Oligodendrocyte precursor cell-derived exosomes combined with cell therapy promote clinical recovery by immunomodulation and gliosis attenuation.

Multiple sclerosis is a chronic inflammatory disease of the central nervous system characterized by autoimmune destruction of the myelin sheath, leading to irreversible and progressive functional deficits in patients. Pre-clinical studies involving the use of neural stem cells (NSCs) have already demonstrated their potential in neuronal regeneration and remyelination. However, the exclusive application of cell therapy has not proved sufficient to achieve satisfactory therapeutic levels. Recognizing these limitations, there is a need to combine cell therapy with other adjuvant protocols. In this context, extracellular vesicles (EVs) can contribute to intercellular communication, stimulating the production of proteins and lipids associated with remyelination and providing trophic support to axons. This study aimed to evaluate the therapeutic efficacy of the combination of NSCs and EVs derived from oligodendrocyte precursor cells (OPCs) in an animal model of multiple sclerosis. OPCs were differentiated from NSCs and had their identity confirmed by gene expression analysis and immunocytochemistry. Exosomes were isolated by differential ultracentrifugation and characterized by Western, transmission electron microscopy and nanoparticle tracking analysis. Experimental therapy of C57BL/6 mice induced with experimental autoimmune encephalomyelitis (EAE) were grouped in control, treated with NSCs, treated with OPC-derived EVs and treated with a combination of both. The treatments were evaluated clinically using scores and body weight, microscopically using immunohistochemistry and immunological profile by flow cytometry. The animals showed significant clinical improvement and weight gain with the treatments. However, only the treatments involving EVs led to immune modulation, changing the profile from Th1 to Th2 lymphocytes. Fifteen days after treatment revealed a reduction in reactive microgliosis and astrogliosis in the groups treated with EVs. However, there was no reduction in demyelination. The results indicate the potential therapeutic use of OPC-derived EVs to attenuate inflammation and promote recovery in EAE, especially when combined with cell therapy.

Mesenchymal stem cells in animal reproduction: sources, uses and scenario.

Studies regarding mesenchymal stem cells turned up in the 1960's and this cell type created a great number of questions about its functions and applicability in science and medicine. When used with therapeutic intent, these cells present an inclination to migrate to sites of injury, inflammation or disease, where they secrete bioactive factors that stimulates the synthesis of new tissue. In this context, studies using rodents reported that MSCs promoted positive effects in the ovarian function in mice with premature aging of follicular reserve. In female bovines, experimental stem cell-based therapies have been used to either generate new oocytes with in vitro quality or stimulate such action in vivo. It is also reported, that the intraovarian application of mesenchymal stem cells generates a greater production of embryos in vitro and the production of early and expanded blastocysts. Additionally, analysis of ovarian tissue in animal subjected to treatment showed an increase in the number of developing follicles. Nevertheless, the treatments involving stem cells with different modes of application, different sources and different species were able to act on the hormonal, tissue, cellular and metabolic levels, generating positive results in the recovery and improvement of ovarian functions.

Estudos sobre células-tronco mesenquimais surgiram na década de 1960 e esse tipo de célula gerou muitas dúvidas sobre suas funções e aplicabilidade na ciência e na medicina. Quando utilizadas com intuito terapêutico, essas células apresentam tendência a migrar para locais de lesão, inflamação ou doença, onde secretam fatores bioativos que estimulam a síntese de novos tecidos. Nesse contexto, estudos utilizando roedores relataram que as CTM promoveram efeitos positivos na função ovariana em camundongos com envelhecimento precoce da reserva folicular. Em fêmeas bovinas, terapias experimentais baseadas em células-tronco têm sido utilizadas para gerar novos oócitos com qualidade in vitro ou estimular tal ação in vivo. É relatado também que a aplicação intraovariana de células-tronco mesenquimais gera maior produção de embriões in vitro e produção de blastocistos precoces e expandidos. Além disso, a análise do tecido ovariano em animais submetidos ao tratamento mostrou aumento no número de folículos em desenvolvimento. Apesar disso, os tratamentos envolvendo células-tronco com diferentes modos de aplicação, diferentes fontes e diferentes espécies foram capazes de atuar nos níveis hormonal, tecidual, celular e metabólico, gerando resultados positivos na recuperação e melhora das funções ovarianas.

Gene editing in small and large animals for translational medicine: a review.

