Investigation on the role of red fox in tuberculosis maintenance community ¿ second opus: experimental infection with a virulent field Mycobacterium bovis strain

Trabajo presentado al: 69th Wildlife Disease Association and 14th European Wildlife Disease Association Conference. Cuenca, Spain. p. 135. 31 agosto-2 septiembre

Fucoidan-based hydrogels particles as versatile carriers for diabetes treatment strategies

There is a current lack of fully efficient therapies for diabetes mel-litus, a chronic disease where the metabolism of blood glucose isseverely hindered by a deficit in insulin or cell resistance to thishormone. Therefore, it is crucial to develop new therapeutic strat-egies to treat this disease, including devices for the controlleddelivery of insulin or encapsulation of insulin-producing cells. Inthis work, fucoidan (Fu)â a marine sulfated polysaccharide exhib-iting relevant properties on reducing blood glucose and antioxi-dant and anti-inflammatory effectsâ was used for thedevelopment of versatile carriers envisaging diabetes advancedtherapies. Fu was functionalized by methacrylation (MFu) using8% and 12% (v/v) of methacrylic anhydride and further photo-crosslinked using visible light in the presence of triethanolamineand eosin-y to produce hydrogel particles. Degree of methacryla-tion varied between 2.78 and 6.50, as determined by1HNMR, andthe produced particles have an average diameter ranging from0.63 to 1.3mm (dry state). Insulin (5%) was added to MFu solutionto produce drug-loaded particles and the release profile wasassessed in phosphate buffer solution (PBS) and simulated intes-tinal fluid (SIF) for 24h. Insulin was released in a sustained man-ner during the initial 8 h, reaching then a plateau, higher in PBSthan in SIF, indicating that lower pH favors drug liberation.Moreover, the ability of MFu particles to serve as templates forthe culture of human pancreatic cells was assessed using 1.1B4cell line during up to 7 days. During the culture period studied,pancreatic beta cells were proliferating, with a global viabilityover 80% and tend to form pseudo-islets, thus suggesting thatthe proposed biomaterial could be a good candidate as versatilecarrier for diabetes treatment as they sustain the release of insulinand support pancreatic beta cells viability.We acknowledge ERDF for the financial support through POCTEP Project 0687_NOVOMAR_1_P, under the scope of INTERREG 2007-2013, and project 0302_CVMAR_I_1_P, under the scope of INTERREG Espana-Portugal 2014-2020, and Structured Projects NORTE-01-0145-FEDER-000021, NORTE-01-0145-FEDER-000023 and ATLANTIDA (ref. NORTE-01–0145-FEDER-000040), under the scope of Programa Operacional Regional do Norte (Norte 2020). Funding from the Portuguese Foundation for Science and Technology for doctoral grant (SFRH/BD/112139/2015) and post-doctoral grant (SFRH/BPD/85790/2012) is also acknowledge

Cartilage regeneration approach based on squid chitosan scaffolds : in-vitro assessment

During the past decades, marine organisms have been the focus of considerable attention as potential source of valuable materials. For instance, chitosan is a biopolymer with high potential in the biomedical field and can be produced from crustacean shells and squid pens [1]. In this sense, we propose the use of chitosan to produce scaffolds for regenerative medicine purposes. An alkaline solution was used to deproteinize squid pens and isolate β- chitin (Chaussard 2004), which was further converted into chitosan through a deacetylation reaction. Chitosan was then processed into porous structures by freeze-drying [3], where chitosan solutions (4%) were submitted to different freezing temperature of -80ºC and - 196ºC. The produced structures were further submitted to neutralization methods with 4% NaHO, including in some cases a pre-washing step using ethanol/water solutions (100:0; 90:10, 80:20; 70:30 and 50:50) [4]. The morphology of scaffolds produced using either squid or commercial chitosan revealed a lamellar structure, independent of the source and/or freezing temperature. All chitosan scaffolds produced exhibited no-cytotoxic behaviour over L929 cells. To test the in vitro functionality of the scaffolds, cells from the mouse chondrogenic cell line ATDC-5 were seeded in the scaffolds and cultured for different time periods. Scaffolds made from squid chitosan were shown to promote better cell adhesion than commercial chitosan scaffolds and comparable or better cell proliferation. This demonstrates that squid chitosan is a valuable alternative to produce scaffolds for different applications in regenerative medicine, namely the regeneration of cartilage

