Bioactive starch-based scaffolds and human adipose stem cells are a good combination for bone tissue engineering

Silicon is known to have an influence on calcium phosphate deposition and on the differentiation of bone precursor cells. This study explores the effect of the incorporation of silanol (Si–OH) groups into poly- meric scaffolds on the osteogenic differentiation of human adipose stem cells (hASC) cultured under dynamic and static conditions. A blend of corn starch with polycaprolactone (30/70 wt.%, SPCL) was used to produce three-dimensional fibre meshes scaffolds by the wet-spinning technique, and a calcium sili- cate solution was used as a non-solvent to develop an in situ functionalization with Si–OH groups. In vitro assessment, using hASC, of functionalized and non-functionalized scaffolds was evaluated in either a- MEM or osteogenic medium under static and dynamic conditions (provided by a flow perfusion bioreac- tor). The functionalized materials, SPCL–Si, exhibit the capacity to sustain cell proliferation and induce their differentiation into the osteogenic lineage. The formation of mineralization nodules was observed in cells cultured on the SPCL–Si materials. Culturing under dynamic conditions using a flow perfusion bioreactor was shown to enhance the hASC proliferation and differentiation and a better distribution of cells within the material. The present work demonstrates the potential of these functionalized mate- rials for future applications in bone tissue engineering. Additionally, these results highlight the simplicity, economic and reliable production process of those materials.Fundação para a Ciência e a Tecnologia (FCT) - (Grant SFRH/BD/69962/2010, SFRH/BPD/26648/2006, PTDC/CTM/67560/2006European NoE EXPERTISSUES - (NMP3-CT-2004- 50028

Stimulatory effects of inorganic ions on osteogenesis in vitro

Introduction: Several studies demonstrated the effect of silicate ions (Si) on differentiation of bone precursor cells1,2, although its exact role in processes related to bone formation and remodeling is still incompletely understood. The focus of this work is to explore the effect of calcium and silicate ions on growth and osteogenic differentiation of human mesenchymal stem cells (hMSCs). This strategy may reduce the need for growth factors required to stimulate bone formation in regenerative approaches, decreasing the associated costs and overcoming stability issues. Materials and Methods In order to define the range of Si concentrations that are not toxic to cells, we performed a preliminary study varying Si concentrations from 0.00357mM to 4mM. The concentration of the Ca ions was selected based on the earlier study by Barradas et. al3. Cell culture media were supplemented by using sodium silicate (Na2SiO3) and/or calcium chloride dehydrate (CaCl2*2H2O) as Si and Ca precursors, respectively. hMSCs derived from bone marrow were seeded at a seeding density of 2.000 cells/cm2 and allowed to adhere overnight. Then, the medium was replaced by the appropriate supplemented medium and cells were cultured for 3, 7, 14 and 18 days. Basic and osteogenic media were used as negative and positive controls. Cell proliferation was evaluated by DNA quantification. hMSCs osteogenic gene expression was evaluated by Q-PCR. Results DNA quantification indicated an increase in cell number during the culture time for all the conditions. Results obtained by Q-PCR revealed a significantly higher expression of osteocalcin (OC) and bone morphogenetic protein-2 (BMP2) in cells cultured in media supplemented by both ions, as compared to media containing either Ca or Si alone. Discussion and Conclusions DNA quantification studies indicated that none of the selected concentrations had a negative influence on cell proliferation. The increase in osteogenic gene expression for cells cultured with both Ca and Si suggested a synergistic effect of the two ions on osteogenic differentiation of hMSCs. We further showed that cells cultured in the medium with the highest concentration of Ca (7.8mM) revealed a higher expression of the selected genes, which is in accordance with the earlier results by Barradas et al3. The obtained results suggest the importance of combining both ions, Ca and Si, for promoting the osteogenic differentiation of hMSCs. References 1. Hoppe A, Biomaterials 32: 2757-2774, 2011. 2. Beck Jr GR, Nanomedicine: Nanotechnology, Biology, and Medicine,1-11, 2011 3. Barradas AMC et al., Biomaterials 3205-3215, 2012. Acknowledgments The author thanks the Portuguese Foundation for Science and Technology (FCT) for the grant (SFRH/BD/69962/2010). Disclosures The authors have nothing to disclose

