Intrathecal Transplantation of Autologous and Allogeneic Bone Marrow-Derived Mesenchymal Stem Cells in Dogs.

The route used in the transplantation of mesenchymal stem cells (MSCs) can directly affect the treatment success. The transplantation of MSCs via the intrathecal (IT) route can be an important therapeutic strategy for neurological disorders. The objective of this study was to evaluate the safety and feasibility of the IT transplantation of autologous (Auto-MSCs) and allogeneic (Allo-MSCs) bone marrow mesenchymal stem cells (BM-MSCs) in healthy dogs. Based on neurodisability score, cerebrospinal fluid (CSF) and magnetic resonance imaging (MRI), no significant differences from the control group were observed on day 1 or day 5 after IT Auto- or Allo-MSCs transplantation (P > 0.05). In addition, analysis of matrix metalloproteinase (MMP)-2 and MMP-9 expression in the CSF revealed no significant differences (P > 0.05) at 5 days after IT transplantation in the Auto- or Allo-MSCs group when compared to the control. Intrathecal transplantation of BM-MSCs in dogs provides a safe, easy and minimally invasive route for the use of cell-based therapeutics in central nervous system diseases.

Prevalence of behavioral changes in senile dogs / Prevalência de alterações comportamentais em cães idosos

ABSTRACT: Owing to improvements in animal welfare, such as in nutrition, preventive medicine, and diagnostic tools, older dogs have become increasingly more prevalent than before. Unfortunately, there is an increasing number of dogs presenting behavioral changes correlated with this growth in the geriatric population. However, many of these owners consider these signs as normal for old age. This study aimed to determine the prevalence of clinical signs related with behavioral changes in the canine geriatric population admitted to the Veterinary Hospital of the Veterinary Medicine and Animal Science College of Universidade Estadual Paulista. The owners of 800 old dogs completed a questionnaire about disorientation, changes in social interaction, loss of training (house soiling), changes in the wake-sleep cycle, physical activity, memory, learning, consciousness, and perception. The results showed that the prevalence of behavioral changes in the canine geriatric population was 90.7% of dogs that presented at least one of these behavioral changes, and 22.3% with at least six changes, and with changes in physical activity (58.5%) and loss of acquired training (30.2%) being the highest percentage groups that were reported by the owners. We concluded that the prevalence of behavioral changes related with aging in the canine geriatric population was high, and that it was important to include questions about behavioral changes during regular veterinary clinical appointments to identify these clinical signs earlier and investigate the possible causes of the cognitive impairment, including neurological diseases, such as Canine Cognitive Dysfunction Syndrome.

RESUMO: Devido à melhora na nutrição e na medicina veterinária, há mais cães idosos do que antes. Infelizmente, acompanhando este crescimento na população geriátrica, houve um aumento no número de cães demonstrando distúrbios de comportamento e senilidade aparente. Entretanto, muitos dos proprietários têm considerado todos esses sinais decorrentes de uma inevitável velhice. O objetivo deste estudo foi obter a prevalência de sinais clínicos relacionados com alterações comportamentais na população canina idosa, atendida no Hospital Veterinário da Faculdade de Medicina Veterinária e Zootecnia da UNESP. Proprietários de 800 cães idosos responderam um questionário sobre desorientação, alterações na interação social, perda de treinamento (urina e/ou defeca em locais não habituais), alterações no ciclo sono-vigília, atividade física, memória, aprendizagem, consciência e percepção. Os resultados obtidos neste estudo apresentaram uma prevalência de alterações comportamentais na população canina idosa significativa, variando de 22,3% dos cães, com pelo menos seis alterações comportamentais e, chegando a 90,7% com pelo menos uma alteração de comportamento, sendo alterações na atividade física (58,5%) e perda de treinamentos adquiridos (30,2%) os grupos de maior percentual relatado pelos proprietários. Conclui-se que a prevalência de alterações comportamentais relacionadas com o envelhecimento da população canina é alta, sendo importante incluir questões sobre as alterações comportamentais durante as consultas clínicas veterinárias regulares para identificar, o mais precocemente possível, estes sinais clínicos e realizar ações para investigar as possíveis causas de déficit cognitivo, incluindo doenças neurológicas, como a Síndrome de Disfunção Cognitiva canina.

