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Understanding the immune response to hydrogel implantation is critical for the design of immunomodulatory biomaterials. To study the progression of inflammation around poly(ethylene glycol) hydrogels presenting Arg-Gly-Asp (RGD) peptides and vascular endothelial growth factor, we used temporal analysis of high-dimensional flow cytometry data paired with intravital imaging, immunohistochemistry, and multiplexed proteomic profiling. RGD-presenting hydrogels created a reparative microenvironment promoting CD206 + cellular infiltration and revascularization in wounded dorsal skin tissue. Unbiased clustering algorithms (SPADE) revealed significant phenotypic transition shifts as a function of the cell-adhesion hydrogel properties. SPADE identified an intermediate macrophage subset functionally regulating in vivo cytokine secretion that was preferentially recruited for RGD-presenting hydrogels, whereas dendritic cell subsets were preferentially recruited to RDG-presenting hydrogels. Last, RGD-presenting hydrogels controlled macrophage functional cytokine secretion to direct polarization and vascularization. Our studies show that unbiased clustering of single-cell data provides unbiased insights into the underlying immune response to engineered materials.

Trabajo presentado al Deerbreeding - Technologies, Products and Economics Congress, celebrado del 21 al 22 de julio de 2021.

Mesenchymal stem cells (MSCs) exhibit multiple beneficial properties, most notably potentimmunomodulation via their secretory factors that render them as an attractive cell source for cell therapy. However, theirclinical application remains limited due to their low survival and persistence in vivo, highlighting the need for new cell-carriers1. Chitosan is a widely applied biomaterial, but its poor solubility has constrained its use as a cell carrier2. Weexamined the fabrication of chitosan-based MSCs-microcarriers using a water-soluble chitosan lactate (ChLA) derivativeand in situ crosslinking in a microfluidic device using tripolyphosphate (TPP) and glycerylphytate (G1Phy) as ioniccrosslinkers. G1Phy is a powerful antioxidant,3 whose crosslinking ability has been evinced4. Our microgel synthesisallows for cell encapsulation, and G1Phy provided beneficial features to microgels in terms of MSCs survival, persistenceand secretome modulation.

[Background]: Deer-meat consumption is increasing in several countries based on claims that it is a healthy red meat. This study assessed the effect of slaughter age (low – 26 months and younger; medium – from 27 months to younger than 42 months; and high – 42 months and older) on carcass characteristics and meat physicochemical properties of 150 Iberian wild red deer., [Results]: Whole carcass yields of neck, backbone and flank increased (P < 0.001) with age, while yields of shoulder, tenderloin and leg decreased (P < 0.05) with age. The pH measured at 48 h post mortem at 9th rib level was not affected by age, while pH measured at 72 h post mortem at the longissimus thoracis et lumborum muscle decreased (P < 0.05) with age. Meat chemical composition was not affected by slaughter age, except for intramuscular fat (IMF), which increased (P < 0.05) with slaughter age. Finally, cooking loss and shear force were higher (P < 0.05) in meat from older deer., [Conclusion]: Most characteristics of meat from wild red deer depend on age at slaughter. The greater IMF of meat from older animals may render it more palatable. However, meat shear force also increased with age. Additional studies testing palatability seem to be necessary to evaluate the effects of slaughter age on the sensorial quality of meat from Iberian wild red deer., The authors are grateful to the Ministerio de Economía y Competitividad (Retos-Colaboración program) (Project INCYDEN, grant number RTC-2016-5327-2) for financial support for the study. Special thanks to Cárnicas Dibe S.L. for the samples supplied for this research. Jose M. Lorenzo is a member of the MARCARNE network, funded by CYTED (ref. 116RT0503).

The deer population present in Sardinia and Corsica represents an endemic subspecies Cervus elaphus corsicanus. We describe for the first time the characteristics of cast antlers of Sardinian red deer from the forest complex Sette Fratelli, south-east part of the island. Moreover, we describe the material mechanical properties, the structural ones, and the mineral profile of antlers from adults, comparing them with the antler characteristics of the subspecies C. e. hispanicus examined exactly with the same methodology. Sixty-one deer casted antlers were collected and classified as belonging to adults (35) or sub-adults (26). A first part of the study described the common features of the antlers of sub-species C. e. corsicanus through the analysis of morphology in all deer antlers. Subsequently, a more detailed study used 12 adult deer antlers for a destructive analysis. Statistical comparisons were conducted using ANOVAs between characteristics of the two age subgroups, and using Pearson¿s correlation coefficients between the antlers morphological variables. In general, morphological antler measures had greater values in adults than in sub-adults. In comparison with Iberian deer, Sardinian adult antlers have a more simple structure with lower values in morphological features, mechanical properties and structural characteristics.