Short-term study of effects of fertilisation and cutting treatments on the vegetation dynamics of mountain heathlands in Spain

P. 181-191La influencia de la gestión y la disponibilidad de nutrientes en la dinámica de la vegetación de páramos caracterizados por Calluna vulgaris y Erica tetralix fueron estudiados en tres sitios de la montaña en el norte de España. En un total de 90 parcelas (1 m2 cada uno) recibieron diferentes combinaciones de corte y dos veces el fondo deposición atmosférica estimada de nitrógeno (56 kg yr_1 ha_1). Una de las dos especies dominantes ericáceas fueron cortados selectivamente a mano a nivel del suelo y su regeneración comparan en la presencia o ausencia de la otra. Los resultados después de 2 años mostraron efectos significativos del fertilizante en la cubierta vegetal, principalmente favoreciendo especies herbáceas perennes. No hubo efectos menos notables en el número de flores y sobre el crecimiento anual de las especies de ericáceas. Se concluye que, en el corto plazo, los nutrientes aumentaron solo, al doble de la deposición atmosférica corriente estimado para la zona, no va a alterar significativamente la composición de los brezales de montaña. Sin embargo, una vez que las gradas llegan a la fase de madurez, la capacidad de la comunidad para regenerar después disminuye una perturbación grave. Un impacto drástico, como el corte no puede dar lugar a la re-crecimiento de las mismas especies de arbustos, sino en sustitución por especies herbáceas, que también se beneficiarán de la mayoría de los nutrientesS

Platelet lysate-loaded photocrosslinkable hyaluronic acid hydrogels for periodontal endogenous regenerative technology

The integrity and function of the periodontium can be compromised by traumatic injuries or periodontitis. Currently available clinical therapies are able to stop the progression of periodontitis and allow the healing of periodontal tissue. However an optimal strategy capable of restoring the anatomy and functionality of the lost periodontal tissue is still to be achieved. Herein is proposed the development of an injectable hydrogel system able to release a growth factors and cells to the periodontal defect. This injectable system is based on a photocrosslinkable hydrogel, prepared from methacrylated Hyaluronic Acid (me-HA) and incorporating Platelet Lysate (PL). The delivery of growth factors and cells in situ is expected to enhance regeneration of the periodontium. Various formulations of me-HA containing increasing PL concentrations were studied for achieving the formation of stable photocrosslinkable hydrogels. The produced hydrogels were subsequently characterized to assess mechanical properties, degradation, protein/growth factor release profile, antimicrobial activity and response towards human Periodontal Ligament fibroblasts (hPDLFs). The results demonstrated that it was possible to obtain stable photocrosslinkable hydrogels incorporating different amounts of PL that can be released in a sustained manner. Furthermore, the incorporation of PL improved (p<0.02) the viscoelastic properties of the hydrogels and enhanced their resilience to the degradation by hyaluronidase (HAase). Additionally, the PL showed to provide antimicrobial properties. Finally, hPDLFs, either seeded or encapsulated into the developed hydrogels, showed enhanced proliferation over time (p<0.05), proportionally to the increasing amounts of PL present in the hydrogel formulations

Structural, UV-VIS-NIR luminescence and decay associated spectral profiles of Sm3+ doped calcium phosphate glass

Rare-Earths or Lanthanide ions (Ln3+) are a group of elements from lanthanum to lutetium (Z=57 to 71), plus scandium (Z=21) and Yttrium (Z=39). Ln3+ doped glasses have been included within the Hydroxyapatite (HA) matrix, inducing promising changes in their physicochemical and biological properties. Because of the lanthanides' ability to modulate bone metabolism, inclusion of Ln3+ in the composition of calcium phosphate biomaterials for bone tissue regeneration has been considered. Earlier, the authors studied Samarium (Sm3+) doped glass-reinforced hydroxyapatite with enhanced osteoblastic performance and antibacterial properties for bone tissue regeneration. Moreover, Sm3+ substituted calcium phosphate is a non-hazardous material that luminesces under UV-visible light. In this work, we study a calcium phosphate host glass doped with samarium oxide, which has been prepared and characterized by FTIR, SEM, EDS analysis, and X-ray mapping. Pumping with two visible excitation sources at 405nm and 423nm, we observed intense, sharp Green, yellow, orange emission peaks (4G5/2â 6H5/2,7/2, 9/2) at 560nm, 596nm and 643nm respectively. A weak red emission was also observed at 704nm. Two NIR peaks at 1134nm (4G5/2â 6F11/2) and 1310 nm (4G5/2â 6F9/2) are monitored by using an excitation at 1060nm. Furthermore, by making use of time-resolved emission spectroscopy (TRES) measurements, the decay associated spectra were obtained allowing the kinetic parameters for the different emission bands to be elucidated and compared with steady state emission spectra.European Union's Seventh Framework Programme (FP7/2007-2013) under grant agreement nº REGPOT-CT2012-316331-POLARIS. Also, SHN would like to thank to Cost Action, ref. # MP 1205 under materials (soft, bio & nano) and technologies for optofluidic devices, and further acknowledge to the FCT -Fundação para a Ciência e a Tecnologia, Portugal, through the project PTDC/SAU-BEB/103034/2008