The CRISPR/Cas9 system is a simpler and more versatile method compared to other engineered nucleases such as Zinc Finger Nucleases (ZFNs) and Transcription Activator-Like Effector Nucleases (TALENs), and since its discovery, the efficiency of CRISPR-based genome editing has increased to the point that multiple and different types of edits can be made simultaneously. These advances in gene editing have revolutionized biotechnology by enabling precise genome editing with greater simplicity and efficacy than ever before. This tool has been successfully applied to a wide range of animal species, including cattle, pigs, dogs, and other small animals. Engineered nucleases cut the genome at specific target positions, triggering the cell's mechanisms to repair the damage and introduce a mutation to a specific genomic site. This review discusses novel genome-based CRISPR/Cas9 editing tools, methods developed to improve efficiency and specificity, the use of gene-editing on animal models and translational medicine, and the main challenges and limitations of CRISPR-based gene-editing approaches.

Impact of ZrO2 Content on the Formation of Sr-Enriched Phosphates in Al2O3/ZrO2 Nanocomposites for Bone Tissue Engineering.

This study investigates the profound impact of the ZrO2 inclusion volume on the characteristics of Al2O3/ZrO2 nanocomposites, particularly influencing the formation of calcium phosphates on the surface. This research, aimed at advancing tissue engineering, prepared nanocomposites with 5, 10, and 15 vol% ZrO2, subjecting them to chemical surface treatment for enhanced calcium phosphate deposition sites. Biomimetic coating with Sr-enriched simulated body fluid (SBF) further enhanced the bioactivity of nanocomposites. While the ZrO2 concentration heightened the oxygen availability on nanocomposite surfaces, the quantity of Sr-containing phosphate was comparatively less influenced than the formation of calcium phosphate phases. Notably, the coated nanocomposites exhibited a high cell viability and no toxicity, signifying their potential in bone tissue engineering. Overall, these findings contribute to the development of regenerative biomaterials, holding promise for enhancing bone regeneration therapies.

Comparative Analysis of Fluorescent Labeling Techniques for Tracking Canine Amniotic Stem Cells.

The utmost aim of regenerative medicine is to promote the regeneration of injured tissues using stem cells. Amniotic mesenchymal stem cells (AmMSCs) have been used in several studies mainly because of their easy isolation from amniotic tissue postpartum and immunomodulatory and angiogenic properties and the low level of rejection. These cells share characteristics of both embryonic/fetal and adult stem cells and are particularly advantageous because they do not trigger tumorigenic activity when injected into immunocompromised animals. The large-scale use of AmMSCs for cellular therapies would greatly benefit from fluorescence labeling studies to validate their tracking in future therapies. This study evaluated the fluorophore positivity, fluorescence intensity, and longevity of canine AmMSCs. For this purpose, canine AmMSCs from the GDTI/USP biobank were submitted to three labeling conditions, two commercial fluorophores [CellTrace CFSE Cell Proliferation kit - CTrace, and CellTracker Green CMFDA - CTracker (CellTracker Green CMFDA, CT, #C2925, Molecular Probes®; Life Technologies)] and green fluorescent protein (GFP) expression after lentiviral transduction, to select the most suitable tracer in terms of adequate persistence and easy handling and analysis that could be used in studies of domestic animals. Fluorescence was detected in all groups; however, the patterns were different. Specifically, CTrace and CTracker fluorescence was detected 6 h after labeling, while GFP was visualized no earlier than 48 h after transduction. Flow cytometry analysis revealed more than 70% of positive cells on day 7 in the CTrace and CTracker groups, while fluorescence decreased significantly to 10% or less on day 20. Variations between repetitions were observed in the GFP group under the present conditions. Our results showed earlier fluorescence detection and more uniform results across repetitions for the commercial fluorophores. In contrast, fluorescence persisted for more extended periods in the GFP group. These results indicate a promising direction for assessing the roles of canine AmMSCs in regenerative medicine without genomic integration.

Addition of synthetic polymer in the freezing solution of mesenchymal stem cells from equine adipose tissue as a future perspective for reducing of DMSO concentration.

The regenerative therapies with stem cells (SC) has been increased by the cryopreservation, permitting cell storage for extended periods. However, the permeating cryoprotectant agents (CPAs) such as dimethylsulfoxide (DMSO) can cause severe adverse effects. Therefore, this study evaluated equine mesenchymal stem cells derived from adipose tissue (eAT-MSCs) in fresh (Control) or after slow freezing (SF) in different freezing solutions (FS). The FS comprise DMSO and non-permeating CPAs [Trehalose (T) and the SuperCool X-1000 (X)] in association or not, totalizing seven different FS: (DMSO; T; X; DMSO+T; DMSO+X; T+X, and DMSO+T+X). Before and after cryopreservation were evaluated, viability, colony forming unit (CFU), and cellular differentiation capacity. After freezing-thawing, the viability of the eAT-MSCs reduced (P< 0.05) in all treatments compared to the control. However, the viability of frozen eAT-MSCs in DMSO (80.3 ± 0.6) was superior (P<0.05) to the other FS. Regarding CFU, no difference (P>0.05) was observed between fresh and frozen cells. After freezing-thawing, the eAT-MSCs showed osteogenic, chondrogenic, and adipogenic lineages differentiation potential. Nonetheless, despite the significative reduction in the osteogenic differentiation capacity between fresh and frozen cells, no differences (P > 0.05) were observed among FS. Furthermore, the number of chondrogenic differentiation cells frozen in DMSO+X solution reduced (P<0.05) comparing to the control, without differ (P>0.05) to the other FS. The adipogenic differentiation did not differ (P>0.05) among treatments. In conclusion, although these findings confirm the success of DMSO to cryopreserve eAT-MSCs, the Super Cool X-1000 could be a promise to reduce the DMSO concentration in a FS.