Valorization of chitosan from squid pens and further use on the development of scaffolds for biomedical applications

Objectives: The aim of the present work is the valorization of squid pens through the production of chitosan that can be used for the development of biomedical applications. The present work is focused on !-chitin extraction from squid pens of the species Dosidicus gigas and its further conversion into chitosan. The biomedical potential of the isolated squid chitosan was assessed by processing this polymer as scaffolds for tissue engineering strategies. Methods: Alkali solution was used to deproteinized squid pens and thus isolate !-chitin, which was further converted into chitosan through a deacetylation reaction. The chitosan scaffolds were developed using a freeze-drying process, from 3% and 4% chitosan solutions in acetic acid and freezing at temperatures of -80ºC and -196ºC. Chitosan scaffolds were neutralized using two different methods: M1 – NaHO solution; and M2 – ethanol/water and NaHO solution. Morphology, Mechanical properties, degradation, cytotoxicity (L929 cells) and cellular adhesion (ATDC5 Chondrocytes like cells) of squid chitosan scaffolds were assessed and compared with the properties of scaffolds produced with commercial chitosan. Results: The morphology of scaffolds revealed a lamellar structure for all produced scaffolds, independent of the origin and concentration of chitosan. The treatment with sodium hydroxide and ethanol caused the formation of larger pores and loose of some lamellar features. Different freezing temperatures gave different pore morphology and the lower temperature a smaller pore size. The in vitro cell culture and cell adhesion studies showed that all chitosan scaffolds exhibited a non-cytotoxic effect over the mouse fibroblast-like cell line, L929 cells. Conclusions: The chitosan produced from the endoskeletons of giant squid Dosidicus Gigas has proven to be a valuable alternative to the commercial one when considering its use as biomaterial for different biomedical applications

Influence of freezing temperature and deacetylation degree on the performance of freeze-dried chitosan scaffolds towards cartilage tissue engineering

Chitosan-based porous structures have been significantly studied across the world as potential tissue engineering scaffolds. Despite the differences in chitosan produced from squid pens or crustacean shells, with the former being more reactive and easily available with a higher degree of deacetylation (DD), most of the studies report the use of crab or shrimp chitosan as they are readily available commercial sources. The aim of this work was to highlight the great potential of chitosan produced from squid pens for biomedical application. From freeze-dried scaffolds for soft tissue engineering, we investigated the influence of the DD of chitosan and the freezing temperature during processing on their performance. Chitosan was obtained by deacetylation of β-chitin previously isolated from endoskeleton of giant squid Dosidicus gigas (DD 91.2%) and compared with a commercially available batch obtained from crab shells (DD 76.6%). Chitosan solutions were frozen at â 80° C or â 196° C and further freeze-dried to obtain 3D porous structures (scaffolds). Scaffolds prepared at â 196° C have a compact structure with smaller pores, while those prepared at â 80° C showed a lamellar structure with larger pores. The compressive modulus varied from 0.7 up to 8.8 MPa. Both types of scaffolds were stable on PBS, including in the presence of lysozyme, up to 4 weeks. Furthermore, the squid chitosan scaffolds processed at â 80° C promoted ATDC5 chondrocyte-like cells adhesion and proliferation. The results suggest that the developed squid chitosan scaffolds might be further exploited for ap- plications in cartilage tissue engineering.This work was partially funded by ERDF through POCTEP Projects 0330_IBEROMARE_1_P and 0687_NOVOMAR_1_P, Atlantic Area Project 2011-1/164 MARMED and by European Union through European Research Council – Project ComplexiTE (ERC-2012-ADG 20120216-321266). Portuguese Foundation for Science and Technology is gratefully acknowledged for post-doc grants of R.P. Pirraco (SFRH/BPD/101886/2014) and S.S. Silva (SFRH/BPD/112140/2015) and PhD grant of Lara L. Reys (SFRH/BD/112139/2015). The authors would also like to acknowledge to Dr. Julio Maroto, from Fundación CETMAR (Spain) and Roi Vilela, from PESCANOVA S.A. (Spain), for the kind offer of squid pens.info:eu-repo/semantics/publishedVersio

