovehicles within LPS stimulated J774A.1, resulted in a significant provocation of reactive oxygen species (ROS) (p < 0.01) in comparison to CpG ODN 1826 alone. Moreover, the formulated nanovehicles succeeded in generating a profound Th1-based cytokine profile constituted by enhanced expression of IFN-γ (p < 0.001) and IL-1ß (p < 0.001) inflammatory cytokines. These findings clearly indicated the immunostimulatory potential of SPG-ODN 1826 nanovehicles for inducing the Th1-type phenotype, which would certainly assist in skewing M2 phenotype into the much-desired M1 type during lung cancer.

Enhancement of schizophyllan production in Schizophyllum commune using microparticles in medium.

Schizophyllum commune is a wood-rotting filamentous fungus that secrets a homopolysaccharide called as schizophyllan. Schizophyllan has several applications such as enhanced oil recovery, pharmaceutical materials and an anti-cancer drug carrier. Biomass growth and schizophyllan production increase the viscosity of the cultivation medium, thus resulting in mass transfer limitation for the substrate. In this study, adding talc and aluminium oxide microparticles into the cultivation medium was studied to improve the fungal growth and morphology. The response surface methodology and one factor at a time were applied to find the effects of microparticles with different sizes and concentrations on the schizophyllan production. The optimum concentration and size of aluminium oxide microparticles were obtained as 20 g L-1 and < 30 µm, respectively. Aluminium oxide microparticles in shake flask culture caused to increase the schizophyllan production from 10 to 15 g L-1 and decrease the cultivation time from 10 to 7 days. The production yield also increased from 0.11 to 0.30 g of schizophyllan/g glucose. Bioreactor cultivation showed a twofold increase in schizophyllan production from 1.5 to 3 g L-1. The results of this study suggested a significant increase in the production of schizophyllan using a low-cost "microparticle-enhanced cultivation" without any further optimization of the culture medium.

Repolarization of glioblastoma macrophage cells using non-agonistic Dectin-1 ligand encapsulating TLR-9 agonist: plausible role in regenerative medicine against brain tumor.

AIM OF THE STUDY: Glioblastoma multiforme (GBM) is the most severe forms of brain cancer, eventually becoming the leading cause of brain cancer-related death worldwide. Owing to the bleak surgical interventions and resistance to the different treatment regime, GBM is a parlous disease demanding newer therapeutical perspective for its treatment. Toll-like receptors (TLRs) are well-known members of pathogen recognition receptors (PRRs) and have been extensively explored for their therapeutic and prophylactic potential in an array of disease including cancer. Recent trends in drug delivery research has shown shift towards delivering short DNA sequences (CpG DNA) to endosomal TLR9 within immune cells (macrophages, dendritic cells, etc.) for the activation of desired inflammatory response using non-agonistic ß-glucan particles; a well-known ligand for Dectin-1 receptors. Our study is therefore focused to explore the role of nano-encapsulated CpG ODN as critical players in polarizing M2 scavenging to much desired pro-inflammatory type. MATERIALS AND METHODS: The nanoparticles entrapping CpG ODN 1826 were prepared by using a fungal polymer Schizophyllan (SPG). The constructed nanoparticles were characterized and assessed for their efficacy on rat glioblastoma cells (C6). RESULTS: The constructed Schizophyllan (SPG) nanoparticles entrapping CpG ODN 1826 (95.3%) were of 25.49 nm in diameter and thus capable of crossing blood-brain barrier. The rat glioblastoma (C6) cells evaluated for intracellular oxidative burst and cytokine levels pre- and post-incubation with nanoparticles exhibited marked elevation in the expression of intracellular ROS and IFN-γ as well as IL-1ß post treatment. CONCLUSION: The findings indicate towards potentiality of repolarizing the M2 macrophages to much desired M1 phase by inducing higgh levels of oxidative burst and inflammatory cytokines. Consequently, the apoptosis was induced in glioblastoma cells establishing the suitablity of CpG ODN carrying nanoformulations as emerging therapeutic intervention for GBM.

Leishmanial CpG DNA nanovesicles: A propitious prophylactic approach against visceral leishmaniasis.