As terapias regenerativas com células-tronco (CT) têm sido incrementadas pela criopreservação, permitindo o armazenamento celular. No entanto, os agentes crioprotetores (ACPs) penetrantes, como DMSO, podem causar efeitos adversos graves. Portanto, este estudo avaliou células-tronco mesenquimais equinas derivadas de tecido adiposo (CTM-TAe) in natura (Controle) ou após congelamento lento (CL) em diferentes soluções de congelamento (SC). As SCs compreendem DMSO e ACPs não permeáveis [Trealose (T) e o SuperCool X-1000 (X)] associados ou não: (DMSO; T; X; DMSO+T; DMSO+X; T +X e DMSO+T+X). Antes e após a criopreservação foram avaliados, viabilidade, unidade formadora de colônia (UFC) e capacidade de diferenciação celular. Após o congelamento-descongelamento, a viabilidade das CTM-TAe reduziu (P< 0,05) em todos os tratamentos em relação ao controle. Entretanto, a viabilidade das CTM-TAe congeladas em DMSO (80,3 ± 0,6) foi superior (P<0,05) às demais SC. Em relação às UFC, não houve diferença (P>0,05) entre células frescas e congeladas. Após congelamento-descongelamento, as CTM-TAe apresentaram potencial de diferenciação de linhagens osteogênicas, condrogênicas e adipogênicas. No entanto, apesar da redução significativa na capacidade de diferenciação osteogênica entre células frescas e congeladas, não foram observadas diferenças (P > 0,05) entre SCs. Além disso, o número de células de diferenciação condrogênica congeladas em solução de DMSO+X reduziu (P<0,05) em relação ao controle, sem diferir (P>0,05) das demais SCs. A diferenciação adipogênica não diferiu (P>0,05) entre os tratamentos. Em conclusão, embora esses achados confirmem o sucesso do DMSO para criopreservar CTM-TAe, o Super Cool X-1000 pode ser uma promessa para reduzir a concentração de DMSO.

Integration of heterologous bone matrix associated with polymethylmethacrylate in induced tibial bone defects. An experimental study in rabbits

ABSTRACT Purpose To analyze and compare the reactions at the interface between the composite, composed of fragmented heterologous mineralized bone matrix (MOMHF) and polymethylmethacrylate (PMMA), and the rabbit's tibias, through macroscopic evaluation and scanning electron microscopy (SEM) in different periods. Methods In this study, 12 New Zealand adult rabbits were used (E1: n = 3, E2: n = 3, E3: n = 3 and E4: n = 3). They had the right tibial defects filled with composite and were evaluated immediately after surgery and at 30, 60, 90, and 120 days. Results The composites were incorporated and integrated into the recipient beds in 100% of the cases, defined by the MOMHF osseointegration and the PMMA fibrointegration, with no sign of infection, migration, or rejection. Conclusions The behavior of the composites in the recipient beds demonstrates that these biomaterials have the potential to be used in bone defect repairs, offering, thus, better quality of life to the orthopedic patient.

Região metafisária da tíbia de coelhos: uma opção para estudos dos substitutos ósseos / Metaphyseal region of the tibia of rabbits: an option for bone substitutes studies

Apresenta-se um modelo experimental de ostectomia de tíbia em coelhos para estudo de biomateriais em processos de reparação óssea. Foi realizada falha segmentar de 6mm de diâmetro na região metafisária medial proximal de tíbia de 72 coelhos as quais foram preenchidas com substituto ósseo e avaliadas clinicamente, por exame radiográfico e por meio de tomografia computadorizada feixe cônico, em diferentes tempos. Conclui-se que a metáfise medial proximal de tíbia de coelhos é adequada como modelo para estudos que avaliem o comportamento de enxertos e/ou biomateriais em falhas ósseas.