Development of marine-based nanocomposite scaffolds for biomedical applications

Despite the increasing attention that marine organisms are receiving, many of those are not efficiently exploited and subproducts with valuable compounds are being discarded. Two examples of those subproducts are the endoskeleton of squid, from which β-­‐chitin and consecutively chitosan can be obtained; and fish-­‐bones, as a source for the production of nano-­‐ hydroxyapatite. In this work, inspired in the nanocomposite structure of human bone, marine-­‐ based nanocomposite scaffolds composed by chitosan and nano-­‐hydroxyapatite (nHA) were developed using particle aggregation methodology. Chitosan was obtained from endoskeleton of giant squid Dosidicus Gigas while fish hydroxyapatite nanoparticles were synthesized from fish-­‐bones by pulsed laser in deionized water. An innovative methodology was used based on the agglomeration of prefabricated microspheres of chitosan/nHA, generally based on the random packing of microspheres with further aggregation by physical or thermal means to create a marine nanocomposite (CHA) .The morphological analysis of the developed nanocomposites revealed a low porosity structure, but with high interconnectivity, for all produced scaffolds. Furthermore, the nanocomposite scaffolds were characterized in terms of their mechanical properties, bioactivity, crystallinity and biological behavior. The obtained results highlight that the chitosan/nHA-­‐based marine nanocomposite can be a good candidate for biomedical applications, namely on bone regeneration

Semiconductor gellan gum based composite hydrogels for tissue engineering applications

Publicado em "Journal of Tissue Engineering and Regenerative Medicine", vol. 7, supp. 1 (2013)Semiconductor hydrogels can be developed by combining the intrinsic electrical properties of semiconductors with the specific characteristics of hydrogels. These hydrogels have recently attracted much attention for applications in tissue engineering, especially formulations incorporating pyrrole and excellent biocompatibility. Several studies have reported that electrical stimulation influences the migration, proliferation and differentiation of stem cells and other cell lines [1]. The goal of this work is to use in situ chemical polymerization of polypyrrole (PPy) with gellan gum (GG) in order to obtain a new generation of semiconductor composite hydrogels. For the synthesis of GG/PPy composites, GG at 1.25% (w/v) final concentration was prepared in distilled water at room temperature. The solution was then heated under stirring at 90°C for 20 min. Temperature was decreased to 65°C and Py was added under vigorous agitation. The crosslinker solution (CaCl2, 0.18%) was added at 50°C. After 2 h, GG/Py composite hydrogels were obtained. In a further step, GG/Py samples were immersed in a solution of oxidizing agent in PBS and the reaction was carried out for 18 h under agitation at room temperature. Finally, the samples were frozen at -80°C for 48 h and lyophilized. The characterization of GG, GG/PPy and PPy samples was performed by scanning electron microscopy (SEM). The incorporation of PPy in the gellan gum was confirmed by SEM analysis. The coating with PPy increases the thickness of each sheet in 3 fold when compared with GG samples. Conductivity tests were also performed. For cytotoxicity assay, the samples were rehydrated with complete culture medium. MTS and DNA quantification assays were performed to evaluate the metabolic activity and proliferation of L929 fibroblast cells after 1, 3 and 7 days in culture with GG, GG/PPy and PPy samples. MTS assays clearly indicate a proportional relation between the cell viability and the PPy concentration: higher concentrations of PPy resulted in lower cell viability. These results show that lower concentration of PPy incorporated in the GG hydrogels can provide an adequate electrical stimulus to improve cell behavior. In conclusion, semiconductor hydrogels can be an excellent platform for tissue engineering and electrochemical therapy application

