Developing Biomimetic Hydrogels of the Arterial Wall as a Prothrombotic Substrate for In Vitro Human Thrombosis Models.

Current in vitro thrombosis models utilise simplistic 2D surfaces coated with purified components of the subendothelial matrix. The lack of a realistic humanised model has led to greater study of thrombus formation in in vivo tests in animals. Here we aimed to develop 3D hydrogel-based replicas of the medial and adventitial layers of the human artery to produce a surface that can optimally support thrombus formation under physiological flow conditions. These tissue-engineered medial- (TEML) and adventitial-layer (TEAL) hydrogels were developed by culturing human coronary artery smooth muscle cells and human aortic adventitial fibroblasts within collagen hydrogels, both individually and in co-culture. Platelet aggregation upon these hydrogels was studied using a custom-made parallel flow chamber. When cultured in the presence of ascorbic acid, the medial-layer hydrogels were able to produce sufficient neo-collagen to support effective platelet aggregation under arterial flow conditions. Both TEML and TEAL hydrogels possessed measurable tissue factor activity and could trigger coagulation of platelet-poor plasma in a factor VII-dependent manner. Biomimetic hydrogel replicas of the subendothelial layers of the human artery are effective substrates for a humanised in vitro thrombosis model that could reduce animal experimentation by replacing current in vivo models.

A Biomimicking and Multiarm Self-Indicating Nanoassembly for Site-Specific Photothermal-Potentiated Thrombolysis Assessed in Microfluidic and In Vivo Models.

Thrombolytic and antithrombotic therapies are limited by short circulation time and the risk of off-target hemorrhage. Integrating a thrombus-homing strategy with photothermal therapy are proposed to address these limitations. Using glycol chitosan, polypyrrole, iron oxide and heparin, biomimicking GCPIH nanoparticles are developed for targeted thrombus delivery and thrombolysis. The nanoassembly achieves precise delivery of polypyrrole, exhibiting biocompatibility, selective accumulation at multiple thrombus sites, and enhanced thrombolysis through photothermal activation. To simulate targeted thrombolysis, a microfluidic model predicting thrombolysis dynamics in realistic pathological scenarios is designed. Human blood assessments validate the precise homing of GCPIH nanoparticles to activated thrombus microenvironments. Efficient near-infrared phototherapeutic effects are demonstrated at thrombus lesions under physiological flow conditions ex vivo. The combined investigations provide compelling evidence supporting the potential of GCPIH nanoparticles for effective thrombus therapy. The microfluidic model also offers a platform for advanced thrombolytic nanomedicine development.

Inovação e ética: a adoção de técnicas substitutivas ao uso de animais no diagnóstico laboratorial da raiva / Innovation and ethics; the adoption of alternative techniques to the use of animals in the laboratory diagnosis of rabies

Abordagens interdisciplinares vêm ganhando maior reconhecimento e destaque nas comunidades de saúde humana e animal, principalmente pela (re)emergência de diversas doenças infecciosas que emanam da interface humano-animal-ambiente. A raiva, zoonose grave, considerada endêmica no Brasil e globalmente negligenciada, é um exemplo. Tanto a vigilância epidemiológica quanto a confirmação dessa doença dependem do diagnóstico laboratorial, que é realizado, frequentemente, por meio dos testes de Imunofluorescência Direta (IFD) e de Isolamento Viral em Camundongo (IVC), via inoculação intracerebral da amostra suspeita em camundongos lactentes ou desmamados. Entretanto, recentemente, a Organização Mundial da Saúde reconheceu a Transcrição Reversa seguida da Reação em Cadeia da Polimerase (RT-PCR) como uma técnica primária válida para esse diagnóstico, podendo ser empregada como alternativa ao uso de animais, evitando sofrimento e morte. Esta dissertação apresenta uma discussão sobre as implicações técnicas e éticas da (não) adoção desse método substitutivo, considerando que todos os animais devem ser respeitados e entendidos como sujeitos singulares em suas percepções do mundo, não como objetos de pesquisa. Esse fato corrobora a necessidade de novas perspectivas que ressignifiquem nossas relações com os animais não humanos, o que é primordial para o estabelecimento de mudanças sistêmicas, de caráter ético-político, que visem o fim da instrumentalização animal e de seu uso no âmbito científico, bem como de qualquer forma de opressão.