Is presented an experimental model of tibial ostectomy in rabbits to study biomaterials during bone repair process. Segmental failure of 6 mm diameter was performed in the medial proximal tibial metaphyseal region of 72 rabbits, which were filled with bone substitute and evaluated by clinical exam, X-ray, and cone beam computed tomography at different times. It is concluded that the medial proximal tibial metaphysis region of rabbits is suitable as a model for studies that evaluate the behavior of grafts and/or biomaterials on bone defects.

Região metafisária da tíbia de coelhos: uma opção para estudos dos substitutos ósseos / Metaphyseal region of the tibia of rabbits: an option for bone substitutes studies

Apresenta-se um modelo experimental de ostectomia de tíbia em coelhos para estudo de biomateriais em processos de reparação óssea. Foi realizada falha segmentar de 6mm de diâmetro na região metafisária medial proximal de tíbia de 72 coelhos as quais foram preenchidas com substituto ósseo e avaliadas clinicamente, por exame radiográfico e por meio de tomografia computadorizada feixe cônico, em diferentes tempos. Conclui-se que a metáfise medial proximal de tíbia de coelhos é adequada como modelo para estudos que avaliem o comportamento de enxertos e/ou biomateriais em falhas ósseas.

Is presented an experimental model of tibial ostectomy in rabbits to study biomaterials during bone repair process. Segmental failure of 6 mm diameter was performed in the medial proximal tibial metaphyseal region of 72 rabbits, which were filled with bone substitute and evaluated by clinical exam, X-ray, and cone beam computed tomography at different times. It is concluded that the medial proximal tibial metaphysis region of rabbits is suitable as a model for studies that evaluate the behavior of grafts and/or biomaterials on bone defects.

Evaluation of polyamide surgical mesh as an abdominal ventral implant in rabbits

Abstract Purpose: To evaluate abdominal ventral wound healing by using a specific biomaterial, a handmade polyamide surgical mesh. Methods: A surgical incisional defect was made in ten rabbits to simulate a hernia in the ventral abdominal musculature. A polyamide surgical mesh was used in hernioplasty. They were monitored for surgical wound healing, and macroscopically and histologically evaluated at the end of the experiment. The polyamide surgical mesh did not cause foreign body reaction, pain, edema, or infection in the surgical site. The manure production was not affected by intestinal tissue adherences to the mesh, consistent with the ultrasonography result where adherences were not observed and organized scarring tissue formed in the incisional defect. The polyamide mesh was fixed over the abdominal wall, and its external and internal sides were surrounded by a vascularized connective tissue. Results: None of the experimental animals developed adherences from internal organs to the polyamide mesh, except two rabbits where the omentum formed adherence to the internal scarring tissue without present herniation or compromise of the rabbit's health. Conclusion: Polyamide surgical mesh for hernioplasty presents, in rabbits, excellent biocompatibility, with minimal body adverse reactions and low cost.

Bone marrow stem cell applied in the canine veterinary clinics / Células tronco de medula óssea utilizadas na clínica veterinária canina

Cell therapy represents an old therapeutic practice initiated with the transfusion of whole blood in different clinical situations. There is now a breakthrough in the study of multipotent stem cell therapy because of its functionality in regeneration of tissues, which promotes attention of the scientific community. Bone marrow is one of the main sources of multipotent stem cells, composed by hematopoietic stem cells responsible for the renewal of the cellular components of the blood, and mesenchymal stem cells that aid in the regeneration of tissues. These cells have a strong potential for the treatment of several diseases, due their main characteristics such as high plasticity, capacity for self-renewal and immunomodulation. Although, there are many studies that show good results with the use of cell therapy as a form of treatment for several diseases, some studies still show inconclusive or unsatisfactory results. Therefore, the objective of this study was to review the application of bone marrow stem cells in the canine model since improvements on the knowledge of the technique are necessary to enable its applicability with safety and efficacy.(AU)

A terapia celular representa uma antiga prática terapêutica iniciada com a transfusão de sangue total em diferentes situações clínicas. Atualmente há um avanço no estudo da terapia com células-tronco mesenquimais por conta de sua funcionalidade na regeneração de tecidos, o que promove uma crescente atenção do meio científico. A medula óssea é uma das principais fontes de células-tronco multipotentes, no qual se encontram as células-tronco hematopoiéticas, responsável pela renovação dos componentes celulares do sangue, e as células-tronco mesenquimais que auxiliam na regeneração de tecidos. Essas células têm um forte potencial para o tratamento de diversas enfermidades, uma vez que possuem como principais características alta plasticidade, capacidade de auto renovação e imunomodulação. Apesar de haver muitos trabalhos que apresentam bons resultados com a utilização da terapia celular como forma de tratamento para diversas enfermidades, alguns estudos ainda demonstram resultados inconclusivos ou não satisfatórios, por isso, objetivou-se com este trabalho revisar a aplicação das células-tronco derivadas da medula óssea no modelo canino uma vez que é necessário melhorias sobre o conhecimento da técnica para que possibilite a sua aplicabilidade com segurança e eficácia.(AU)

