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Chitosan (CS) is a natural biopolymer that shows promise as a biomaterial for bone-tissue regeneration. However, because of their limited ability to induce cell differentiation and high degradation rate, among other drawbacks associated with its use, the creation of CS-based biomaterials remains a problem in bone tissue engineering research. Here we aimed to reduce these disadvantages while retaining the benefits of potential CS biomaterial by combining it with silica to provide sufficient additional structural support for bone regeneration. In this work, CS-silica xerogel and aerogel hybrids with 8 wt.% CS content, designated SCS8X and SCS8A, respectively, were prepared by sol-gel method, either by direct solvent evaporation at the atmospheric pressure or by supercritical drying in CO2, respectively. As reported in previous studies, it was confirmed that both types of mesoporous materials exhibited large surface areas (821 m2g−1–858 m2g−1) and outstanding bioactivity, as well as osteoconductive properties. In addition to silica and chitosan, the inclusion of 10 wt.% of tricalcium phosphate (TCP), designated SCS8T10X, was also considered, which stimulates a fast bioactive response of the xerogel surface. The results here obtained also demonstrate that xerogels induced earlier cell differentiation than the aerogels with identical composition. In conclusion, our study shows that the sol-gel synthesis of CS-silica xerogels and aerogels enhances not only their bioactive response, but also osteoconduction and cell differentiation properties. Therefore, these new biomaterials should provide adequate secretion of the osteoid for a fast bone regeneration.

Silica (SiO2)/chitosan (CS) composite aerogels are bioactive when they are submerged in simulated body fluid (SBF), causing the formation of bone-like hydroxyapatite (HAp) layer. Silica-based hybrid aerogels improve the elastic behavior, and the combined CS modifies the network entanglement as a crosslinking biopolymer. Tetraethoxysilane (TEOS)/CS is used as network precursors by employing a sol-gel method assisted with high power ultrasound (600 W). Upon gelation and aging, gels are dried in supercritical CO2 to obtain monoliths. Thermograms provide information about the condensation of the remaining hydroxyl groups (400-700 degrees C). This step permits the evaluation of the hydroxyl group's content of 2 to 5 OH nm(-2). The formed Si-OH groups act as the inductor of apatite crystal nucleation in SBF. The N-2 physisorption isotherms show a hysteresis loop of type H3, characteristic to good interconnected porosity, which facilitates both the bioactivity and the adhesion of osteoblasts cells. After two weeks of immersion in SBF, a layer of HAp microcrystals develops on the surface with a stoichiometric Ca/P molar ratio of 1.67 with spherulite morphology and uniform sizes of 6 mu m. This fact asserts the bioactive behavior of these hybrid aerogels. Osteoblasts are cultured on the selected samples and immunolabeled for cytoskeletal and focal adhesion expression related to scaffold nanostructure and composition. The initial osteoconductive response observes points to a great potential of tissue engineering for the designed composite aerogels.

Background Currently available treatments for secondary progressive multiple sclerosis(SPMS) have limited efficacy and/or safety concerns. Adipose-mesenchymal derived stem cells(AdMSCs) represent a promising option and can be readily obtained using minimally invasive procedures. Go to: Patients and methods In this triple-blind, placebo-controlled study, cell samples were obtained from consenting patients by lipectomy and subsequently expanded. Patients were randomized to a single infusion of placebo, low-dose(1x106cells/kg) or high-dose(4x106cells/kg) autologous AdMSC product and followed for 12 months. Safety was monitored recording adverse events, laboratory parameters, vital signs and spirometry. Expanded disability status score (EDSS), magnetic-resonance-imaging, and other measures of possible treatment effects were also recorded. Go to: Results Thirty-four patients underwent lipectomy for AdMSCs collection, were randomized and thirty were infused (11 placebo, 10 low-dose and 9 high-dose); 4 randomized patients were not infused because of karyotype abnormalities in the cell product. Only one serious adverse event was observed in the treatment arms (urinary infection, considered not related to study treatment). No other safety parameters showed changes. Measures of treatment effect showed an inconclusive trend of efficacy. Conclusion Infusion of autologous AdMSCs is safe and feasible in patients with SPMS. Larger studies and probably treatment at earlier phases would be needed to investigate the potential therapeutic benefit of this technique.