Interdisciplinary approaches have been gaining greater recognition and prominence in the human and animal health communities, mainly due to the (re)emergence of several infectious diseases that emanate from the human-animal-environment interface. Rabies is an example, considered a serious zoonosis endemic in Brazil and globally neglected. Both epidemiological surveillance and confirmation of this disease depend on laboratory diagnosis, which is usually performed by the direct fluorescent antibody test (DFAT) and the mouse inoculation test (MIT) via intracranial inoculation of the suspected sample into suckling or weanling mice. However, the World Health Organization recently recognized the reverse transcription polymerase chain reaction (RT-PCR) as a valid primary technique for this diagnosis, which can replace the use of animals, avoiding suffering and death. This study presents a discussion about the technical and ethical implications of (not) adopting this alternative method, considering that all animals must be respected and understood as unique beings with their perceptions of the world, not as objects of research. It also further corroborates the need for new perspectives that reframe our relationships with non-human animals, which is fundamental for the implementation of systemic ethical-political changes, aiming at the end of animal instrumentalization and use in scientific research, as well as all forms of oppression.

Adapting to Changes in Publishing When Searching for Alternatives and Reporting on Animal Research: A Librarian's Perspective.

Since the inaugural issue of ATLA, many changes within publishing have occurred, impacting when, where, and how researchers conduct literature searches for non-animal alternatives. Such changes include increased rate of growth in scientific publications, greater number of databases and online resources available to search, opportunities for open and almost immediate dissemination of research outputs such as preprints and method protocols, and the development of reporting guidelines for animal research. Here we offer a librarian's perspective on these changes and advice on how to manage them to enable robust and diverse alternatives to be implemented in future research.

In Vivo Microdialysis in Mice Captures Changes in Alzheimer's Disease Cerebrospinal Fluid Biomarkers Consistent with Developing Pathology.

BACKGROUND: Preclinical models of Alzheimer's disease (AD) can provide valuable insights into the onset and progression of the disease, such as changes in concentrations of amyloid-ß (Aß) and tau in cerebrospinal fluid (CSF). However, such models are currently underutilized due to limited advancement in techniques that allow for longitudinal CSF monitoring. OBJECTIVE: An elegant way to understand the biochemical environment in the diseased brain is intracerebral microdialysis, a method that has until now been limited to short-term observations, or snapshots, of the brain microenvironment. Here we draw upon patient-based findings to characterize CSF biomarkers in a commonly used preclinical mouse model for AD. METHODS: Our modified push-pull microdialysis method was first validated ex vivo with human CSF samples, and then in vivo in an AD mouse model, permitting assessment of dynamic changes of CSF Aß and tau and allowing for better translational understanding of CSF biomarkers. RESULTS: We demonstrate that CSF biomarker changes in preclinical models capture what is observed in the brain; with a decrease in CSF Aß observed when plaques are deposited, and an increase in CSF tau once tau pathology is present in the brain parenchyma. We found that a high molecular weight cut-off membrane allowed for simultaneous sampling of Aß and tau, comparable to CSF collection by lumbar puncture in patients. CONCLUSION: Our approach can further advance AD and other neurodegenerative research by following evolving neuropathology along the disease cascade via consecutive sampling from the same animal and can additionally be used to administer pharmaceutical compounds and assess their efficacy.

