Are hydroxyapatite-based biomaterials free of genotoxicity? A systematic review.

Hydroxyapatite (HA) is a biomaterial widely used in clinical applications and pharmaceuticals. The literature on HA-based materials studies is focused on chemical characterization and biocompatibility. Generally, biocompatibility is analyzed through adhesion, proliferation, and differentiation assays. Fewer studies are looking for genotoxic events. Thus, although HA-based biomaterials are widely used as biomedical devices, there is a lack of literature regarding their genotoxicity. This systematic review was carried out following the PRISMA statement. Specific search strategies were developed and performed in four electronic databases (PubMed, Science Direct, Scopus, and Web of Science). The search used "Hydroxyapatite OR Calcium Hydroxyapatite OR durapatite AND genotoxicity OR genotoxic OR DNA damage" and "Hydroxyapatite OR Calcium Hydroxyapatite OR durapatite AND mutagenicity OR mutagenic OR DNA damage" as keywords and articles published from 2000 to 2022, after removing duplicate studies and apply include and exclusion criteria, 53 articles were identified and submitted to a qualitative descriptive analysis. Most of the assays were in vitro and most of the studies did not show genotoxicity. In fact, a protective effect was observed for hydroxyapatites. Only 20 out of 71 tests performed were positive for genotoxicity. However, no point mutation-related mutagenicity was observed. As the genotoxicity of HA-based biomaterials observed was correlated with its nanostructured forms as needles or rods, it is important to follow their effect in chronic exposure to guarantee safe usage in humans.

Relationship between boldness and exploratory behavior in adult zebrafish.

Behavioral responses vary between individuals and may be repeated in different contexts over time. When a behavioral response set is linked and present regardless of the context, it characterizes a behavioral syndrome. By evaluating how bold and shy (profiles related to risk-taking) individuals perform about exploration and anxiety, we can predict relationships of behavioral syndromes and better understand how different axis of personality is formed. Here we classified the profiles by risk-taking and evaluated their exploration behavior in the open field test. In this context, the two groups showed significant differences in thigmotaxis behavior: bold individuals habituate faster and show decreased thigmotaxis (less anxiety), while shy ones are less prone to leave the security of the side areas of the open tank and present higher anxiety. We emphasized the importance of further investigating the behavior of these profiles in other contexts and the importance of each one for the evolution and fitness of the species, in addition to a better understanding of which behaviors are involved in the behavioral syndromes in zebrafish.

Variation in nest building, aggression, learning, and steroid hormone levels in Betta splendens.

Individuals within a population present behavioral responses that vary according to intrinsic and extrinsic factors such as ontogenetic phase, nutritional status, reproductive stage, and previous experiences. These differences can be explained by endogenous changes, such as hormone release, that can modulate reproductive behaviors, stress response, and cognitive processes. In order to investigate the relationship between behavior and hormonal levels in the fighting fish Betta splendens, the present study characterized nest building, aggressive behavior, learning of a task, and levels of cortisol and 11-ketotestosterone (11-KT) in 86 male Fighting fish. At the beginning of the experiment (days 1-4), fish were characterized as nest builders, intermediate builders, or non-builders. They were then sequentially tested for aggression (days 7-8), learning performance (days 11-21), and circulating hormone levels (day 23). Nest builders showed the lowest hormonal levels at the end of the experiment and low aggressiveness; Intermediate builders presented low cortisol, but high KT levels and best learned the task; Non-builders were the most aggressive animals with higher cortisol levels (at day 23). Our data suggest that in B. splendens, aggressive behavior and learning performance are related to the relative investment in reproduction and variation in circulating levels of corticosteroids and androgens.

Individual differences guide mate choice in the fighting fish (Betta splendens).

Individual differences in behavior are observed in the variety of ways animals respond to environmental challenges, interact in a social group, take risks to access a resource, and so forth. In the present study, we investigated the behavioral reactions of male and female Siamese fighting fish (Betta splendens) in different contexts and how they affect female mate choice. A total of 100 females and 50 males were classified according to boldness, sociability, and aggression, and then female mate choice was observed based on male profile and body color. Our results showed sex-related differences in Siamese fighting fish behavioral profiles: Males exhibited a higher correlation with aggression and females with boldness. Both male and female behavioral profile affected female mate choice. Females preferred bold red and nonaggressive red males. Sex-related differences in behavioral profiles may reflect ecological differences between male and female Siamese fighting fish. Female mate choice could be related to behavioral profiles that indicate better parental care, as male features may influence parental care decisions and the development of offspring behavioral profiles. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Coffee time: Low caffeine dose promotes attention and focus in zebrafish.

