A three-dimensional cell culture system as an in vitro canine mammary carcinoma model for the expression of connective tissue modulators.

In this study, derived complex carcinoma (CC) and simple carcinoma (SC) cell lines were established and cultured under two-dimensional (2D) and three-dimensional (3D) conditions. The 3D was performed in six-well AlgiMatrix™ (LifeTechnologies®, Carlsbad, CA, USA) scaffolds, resulting in spheroids sized 50-125 µm for CC and 175-200 µm for SC. Cell viability was demonstrated up to 14 days for both models. Epidermal growth factor receptor (EGFR) was expressed in CC and SC in both systems. However, higher mRNA and protein levels were observed in SC 2D and 3D systems when compared with CC (P < 0.005). The connective tissue modulators, metalloproteinases-1, -2, -9 and -13 (MMPs), relaxin receptors 1 and 2 (RXR1 and RXR2) and E-cadherin (CDH1) were quantitated. All were upregulated similarly when canine mammary tumour (CMT)-derived cell lines were cultured under 3D AlgiMatrix, except CDH1 that was downregulated (P < 0.005). These results are promising towards the used of 3D system to increase a high throughput in vitro canine tumour model.

Cardiovascular and respiratory alterations in dogs during cervical myelography using ioversol / Alterações cardiovasculares e respiratórias durante a mielografia cervical com ioversol em cães

Subarachnoid infusion of most contrast mediums and the steps involved in performing a cervical myelography have adverse affects that can discourage its use in the radiographic diagnosis of spinal cord diseases. Thus, the cardiovascular and respiratory alterations associated with neck flexion, subarachnoid puncture, and cerebrospinal fluid drainage during subarachnoid infusion of ioversol (320mgI/mL) in dogs under general anesthesia using isoflurane were evaluated. The dogs received subarachnoid infusion of autologous cerebrospinal fluid kept at 38°C - control group (GC); ioversol 0.3mL/kg at 25°C (GI25) and ioversol 0.3mL/kg heated to 38°C (GI38). Each dog had its heart rate (HR), systolic arterial pressure (SAP), diastolic arterial pressure (DAP), respiratory rate (RR), oxyhemoglobin saturation (SaO2) and electrocardiography readings (PR and QT intervals) recorded. Group comparisons showed no statistical difference regarding neck positioning, subarachnoid puncture, and subarachnoid infusion of contrast medium on HR, RR and SaO2, cardiac rhythm or conduction. However, isoflurane significantly increased PR and QT intervals. Based on these findings, it is concluded that the steps involved in cervical myelography and the use of ioversol 320mgI/mL at 0.3mL/kg (25ºC and 38ºC) during cervical myelography did not result in relevant cardiovascular and respiratory alterations, except for an elevation in arterial pressure after injection of ioversol.

Os efeitos adversos provocados pela infusão subaracnoidea de meios de contraste e da mielografia cervical podem desencorajar sua aplicação como alternativa para o diagnóstico radiológico de doenças da medula espinhal. Assim, avaliaram-se os efeitos cardiovascular e respiratório dos procedimentos técnicos necessários à mielografia cervical e da infusão subaracnoidea do ioversol - 320mgI/mL - em cães anestesiados com isofluorano. Foi realizada infusão do líquido cefalorraquidiano autólogo mantido a 38°C - grupo-controle (GC); infusão do ioversol 0,3mL/kg a 25°C (GI25); e infusão do ioversol 0,3mL/kg aquecido a 38°C (GI38). Estudaram-se: pressões arteriais sistólica (PAS) e diastólica (PAD), frequências cardíaca (FC) e respiratória (FR), saturação de oxi-hemoglobina (SaO2) e medidas eletrocardiográficas (intervalos PR e QT). Não houve influência significativa do posicionamento, da punção ou da infusão subaracnoidea do ioversol sobre FC, FR, SaO2, ritmo e condução cardíaca, entretanto o isofluorano aumentou significativamente os intervalos PR e QT. Concluiu-se que a mielografia cervical com ioversol 320mgI/mL, nas temperaturas de 25ºC e 38ºC e na dose de 0,3mL/kg, não causa arritmias cardíacas nem alterações cardiovasculares e respiratórias relevantes, exceto elevação da pressão arterial após a injeção do contraste.

Healing and expression of growth factors (TGF-ß and PDGF) in canine radial ostectomy gap containing platelet-rich plasma.