Eczema Crónico das Mãos Relacionado com Alergia de Contacto a Lactonas Sesquiterpénicas e Partenolido

Allergic contact dermatitis to plants can assume different clinical patterns. We report a 63-year-old female patient with chronic hand dermatitis. Given the clinical suspicion of allergic contact dermatitis, patch tests were performed with the Baseline series of the GPEDC and a plants series which showed positivity to sesquiterpene lactone mix 0.1% pet (++) and parthenolide 0.1% pet (++) at day D2 and D4. These test results allowed the diagnosis of contact allergy to parthenolide that was related to occupational exposure Chamaemelum fuscatum during gardening. We emphasize the importance of a meticulous clinical history to increase the level of suspicion of Allergic contact dermatitis and evaluate the relevance of a positive patch test to sesquiterpene lactone and parthenolide as the etiology in chronic hand dermatitis.As dermatites de contacto alérgicas por plantas podem manifestar-se sob padrões clínicos diversos. Descrevemos o caso clínico de doente do sexo feminino, de 63 anos com eczema das mãos. Perante a suspeita clínica de dermatite de contacto alérgica efectuaram-se provas epicutâneas com as séries básica (Grupo Português de Estudo das Dermites de Contacto - GPEDC) e de plantas, que revelaram positividades para a mistura de lactonas sesquiterpénicas 0,1% vas (++) e partenolido 0,1% vas. (++) em D2 e D4, o que permitiu o diagnóstico de alergia do contacto ao partenolido relacionada com a exposição ocupacional à planta margaça-de-inverno (Chamaemelum fuscatum) nas atividades de jardinagem. Salienta-se a importância de uma anamnese exaustiva para aumentar o nível de suspeição de dermatite de contacto alérgica e avaliar a relevância dos testes positivos a lactonas sesquiterpénicas e ao partenolido, como agentes etiológicos em casos de eczema crónico das mãos

Mineralization of chitosan membrane using a double diffusion system for bone related applications

Chitosan membranes were subjected to a pre-treatment in a double diffusion system, with a calcium solution in one chamber and a phosphate solution in the other chamber. Both chambers were separated by the chitosan membrane and subject to three mineralization periods (5, 10 and 15 minutes). After this pre-treatment the bioactivity of the different calcium phosphate coatings formed was tested for different periods of immersion time, 7, 14 and 21 days at room temperature and 37ºC, in acellular simulated body fluid (1.0x). The results obtained demonstrated that the calcium phosphate coatings formed during the pre-treatment process are bioactive. It was found that the calcification is effective just in the side of the membrane exposed to the calcium solution chamber. This enabled to develop membranes with asymmetric osteoinductive properties that can be useful in different orthopedic applications

Silk-based biomaterials functionalized with fibronectin type II promotes cell adhesion