In this study we investigated the ability of zebrafish to discriminate visual signs and associate them with a reward in an associative-learning protocol including distractors. Moreover, we studied the effects of caffeine on animal performance in the task. After being trained to associate a specific image pattern with a reward (food) in the presence of other, distractor images, the fish were challenged to locate the exact cue associated with the reward. The distractors were same-colored pattern images similar to the target. Both the target and distractors were continually moved around the tank. Fish were exposed to three caffeine concentrations for 14 days: 0 mg/L (control, n = 12), 10 mg/L (n = 14), and 50 mg/L (n = 14). Zebrafish spent most of the time close to the target (where the reward was offered) under the effects of 0 and 10 mg/L caffeine, and the shortest latency to reach the target was observed for the 10-mg/L caffeine group. Both caffeine treatments (10 and 50 mg/L) increased the average speed and distance traveled when compared to the control group. This study confirms previous results showing that zebrafish demonstrate conditioned learning ability; however, low-dose caffeine exposure seems to favor visual cue discrimination and to increase zebrafish performance in a multicue discrimination task, in which primarily focus and attention are required in order to obtain the reward.

Effects of Alcohol on Inhibitory Avoidance Learning in Zebrafish (Danio rerio).

The zebrafish (Danio rerio) can be used in studies addressing the effects of drugs on learning, memory, and anxiety. In the present study, we investigated the effect of different alcohol treatments (chronic and acute) on the learning and anxiety response of zebrafish in an inhibitory avoidance paradigm. Zebrafish were initially exposed to different alcohol treatments and submitted to an inhibitory avoidance protocol, where an electroshock was applied to the fish as they swam from the white to the black side of a shuttle box tank (naturally preferred environment of zebrafish). Animals from the control and 0.5% acute alcohol groups exhibited high latency to enter the black side of the tank after the first exposure to electroshock, in addition to higher freezing and a shorter distance from the bottom of the tank, suggesting acute alcohol exposure did not affect aversive learning in zebrafish. However, chronic exposure and alcohol withdrawal impaired the fish's capacity to properly respond to the aversive stimulus. Overall, our results show the harmful effects of chronic alcohol exposure, both continued intake and its cessation, but avoidance behavior persisted and anxiety increased following acute alcohol exposure.

Good night, sleep tight: The effects of sleep deprivation on spatial associative learning in zebrafish.

Learning and memory are vital to an animal's survival, and numerous factors can disrupt cognitive performance. Sleep is an evolutionarily conserved physiological process known to be important for the consolidation of learning and memory. The zebrafish has emerged as a powerful model organism sharing organizational and functional characteristics with other vertebrates, providing great translational relevance. In our study, we used a simple spatial associative learning task to quantify the effects of sleep deprivation (partial vs. total) on learning performance in zebrafish, using an animated conspecific shoal image as a reward. Control animals maintained on a regular light:dark cycle were able to acquire the association between the unconditioned and conditioned stimulus, reinforcing zebrafish as a valid and reliable model for appetitive conditioning tasks. Notably, sleep deprivation did not alter the perception of and response to the conspecific image. In contrast, although partial sleep deprivation did not impair cognitive performance, total sleep deprivation significantly impaired performance on the associative learning task. Our results suggest that sleep is important for learning and memory, and that the effects of sleep deprivation on these processes can be investigated in zebrafish.

Sleep deprivation effects on object discrimination task in zebrafish (Danio rerio).

The zebrafish is an ideal vertebrate model for neurobehavioral studies with translational relevance to humans. Many aspects of sleep have been studied, but we still do not understand how and why sleep deprivation alters behavioral and physiological processes. A number of hypotheses suggest its role in memory consolidation. In this respect, the aim of this study was to analyze the effects of sleep deprivation on memory in zebrafish (Danio rerio), using an object discrimination paradigm. Four treatments were tested: control, partial sleep deprivation, total sleep deprivation by light pulses, and total sleep deprivation by extended light. The control group explored the new object more than the known object, indicating clear discrimination. The partially sleep-deprived group explored the new object more than the other object in the discrimination phase, suggesting a certain degree of discriminative performance. By contrast, both total sleep deprivation groups equally explored all objects, regardless of their novelty. It seems that only one night of sleep deprivation is enough to affect discriminative response in zebrafish, indicating its negative impact on cognitive processes. We suggest that this study could be a useful screening tool for cognitive dysfunction and a better understanding of the effect of sleep-wake cycles on cognition.