Métodos eletrocardiográficos em equinos American Miniature Horse / Electrocardiographic methods in equines American Miniature Horse

O objetivo deste estudo foi analisar e comparar os registros eletrocardiográficos de 50 equinos American Miniature Horse, clinicamente saudáveis, obtidos pelos métodos convencional e computadorizado, para a padronização do método informatizado. As medidas de duração e amplitude das ondas e intervalos e frequência cardíaca foram analisadas nas derivações do plano frontal e na derivação base-ápice. Diferenças quanto à duração da onda P, foram encontradas nas derivações I e aVL, e do complexo QRS, em todas as derivações do plano frontal. Quanto à derivação base-ápice, houve diferença na duração do intervalo QT e da onda T, assim como na amplitude da onda S. Concluiu-se, portanto, que ocorre superestimação da duração do complexo QRS no método computadorizado e que, devido às diferenças inerentes de cada método, torna-se importante a utilização de valores de referência para o método computadorizado também na espécie equina.

The aim of this study was to analyze and compare electrocardiographic records of 50 horses American Miniature Horse, clinically healthy, obtained by conventional methods and computerized, to the standardization of the computerized method. The measures of duration and amplitude of waves and intervals and heart rate were analyzed in the leads of the frontal plane and base-apical lead. Differences about the duration of the P wave were found in I and aVL derivations, and QRS complex, in all derivations of the frontal plane. As for basis-apical derivation, there was no difference in the duration of the QT interval and the T wave and the amplitude of the S wave, therefore concluding that overestimation occurs in the duration of the QRS complex in computerized method and that due to inherent differences of each method, it becomes important the use of reference values for the computerized method also in the equine species.

Equine mesenchymal stem cells from bone marrow, adipose tissue and umbilical cord: immunophenotypic characterization and differentiation potential.

INTRODUCTION: Studies with mesenchymal stem cells (MSCs) are increasing due to their immunomodulatory, anti-inflammatory and tissue regenerative properties. However, there is still no agreement about the best source of equine MSCs for a bank for allogeneic therapy. The aim of this study was to evaluate the cell culture and immunophenotypic characteristics and differentiation potential of equine MSCs from bone marrow (BM-MSCs), adipose tissue (AT-MSCs) and umbilical cord (UC-MSCs) under identical in vitro conditions, to compare these sources for research or an allogeneic therapy cell bank. METHODS: The BM-MSCs, AT-MSCs and UC-MSCs were cultured and evaluated in vitro for their osteogenic, adipogenic and chondrogenic differentiation potential. Additionally, MSCs were assessed for CD105, CD44, CD34, CD90 and MHC-II markers by flow cytometry, and MHC-II was also assessed by immunocytochemistry. To interpret the flow cytometry results, statistical analysis was performed using ANOVA. RESULTS: The harvesting and culturing procedures of BM-MSCs, AT-MSCs and UC-MSCs were feasible, with an average cell growth until the third passage of 25 days for BM-MSCs, 15 days for AT-MSCs and 26 days for UC-MSCs. MSCs from all sources were able to differentiate into osteogenic (after 10 days for BM-MSCs and AT-MSCs and 15 days for UC-MSCs), adipogenic (after 8 days for BM-MSCs and AT-MSCs and 15 days for UC-MSCs) and chondrogenic (after 21 days for BM-MSCs, AT-MSCs and UC-MSCs) lineages. MSCs showed high expression of CD105, CD44 and CD90 and low or negative expression of CD34 and MHC-II. The MHC-II was not detected by immunocytochemistry techniques in any of the MSCs studied. CONCLUSIONS: The BM, AT and UC are feasible sources for harvesting equine MSCs, and their immunophenotypic and multipotency characteristics attained minimal criteria for defining MSCs. Due to the low expression of MHC-II by MSCs, all of the sources could be used in clinical trials involving allogeneic therapy in horses. However, the BM-MSCs and AT-MSCs showed fastest ''in vitro'' differentiation and AT-MSCs showed highest cell growth until third passage. These findings suggest that BM and AT may be preferable for cell banking purposes.

Immunophenotypic, immunocytochemistry, ultrastructural, and cytogenetic characterization of mesenchymal stem cells from equine bone marrow.

The aim of this study was to isolate, culture, and characterize mesenchymal stem cells (MSCs) from horse bone marrow (BM) using the techniques of flow cytometry, immunocytochemistry, cytogenetics, and electron microscopy. Immunophenotypic analysis revealed the presence of MSCs with high expression of the CD90 marker, lower expression of the CD44 marker, and absent expression of the CD34 marker. In assays of differentiation, the positive response to osteogenic (OST), chondrogenic (CDG), and adipogenic (ADP) differentiation signals was observed and characterized by deposition of calcium-rich extracellular matrix (OST), proteoglycans and collagen II (CDG) and intracellular deposition of fat drops (ADP). In immunocytochemical characterization, MSCs were immunopositive for CD44, vimentin, and PCNA, and they were negative for CD13. In the ultrastructural analysis of MSCs, the most outstanding characteristic was the presence of rough endoplasmic reticulum with very dilated cisterns filled with a low electrodensity material. Additionally, MSCs had normal karyotypes (2n = 64) as evidenced by cytogenetic analysis, and aneuploidy in metaphase was not observed. The protocols for isolating, culturing, and characterizing equine MSCs used in this study were shown to be appropriate for the production of a cell population with a good potential for differentiation and without aneuploidy that can be used to study future cellular therapies.

