Collagen and chitosan blends for 3D bioprinting: A rheological and printability approach

Collagen and chitosan are widely employed as biomaterials, including for 3D-bioprinting. However, the use of collagen and chitosan (col:chi) blends as bioinks is still scarce. In this work, the rheology of different hydrogel precursors (0.5–1.50% w/v chi: 0.18–0.54% w/v col) was analyzed through frequency and strain sweeps, as well as at different shear rates. Col:chi blends showed a shear-thinning behavior, with viscosity values at low shear rates between 0.35 and 2.80 Pa s. Considering the strain rate determined by the applied flow in a 3D-bioprinter, precursor viscosities during the extrusion were in the interval 0.5–0.8 Pa s. Printability (Pr) was measured comparing images of the printed meshes and the corresponding CAD grid design, using photograph analysis. Col:chi 0.36:1.00 was chosen to print mono-layered scaffolds for tissue engineering (TE) because of its suitable viscosity, printability and polymer ratio content. Hydrogels were obtained through NaHCO3 nebulization and 37° incubation, and NHS/EDC were added to obtain scaffolds with improved mechanical behavior. They were stable after 44 h in PBS with collagenase at physiological level and showed no cytotoxic effect in NIH-3T3 fibroblasts.Fil: Heidenreich, Ana Carolina. Universidad Nacional de San Martin. Escuela de Ciencia y Tecnologia. Laboratorio de Biomateriales, Biomecanica y Bioinstrumentacion.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pérez Recalde, Mercedes. Universidad Nacional de San Martin. Escuela de Ciencia y Tecnologia. Laboratorio de Biomateriales, Biomecanica y Bioinstrumentacion.; ArgentinaFil: González Sánchez Wusener, Ana Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Hermida, Elida Beatriz. Universidad Nacional de San Martin. Escuela de Ciencia y Tecnologia. Laboratorio de Biomateriales, Biomecanica y Bioinstrumentacion.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Lavandin essential oil combined with the biopolymer PHBV, poly(3-hydroxybutyrate-co-3-hydroxyvalerate), for wound treatment

In this work, poly(3-hydroxybutyrate-co-3-hydroxyvalerate), PHBV, was combined with lavandin essential oil (LavEO), from Lavandula hybrida grown in Argentina. Linalool (36%), linalyl acetate (29%) 1,8-cineole (6%) and camphor (6%) were its main compounds as analyzed by GC-MS. PHBV is a biodegradable copolyester from a bacterial source; PHBV porous scaffolds have been shown to be suitable for fibroblast and keratinocyte proliferation [1,2] and thus for skin regeneration. In addition, LavEO exhibits strong anti-inflammatory and antibacterial activity, what improves wound healing [4,5]. Purified monoterpenes and entire EOs have been combined with other biopolymers for different applications but scarcely for wound healing and never previously with PHBV [6]. PHBV porous membranes, 100-200 µm thickness, containing LavEO at 2, 4 or 8% m/m were obtained by an emulsion-solvent evaporation method. LavEO-PHBV membranes did not show cytotoxicity when tested on NIH/3T3 fibroblasts (according to the ISO 10993-5 standard). SEM membrane analysis showed, in all cases, a high level of porosity, ̴ 20% of the surface. Keratinocytes (KC) adhesion and proliferation were evaluated with the HaCaT cell line. Membranes containing 2 or 4% of LavEO, allowed KC proliferation of the same level as that of PHBV membrane controls. However, cells neither adhered to nor proliferated on membranes with 8% LavEO although their hydrophobicity, estimated by the contact angle, was similar to that of 4%-LavEO membranes. The reduction in the elastic modulus–determined by the dynamic mechanical analysis–due to the presence of LavEO, suggests its plasticizer effect (control E=305±10 MPa; 8%-LavEO membrane E=150±20 MPa). According to these results, 2 or 4% LavEO-PHBV membranes seem as a promising treatment, especially in the case of infected and chronic wounds, often arrested at the inflammatory phase

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

Polysaccharides from the red seaweed nemalion helminthoides: characterization, chemical modification and biological activity