Figures showing reproducibility data for our nature cell biology manuscript NCB-S33674B

Implant-associated inflammation and bacterial infection severely limit the functional performance of medical devices and are a major cause of implant failure. Therefore, it is crucial to develop methodologies to monitor/image implant-associated aseptic inflammation and bacterial infection in a minimally invasive manner. Here, we exploited near-infrared fluorescence (NIRF) molecular probes injected locally at the implant site to perform minimally invasive, simultaneous imaging of inflammation, and infection associated with implanted polymer disks. The hydro-sulfo-Cy5 (H-s-Cy5) probe detected reactive oxygen species associated with inflammatory responses to both aseptic and biofilm-containing implants, whereas diaminocyanine sulfonate selectively detected nitric oxide associated with a biofilm on the biomaterial at acute time points (

Resumen del trabajo presentado al XX Congreso de la Sociedad Española de Investigación Osea y del Metabolismo Mineral, celebrado en Bilbao del 21 al 23 de octubre de 2015., [Introducción]: La cuerna de los ciervos es un apéndice craneal externo constituido por un hueso primario que está muerto y seco. Como material óseo, la cuerna tiene algunas propiedades mecánicas sobresalientes. En comparación con otros huesos del esqueleto, p.ej. el fémur, la cuerna posee mayor tenacidad y mayor energía de impacto; circunstancias que hacen de la cuerna un hueso difícil de romper, y apropiado para su función en la berrea cuando los machos luchan entre sí. [Objetivo]: Analizar la microdureza, la microestructura, el contenido de calcio-mineral, y la secuencia de mineralización de la cuerna en comparación con el fémur. [Material]: Tres cuernas de ciervos adultos y un fémur de vareto. [Método]: La microdureza se determinó con microindentación Vickers (IndentaMet 11100, Buehler) usando una carga de 25g/10s aplicada en la superficie pulida de muestras encastradas en plástico. La microestructura se estudió en cortes mineralizados de 100 μm con microscopía de polarización circular (mapas de orientación colágena en falso color), y en cortes teñidos. El contenido de calcio (g/100g) se determinó mediante espectrometría de emisión atómica con plasma acoplado inductivamente (ICP). La secuencia de mineralización se dedujo tras inyectar a los ciervos con calceína (5 mg/kg s.c.) en el día 117 del desarrollo de sus cuernas, y posteriormente localizando el marcaje en cortes mineralizados con microscopía de epifluorescencia y calculando el tiempo transcurrido desde la formación de las osteonas primarias a la caída del terciopelo. [Resultados]: La microdureza era mucho menor en las cuernas respecto del fémur (48%), y también el contenido de calcio (70%). En una misma cuerna ambos valores disminuían algo en dirección próximo-distal según la secuencia de mineralización. La microarquitectura de la cuerna difería notablemente respecto de la del fémur (fibrolaminar) al estar constituida por dos tipos de tejido óseos (no laminar y laminar) que se engarzan para formar un hueso compacto. [Conclusiones]: La microdureza de la cuerna es muy inferior a la del fémur. Dado que la microdureza representa una propiedad mecánica relacionada con la máxima carga elástica de un material, y en el hueso en parte también con el contenido de calcio, se apunta que las excepcionales propiedades mecánicas de la cuerna que hace difícil su fractura han de basarse sobre todo en su microestructura, baja mineralización, y en su composición orgánica de cemento (riqueza de uniones de sacrificio).

The preorbital gland plays not only an olfactory role in cervids but also a visual one. Opening this gland is an easy way for the calf to communicate with the mother, indicating hunger/satiety, stress, pain, fear, or excitement. This information can be also useful for farm operators to assess how fast the calves habituate to handling routines and to detect those calves that do not habituate and may suffer chronic stress in the future. Thirty-one calves were subjected to 2 consecutive experiments to clarify if observing preorbital gland opening is related to habituation to handling in red deer calves (Cervus elaphus). Calves were born in 3 different paddocks, handled as newborns (Exp. 1), and then subjected to the same routine handling but with different periodicity: every 1, 2, or 3 wk (Exp. 2). In Exp. 1, preorbital gland opening was recorded in newborns during an initial handling (including weighing, ear tagging, and sex determination). Preorbital gland opening occurred in 93% of calves during this procedure and was not affected by sex, time since birth, or birth weight. Experiment 2 consisted of measuring preorbital opening during the same routine handling (weighing, blood sampling, and rump touching to assess body condition) when calves were 1, 3, and 5 mo old. Binary logistic regression showed that gland opening was associated with habituation to handling, since at 1 and 3 mo the probability of opening the gland decreased with the number of handlings that a calf experienced before (P = 0.008 and P = 0.028, respectively). However, there were no further changes in preorbital gland opening rate in the 5-mo-old calves (P = 0.182). The significant influence of the number of previous handlings on the probability of opening the preorbital gland was confirmed through generalized linear model with repeated measures (P = 0.007). Preorbital gland opening decreased along the phases of the study. Nevertheless, we found a significant trend in individuals to keep similar opening patterns (intraclass correlation coefficient = 0.807, P < 0.001), which suggests that the more stressed individuals can be detected with this method. Therefore, we conclude that preorbital gland opening during routine handlings is related to the number of previous handlings, and thus it can be used as an indicator of lack of habituation to handling in farmed cervids., This study was supported by projects AGL2012-38898 (Ministry of Economy and Competitiveness, Spain), MZERO0714‏ (Ministry of Agriculture, Czech Republic), CIGA20145001 (Czech University of Life Sciences, Czech Republic), and IGA no. 20145026 (Faculty of Tropical AgriSciences, Czech Republic).