Homoimplante de costela conservada em solução supersaturada de açúcar a 300 por cento ou em açúcar in natura na reconstituição experimental de costelas em gatos / Homoimplants rib preserved in 300 percent hyper saturated sugar solution or sugar in natura for experimental reconstitution of ribs in cats

O objetivo deste estudo foi avaliar a eficiência do açúcar em diferentes formas de conservação para ossos, verificar a eficiência do implante de costela conservada na reconstituição de defeito torácico, avaliar a fixação da costela com agulha de cateter, e monitorar a estabilidade e a dinâmica do tórax após o implante. Além disso, buscou-se avaliar clinicamente, macroscopicamente, histologicamente e radiograficamente o material implantando. Os 16 implantes utilizados foram costelas homólogas conservadas em açúcar in natura (Ga) ou a 300 por cento (Gb), na reconstituição de costelas de gatos. Os animais foram separados em dois grupos: Ga (8), utilizando açúcar "in natura" e o Gb (8), solução a 300 por cento. Foi realizado defeito no terço médio do lado direito do tórax de 16 gatos adultos, os quais foram fixados no leito receptor por meio de agulha de cateter. A porcentagem de incorporação dos implantes foi de 91,7 por cento aos 60 dias no Ga e 73,9 por cento no Gb. Foram verificadas, em diferentes etapas, leve reabsorção do implante em 29,2 por cento no Ga e de 37,8 por cento no Gb. Não foi observado qualquer caso de eliminação ou de infecção. É possível concluir que o implante ósseo homólogo de costela conservado em açúcar "in natura" ou a 300 por cento é uma opção viável para a reconstituição da parede torácica de gatos.

This study was aimed at evaluating the use of a homologous bone implant preserved sugar in natura (Ga) (8) or in 300 percent sugar solution (Gb) (8) for reconstitution of the thoracic wall in 16 cats, involving three ribs. A costal defect was made on the right side of the thoracic wall and the preserved ribs were used as homologous bone implants for the correction of this defect. These ribs were fixed to the receptor by means of a catheter needle. The animals were clinically evaluated by X-rays and histology. The percentual of implant incorporation at 60 days was 91.7 percent in the Ga and 73.9 percent in the Gb. A mild resorption was observed at different times, 29.2 percent in the in natura sugar group, and 37.8 percent in the 300 percent sugar group. No case of infection or rejection was observed in the evaluated period. The homologous rib bone implant preserved in sugar in natura or in 300 percent solution is efficient for the reconstitution of costal wall in cats.

Pós-operatório de homoimplante de costela avaliado por toracoscopia paraxifóide transdiafragmática modificada em gatos / Post-operative of toracoscopy paraxiphoid transdiaphragmatic modificed approach, in cats

O objetivo deste estudo foi de avaliar a exeqüibilidade do acesso paraxifóide transdiafragmático modificado, em gatos, no período pós-operatório de homoimplante de costela para reconstituição de parede torácica. Para tanto foram avaliados 16 gatos após a reconstituição de parede torácica com o uso de homoimplante de costela conservado em açúcar. A cavidade torácica dos animais foi acessada por via transdiafragmática modificada, por meio de uma incisão de 2cm na linha mediana paraxifóide para posterior introdução da cânula ao diafragma utilizando uma óptica de 10mm de diâmetro. Foi identificado o terço médio da parede torácica, sendo necessário que os animais fossem posicionados em decúbito lateral obliquado. Foram constatadas aderências e visibilizado o aspecto dos implantes. Foi possível avaliar a parede torácica de gatos com óptica de 10mm de diâmetro através do acesso transdiafragmático modificado.

The purpose of this study was to evaluate a modified thoracic access to test the rib homoimplant in cats. Sisteen adults cats was evaluated ofter thoracic wall reconstituition using a rib homoimplant preserved in sugar. The animals were submitted to a modified trans-diaphragmatic paraxiphoid access, using a optical of 10mm diameter through the midline. The lateral oblique recumbency made become possible the identification of the medial third of the thorax, and it also allowed the identification the adhesions and the aspect of the implants. The evaluation thoracic cavity of the cats with an optic of 10mm of diameter through the modified trans-diaphragmatic paraxiphoid access.