The objective of this work was to exploit the fibronectin type II (FNII) module from human matrix metalloproteinase-2 as a functional domain for the development of silk-based biopolymer blends that display enhanced cell adhesion properties. The DNA sequence of spider dragline silk protein (6mer) was genetically fused with the FNII coding sequence and expressed in Escherichia coli. The chimeric protein 6mer + FNII was purified by non-chromatographic methods. Films prepared from 6mer + FNII by solvent casting promoted only limited cell adhesion of human skin fibroblasts. However, the performance of the material in terms of cell adhesion was significantly improved when 6mer + FNII was combined with a silk-elastin-like protein in a concentration-dependent behavior. With this work we describe a novel class of biopolymer that promote cell adhesion and potentially useful as biomaterials for tissue engineering and regenerative medicine. Statement of Significance This work reports the development of biocompatible silk-based composites with enhanced cell adhesion properties suitable for biomedical applications in regenerative medicine. The biocomposites were produced by combining a genetically engineered silk-elastin-like protein with a genetically engineered spider-silk-based polypeptide carrying the three domains of the fibronectin type II module from human metalloproteinase-2. These composites were processed into free-standing films by solvent casting and characterized for their biological behavior. To our knowledge this is the first report of the exploitation of all three FNII domains as a functional domain for the development of bioinspired materials with improved biological performance. The present study highlights the potential of using genetically engineered protein-based composites as a platform for the development of new bioinspired biomaterials.This work was supported by Fundação para a Ciência e Tecnologia (FCT – Portugal) Funded Project “Chimera” (PTDC/EBB-EBI/109093/2008), by FCT/MEC through Portuguese funds (PIDDAC) – PEst-OE/BIA/UI4050/2014, by the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P. and by the ERDF through COMPETE2020 – Programa Operacional Competitividade e Internacionalização (POCI). TC is thankful to the FCT, ESF and POPH for its support through the Investigador FCT Programme (IF/01635/2014). ARibeiro thanks FCT for the SFRH\BPD\98388\2013 grant. AMPereira, RMachado and AdaCosta acknowledge FCT for PD/BD/113811/2015, SFRH-BPD/86470/2012 and SFRH/BD/75882/2011 grants, respectively

Time resolved emission spectra and electron paramagnetic resonance studies of Gd3+ doped calcium phosphate glasses

Gadolinium, Gd3+ (4f7 ), containing glasses are particularly interesting owing to their magnetic and optical properties. Gd3+ has an electron paramagnetic resonance (EPR) centre, which is often used as the EPR probe in different matrices. Another important feature of Gd3+ is that emits ultraviolet radiation (UVR) at around 310 â 313 nm from the 6 P7/2 ï  8 S7/2 transition and is suitable for the treatment of skin disease. Here, we report on Gd3+ doped calcium phosphate host glasses with four different concentrations; 0.5 - 4 mol % and labelled as 0.5Gd, 1Gd, 2Gd and 4Gd. A strong emission peak at 310 nm was observed in the reported glasses with excitation at 274 nm. The emission spectra can also be obtained at specific times during the decay. The time resolved emission data was analyzed with three different approaches namely, simple exponential, NED distribution and stretched exponential. The other approaches confirm the single exponential behaviour for all samples with deviation for the 0.5 Gd. The decay associated spectra (DAS) for the 0.5 Gd indicate that both decays have the same origin. The EPR spectrum exhibits several resonance signals characteristic of Gd3+ ions. The stimulated emission cross sections (Ï P EÃ 10â 21 cm2 ) of the UV emission transition 6 P7/2 ï  8 S7/2 were also reported in the above three approaches.The authors would like to acknowledge the financial support from the European Union's Seventh Framework Programme (FP7/2007-2013) under grant agreement nº REGPOT-CT2012-316331-POLARIS. Also, SHN would like to thank to Cost Action, ref. # MP 1205 under materials (soft, bio & nano) and technologies for ptofluidic devices, and further acknowledge to the FCT-Fundação para a Ciência e a Tecnologia, Portugal, hrough the project PTDC/SAU-BEB/103034/2008.info:eu-repo/semantics/publishedVersio

Diferencias transculturales en la manifestación del bullying en estudiantes de escuela secundaria

Con un diseño de grupos apareados por edad y grado escolar, comparamos los resultados de dos investigaciones sobre bullying, realizadas en Colombia y Costa Rica. El propósito fue identificar los comportamientos de los agresores y agresoras para configurar su perfil, y explorar la variable género en este fenómeno. Utilizamos el Cuestionario Paredes, Lega Vernon para Deteccion del Bullying en 824 estudiantes de 6o a 8o. El resultado de un MANOVA de dos vías mostró diferencias significativas en las categorías género y país, y una relación significativa en la interacción género y país.-1. Introducción. -2. Descripción del estudio. -3. Método. -4. Resultados. -5. Discusión. -Lista de referencia