Resumen del trabajo presentado al 8th lnternational Deer Biology Congress & International Wildlife Management Symposium, celebrado en Harbin (China) del 27 al 31 de julio de 2014., In Ungulates, maternal effects occur when the phenotype of the mother influence that of the calf, with a long term relevance in terms of survival, growth or fitness. lt is common a strong effect of the maternal identity, but there is also certain plasticity in these maternal effects related to the maternal condition, experience or senescence. Trivers-Willard and Local Resources Competition are two hypothesis that has repeatedly demonstrate that different relevant aspects of the maternal phenotype (like age, weight, body condition, and dominance status) may affect the offspring sex-ratio: depending on these combination of factors in the mother, it m ay be more interesting (from the point of view of long-term reproductive fitness) to invest preferentially in one sex or the other. For the same reason, once that the sex of the calf is defined the hind m ay make different investment decisions depending on the sex of the calf. We studied how these aspects of the maternal phenotype influenced different relevant aspects of the calves at birth and during the early growth: birth date, birth weight, weight at weaning, and body condition at weaning. The analyses were performed with the objective of finding differences in the maternal investment related to the sex of the calf, and were conducted on 188 births from 75 hinds aged from 2 to 19. Data was collected along S seasons. All the animals involved were born in captivity under natural mating. For each possible effect (on each calf trait), 4 single GLMMs were built (one for each maternal trait) including hind ID and year as random factors, and the best models were selected using Akaike's lnformation Criterion. As expected, hind weight influences calf birth weight (p

Trabajo presentado a The International Scientific Conference on Deer Genetics and Management, celebrada en Sigulda (Latvia) el 6 y 7 de agosto de 2014., Mechanical properties, and in general, growth of antlers, has been thought to be mainly depending on Ca and P input. However, many other minerals can influence growth and mechanical performance in bones. Our research group examines mineral composition of antlers as a diagnostic tool to detect nutrition deficiencies, and also their effect in mechanical quality of the antler material, internal structure, density of cortical wall, and other antler characteristics. One of the most interesting minerals, although not the only one, seems to be manganese. In this presentation we remind early studies where a deficiency in manganese (and also phosphorus) detected in broken antlers naturally occurring in 2005, produced as a result a 30% reduction in antler weight, and a similar reduction in resistance to impact (impact energy), other mechanical properties, and thinner cortical wall. As a result of this experiment, we decided to examine the effects of manganese supplementation in spikers and adults of red deer under a balanced diet. Subjects were 29 deer of different classes of age (adult n=16, subadult n=3 , spiker n=10) that were divided into a control group (n=15) and a group subjected to injection (n=14). Antler mineral content (ashes, micro and macro minerals), mechanical properties (Work to peak force, Bending strength, Young's Modulus of elasticity) and structure (average cortical thickness, cortical bone's specific gravity, cortical bone ratio) were examined in to 4 different test points along the antler beam. Each variable has been related to treatment, class of age, weight and year of growth of the antlers in a statistical test based on General Linear Model. The results show that manganese supplementation did not produce any effects in spiker antlers beyond an increase in manganese content. However, in adults results on the overall mean of the antler show a clear effect of manganese supplementation increasing the content of Ca, Na, P, B, Co, Cu, K, Mn, Ni, Se, Si, but no effect on mechanical properties. An analysis on the top portion of the antlers, where the effects are shown more clearly, showed the same effect in mineral composition, but also a 16% increase in work to fracture, much greater than in the mentioned study in wild deer. Thus, manganese supplementation can improve mineral composition of antlers, structure and some mechanical properties despite animals having a balance diet. The results are important for mineral and feed supplements in deer, but they may also have implications for human health and bone biology.