Optimized Electrophoretic Deposition of Chitosan/Mesoporous Glass Nanoparticles with Gentamicin on Titanium Implants: Enhancing Hemocompatibility and Antibacterial Activity.

Titanium-based implants have long been studied and used for applications in bone tissue engineering, thanks to their outstanding mechanical properties and appropriate biocompatibility. However, many implants struggle with osseointegration and attachment and can be vulnerable to the development of infections. In this work, we have developed a composite coating via electrophoretic deposition, which is both bioactive and antibacterial. Mesoporous bioactive glass particles with gentamicin were electrophoretically deposited onto a titanium substrate. In order to validate the hypothesis that the quantity of particles in the coatings is sufficiently high and uniform in each deposition process, an easy-to-use image processing algorithm was designed to minimize human dependence and ensure reproducible results. The addition of loaded mesoporous particles did not affect the good adhesion of the coating to the substrate although roughness was clearly enhanced. After 7 days of immersion, the composite coatings were almost dissolved and released, but phosphate-related compounds started to nucleate at the surface. With a simple and low-cost technique like electrophoretic deposition, and optimized stir and suspension times, we were able to synthesize a hemocompatible coating that significantly improves the antibacterial activity when compared to the bare substrate for both Gram-positive and Gram-negative bacteria.

Short-Term In Vivo Response to Anodized Magnesium Alloy as a Biodegradable Material for Bone Fracture Fixation Devices.

Biodegradable materials based on magnesium alloys have a huge potential for bone fracture fixation devices due to their adequate mechanical properties and biocompatibility. However, their fast degradation and the consequent liberation of hydrogen gas at the initial stages of implantation is the major limitation for their use. In this study, the AZ91D magnesium alloy was surface treated by an environment-friendly, nontoxic, and low-cost anodizing process and the early in vivo response was studied in a rat transcortical model. Adequate maturation of woven bone around implants-detected at day 7 post implantation-to lamellar bone was observed from day 15. Lamellar bone after 15 and 30 days of implantation presented similar volume, mineralization pattern, mineral to protein content, and estimated bone maturity between anodized AZ91D and polylactic acid (control) implants. Histology observation showed neither release of hydrogen bubbles in the region closed to the anodized AZ91D implant nor systemic effects on liver, kidney, and spleen. Thus, anodizing of AZ91D in the conditions stated here induced an adequate short-term in vivo response, which postulates their use as potential biodegradable fracture fixation devices for bone healing.

Assessment of Trade-Offs between Simultaneous Immune Challenges in a Slow-Living Subterranean Rodent.

The coexistence of two or more infectious agents in the same host is common in nature. Given this, the study of trade-offs within the immune system itself is key to understanding how immune defenses act in wild species in their natural environment. Here we assessed the possible trade-off between an inflammatory response (induced by phytohemagglutinin [PHA]; involving innate and adaptive responses in the study species) and an antibody response (induced by sheep red blood cells [SRBC]; adaptive response) in a slow-living subterranean rodent, the Talas tuco-tuco (Ctenomys talarum Thomas, 1898). According to life-history theory, slow-living species should rely more heavily on adaptive immunity, which develops more slowly than an innate response but is beneficial against repeated infections. Individual physiological condition (estimated by measuring levels of infection and immune, nutritional, and stress parameters) was analyzed during immune challenges. Contrary to what was expected, we found that the magnitude and energetic costs of both immune responses were similar when stimulated alone or simultaneously. Variation in natural antibodies, neutrophils, basophils, total leukocytes, and the ratio of neutrophils to lymphocytes in relation to the different treatments was also detected. In particular, natural antibodies were negatively affected by the induction of both immune challenges simultaneously and an increase of neutrophil counts was detected in all animals with the exception of those challenged with SRBC, while the pattern of variation of basophils, total leukocytes, and ratio of neutrophils to lymphocytes was not clearly associated with any triggered immune response. In general, our results suggest the absence of an energetic or resource-based trade-off between the immune responses triggered by PHA and SRBC in C. talarum.

Prenatal predatory stress in a wild species of subterranean rodent: Do ecological stressors always have a negative effect on the offspring?

In pregnant females, a failed predatory event not only induces individual responses but also represents a significant change in the developmental environment of the offspring, which may lead to modifications in their phenotype that may persist at different stages of life. We evaluate whether prenatal exposure to predatory cues affects anxiety behavior, behavioral response to predator cues, stress response to immobilization, and immune response to sheep red blood cells (SRBC) and phytohemagglutinin (PHA) in juveniles of the subterranean rodent Ctenomys talarum. We found that prenatal predator stress (PPS) (1) increased juvenile anxiety-like behaviors and the appearance of antipredator behaviors, (2) did not affect the response of offspring to predatory stressors, and (3) did not influence the physiological response of juveniles to stressors (immobilization) nor the immunological responses to SRBC and PHA challenges. This work shows the influence of PPS on the development of behavioral responses in the offspring, whom displayed a state of anxiety and behavioral changes associated with decreased locomotor activity and avoidance behaviors. Thus, these individuals prenatally exposed to predatory cues show behavioral adaptations that may contribute to avoid predators in the adult life.