Transplantation of human immature dental pulp stem cell in dogs with chronic spinal cord injury

Abstract Purpose: To investigate the therapeutic potential of human immature dental pulp stem cells in the treatment of chronic spinal cord injury in dogs. Methods: Three dogs of different breeds with chronic SCI were presented as animal clinical cases. Human immature dental pulp stem cells were injected at three points into the spinal cord, and the animals were evaluated by limb function and magnetic resonance imaging (MRI) pre and post-operative. Results: There was significant improvement from the limb function evaluated by Olby Scale, though it was not supported by the imaging data provided by MRI and clinical sign and evaluation. Conclusion: Human dental pulp stem cell therapy presents promising clinical results in dogs with chronic spinal cord injuries, if used in association with physical therapy.

Avaliação da transmissibilidade ultrassônica do gel fitoterápico de Copaifera duckei Dwyer / Evaluation of ultrasonic transmission of Copaifera duckei Dwyer herbal gel

Esse estudo teve por objetivo avaliar o potencial de transmissibilidade ultrassônica de um gel fitoterápico de copaíba da espécie Copaifera duckei Dwyer na concentração de 10%. A pesquisa foi registrada junto ao Sistema Brasileiro de Biodiversidade. O gel fitoterápico de Copaifera duckei 10% foi manipulado de acordo diretrizes da Farmacopéia Brasileira e testado em um aparelho de Ultrassom (US) operado na faixa de 1 MHz. Como grupos controle foram selecionados a água destilada e o gel hidroalcoólico. A análise ocorreu de forma qualitativa e quantitativa através do modelo proposto na literatura. O US foi programado para modos de corrente pulsado/contínuo e testados nas intensidades (0,2/0,4/0,6/0,8/1,0W/cm2), em 1mim/cm2. Os dados receberam tratamento estatístico pelo software BioEstat 5.3 e foi admitido um nível de significância de ≥0,05. No modo contínuo e na intensidade de 0.2W/cm2 o gel foi considerado "Bom transmissor", na intensidade de 0.4 W/cm2 e 0.6W/cm2 um "Transmissor moderado" e nas intensidades de 0.8W/cm2 e 1.0W/cm2 um "Transmissor pobre". Concluiu-se que o gel de C. duckei 10% não atenuou as ondas de US em nenhum modo ou intensidade testado. E pode assim ser adicionado a esse para tratamentos sendo considerado transmissor bom ou moderado de acordo com a intensidade do US.(AU)

This study aimed to evaluate the potential of transmissibility of an herbal gel of Copaifera duckei Dwyer at a concentration of 10%. The research was registered with the Brazilian Biodiversity System. The gel of Copaifera duckei 10% was guidelined by the Brazilian Pharmacopoeia and tested on an Ultrasound device (US) operated in the range of 1 MHz. The control groups were selected distilled water and hydro alcoholic gel. The analysis was qualitatively and quantitatively using the model proposed in the literature. The US was scheduled to current modes pulsed/continuous and tested in the intensities (0.2/0.4/0.6/0.8/1.0W/cm2) in 1mim/cm2. The data received statistical treatment by BioEstat software 5.3 and was admitted to a significance level of ≥0.05. In continuous mode and intensity of 0.2W/cm2 gel was considered "good transmitter" at the intensity of 0.4W/cm2 and 0.6W/cm2 a "moderate Transmitter" and the intensity of 0.8W/cm2 and 1.0W/cm2 a "poor transmitter". It was concluded that the 10% gel C. duckei US did not attenuate US waves in any form or intensity tested; it can thus be used for this treatment being considered a good or moderate transmitter according to the intensity of US.(AU)

Glial scar-modulation as therapeutic tool in spinal cord injury in animal models

Abstract Purpose: Spinal Cord injury represents, in veterinary medicine, most of the neurological attendances and may result in permanent disability, death or euthanasia. Due to inflammation resulting from trauma, it originates the glial scar, which is a cell interaction complex system. Its function is to preserve the healthy circuits, however, it creates a physical and molecular barrier that prevents cell migration and restricts the neuroregeneration ability. Methods: This review aims to present innovations in the scene of treatment of spinal cord injury, approaching cell therapy, administration of enzyme, anti-inflammatory, and other active principles capable of modulating the inflammatory response, resulting in glial scar reduction and subsequent functional improvement of animals. Results: Some innovative therapies as cell therapy, administration of enzymes, immunosuppressant or other drugs cause the modulation of inflammatory response proved to be a promising tool for the reduction of gliosis. Conclusion: Those tools promise to reduce gliosis and promote locomotor recovery in animals with spinal cord injury.