O oftalmologista e as uvas: Um modelo de treinamento microcirúrgico / The ophthalmologist and the grapes: A microsurgical training model

Resumo Objetivo: Desenvolver um modelo de treinamento de cirurgias corneanas utilizando uvas. Métodos: Foram empregadas uvas como estruturas que mimetizam o tamanho do globo ocular humano, recobertas com materiais de látex, simulando a pratica de cirurgias de córnea utilizando um sistema de videomagnificação. Foram realizados oito pontos simples. Foi avaliado o tempo de confecção do procedimento. Resultados: Foram realizadas 25 simulações como o modelo descrito. O tempo médio de realização da rafia foi de 34,56 ±5,79 minutos. A análise da correlação entre o tempo e a ordem das cirurgias mostrou uma redução no tempo de confecção. Conclusão: O modelo de treinamento oftalmológico utilizando uvas mostrou-se capaz de simular as etapas básicas do treinamento de suturas microcirúrgicas.

Abstract Objective: Develop a training model for corneal surgery using grapes. Methods: Grapes were used as structures that mimic the size of the human eyeball, covered with latex materials, simulating the practice of corneal surgery using a videomagnification system. Eight simple stitches were performed. The surgical time was evaluated. Results: 25 simulations were carried out as the model described. The mean time taken for the raffia was 34.56 ± 5.79 minutes. The analysis of the correlation between the time and the order of the surgeries showed a reduction in the confection time. Conclusion: The ophthalmic training model using grapes proved to be capable of simulating the basic stages of microsurgery suture training.

The Contribution of Rat Studies to Current Knowledge of Major Depressive Disorder: Results From Citation Analysis.

Major depressive disorder (MDD) is the most severe depression type and one of the leading causes of morbidity worldwide. Animal models are widely used to understand MDD etiology, pathogenesis, and treatment, but the efficacy of this research for patients has barely been systematically evaluated. Such evaluation is important given the resource consumption and ethical concerns incurred by animal use. We used the citation tracking facilities within Web of Science and Scopus to locate citations of original research papers on rats related to MDD published prior to 2013-to allow adequate time for citations-identified in PubMed and Scopus by relevant search terms. Resulting citations were thematically coded in eight categories, and descriptive statistics were calculated. 178 publications describing relevant rat studies were identified. They were cited 8,712 times. More than half (4,633) of their citations were by other animal studies. 794 (less than 10%) were by human medical papers. Citation analysis indicates that rat model research has contributed very little to the contemporary clinical understanding of MDD. This suggests a misuse of limited funding hence supporting a change in allocation of research and development funds targeting this disorder to maximise benefits for patients.

Acute toxicity "six-pack" studies supporting approved new drug applications in the U.S., 2015-2018.

The acute toxicity "six-pack" is a battery of animal tests used to evaluate acute systemic toxicity by three routes of exposure, skin and eye irritation/corrosion, and skin sensitization. A perception exists that these tests are not required for pharmaceuticals. For the four years from 2015 through 2018, we tallied the number of corresponding tests submitted by sponsors in approved, original new drug applications, as reported by the U.S. Food and Drug Administration (FDA) in publicly available reviews. In 125 reviews, we identified 228 single dose acute toxicity studies, 62 in vivo local tolerance studies, and 32 in vivo skin sensitization studies, as well as 37 in vitro or ex vivo local tolerance studies. A total of 4798 animals were used in these studies; however, FDA's reporting was inconsistent, and we estimate the actual number of animals used to be 8998. For the evaluation of single dose acute toxicity, we accessed two guidance documents with conflicting recommendations regarding routes of administration and number of species to be used. For the evaluation of local tolerance and skin sensitization, most studies examined were conducted by routes other than that intended for human administration. Non-animal methods used to evaluate skin sensitization were not reported.

The Relevance of In Silico, In Vitro and Non-human Primate Based Approaches to Clinical Research on Major Depressive Disorder.

Major depressive disorder (MDD) is the most severe form of depression and the leading cause of disability worldwide. When considering research approaches aimed at understanding MDD, it is important that their effectiveness is evaluated. Here, we assessed the effectiveness of original studies on MDD by rating their contributions to subsequent medical papers on the subject, and we compared the respective contribution of findings from non-human primate (NHP) studies and from human-based in vitro or in silico research approaches. For each publication, we conducted a quantitative citation analysis and a systematic qualitative analysis of the citations. In the majority of cases, human-based research approaches (both in silico and in vitro) received more citations in subsequent human research papers than did NHP studies. In addition, the human-based approaches were considered to be more relevant to the hypotheses and/or to the methods featured in the citing papers. The results of this study suggest that studies based on in silico and in vitro approaches are taken into account by medical researchers more often than are NHP-based approaches. In addition, these human-based approaches are usually cheaper and less ethically contentious than NHP studies. Therefore, we suggest that the traditional animal-based approach for testing medical hypotheses should be revised, and more opportunities created for further developing human-relevant innovative techniques.

Adaptation of Mammalian Cells to Chemically Defined Media.

In order to circumvent ethical, technical, and economic drawbacks regarding the use of animal serum in cell culturing, it is possible to adapt mammalian cells to serum-free media. Nowadays, there are several serum-free formulations available, including fully animal derived-free and chemically defined media, and different adaptation techniques. This article focuses on the gradual adaptation of a mammalian suspension cell culture to a chemically defined medium. The first step is to transfer the cells cultured in medium supplemented with fetal bovine serum (FBS) to a chemically defined medium of your choice, containing the same amount of FBS. The next steps consist of progressively reducing the amount of FBS, while monitoring cell growth and viability up to the complete elimination of FBS. This protocol has been successfully used to adapt THP-1 cells to a chemically defined medium with similar maximum specific growth rate as those cultured with FBS. © 2019 by John Wiley & Sons, Inc. Basic Protocol: Gradual adaptation to chemically defined medium Alternate Protocol: Direct adaptation to chemically defined medium Support Protocol 1: Determining maximum specific growth rate of a cell culture Support Protocol 2: Cell freezing and thawing.

Conhecimento sobre a bioética e a Lei 11. 794/2008 na graduação / The knowledge of bioethics and Law 11, 794/2008 in undergraduate courses / Conocimiento sobre bioética y la Ley 11. 794/2008 en el grado

Resumo O uso de animais para fins didáticos e de pesquisa requer cuidados específicos. Atualmente, vigora no Brasil a Lei 11.794/2008, que rege parâmetros legais de manejo e conduta neste caso. Esta lei foi acompanhada da instalação ou adequação de comissões de ética em instituições que utilizam animais para ensino e investigação, bem como da criação do Conselho Nacional de Controle de Experimentação Animal. No entanto, apesar dos avanços, especialmente na legislação, ainda não foi consolidada nenhuma grande mudança de comportamento de pesquisadores e alunos de graduação que manuseiam animais em laboratório. A divulgação de informações deixa a desejar, e a prática acaba por repercutir a carência de reflexão ética. Este artigo busca averiguar o atual conhecimento bioético de alunos de graduação e professores com o objetivo de estimular mudanças de conduta. Aprovado CEP-Unioeste CAAE 8563417.8.0000.0107

Abstract Used for education and research, laboratory animals require special care on their handling. Brazilian Law 11,794/2008 establishes the legal parameters for animal manipulation and welfare. It was accompanied by the obligatory installation of the Institutional Ethics Committees on the Use of Animals and the creation of the National Council for Animal Experimentation Control. There have been advances in the field of animal bioethics legislation. However, considering the behavior of those who handle the animals in laboratory environment, especially undergraduate students, these advances are insufficient: the information does not reach them and their attitudes remain in need of ethical reflection. This article seeks to investigate the current bioethical knowledge of undergraduate students and teachers in order to stimulate changes in conduct. Aprovado CEP-Unioeste CAAE: 78563417.8.0000.0107

Resumen El uso de animales con fines didácticos y de investigación requiere cuidados específicos. Actualmente, rige en Brasil la Ley 11.794/2008 que regula los parámetros legales de manejo y conducta en estos casos. Esta ley estuvo acompañada de la instalación o adecuación de comisiones de ética en instituciones que utilizan animales para enseñanza e investigación, así como de la creación del Consejo Nacional de Control de Experimentación Animal. No obstante, a pesar de los avances, especialmente en la legislación, aún no se ha consolidado ninguna gran transformación en el comportamiento de los investigadores y alumnos que manipulan animales en el laboratorio. La divulgación de informaciones es insuficiente, y la práctica acaba reflejando la falta de reflexión ética. Este artículo procura identificar el conocimiento bioético actual de los alumnos de grado y de los profesores, con el objetivo de estimular cambios en la conducta. Aprovado CEP-Unioeste CAAE: 78563417.8.0000.0107

Students' perception of animal or virtual laboratory in physiology practical classes in PBL medical hybrid curriculum.

Over the years, much criticism against animal use for physiology teaching has been made. Hence, replacement by suitable alternatives has increased in several pedagogical approaches. This study examined students' perceptions of animal versus virtual (video/computer) laboratory classes in physiological sciences associated with the effectiveness of the problem-based learning (PBL) hybrid curriculum. Three cohorts of medical students from the University of Ribeirão Preto, who participated in animal or virtual physiology classes or both, were asked to fill out a 5-point Likert questionnaire about knowledge acquisition/motivation, importance to PBL learning goals, skills acquired, need for animal use, academic formation, learning impairment, and alternative methods. We also assessed their grades in the final exam. A total of 350 students were included, in which 108 participated only in virtual classes, 120 only in practical animal laboratory classes, and 122 in both approaches. The majority agreed that the two methods improved their knowledge acquisition/motivation and helped to reinforce tutorial goals and to acquire skills. However, the cohort who experienced both approaches favored animal laboratory. Students believe animal use is needed and did not impair their learning. Conversely, their opinion about academic formation without animal laboratory classes was divided, as was whether this approach inspired them to seek alternative methods. Despite the different perceptions, there was no difference among the groups' final grades (7.3 ± 1 vs. 7.2 ± 1 vs. 7.2 ± 2 for virtual or practical animal laboratory classes or both, respectively). Therefore, virtual activities are not as effective as animal use in the opinions of the students, but they are successful strategies in physiology learning that can be used in practical classes in a hybrid PBL curriculum.

Refining humane endpoints in mouse models of disease by systematic review and machine learning-based endpoint definition.

Ideally, humane endpoints allow for early termination of experiments by minimizing an animal's discomfort, distress and pain, while ensuring that scientific objectives are reached. Yet, lack of commonly agreed methodology and heterogeneity of cut-off values published in the literature remain a challenge to the accurate determination and application of humane endpoints. With the aim to synthesize and appraise existing humane endpoint definitions for commonly used physiological parameters, we conducted a systematic review of mouse studies of acute and chronic disease models, which used body weight, temperature and/or sickness scores for endpoint definition. In the second part of the study, we used previously published and unpublished data on weight, temperature and sickness scores from mouse models of sepsis and stroke and applied machine learning algorithms to assess the usefulness of this method for parameter selection and endpoint definition across models. Studies were searched for in two electronic databases (MEDLINE/Pubmed and Embase). Out of 110 retrieved full-text manuscripts, 34 studies were included. We found large intra- and inter-model variance in humane endpoint determination and application due to varying animal models, lack of standardized experimental protocols and heterogeneity of performance metrics (part 1). Machine learning models trained with physiological data and sickness severity score or modified DeSimoni neuroscore identified animals with a high risk of death at an early time point in both mouse models of stroke (male: 93.2% at 72h post-treatment; female: 93.0% at 48h post-treatment) and sepsis (96.2% at 24h post-treatment), thus demonstrating generalizability in endpoint determination across models (part 2).

Global Collaboration to Modernize Advanced Trauma Life Support Training.

BACKGROUND: Each year, thousands of surgeons and other trauma health care providers participate in the American College of Surgeon's Advanced Trauma Life Support (ATLS) program, which historically has allowed trainees to practice cricothyroidotomy, chest tube insertion, pericardiocentesis, venous cutdown, and diagnostic peritoneal lavage on live dogs, pigs, sheep, and goats. However, more than 99% of ATLS programs in the United States and Canada have now ended animal use, driven primarily by simulation technology advancements. OBJECTIVE: This review details an international survey of animal versus simulation use in ATLS programs and summarizes the surgical training impact of a novel collaboration between the industry manufacturer of the TraumaMan human simulator, Simulab Corporation (Seattle, Washington), and an animal protection nongovernmental organization (NGO) based in Norfolk, Virginia, to replace animal use in ATLS programs with human simulators. METHODS: From 2012 through 2017, the NGO e-mailed formal surveys concerning program statistics and animal use practices to ATLS officials in various countries (N = 64). The survey response rate was 87.5% and included pre- and post-comparison surveys relative to the industry-NGO simulation collaboration. RESULTS: Eighteen ATLS programs (32.1%) initially replied that they use nonanimal training methods, whereas 38 ATLS programs (67.8%) replied that they use animals for surgical skills training and cited financial constraints as the primary barrier to adopting human simulation methods. Through the industry-NGO collaboration, the NGO donated 119 TraumaMan models valued at nearly $3 million (USD) to ATLS programs in 22 countries, such that 75% of those ATLS programs surveyed by the NGO now use exclusively nonanimal simulation models. CONCLUSIONS: The industry-NGO collaboration successfully transformed the surgical skills laboratories of 22 international ATLS programs to replace animal use with nonanimal simulation models that are more anatomically realistic, cost less, and allow trainees to repeat surgical skills until proficiency.

Adaptation of a skin sensitization assay to a chemically defined culture.

There are currently three in vitro methods adopted by the Organization for the Economic Co-operation and Development for testing chemicals based on the third key event of the skin sensitization adverse outcome pathway, the activation of dendritic cells. All of them use culture medium supplemented with fetal bovine serum (FBS), which brings technical disadvantages and animal welfare concerns. The objective of this study was to analyze the possibility of eliminating the use of FBS in the human Cell Line Activation Test (h-CLAT). After successful implementation of the h-CLAT using THP-1 cells cultured in FBS-containing medium, several attempts to adapt THP-1 cells to four different serum-free media were made. The best results were obtained with gradual adaptation to RPMI-1640 medium with HL-1™ Supplement and to X-VIVO™ 10. Adapted cells were cryopreserved and submitted to the reactivity check. After being approved, they were used in dose finding and proficiency assays. Despite minor adjustments in the original protocol, it was possible to correctly predict the sensitizing potential of the ten proficiency substances using THP-1 cells adapted to X-VIVO™ 10, which indicates that it is possible to eliminate the use of FBS in the h-CLAT, using a chemically defined medium.

Characterization and In Vivo Validation of a Three-Dimensional Multi-Cellular Culture Model to Study Heterotypic Interactions in Colorectal Cancer Cell Growth, Invasion and Metastasis.

Colorectal cancer (CRC) is the third cause of cancer-related mortality in industrialized countries. Local invasion and metastasis formation are events associated with poor prognosis for which today there are no effective therapeutic options. Invasion and metastasis are strongly modulated by cells of the tumor microenvironment (TME), in particular fibroblasts and endothelial cells. Unraveling interactions between tumor cells and cells of the TME may identify novel mechanisms and therapeutic targets to prevent or treat metastasis. We report here the development and in vivo validation of a 3D tumor spheroid model to study the interactions between CRC cells, fibroblasts and endothelial cells in vitro. Co-cultured fibroblasts promoted SW620 and HCT116 CRC spheroid invasion, and this was prevented by the SRC and FGFR kinase inhibitors Dasatinib and Erdafitinib, respectively. To validate these findings in vivo, we injected SW620 cells alone or together with fibroblasts orthotopically in the caecum of mice. Co-injection with fibroblasts promoted lung metastasis growth, which was fully reversed by treatment with Dasatinib or Erdafitinib. Co-culture of SW620 or HCT116 CRC spheroids with endothelial cells suppressed spheroid growth while it had no effect on cancer cell migration or invasion. Consistent with this in vitro effect, co-injected endothelial cells significantly inhibited primary tumor growth in vivo. From these experiments we conclude that effects on cancer cell invasion and growth induced by co-cultured TME cells and drug treatment in the 3D spheroid model in vitro, are predictive of in vivo effects. The 3D spheroid model may be considered as an attractive model to study the effect of heterotypic cellular interactions and drug activities on cancer cells, as animal testing alternative. This model may be adapted and further developed to include different types of cancer and host cells and to investigate additional functions and drugs.

Ethical and Scientific Pitfalls Concerning Laboratory Research with Non-Human Primates, and Possible Solutions.

Basic and applied laboratory research, whenever intrusive or invasive, presents substantial ethical challenges for ethical committees, be it with human beings or with non-human animals. In this paper we discuss the use of non-human primates (NHPs), mostly as animal models, in laboratory based research. We examine the two ethical frameworks that support current legislation and guidelines: deontology and utilitarianism. While human based research is regulated under deontological principles, guidelines for laboratory animal research rely on utilitarianism. We argue that the utilitarian framework is inadequate for this purpose: on the one hand, it is almost impossible to accurately predict the benefits of a study for all potential stakeholders; and on the other hand, harm inflicted on NHPs (and other animals) used in laboratory research is extensive despite the increasing efforts of ethics committees and the research community to address this. Although deontology and utilitarianism are both valid ethical frameworks, we advocate that a deontological approach is more suitable, since we arguably have moral duties to NHPs. We provide suggestions on how to ensure that research currently conducted in laboratory settings shifts towards approaches that abide by deontological principles. We assert that this would not impede reasonable scientific research.

* Ethical Issues in the Use of Animal Models for Tissue Engineering: Reflections on Legal Aspects, Moral Theory, Three Rs Strategies, and Harm-Benefit Analysis.

Animal experimentation requires a solid and rational moral foundation. Objective and emphatic decision-making and protocol evaluation by researchers and ethics committees remain a difficult and sensitive matter. This article presents three perspectives that facilitate a consideration of the minimally acceptable standard for animal experiments, in particular, in tissue engineering (TE) and regenerative medicine. First, we review the boundaries provided by law and public opinion in America and Europe. Second, we review contemporary moral theory to introduce the Neo-Rawlsian contractarian theory to objectively evaluate the ethics of animal experiments. Third, we introduce the importance of available reduction, replacement, and refinement strategies, which should be accounted for in moral decision-making and protocol evaluation of animal experiments. The three perspectives are integrated into an algorithmic and graphic harm-benefit analysis tool based on the most relevant aspects of animal models in TE. We conclude with a consideration of future avenues to improve animal experiments.

Utilização de animais em pesquisas: breve revisão da legislação no Brasil / Use of animals in research: a brief review of legislation in Brazil / Utilización de animales en la investigación: breve revisión de la legislación en Brasil

A utilização de animais para fins científicos configura prática histórica na civilização humana, mas gera polêmica em sociedades preocupadas com proteção dos animais. No Brasil, até 2008, não havia norma ou lei que regulamentasse especificamente a experimentação animal. Este trabalho discute a utilização de animais em experimentos científicos, considerando o delineamento da Lei Arouca, por meio da leitura de artigos científicos que contemplam o histórico da experimentação no contexto mundial e brasileiro, incluindo a regulamentação do uso de animais do filo Chordata, subfilo Vertebrata, em pesquisas no Brasil. A Lei Arouca pode representar avanço na legislação brasileira quanto à utilização de animais para fins científicos, sobretudo pela criação das comissões de ética para uso de animais em instituições de pesquisa e do Conselho Nacional de Controle de Experimentação Animal, que examinam o cumprimento da legislação aplicável em projetos científicos que envolvem a utilização de animais.

The use of animals for scientific purposes is a historical procedure in human civilization, but is controversial for societies concerned with the protection of animals. In Brazil, until 2008, there was no rule or law that specifically regulated animal testing. This paper discusses the use of animals in scientific experiments, considering the Brazilian Arouca Law, through the analysis of scientific articles that consider the history of experimentation in the world and in Brazil, including the regulation of the use of animals of the phylum Chordata, subphylum Vertebrata, in Brazilian research. The Arouca Law may represent an advance in Brazilian law regarding the use of animals for scientific purposes, particularly given the creation of the Ethics Committees for Animal Use in research institutions and the National Council for Animal Experimentation Control, which examine the compliance of scientific projects involving the use of such animals to applicable law.

El uso de animales para fines científicos configura una práctica histórica en la civilización humana, pero genera controversia en las sociedades preocupadas por la protección de éstos. En Brasil, hasta 2008, no había una norma o una ley que regulara la experimentación animal. Este trabajo discute acerca del uso de animales en experimentos científicos, teniendo en cuenta los lineamientos de la Ley Arouca, a partir de la lectura de artículos científicos que abordan la historia de la experimentación animal en el mundo y en el contexto brasilero, incluyendo la regulación del uso de animales del filo Cordados, subfilo Vertebrados, en investigaciones en Brasil. La Ley Arouca puede representar un avance en la legislación brasilera con respecto al uso de estos animales para fines científicos, sobre todo por la creación de las comisiones de ética para el uso de animales (Ceua) en instituciones de investigación y del Consejo Nacional de Control de la Experimentación Animal (Concea), que son los responsables de examinar el cumplimiento de la legislación aplicable a proyectos científicos que involucran la utilización de animales.

El 1, 2, 3 de la experimentación con animales de laboratorio / The 1, 2, 3 of laboratory animal experimentation

RESUMEN El lento desarrollo científico experimentado en América Latina en las últimas décadas ha retrasado la incorporación de la experimentación con animales de laboratorio, sin embargo, esta realidad ha comenzado a cambiar. En la actualidad, se evidencia un extraordinario progreso científico que ha promovido la introducción e incremento del uso de animales de laboratorio como una importante herramienta para el avance de las ciencias biomédicas. A raíz de este auge, surge la necesidad de proporcionar a la comunidad científica la formación y directrices en todos los aspectos relacionados con la experimentación animal. Es responsabilidad de cada país legislar esta práctica para que no solo por razones bioéticas, sino también legales, se considere el derecho de los animales y con ello su bienestar. El siguiente manuscrito es el resultado de las comunicaciones presentadas en el Taller Internacional en Ciencias de Animales de Laboratorio celebrado en la Universidad Técnica de Ambato, Ecuador; contiene la actualidad en la ciencia de animales de laboratorio, haciendo énfasis en aspectos fundamentales, tales como: principales especies utilizadas y su biología, principios éticos y legales, diseño experimental y alternativas al uso de animales, todas ellas encaminadas a garantizar las buenas prácticas que deben caracterizar el quehacer científico. Sin duda, este documento será relevante tanto para aquellos investigadores que pretenden incorporar la experimentación animal a sus investigaciones, como para aquellos que, aun teniendo experiencia, pretendan actualizar sus conocimientos.

ABSTRACT The slow scientific development in Latin America in recent decades has delayed the incorporation of laboratory animal experimentation; however, this situation has started to change. Today, extraordinary scientific progress is evident, which has promoted the introduction and increased use of laboratory animals as an important tool for the advancement of biomedical sciences. In the aftermath of this boom, the need to provide the scientific community with training and guidance in all aspects related to animal experimentation has arisen. It is the responsibility of each country to regulate this practice, for both bioethical and legal reasons, to ensure consideration of the animals' rights and welfare. The following manuscript is the result of papers presented at the International Workshop on Laboratory Animal Testing held at the Technical University of Ambato, Ecuador; it contains information regarding the current state of affairs in laboratory animal testing and emphasizes critical aspects such as main species used, ethical and legal principles, and experimental and alternative designs for animal use. These works aim to ensure good practices that should define scientific work. This document will be relevant to both researchers who aim to newly incorporate animal testing into their research and those who seek to update their knowledge.