Parasite infection negatively affects PHA-triggered inflammation in the subterranean rodent Ctenomys talarum.

Magnitude and effectiveness of immune responses vary greatly between and within species. Among factors reported to determine this variation, parasitism is a critical one, although controversial effects of parasites over immunological indices have been reported. Information regarding immune strategies in species with different life histories is crucial to better understand the role of immune defenses in an ecological and evolutionary context. Here, we examine the influence of the parasite community on immune responsiveness of a solitary subterranean rodent, Ctenomys talarum. To do this, we assessed the impact of the natural parasite community and the experimental infection with Eimeria sp. on the phytohemagglutinin (PHA)-response, as well as other immune, condition, nutrition, and stress parameters. PHA-triggered inflammation was similarly impaired by Eimeria sp. infection alone or co-occurring with a number of gastrointestinal nematodes. None of the other physiological parameters studied were affected by parasitism. This indicates that parasitism is a general key factor modulating immune responsiveness of the host, and in particular for C. talarum, it could explain the great inter-individual variation previously observed in the PHA-response. Thus, our results highlight the importance of taking the parasite community into account in ecoimmunological studies, particularly when using immunological indices.

Food Restriction Affects Inflammatory Response and Nutritional State in Tuco-tucos (Ctenomys talarum).

Insufficient or unbalanced food intake typically has a negative impact on immune responses. The understanding of this effect is, however, hampered by the effect that food has on general condition, which, in turn, affects immunity, and the interaction among general condition, immunocompetence, and concurrent infections. The goal of this study was to determine the effects of food restriction and methionine supplementation on immunity in tuco-tucos (Ctenomys talarum). Effects of diet manipulations on nutritional state, inflammatory response to phytohemagglutinin (PHA), and other immune parameters (bacterial killing capacity, natural antibodies, and leukocyte profile) were evaluated. Health and stress parameters and endoparasite loads were assessed to understand more deeply potential effects of treatments on immune status. Individuals under food restriction presented an altered nutritional state as well as increased stress levels (higher N: L ratios) compared with individuals fed ad libitum, and a marked reduction in the inflammatory response to PHA. Supplementation with methionine did not affect any of the parameters analyzed. Endoparasite loads were not affected by treatments. Our results support the idea that food insufficiency can modulate the individual's immune responsiveness through the lack of adequate essential nutrients, metabolic fuel and energetic reserves, or by a detrimental effect of the stress caused by nutrient limitation. We show that the response to PHA previously reported as nonenergetically costly for C. talarum, implies a nutritional cost; an opposite pattern to that previously found for the adaptive antibody response to sheep red blood cells in the same species.

PHA-induced inflammation is not energetically costly in the subterranean rodent Ctenomys talarum (tuco-tucos).

Immune activity has been proposed to be associated with substantial costs, due to trade-offs with other functions or activities that share common resources and contribute to an animal's fitness. However, direct estimates of the cost of mounting an immune response are few and have been performed mainly in birds. Thus, further work is needed to clarify the relative costs of different components of the immune system and the role of environmental and life-history traits in modulating the costs of resistance. Within the components of immunity, inflammation is considered to be associated with a larger energetic expenditure. Here, we evaluated the energetic cost of the inflammatory response to phytohemagglutinin (PHA) in a wild population of a subterranean rodent, Ctenomys talarum, and the trade-offs between immune activity and reproduction. C. talarum develops an inflammatory response to PHA, but contrary to our predictions, this response was not associated with an increase in oxygen consumption regardless of reproductive status or sex. Our study shows that an immune challenge may not always result in a detectable energetic cost. We discuss the possibility that other currencies could be underlying the cost, such as micro-or macronutrients requirements, autoimmunity or oxidative stress.

Assessing the energetic costs and trade-offs of a PHA-induced inflammation in the subterranean rodent Ctenomys talarum: immune response in growing tuco-tucos.

A traditional approach used to assess whether immune defense is costly is to explore the existence of trade-offs between immunity and other functions; however, quantitative studies of the energetic costs associated with the activation of the immune system are scarce. We assessed the magnitude of a PHA-triggered immune response and the associated energetic costs in 60-day old Ctenomys talarum. We expected that the magnitude of the macroscopic inflammatory response to PHA is lower in young tuco-tucos compared with that of adults, given the allocation of substantial energy to growth, and that the magnitude of the inflammation is lower in male pups compared to females, due to the higher investment in growth of the larger sex. Concomitantly, we expected that the pups challenged with PHA show an increase in oxygen consumption compared to control animals and that a positive association exists between magnitude of the PHA-induced inflammation and oxygen consumption. Contrary to what was expected, young tuco-tucos mounted a higher inflammatory response compared with adults and there were no differences in the magnitude of this response between sexes. The inflammatory response induced by a PHA injection did not represent a significant energetic cost for young tuco-tucos. There were no differences in oxygen consumption between PHA-injected and control animals, and tuco-tucos that mounted a higher inflammatory response to PHA did not show higher oxygen consumption. Energy expenditure, however, is not the only physiological cost involved in trade-offs between immune response and various functions of the organism, and other currencies are discussed.