Dimorfismo sexual e estrutura perineal em Coendou prehensilis (Ouriço-cacheiro) / Sexual dimorphism and perineal structure in Coendou prehensilis (Prehensile-tailed porcupine)

Coendou prehensilis é um mamífero da ordem Rodentia, que é composta por cinco subordens: Sciuromorfos, a qual pertencem os esquilos; Castorimorfos, onde encontramos os castores; Myomorfos, a qual encontramos os ratos; Anomaluromorfos, Lebre-saltadora, e os Histricomorfos, onde observamos a capivara. Utilizamos nesta pesquisa o ouriço-cacheiro (Coendou prehensilis) sendo um macho e uma fêmea, ambos adultos. Estes animais foram encontrados em óbito por atropelamento nas rodovias da região de Pirassununga-SP e cedidos para o Laboratório de Anatomia do Departamento de Medicina Veterinária da Faculdade de Zootecnia e Engenharia de Alimentos da Universidade de São Paulo FZEA/USP devido a permissão do Ibama para estudar os animais (Registro SISBIO no. 43274-1), os quais foram fixados com formaldeído a 10% e dissecados na região do períneo. Os resultados mostraram que a única característica que nos permite diferenciar machos de fêmeas, é o espaço entre o ânus e as genitais dos animais, sendo que o macho apresenta essa distância maior que a fêmea. O períneo é uma região extra cavitária de delimitação losangular, sendo essa delimitação feita pelos ossos interpubiano, púbico e pelos processos do púbis e ísquio. O períneo do Coendou prehensílis macho é composto por cinco principais músculos, sendo três encontrados dispostos no diafragma urogenital superficial, (Músculos isquiocavernosos, M. bulbocavernoso e Mm. Bulboesponjosos) e dois no diafragma pélvico (M. levantador do ânus e M. retrator do pênis). O períneo da fêmea é composto pelos músculos (isquiocavernoso, o bulboesponjoso, levantador do ânus e esfíncter anal externo). Conclui-se que ambos os sexos possuem os músculos isquiocavernosos, Mm. Bulboesponjosos, M. levantador do ânus e esfíncter anal externo, além de que o estudo da região perineal dos animais é de grande importância, pois através do entendimento da função e localização de cada músculo, podemos contribuir para futuras pesquisas sobre reprodução e comportamento animal durante a cópula auxiliando para um melhor entendimento do potencial reprodutivo de roedores da família Erethizontidae, além de contribuir na base do desenvolvimento da biotecnologia aplicada à reprodução visando até mesmo à reprodução desses animais em cativeiro além do meio ambiente livre.(AU)

Coendou prehensilis is a mammal that belongs to the Rodentia order, which consists of five suborders: Sciuromorfos, to which the squirrels belongs; Castorimorfos where we have the beavers; Myomorfos, which include the mice; Anomaluromorfos, Hare jumper, and Hystricomorfos, where we have the capybara. We used the porcupine clerk in the research (Coendou prehensilis), one male and one female, both adults. These animals were transferred to the Laboratory of Anatomy, Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo (FZEA/USP), originating from roadaccidents in Pirassununga, SP (Registration n. 43274-1 SISBIO). The animals were fixed with 10% formaldehyde and dissected in the perineum region. The results showed that the only feature which allows us to differentiate males from females is the space between anus and the genitals of the animals; the male has a larger space than the female. The perineum is an extra cavity region with lozenge definition, and this definition is formed by interpubic bones, the pubic and processes of the pubic and ischium. The male Coendou prehensilis perineum consists of five major muscles, three arranged on the urogenital diaphragm surface (ischiocavernosus muscles, M. bulbocavernosus and Mm. bulboesponjosos) and two in the pelvic diaphragm (M. levator ani and M. penis retractor). The female's perineum is composed by ischiocavernosus, bulbospongiosus, levator ani and external anal sphincter muscles. We conclude that both sexes have the ischiocavernosus muscles, Mm. bulboesponjosos, M. levator ani and external anal sphincter, and it is very important to study the perineal region of the animals, because understanding the function and location of each muscle we can contribute to future research on reproduction and animal behavior during copulation, helping to a better knowledge about reproductive potential of Erethizontidae rodents, and to contribute in the applied biotechnology development of reproduction, what can be helpful even in captivity reproduction of these animals besides the ones in free environment.(AU)

Uso da glicerina para a substituição do formaldeído na conservação de peças anatômicas / Gliceryn can replace formaldehyde for anatomic conservation

A formalização é a principal forma de conservação de corpos e peças anatômicas, devido principalmente ao seu baixo custo. A técnica baseia-se na utilização do formaldeído (5-20%) como fixador e conservador. Entretanto existem vários fatores negativos à sua utilização, como odor desagradável, escurecimento, aumento de peso e rigidez das peças e sérios problemas ambientais quando descartada de forma incorreta, além de ser um produto classificado pela Agência Internacional de Pesquisas em Câncer como altamente cancerígeno. Existem várias opções para substituir o formaldeído, a glicerinação é uma delas, cujo será tratada neste trabalho. A glicerina atua como antifúngico e bactericida, além de ter muitas vantagens em relação ao formol, tratando-se de odor, textura e coloração, além de não ser prejudicial a saúde. Porém o seu custo ainda é elevado e isso explica sua pouca utilização em laboratórios de anatomia. Este trabalho tem como objetivo mostrar as vantagens do uso da glicerina em relação ao formol e propor uma possível substituição deste produto cancerígeno para preservar a saúde dos alunos, funcionários e professores dentro do laboratório, além de permitir um ambiente mais agradável para o aprendizado.(AU)

The formalization is the main form of body and anatomic conservation, mainly due to its low cost. The technique is based on the use of formaldehyde (5-20%) as fixative and preservative. However there are several negative factors on their use, unpleasant smell, browning, weight gain and stiffness of parts and serious environmental problems if disposed of incorrectly, and is a product classified by the International Agency for Research on Cancer as highly carcinogenic. There are several options to replace formaldehyde and glicerinação is one of them, whose which will be treated in this work. Glycerin dehydrates the cell and acts as antifungal and antibacterial, and has many advantages compared to formaldehyde. But its cost is still high and this explains their limited use in the anatomy lab. This work aims to show the advantages of using glycerin in relation to formaldehyde and propose a possible replacement of this carcinogen to preserve the health of students, staff and teachers within the laboratory, and allows a more pleasant environment for learning.(AU)

Animal-assisted therapy in early childhood schools in São Paulo, Brazil / Terapia assistida por animais em escolas de educação infantil de São Paulo

Since ancient times, humans and animals have interacted for different purposes. Animal Assisted Therapy (AAT) is used for the assistance and treatment in humans and educational projects where animals are used as co-therapists or co-educators. The use of animals facilitates the process of teaching and learning, and stimulates physical and therapeutic activities. So that knowledge on AAT could be expanded, current study analyzes the opinion of people directly involved in education on AAT implementation as an educational model in early childhood schools in São Paulo, Brazil. Questionnaires were handed out to 10 pedagogical coordinators, 32 teachers, 23 parents and 26 children aged 3-6 years. Results revealed that AAT is not well-known for most interviewees, including pedagogical coordinators, teachers and parents. However, interviewees believe in the benefits of child-pet interactions and are favorable to the implementation of AATs in schools. Projects should be interdisciplinary and must involve professionals from other areas, such as psychologists and veterinarians. Regarding the educational model, interviewees believe in the innovation capacity of AAT and in the possibilities of interdisciplinarity among teachers in the use of animals. Research also demonstrated that children like and support the use of animals in the school.(AU)

Desde a pré-história já existia a interação dos humanos com os animais com diferentes finalidades. A terapia assistida por animais (TAA) é utilizada para assistências e tratamentos em humanos e projetos pedagógicos, na qual os animais são utilizados como co-terapeutas ou co-educadores. O emprego de animais facilita o processo de ensino-aprendizagem e estimula atividades físicas e terapêuticas. Para contribuir com o conhecimento da TAA, o presente trabalho objetivou abordar a opinião de pessoas diretamente relacionadas à escola, a respeito da implantação da TAA, como modelo educacional nas escolas de educação infantil da cidade de São Paulo. Foram aplicados questionários em 10 coordenadores pedagógicos, 32 professores, 23 pais e 26 crianças de 3 a 6 anos de idade. Os resultados encontrados demonstraram que a TAA ainda não é bem conhecida por uma parcela dos entrevistados, incluindo coordenadores pedagógicos, professores e pais de alunos. No entanto, os entrevistados acreditam nos benefícios da interação criança-animal e defendem projetos voltados a implantação da TAA nas escolas, embora estes projetos devam ser interdisciplinares e envolver profissionais de outras áreas, tais como, pedagogos, psicólogos e médicos veterinários. Em relação ao modelo educacional, nossos entrevistados acreditam na capacidade inovadora da TAA, assim como, nas possibilidades de interdisciplinaridade entre os professores no uso dos animais. Também ficou demonstrado que as crianças gostam e apoiam o uso de animais na escola.(AU)

Rabbit olfactory stem cells. Isolation protocol and characterization

PURPOSE: To describe a new technique for isolation of a mesenchymal stem cells (MSCs) population from the olfactory mucosa in rabbits. METHODS: Olfactory stem cells (OSCs) were retrieved from under the cribriform plate of the Ethmoid bone. Several assays were accomplished to characterize the cell population and attest its viability in vitro. The cells were submitted to flow cytometry with the antibodies CD34, CD45, CD73, CD79, CD90 and CD105 and also they were induced to differentiate in three lineages. Functional evaluation involved analysis of in vitro growth behavior, colony forming unit like fibroblasts (CFU-f) and cryopreservation response. Further transduction with Green Fluorescent Protein (GFP) was also performed. RESULTS: The OSCs showed mesenchymal features, as positive response to CD34, CD73 and CD90 antibodies and plasticity. Additionally, these cells have high proliferated rate, and they could be cultured through many passages and kept the ability to proliferate and differentiate after cryopreservation. The positive response to the transduction signalizes the possibility of cellular tracking in vivo. This is a desirable feature in case those cells are used for pre-clinical trials. CONCLUSION: The cells harvested were mesenchymal stem cells and the technique described is therefore efficient for rabbit olfactory stem cells isolation.

Morphological and ultrastructural analysis of the tongue of Gracilinanus microtarsus (Wagner, 1842) / Análise morfológica e ultraestrutural da língua da Gracilinanus microtarsus (Wagner, 1842)

The G. microtarsus is a small marsupial belonging to the Didelphidae family and may be found mainly in the Atlantic Rainforest of the Brazilian Southeastern region. G. microtarsus has important ecological importance in the dispersion of seeds of several plant species. Current research characterizes morphologically six tongues of adult male and female specimens, which are fixed in 10% formaldehyde. The tongues were analyzed, measured and photographed "in situ" and "ex situ". Anatomy, light microscopy and ultrastructural analysis were performed. Macroscopically the tongue has root, a fixed body and a free apex. The tongue has fungiform papillae and high conical and acute filiform papillae across the dorsal surface. It also featured three vallate papillae located at the root and marginal fungiform papillae at the apex. Microscopically, the tongue is composed of longitudinal and transversal muscle fibers. Blood vessels, nerves, serous, mucous and mixed acini are found in non-modeled connective tissue. Ventrally, the tongue is lined by a keratinized stratified pavement epithelium. The morphology of the tongue of G. microtarsus resembles that described in carnivores and marsupials Didelphidae regarding its microscopic structure, differing only in the distribution and number of papillae.

O G. microtarsus é um pequeno marsupial pertencente à família Didelphidae, que pode ser encontrado principalmente na Mata Atlântica da região sudeste brasileira. O G. microtarsus possui importância ecológica na dispersão de sementes de diversas espécies de plantas. A pesquisa atual caracteriza morfologicamente as línguas de seis animais adultos, fixados em formol a 10%. As línguas foram analisadas, mensuradas e fotografadas. Foram realizadas análises anatômicas, microscópicas de luz e ultraestruturais. Macroscopicamente a língua possui raiz, um corpo fixo e um ápice livre. A língua possui papilas fungiformes, filiformes cônicas altas e agudas em toda a superfície dorsal. Também possui três papilas valadas localizadas na raiz e papilas fungiformes marginais no ápice da língua. Microscopicamente, a língua é composta por fibras musculares longitudinais e transversais. Os vasos sanguíneos, nervos e ácinos serosos, mucosos e mistos são encontrados em tecido conjuntivo não modelado. Ventralmente, a língua é revestida por um epitélio pavimentoso estratificado queratinizado. A morfologia da língua de G. microtarsus se assemelha ao descrito em carnívoros e marsupiais Didelphidae quanto à sua estrutura microscópica, diferindo apenas na distribuição e número de papilas.

Tomographic imaging of fragmented cortical bone heteroimplant and methylmethacrylate in segmental bone defect of rabbit tibia

PURPOSE: To evaluate the performance of composites consisting of fragmented cortical bone heteroimplant in association with methylmethacrylate preserved in 98% glycerin, in segmental bone defect of rabbit tibia medial metaphysis. METHODS: In this study were used twelve adult New Zealand rabbits, divided into three groups of four animals each: G30 (30 days), G60 (60 days) and G90 (90 days). The bone defects previously created in the tibia were filled with composites and both were evaluated by cone-beam computed tomography, immediately after surgery and after 30, 60, and 90 days. RESULTS: The composites fulfilled and remained in the sites of bone defects in all cases and were not registered signals of infection, migration or rejection. CONCLUSIONS: The implanted composites promoted the bone defects repair without signals of infection and/or rejection. The composites are one more option for bone defects repair. .