La presente Tesis comprende cuatro temas principales: Estudio del sistema de polisacáridos del alga roja Nemalion helminthoides, familia Liagoraceae, orden Nemaliales. Se determinó que la matriz de la pared celular estaba compuesta: (i) mayoritariamente por α-D-(1→3)-mananos con unidades monosulfatadas en C-4 ó C-6, con un promedio de un grupo sulfato cada cuatro residuos, y un grado de polimerización de ~200; (ii) en menor proporción por α-D-(1→3)-mananos con ramificaciones simples de β-D-xilosa en C-2, de similar grado de sulfatación también en C-4 ó C-6, y peso molecular promedio de ~10 kDa; (iii) por xilanos neutros β-D- (1→3;1→4), de ~6 kDa. Por último, de pared fibrilar se aislaron (1→4)-β-D-xilanos neutros. Modificaciones químicas de las fracciones de mananos y xilomananos de Nemalion helminthoides. (i) A partir de los mananos nativos se generaron polisacáridos modificados con 43-50% de sulfatación, constituidos mayormente por unidades disulfatadas en C-2 y C-6 y de unidades trisulfatadas. (ii) A partir de las fracciones de xilomananos se obtuvieron, por degradación de Smith, derivados sin ramificaciones de xilosa. Estudio de actividad antiviral de fracciones nativas y modificadas de Nemalion helminthoides. Los xilomananos inhibieron in vitro al virus herpes simplex (HSV-1 y HSV-2) y al virus dengue (DENV-2) agregados durante la adsorción viral. Los mananos sobresulfatados inhibieron DENV-2 agregados durante la adsorción viral, y HSV-1 y HSV-2 colocados durante y/o después de la adsorción viral in vitro. Utilizando un modelo de infección intranasal de ratones BALB/c con HSV-2, los mananos sobresulfatados protegieron de la enfermedad en un 100% con una única dosis, por la misma vía. Evaluación de acción inmunomoduladora de polisacáridos de Nemalion helminthoides. Fracciones de xilomananos indujeron proliferación de linfocitos T humanos (línea H9) y macrófagos murinos (línea RAW). En macrófagos, además, estimularon la secreción de citoquinas proinflamatorias IL-6 y TNF-α y de óxido nítrico. La respuesta de citoquinas también se detectó en plasma de ratones BALB/c inyectados por vía intravenosa. Los animales tratados con xilomananos no desarrollaron la enfermedad al ser infectados, una hora después, con HSV-2.This Thesis comprises four main topics: Study of the polysaccharide system from the red seaweed Nemalion helminthoides family Liagoraceae, order Nemaliales. It was determined that the cell wall matrix was composed: (i) mainly by α-D-(1→3)-mannans with monosulfated units at C-4 or C-6, one sulfate group every four residues, on average, and a polymerization degree of ~200; (ii) in minor amount by α-D-(1→3)-mannans with β-D-xylose at C-2, with similar sulfation degree at C-4 or C-6, and average molecular weigth of ~10 kDa; (iii) by neutral β-D-(1→3;1→4) mixed linkage xylans, of ~6 kDa. Finally, from the fibrillar wall, neutral (1→4)-β-D-xylans were isolated. Chemical modifications of mannan and xylomannan fractions from Nemalion helminthoides. (i) From the native mannans, oversulfated polysaccharides with a 43- 50% sulfation were prepared. These mainly constituted by disulfated units at C-2 and C- 6, and trisulfated units. (ii) From the xylomannan fractions, derivatives without xylose branches were obtained, by Smith degradation. Study of antiviral activity of native and modified fractions of Nemalion helminthoides. Xylomannans were inhibitors of herpes simplex virus (HSV-1 and HSV- 2) and dengue virus (DENV-2) in vitro when were added during viral adsorption. Oversulfated mannans were inhibitors of DENV-2 when were incorporated during viral adsorption, and HSV-1 and HSV-2 during and/or after viral adsorption in vitro. Oversulfated mannans were evaluated in BALB/c mice with intranasal infection with HSV-2, 100% of the mice receiving one dose were protected. Evaluation of immunomodulatory effect by Nemalion helminthoides polysaccharides. Xylomannans fractions were inductors of human T lymphocites (H9 line) and murine macrophages (RAW line) proliferation. Besides, this compounds were able to stimulate secretion of nitric oxide and cytokines (IL-6 and TNF-α) in macrophages. Cytokine response was also observed in BALB/c mice inoculated by intravenous route. Remarkably no illness was developed in mice treated with xylomannans fractions one hour before HSV-2 infection.Fil:Pérez Recalde, María Mercedes. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina

Polisacáridos del alga roja Nemalion helminthoides: caracterización, modificación química y actividad biológica