Marine algae sulfated polysaccharides for tissue engineering and drug delivery approaches

Biomedical field is constantly requesting for new biomaterials, with innovative properties. Natural polymers appear as materials of election for this goal due to their biocompatibility and biodegradability. In particular, materials found in marine environment are of great interest since the chemical and biological diversity found in this environment is almost uncountable and continuously growing with the research in deeper waters. Moreover, there is also a slower risk of these materials to pose illnesses to humans. In particular, sulfated polysaccharides can be found in marine environment, in different algae species. These polysaccharides don’t have equivalent in the terrestrial plants and resembles the chemical and biological properties of mammalian glycosaminoglycans. In this perspective, are receiving growing interest for application on health-related fields. On this review, we will focus on the biomedical applications of marine algae sulfated polymers, in particular on the development of innovative systems for tissue engineering and drug delivery approaches.European Regional Development Fund (ERDF)Fundação para a Ciência e a Tecnologia (FCT

The effect of large-scale agricultural investments on household food security in Madagascar

Large-scale agricultural investments in developing countries have escalated over the past decade. While much is written about the potential adverse effects of these acquisitions on local communities, there is a paucity of evidence of these impacts. This paper explores the effect of large-scale agribusinesses on household food security in two locations in Madagascar. One is plantation area or Location A and the other one is contract farming area or Location B. The sample of 601 households was classified into households (i) in which at least one member was employed or (ii) contracted to the agribusiness, (iii) in the same area that were neither employees nor contractors (non-engaged) and (iv) counterfactual households from another community. Employment opportunities from the agribusinesses seemed to improved food security. Dietary quality, food security and resilience were higher among employed households. Contract households were generally more food insecure than the counterfactual and non-engaged households. Living in the zone of influence did not seem to have major adverse effects on the food security of non-engaged households. However, female-headed households seemed disadvantaged in terms of access to employment and contracting opportunities. Unless attention is paid to women’s access to employment and contracting opportunities, inequality may be exacerbated.The Belmont Forum and the Joint Programming Initiative on Agriculture, Food Security and Climate Change (FACCE-JPI) African Food, Agriculture, Land and Natural Resource Dynamics (AFGROLAND) project funded by the Swiss National Science Foundation, the French National Research Agency and the South African National Research Foundation.http://link.springer.com/journal/125712021-07-13hj2020Agricultural Economics, Extension and Rural Developmen

The use of ionic liquids in the processing of chitosan/silk hydrogels for biomedical applications

Natural polymers are adequate renewable resources for the processability of well-defined architectures for several applications. Combinations of polysaccharides and proteins may mimic the naturally occurring environment of certain tissues. The main goal of this work renders the application of green chemistry principles, namely the use of ionic liquids (ILs) and biorenewable sources, such as chitosan (CHT) and silkfibroin (SF), to process new hydrogel-based constructs. Although the solubilization of both materials in ILs has been studied individually, this work reports, for the first time, the role of ILs as solvent, for the production of hydrogels from blends of chitosan and silkfibroin (CSF). These systems offer the advantage of being homogeneous and presenting easy and short dissolution time of both biomacromolecules. Moreover, the use of chitosan obtained fromα- andβ-chitin allowed the production of blended hydrogels with distinct physical–chemical properties.In vitroassays demonstrated that these hydrogels supported the adhesion and growth of primary human dermalfibroblasts. Taken these properties together, the CSF hydrogels might be promising biomaterials to be explored for skin tissue engineering approaches.Fundação para a Ciência e a Tecnologia FCT - SFRH/BPD/45307/2008, SFRH/BPD/ 34704/2007, SFRH/BD/64601/2009, PTDC/QUI/68804/2006FEDER - POCTEP 0330_IBEROMARE_1_P