Avaliação morfológica do concentrado autólogo de plaquetas em cães sob refrigeração e criopreservação em DMSO 6% / [Morphological evaluation of the autologous platelet concentrate in dogs under refrigeration and criopreservation in 6% DMSO]

The morphological characteristics of the autologous platelet concentrate (APC) of 31 dogs were evaluated after cooling and freezing in 6% DMSO. Blood from the jugular vein of each patient was collected and centrifuged at 191g for six minutes to obtain APC. In the fresh sample, the platelet count, MPV, PDW and cell morphology were evaluated. Four samples of each animal were sent for storage, one refrigerated at 4°C for seven days, another for 30 days and two more stored in a freezer at -80°C in the same time interval, using 6% DMSO as cryoprotectant. The conserved samples were submitted to the same laboratory analysis as the fresh sample. There was a difference between fresh and preserved samples for platelet count, cell concentration, MPV and PDW (P<0.05), except in the 30-day refrigerated group, which showed severe morphological changes. In the frozen group for seven days, no difference was observed in the percentage of activation (P>0.05). The results obtained lead to the conclusion that cryopreservation with 6% DMSO at -80°C for seven days is a favorable option for the maintenance of platelet concentrations and the morphological characteristics of APC in dogs.(AU)

Caudal vena cava collapsibility index in healthy cats by ultrasonography / [Índice de colapsibilidade da veia cava caudal em gatos domésticos Saudáveis, obtido por meio de ultrassonografia]

The objective of this study was to calculate the collapsibility index (CI) in a group of 15 healthy adult mixed breed cats via right hepatic intercostal ultrasound view. The minimal and the maximal diameters of the caudal vena cava (CVC) were obtained during inspiration and expiration, respectively, then CI was calculated. The mean diameter of the CVC was 0.5cm. The mean CI was 28±3% and CI was not significantly associated with gender. As in human medicine, there is a growing need for less invasive monitoring in small animal practice. The CI enables the assessment of estimated volemia without the need for a central venous catheter. This is the first reported study investigating CI in cats.(AU)

O objetivo deste estudo foi calcular o índice de colapsabilidade (IC) em um grupo de 15 gatos adultos, saudáveis e SRD, por meio da janela ultrassonográfica hepática intercostal direita. Os diâmetros mínimo e máximo da veia cava caudal (VCC) foram obtidos durante a inspiração e a expiração, respectivamente. O IC foi calculado, posteriormente. O diâmetro médio da VCC foi de 0,5cm. O diâmetro médio do IC foi de 28±3%, e o IC não foi significativamente associado ao gênero. Como na medicina humana, há uma necessidade crescente de monitoramento menos invasivo na prática de pequenos animais. O IC possibilita a avaliação da volemia estimada sem a necessidade de um cateter venoso central. Este é o primeiro estudo relatado sobre IC em gatos domésticos.(AU)

Dimensões da patela relacionadas à massa corporal do cão para emprego na confecção de prótese: estudo em cadáver / Patellar dimensions related to canine body mass for use in the manufacture of prosthesis: cadaver study

O objetivo deste trabalho foi mensurar as dimensões da patela de cadáveres caninos e avaliar sua relação com a massa corporal. Para a realização das medidas patelares, foram utilizados 70 cadáveres de cães adultos, com massa corpórea de 1 a 50kg, sem evidência clínica de afecção na articulação femorotibiopatelar. Com auxílio de um paquímetro, foram mensurados os comprimentos externo e interno, a largura externa no terço médio, a largura e a espessura internas nos terços proximal, médio e distal das patelas. Também foram mensuradas a largura nos terços proximal, médio e distal, a profundidade nos terços proximal, médio e distal do sulco troclear; essas medidas foram exclusivamente internas. Observou-se, de forma geral, forte correlação entre as medidas patelares e a massa corporal de cadáveres de cães. Nas condições deste estudo, pode-se concluir que o tamanho da patela varia segundo a massa corporal do cão e que essas medidas devem ser consideradas ao se planejar uma substituição protética.(AU)

The aim of this study was to measure the canine cadaver patellar dimensions and evaluate its relationship with body mass. 70 cadavers of adult dogs were used, with a body mass between 1 and 50kg, without clinical evidence of affection in the stifle joint. The external and internal lengths, external width in the middle third, width and thickness of the proximal, middle and distal thirds were measured using a pachymeter. The width was also measured in the proximal third, middle third and distal third, depth in the proximal third, middle third and distal third of the trochlear groove; these measurements were exclusively internal. A strong correlation was observed between the patellar dimension and body mass of canine cadavers. Under this study conditions, it is possible to conclude that the patellar size varies according to the canine body mass and these measures should be considered when planning a prosthetic replacement.(AU)

Matriz porosa do BV60S associada a células osteoprogenitoras alógenas no tratamento de defeitos ósseos críticos em rádios de cães / Porous matrix of BG60S associated with allogenic osteoprogenitor cells in the treatment of critical bone defects in the radius of dogs

O objetivo do estudo foi avaliar o efeito da matriz porosa do biovidro 60S (BV60S) associada a células osteoprogenitoras (CO) alógenas no tratamento de defeitos ósseos críticos de cães. Foram utilizados 20 cães, machos, sem raça definida, com dois anos de idade e massa corporal média de 25kg. Com os cães sob anestesia geral, foram criados defeitos ósseos críticos no terço médio dos ossos rádios. Procedeu-se à fixação óssea com uma placa em ponte, e os defeitos foram tratados de acordo com cada grupo experimental. Constituíram-se três grupos experimentais, em que os defeitos ósseos foram preenchidos com: BV60S associado a CO alógenas (grupo BV60S+CO), osso autógeno (grupo C+), ou não preenchidos (grupo C-). A regeneração óssea foi avaliada por meio de exames radiográficos, densitométricos e histomorfométricos ao longo de 90 dias. Os grupos C- e BV60S+CO mostraram preenchimento ósseo parcial do defeito de, no máximo, 56,68% e 35,23%, respectivamente, sem a formação de ponte óssea entre as extremidades, e o controle positivo (C+) mostrou regeneração óssea completa. Conclui-se que a matriz porosa do BV60S associada às células osteoprogenitoras não é eficiente no tratamento de defeitos ósseos críticos em rádios de cães.(AU)

The objective of this study was to evaluate the effect of the porous matrix of bioglass 60S (BV60S) associated with allogenic osteoprogenitor cells (CO) in the treatment of critical bone defects of dogs. 20 male mongrel dogs at two years old and mean weight of 25kg were used. Dogs were anesthetized and critical bone defects were created in the middle third of the radios bones. With dogs under general anesthesia, critical bone defects were created in the middle third of bone radios. Bone fixation was done with a bridge plate and defects treated according to each experimental group. Three experimental groups were formed according to the treatment. The defects filled with BV60S associated with allogenic CO (Group-BV60S+CO), autogenous bone (Group-C+) or unfilled (Group-C-). Bone regeneration was evaluated by radiography, bone densitometry and histomorphometry over 90 days. The BV60S+CO and C- groups showed partial bone filling of the defect of at most 56.68% and 35.23%, respectively. No bone bridge formation was observed between the extremities in the BV60S+CO and C- groups. Positive control showed complete bone regeneration at 90 days. It was concluded that the porous matrix of BV60S associated with osteoprogenitor cells was not effective in the treatment of critical bone defects in the radius of dogs.(AU)

Patellar luxation and articular lesions in dogs: a retrospective: study research article / Luxação de patela e lesões articulares em cães: estudo retrospectivo

This study describes lesions that occur in the stifle joints of dogs with patellar luxation. These lesions are associated with the animal's age, body weight, and degree of luxation. The rate of redislocation was also evaluated. The patellar lesions found include articular cartilage erosion, subchondral bone exposure, a flattened or concave patellar surface, and enthesophytes. Extra-patellar lesions included synovitis, osteophytes, blunting of the trochlear groove, an absent trochlea, erosion of the condylar margins, capsule thickening, a long digital extensor tendon injury, cranial cruciate ligament rupture, and meniscal prolapse. Such lesions were frequently found in animals with Grade II or III luxation who were aged 24 months or more, and they were more severe in dogs weighing more than 15 kg. Patellar luxation causes changes that favor articular degeneration and should be treated surgically. Conservative treatment relieves pain, but does not address tissue alterations.(AU)

O estudo descreve as lesões articulares em cães com luxação de patela. