Preliminary survey of the attitudes of Brazilian scientists towards pain management and assessment in animals used in science.

OBJECTIVE: To investigate the current scenario in Brazil regarding pain assessment and control in experimental animals. STUDY DESIGN: Prospective survey. METHODS: A confidential questionnaire was available online and sent by e-mail to Brazilian scientists working with animal experimentation in Brazil. Data collection was conducted from October 2016 to October 2017. The exclusion criteria included blank questionnaires or with <80% completed responses, researchers not performing experiments involving animals and foreign scientists. RESULTS: A total of 96 questionnaires from 104 respondents were analyzed. The Fisher's exact test showed a disparity between the proportions of scientists who recognized the importance of analgesia and their application of analgesic techniques in painful procedures (p < 0.0003), and also for the researchers who assumed that experiments inflicted pain and their classification of the degree of invasiveness (p < 0.0001), indicating their insufficient knowledge of these topics. Overall, 77% of institutions did not offer specific training to assess pain in experimental animals, and 24% of respondents had no training to work with animal experimentation. In total, 62% of the studies inflicted pain, 48% of respondents used pain scales, and the drugs administered most frequently for pain management were morphine (44%), meloxicam (43%) and tramadol (37%); 15% of respondents did not include analgesics even though their studies inflicted pain. Commonly used animals were rats (33%), mice (29%) and rabbits (8%). CONCLUSIONS AND CLINICAL RELEVANCE: The results of this preliminary survey indicated that in Brazil there is a gap in the knowledge and training on pain assessment and management of experimental animals. Therefore, there is a necessity for an educational program to prepare and train scientists to assess and manage pain in laboratory or experimental animals. Further studies using a psychometrically validated survey instrument are warranted.

Postoperative Analgesic Effects of Laserpuncture and Meloxicam in Bitches Submitted to Ovariohysterectomy.

BACKGROUND: This prospective, randomised and blind study investigated the efficacy of laserpuncture for postoperative pain management in dogs. METHOD: Sixteen bitches were sedated with acepromazine and randomly treated before ovariohysterectomy with meloxicam 0.2 mg·kg-1 intramuscular or laserpuncture (wavelength 904 mm, frequency 124 Hz, potency 10 Joules, 100 s in each acupoint). Anaesthesia was performed with propofol, isoflurane/O2, and fentanyl. The Glasgow Composite Measure Pain Scale (GCMPS) and Dynamic Interactive Visual Analog Scale (DIVAS) were used to evaluate postoperative pain before and for 24 h after surgery. Morphine was administrated as rescue analgesia when pain scores were ≥3.33 (GCMPS). Differences between treatments, time points, and amount of rescue analgesia between groups were investigated by the Mann-Whitney test and the area under the curve (AUC) for GCMPS, Friedman, and Chi-squared tests, respectively (p < 0.05). RESULTS: Dogs treated with laserpuncture presented lower GCMPS AUC for 24 h and lower GCMPS scores at 2 and 4 h postoperatively (p = 0.04). Three dogs treated with meloxicam required postoperatively rescue analgesia against none treated with laserpuncture. CONCLUSIONS: In this preliminary study, laserpuncture mitigated postoperative pain in dogs following ovariohysterectomy, and the technique is a promising adjunct to perioperative pain management in dogs undergoing soft tissue surgery.

Potencial de transdiferenciação neural das células-tronco mesenquimais da medula óssea de equino / Potential of neural transdifferentiation of mesenchymal stem cells from equine bone marrow

Os primeiros estudos demonstrando o potencial de trandiferenciação neural das células-tronco mesenquimais (CTMs) provenientes da medula óssea (MO) foram conduzidos em camundogos e humanos no início da década de 2000. Após esse período, o número de pesquisas e publicações com o mesmo propósito tem aumentado, mas com raros ou escassos estudos na espécie equina. Nesse sentindo, o objetivo desse trabalho foi avaliar o potencial in vitro da transdiferenciação neural das CTMs provenientes da MO de equinos utilizando-se dois protocolos: P1 (forksolin e ácido retinóico) e P2 (2-βmecarptoetanol). Após a confirmação das linhagens mesenquimais, pela positividade para o marcador CD90 (X=97,94%), negatividade para o marcador CD34 e resposta positiva a diferenciação osteogênica, as CTMs foram submetidas a transdiferenciação neural (P1 e P2) para avaliação morfológica e expressão dos marcadores neurais GFAP e β3 tubulina por citometria de fluxo. Os resultados revelaram mudanças morfológicas em graus variados entre os protocolos testados. No protocolo 1, vinte quatro horas após a incubação com o meio de diferenciação neural, grande proporção de células (>80%) apresentaram morfologia semelhante a células neurais, caracterizadas por retração do corpo celular e grande número de projeções protoplasmáticas (filopodia). Por outro lado, de forma comparativa, já nos primeiros 30 minutos após a exposição ao antioxidante β-mercaptoetanol (P2) as CTMs apresentaram rápida mudança morfológica caracterizada principalmente por retração do corpo celular e menor número de projeções protoplasmáticas. Também ficou evidenciado com o uso deste protocolo, menor aderência das células após tempo de exposição ao meio de diferenciação, quando comparado ao P1. Com relação a análise imunofenotípica foi observado uma maior (P<0,001) expressão dos marcadores GFAP e β3 tubulina ao término do P2 quando comparado ao P1. A habilidade das CTMs em gerar tipos celulares relacionados a linhagem neural é complexa e multifatorial, dependendo não só dos agentes indutores, mas também do ambiente no qual estas células são cultivadas. Desta forma um maior número de estudos é necessário para o melhor entendimento do processo de transdiferenciação neural a partir de CTMs de equinos.

The first studies showing the potential of neural transdifferentiation of mesenchymal stem cells (MSCs) from bone marrow (BM) were conducted in camundogos and humans in the early 2000s. After this period, the number of research and publications with the same purpose increased, but with rare or scarce studies in horses. The aim of this study was to evaluate in vitro neuronal transdifferentiation potential of MSCs from equine BM using two protocols: P1 (forksolin and retinoic acid) and P2 (2-βmecarptoetanol). After confirming the mesenchymal lineages, by positivity for the marker CD90 (X=97.94%), negative for the marker CD34 and positive response for osteogenic differentiation, MSCs were subjected to neural transdifferentiation (P1 and P2) for morphological analysis and expression of neural markers GFAP and β3 tubulin by flow cytometry. The results revealed morphological changes in varying degrees between the tested protocols. In protocol 1, twenty four hours after incubation with the media of neural differentiation, a large proportion of cells (>80%) had similar morphology to neural cells, characterized by retraction of cellular body and a large number of cytoplasmic extension (filopodia). However, comparatively, within the first 30 minutes after exposure to the antioxidant β-mercaptoethanol (P2) MSCs showed rapid morphological changes characterized mainly by retraction of cellular body and less cytoplasmic extension. It was also evidenced with the use of this protocol, lower cellular adhesion after exposure to media when compared to P1. Regarding the immunophenotyping analysis it was observed a higher (P<0.001) expression of the markers GFAP and β3 tubulin at the end of P2 compared to P1. The ability of MSCs to generate cell types related to neural lineage is complex and multifactorial, depending not only of inducing agents, but also the environment in which these cells will be cultivated. Thus a greater number of studies are necessary to better understand the process of neural transdifferentiation of MSCs from equine.