OBJECTIVES: To evaluate bone healing in an experimental radial ostectomy in dogs treated with autologous platelet-rich plasma (PRP), through histological, densitometric, radiographic studies, as well as expression of growth factors in the ostectomy gap. METHODS: Twenty-one dogs were randomly divided into either a control or a PRP group. All underwent unilateral ostectomy of the radius to generate a gap of 2.0 mm, that was stabilized with external skeletal fixation. The ostectomy gap was either filled with PRP or left empty as a control. The radiographic and densitometric studies were performed after surgery, then at intervals until 60 days during the post-surgery period. Histological and immunohistochemical evaluations were performed at seven or 60 days post-surgery. Analyses were performed using a statistical analysis system, and the level of significance was set at p <0.05. RESULTS: The median radiographic healing score in the PRP group increased significantly between day 0 and day 60. Furthermore, at 60 days, the median healing score and the proportion of healed ostectomies in the control group (score 1; 1/6 healed) and the PRP group (score 5; 4/5 healed) were significantly different. There were differences between groups in radiographic and densitometric values at days 45 and 60. The histological evaluation showed advanced bone healing at 60 days in the PRP group and signs of delayed union in the control group. CLINICAL RELEVANCE: Platelet-rich plasma can be used as an adjuvant therapy because it may promote better bone healing of a radial ostectomy treated with external skeletal fixation in dogs.

Influência do butorfanol sobre os períodos de latência e de ação da ropivacaína pela via peridural na ovariossalpingo-histerectomia em cadelas / Influence of butorphanol on latency and duration of ropivacaine epidural anesthesia for ovariosalpingohysterectomy in bitches

Avaliaram-se os períodos de latência e de duração do efeito do butorfanol associado à ropivacaína aplicados pela via peridural e a possibilidade de uso dessa associação como protocolo anestésico para realização de ovariossalpingo-histerectomia (OSH) em cadelas. Utilizaram-se 16 cadelas pré-medicadas com acepromazina e midazolam compondo dois grupos: no grupo 1 (n=8) aplicou-se ropivacaína isolada (0,3mL/kg) e no grupo 2 (n=8), butorfanol (0,1mg/kg) e ropivacaína (até o volume de 0,3mL/kg), pela via peridural. Consideraram-se nove momentos (M): M0 - animal sem anestesia, M1 - 15 minutos após medicação pré-anestésica; M2 - 30 minutos após a aplicação peridural; M3, M4 e M5 - correspondentes ao início do procedimento e ao pinçamento dos pedículos esquerdo e direito, respectivamente; M6 - ligadura da cérvix uterina; e M7 e M8 - início da laparorrafia e fim da sutura de pele, respectivamente. Foram avaliados os períodos de latência e ação do bloqueio, além da viabilidade de realização do procedimento cirúrgico por meio do bloqueio efetuado. O que recebeu ropivacaína + butorfanol apresentou sedação pronunciada e permitiu a realização de OSH em 75 por cento dos animais, sem indução anestésica. Não houve diferença entre os grupos quanto aos períodos de latência e duração. A associação do butorfanol à ropivacaína proporcionou bloqueio anestésico compatível com a realização de OSH e período de latência curto, com duração de efeito suficiente para o procedimento cirúrgico.

The periods of latency and duration of butorphanol associated with ropivacaine used via epidural, and this combination as anesthetic protocol for carrying out ovariosalpingohysterectomy (OSH) in bitches were evaluated. Sixteen animals pre-medicated with acepromazine and midazolam were used composing two groups that received: 1 (n=8) ropivacaine (0.3mL/kg) and 2 (n=8) butorphanol (0.1mg/kg) and ropivacaine (up to the volume of 0.3ml/kg) via epidural. Nine moments were studied: M1 - 15 minutes after pre-anesthetic medication; M2 - 30 minutes after the epidural medication; M3, M4, and M5 - at the beginning of surgery and at clamping left and right pedicles, respectively; M6 - at ligation of the uterine cervix; and M7 and M8 - at laparorhaphy and end of skin suture, respectively. Periods of latency and blocking the action of random double-covert manner were evaluated as well as the feasibility of carrying out the surgery performed by the blockade. It was observed that the group receiving ropivacaine + butorphanol allowed the execution of OSH in 75 percent of animals without the need for anesthesia. There was no statistical difference between the periods of latency and duration. It was concluded that the combination of butorphanol to ropivacaine provides, in bitches, block compatible with the implementation of OSH with short period of latency and duration of effect sufficient for the surgical procedure.