[Background] Currently available treatments for secondary progressive multiple sclerosis(SPMS) have limited efficacy and/or safety concerns. Adipose-mesenchymal derived stem cells(AdMSCs) represent a promising option and can be readily obtained using minimally invasive procedures., [Patients and methods] In this triple-blind, placebo-controlled study, cell samples were obtained from consenting patients by lipectomy and subsequently expanded. Patients were randomized to a single infusion of placebo, low-dose(1x106cells/kg) or high-dose(4x106cells/kg) autologous AdMSC product and followed for 12 months. Safety was monitored recording adverse events, laboratory parameters, vital signs and spirometry. Expanded disability status score (EDSS), magnetic-resonance-imaging, and other measures of possible treatment effects were also recorded., [Results] Thirty-four patients underwent lipectomy for AdMSCs collection, were randomized and thirty were infused (11 placebo, 10 low-dose and 9 high-dose); 4 randomized patients were not infused because of karyotype abnormalities in the cell product. Only one serious adverse event was observed in the treatment arms (urinary infection, considered not related to study treatment). No other safety parameters showed changes. Measures of treatment effect showed an inconclusive trend of efficacy., [Conclusion] Infusion of autologous AdMSCs is safe and feasible in patients with SPMS. Larger studies and probably treatment at earlier phases would be needed to investigate the potential therapeutic benefit of this technique.

Utilización de nanopartículas de metales nobles como inmunomoduladores y composición inmunomoduladora. La presente invención tiene por objeto la utilización de los efectos inmunomoduladores de nanopartículas metálicas de plata funcionalizadas con tiopronina [N-(2-mercaptopropionil)glicina]. Un segundo aspecto objeto de la invención es una composición inmunomoduladora que incluye nanopartículas de plata funcionalizadas con tiopronina.

Nanopartículas metálicas funcionarizadas con el neuro péptido VIP y procedimiento de preparación. Constituye el objeto de la presente invención nanopartí culas metálicas funcionalizadas con el neuropéptido VIP, así como el procedimiento de preparación de dichas na nopartículas. Las nanopartículas objeto de la presente invención pre sentan uniones selectivas de nanopartículas a péptidos en dos orientaciones posibles, grupo NH2 o grupo COOH. En esta última orientación, los péptidos si son reconoci dos por los receptores de membranas celulares, lo que proporciona una herramienta que permite discernir efec tos dependientes e independientes de receptor. El péptido empleado ha sido el VIP, mediante el cual se obtiene un amplio espectro de funciones biológicas, in cluida inmunomodulación, actuando predominantemente como un potente anti-inflamatorio y un agente inhibidor de la respuesta del Th1 en el sistema inmunitario y emer giendo como un importante factor terapéutico para el tra tamientode enfermedades con componentes inflamatorias y autoinmunes