Unmethylated CpG motifs with phosphothioate backbone trigger TLR9 to elicit innate immune response characterized by the production of Th1 cytokines. The use of CpG DNA as an adjuvant has established its role in potentiating the humoral and cell mediated vaccine specific immune response. However, none of the synthetic oligodeoxynucleotides (ODNs) know and used till date are associated with the parasite itself. Our group identified a novel CG rich sequence of 14 base pairs from Leishmania donovani genome (Ld CpG ODN) and established it as a TLR9 agonist. The present study was designed to ascertain the adjuvanticity of Ld CpG ODN with soluble leishmanial antigen in experimental model of L. donovani. During the study Schizophyllan (SPG), a fungal polymer was used for encapsulating Ld CpG ODN for efficient endosomal delivery. The synthesized nanovehicles were of nearly 100 nm and localized within endosomes as confirmed by confocal microscopy. Immunization studies displayed the superior ability of synthesized nanovehicles co-administered with parasite antigen in augmenting innate immune response in comparison to ODN, nanoparticles or soluble antigen alone. The response included generation of ROS, NO and iNOS expression followed by proinflammatory cytokine milieu with reduced parasitic load within liver, spleen and bone marrow. These immune-tailored particles in combination with parasitic antigens elicited significant generation of cell mediated response owing to the presence of high levels of CD8+ T-cells and lymphocyte proliferation. Moreover, vaccination regime with synthesized adjuvant also activated humoral immunity by escalating the levels of IgG2 followed by reduced levels of anti-leishmanial IgG and IgG1 antibodies. The findings support the efficacy of Ld CpG ODN as a potential adjuvant against visceral leishmaniasis.

Schizophyllum commune ß-glucan: Effect on interleukin-10 expression induced by lipopolysaccharide from periodontopathic bacteria.

ß-glucans are potent immunomodulators, with effects on innate and adaptive immune responses via dectin-1 as the main receptor. In this study, we investigated the biological effect of ß-glucan from Schizophyllum commune, called Schizophyllan (SPG) on Interleukin-10 (IL-10) expression induced by a lipopolysaccharide (LPS) from Aggregatibacter actinomycetemcomitans in murine macrophages (J774.1). SPG and dectin-1 interaction up-regulates LPS-induced IL-10 expression. The regulative effect of SPG on IL-10 expression is dependent on prolongation of nuclear translocation activity of nuclear factor-kappa B (NF-κBα) pathway induced by LPS. We also found that LPS-induced phosphorylation of mitogen- and stress-activated protein kinase 1 (MSK1) and cAMP-responsive-element-binding protein (CREB), followed by up-regulation of IL-10, was stimulated by SPG priming via activation of the spleen tyrosine kinase (Syk). Our data indicate that SPG augments the anti-inflammatory response in murine macrophages which can be useful to create an intervention for periodontal disease treatment.

A novel self-assembled micelles based on stearic acid modified schizophyllan for efficient delivery of paclitaxel.

This study was aimed to design a novel amphiphilic carrier based on schizophyllan (SPG) exopolysacharide for drug delivery. Stearic acid (SA) was used for the esterification of SPG with two degrees of substitutions (SA-SPG0.5 and SA-SPG1). The H NMR and FTIR spectroscopies verified the succesfull esterification of SPG. The polymeric micelles easily self-assembled into nanomicelles by ultrasound method. Fluorescence spectroscopy showed that the critical micelle concentrations (CMCs) of SA-SPG0.5 and SA-SPG1 micelles were 0.068 mg/mL and 0.027 mg/mL, respectively. DLS analyses showed that nanomicelles were ranged from 156 to 175 nm. SEM and TEM images showed that nanomicelles were mostly spherical. Paclitaxel (PTX) as a drug model was successfully loaded into SA-SPG nanomicelles with three different drug/polymer weight ratios of 0.1, 0.2 and 0.3. The highest encapsulation efficiency (75 %) was obtained when the PTX/SA-SPG weight ratio was 0.1. The in vitro release of PTX from SA-SPG micelles represented the sustained release profile over 144 h. MTT assay showed that the PTX-loaded SA-SPG nanomicelles had the higher cytotoxicity against MCF-7 cells than free PTX. These results revealed that the synthesized SA-SPG nanomicelles had a promising potential as a new carrier for efficient delivery of hydrophobic drugs.