La presente Tesis comprende cuatro temas principales: Estudio del sistema de polisacáridos del alga roja Nemalion helminthoides, familia Liagoraceae, orden Nemaliales. Se determinó que la matriz de la pared celular estaba compuesta: (i) mayoritariamente por α-D-(1→3)-mananos con unidades monosulfatadas en C-4 ó C-6, con un promedio de un grupo sulfato cada cuatro residuos, y un grado de polimerización de ~200; (ii) en menor proporción por α-D-(1→3)-mananos con ramificaciones simples de β-D-xilosa en C-2, de similar grado de sulfatación también en C-4 ó C-6, y peso molecular promedio de ~10 kDa; (iii) por xilanos neutros β-D- (1→3;1→4), de ~6 kDa. Por último, de pared fibrilar se aislaron (1→4)-β-D-xilanos neutros. Modificaciones químicas de las fracciones de mananos y xilomananos de Nemalion helminthoides. (i) A partir de los mananos nativos se generaron polisacáridos modificados con 43-50% de sulfatación, constituidos mayormente por unidades disulfatadas en C-2 y C-6 y de unidades trisulfatadas. (ii) A partir de las fracciones de xilomananos se obtuvieron, por degradación de Smith, derivados sin ramificaciones de xilosa. Estudio de actividad antiviral de fracciones nativas y modificadas de Nemalion helminthoides. Los xilomananos inhibieron in vitro al virus herpes simplex (HSV-1 y HSV-2) y al virus dengue (DENV-2) agregados durante la adsorción viral. Los mananos sobresulfatados inhibieron DENV-2 agregados durante la adsorción viral, y HSV-1 y HSV-2 colocados durante y/o después de la adsorción viral in vitro. Utilizando un modelo de infección intranasal de ratones BALB/c con HSV-2, los mananos sobresulfatados protegieron de la enfermedad en un 100% con una única dosis, por la misma vía. Evaluación de acción inmunomoduladora de polisacáridos de Nemalion helminthoides. Fracciones de xilomananos indujeron proliferación de linfocitos T humanos (línea H9) y macrófagos murinos (línea RAW). En macrófagos, además, estimularon la secreción de citoquinas proinflamatorias IL-6 y TNF-α y de óxido nítrico. La respuesta de citoquinas también se detectó en plasma de ratones BALB/c inyectados por vía intravenosa. Los animales tratados con xilomananos no desarrollaron la enfermedad al ser infectados, una hora después, con HSV-2

Gelatin–alginate–hyaluronic acid inks for 3D printing: effects of bioglass addition on printability, rheology and scaffold tensile modulus

Natural hydrogels are widely used for 3D-bioprinting because of their qualities fortissue engineering. Recently, hydrogels have been combined with bioactive glasses,due to their angiogenic properties that aid tissue regeneration. In this work, we studiedthe printability and rheological properties of gelatin-alginate-hyaluronic acid inks with 2to 8 % w.w-1 of 45S5 bioglass (BG) that followed a pseudoplastic behavior along the3D-printing process. The reduction of the storage modulus of the inks after adding BGindicates that the microparticles might disrupt the polymeric network; furthermore, areduction of the viscosity was determined at BG concentrations above 6%. Inks withoutBG or up to 2 % evidenced the best printing fidelity on 10 % infill scaffolds. The tensilemodulus of crosslinked 40 %-filled scaffolds increased from 130 kPa (without BG) to160 kPa (6 to 8 % BG). Moreover, scaffolds containing BG 6 % and 8 % w.w-1presented a hydroxyapatite layer after being cultured for 2 days. Attachment andgrowth of fibroblasts on the scaffolds revealed their cytocompatibility, making thesematerials an alternative for further research on soft tissues regeneration.Fil: Bertuola, Marcos. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Tecnologías Emergentes y Ciencias Aplicadas. - Universidad Nacional de San Martin. Instituto de Tecnologías Emergentes y Ciencias Aplicadas; ArgentinaFil: Aráoz, Silvina Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Tecnologías Emergentes y Ciencias Aplicadas. - Universidad Nacional de San Martin. Instituto de Tecnologías Emergentes y Ciencias Aplicadas; ArgentinaFil: Gilabert, Ulises Eduardo. Secretaría de Industria y Minería. Servicio Geológico Minero Argentino; ArgentinaFil: González Sánchez Wusener, Ana Elena. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Pérez Recalde, Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Tecnologías Emergentes y Ciencias Aplicadas. - Universidad Nacional de San Martin. Instituto de Tecnologías Emergentes y Ciencias Aplicadas; ArgentinaFil: Arregui, Carlos Oscar. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Hermida, Élida B.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Tecnologías Emergentes y Ciencias Aplicadas. - Universidad Nacional de San Martin. Instituto de Tecnologías Emergentes y Ciencias Aplicadas; Argentin

Compound Petri nets and alternatives aggregation Petri nets: Two formalisms for decision-making support

Decision-making in technological systems, such as communication networks, manufacturing facilities and supply chains, constitutes a common requirement able to lead companies galore to success or failure. This article presents a decision-making methodology, where the feasible structural configurations to be analysed are chosen heuristically in the frame of a single optimization problem. For stating the optimization problem and solving it efficiently, appropriate formalisms would be used. Compound Petri nets, a particular kind of parametric Petri nets, and alternatives aggregation Petri nets, are two Petri net-based formalisms able to integrate in the same model different alternative structural configurations. Moreover, even having different characteristics that might make them useful for different applications, both formalisms present common features, such as including a set of exclusive entities and the possibility of developing compact Petri net models, by the removal of redundant information. This article is also focused on the transformation algorithm between compound Petri nets and alternatives aggregation Petri nets. This algorithm is devoted to transform a model described by one of the formalisms into an equivalent model, that is, with the same behaviour, represented using the other formalism. Finally, several application examples are given for illustrating the steps of the transformation algorithm. © 2016 The Author(s)