[Background]: Parkinson’s disease (PD) is a progressive neurodegenerative disorder characterized pathologically by the presence in the brain of intracellular protein inclusions highly enriched in aggregated alpha-synuclein (a-Syn). Although it has been established that progression of the disease is accompanied by sustained activation of microglia, the underlying molecules and factors involved in these immune-triggered mechanisms remain largely unexplored. Lately, accumulating evidence has shown the presence of extracellular a-Syn both in its aggregated and monomeric forms in cerebrospinal fluid and blood plasma. However, the effect of extracellular a-Syn on cellular activation and immune mediators, as well as the impact of familial PD-linked a-Syn mutants on this stimulation, are still largely unknown. [Methods and Findings]: In this work, we have compared the activation profiles of non-aggregated, extracellular wild-type and PD-linked mutant a-Syn variants on primary glial and microglial cell cultures. After stimulation of cells with a-Syn, we measured the release of Th1- and Th2- type cytokines as well as IP-10/CXCL10, RANTES/CCL5, MCP-1/CCL2 and MIP-1a/CCL3 chemokines. Contrary to what had been observed using cell lines or for the case of aggregated a-Syn, we found strong differences in the immune response generated by wild-type a-Syn and the familial PD mutants (A30P, E46K and A53T). [Conclusions]: These findings might contribute to explain the differences in the onset and progression of this highly debilitating disease, which could be of value in the development of rational approaches towards effective control of immune responses that are associated with PD.

Nanopartículas metálicas funcionalizadas con el neuropéptido VIP y procedimiento de preparación. Constituye el objeto de la presente invención nanopartículas metálicas funcionalizadas con el neuropéptido VIP, así como el procedimiento de preparación de dichas nanopartículas. Las nanopartículas objeto de la presente invención presentan uniones selectivas de nanopartículas a péptidos en dos orientaciones posibles, grupo NH2 o grupo COOH. En esta última orientación, los péptidos si son reconocidos por los receptores de membranas celulares, lo que proporciona una herramienta que permite discernir efectos dependientes e independientes de receptor. El péptido empleado ha sido el VIP, mediante el cual se obtiene un amplio espectro de funciones biológicas, incluida inmunomodulación, actuando predominantemente como un potente anti-inflamatorio y un agente inhibidor de la respuesta del Th1 en el sistema inmunitario y emergiendo como un importante factor terapéutico para el tratamiento de enfermedades con componentes inflamatorias y autoinmunes.

The invention relates to metal nanoparticles functionalised with neuropeptide VIP as well as to the method for preparing the nanoparticles. Said nanopartcles have selective bonds to peptides in two possible orientations, group NH2 or group COOH. In the latter orientation, the peptides are recognised by the cell membrane receptors, providing a tool that can be used to distinguish dependent and independent receptor effects. Peptide VIP has been used, enabling a wide range of biological functions, including immunomodulation, to be obtained, acting predominantly as a powerful anti-inflammatory and an agent inhibiting the Th1 response in the immune system and emerging as an important therapeutic factor for the treatment of diseases with inflammatory and autoimmune components., La invención relaciona a las nanopartículas del metal funcionalizadas con el neuropeptide VIP así como al método para preparar las nanopartículas. Los nanopartcles dichos tienen enlaces selectivos a los péptidos en dos posibles orientaciones, grupos NH2 o grupos COOH. En la orientación más última, el son de los péptidos reconocido por los receptores de la membrana celular, proporcionando una herramienta que el bote sea utilizado distinguir efectos dependientes y independientes del receptor. El péptido VIP ha sido utilizado, posibilitando una amplia gama de funciones biológicas, incluyendo el immunomodulation, para ser obtenido, actuando predominante como un antiinflamatorio potente y agente que inhibían la respuesta Th1 en el sistema inmune y que emergían como factor terapéutico importante para el tratamiento de enfermedades con los componentes inflamatorios y autoinmunes.

Vasoactive intestinal peptide (VIP) was originally isolated as a vasodilator intestinal peptide, then as a neuropeptide. In the immune system, VIP is described as an endogenous macrophage-deactivating factor. VIP exerts its immunological actions in a paracrine and/or autocrine manner, through specific receptors. However, very little is known about the molecular regulation of VIP type 2 receptor (VPAC(2)) in the immune system. We now report that different toll-like receptor (TLR) ligands selectively regulate the VPAC(2) receptor gene and show a gene repression system controlled by key protein kinase signalling cascades in macrophages. VPAC(2) gene expression is regulated by gram-positive (TLR2 ligands) and gram-negative bacteria wall constituents (TLR4 ligands). Moreover, VPAC(2) is tightly regulated: TLR2- or TLR2/6- but not TLR2/1-mediated mechanisms are responsible for the induction of VPAC(2). TLR stimulation by viral or bacterial nucleic acids did not modify the VPAC(2) mRNA levels. Remarkably, imiquimod--a synthetic TLR7 ligand--led to a potent up-regulation of VPAC(2) gene expression. TLR5 stimulation by flagellin present in gram-positive and gram-negative bacteria did not affect VPAC(2) mRNA. The p38 mitogen-activated protein kinase (MAPK) activity accounted for the TLR4-mediated induction of VPAC(2) gene expression. Surprisingly, our data strongly suggest for the first time a tightly repressed control of VPAC(2) mRNA induction by elements downstream of MAPK kinase 1/2, PI3K/Akt, and particularly Jun-NH(2)-terminal kinase signalling pathways.

We characterized a method to conjugate functional silver nanoparticles with vasoactive intestinal peptide (VIP), which could be used as a working model for further tailor-made applications based on VIP surface functionality. Despite sustained interest in the therapeutic applications of VIP, and the fact that its drugability could be largely improved by the attachament to functionalized metal nanoparticles, no methods have been described so far to obtain them. Materials & methods: VIP was conjugated to tiopronin-capped silver nanoparticles of a narrow size distribution, by means of proper linkers, to obtain VIP functionalized silver nanoparticles with two different VIP orientations (Ag-tiopronin-PEG-succinic-[His]VIP and Ag-tiopronin-PEG-VIP[His]). VIP intermediate nanoparticles were characterized by transmission-electron microscopy and Fourier transform infrared spectroscopy. VIP functionalized silver nanoparticles cytotoxicity was determined by lactate dehydrogenase release from mixed glial cultures prepared from cerebral cortices of 1-3 days-old C57/Bl mice. Cells were used for lipopolysaccharide stimulation at day 18-22 of culture. Results: Two different types of VIP-functionalized silver nanoparticles were obtained; both expose the C-terminal part of the neuropeptide, but in the first type VIP is attached to silver nanoparticle through its free amine terminus (Ag-tiopronin-PEG-succinic-[His]VIP), while in the second type, VIP N-terminus remains free (Ag-tiopronin-PEG-VIP[His]). VIP-functionalized silver nanoparticles did not compromise cellular viability and inhibited microglia-induced stimulation under inflammatory conditions. Conclusion: The chemical synthesis procedure developed to obtain VIP-functionalized silver nanoparticles rendered functional products, in terms of biological activity. The two alternative orientations designed, reduced the constraints for chemical synthesis that depends on the nanosurface to be functionalized. Our study provides, for the

Uso de la melatonina como adyuvante. La presente invención se refiere al uso de la melatonina como adyuvante en preparados vacunales para la prevención de enfermedades infecciosas en mamíferos ungulados. También se refiere al uso de la melatonina en combinación con otros inmunoestimuladores, entre ellos, el hidróxido de aluminio.

Alzheimer's disease (AD) is a complex disease, with no definitive biomarkers available that allow clinical diagnosis; this represents a major problem for the advance of efficient drug discovery programs. A successful approach towards the understanding and treatment of AD should take into consideration this complex nature. In this sense, metabolic networks are subject to severe stoichiometric restrictions. Metabolomics amplifies changes both in the proteome and the genome, and represents a more accurate approximation to the phenotype of an organism in health and disease. In this article, we will examine the current rationale for metabolomics in AD, its basic methodology and the available data in animal models and human studies. The discussed topics will highlight the importance of being able to use the metabolomic information in order to understand disease mechanisms from a systems biology perspective as a non-invasive approach to diagnose and grade AD. This could allow the assessment of new therapies during clinical trials, the identification of patients at risk to develop adverse effects during treatment and the final implementation of new tools towards a more personalized medicine.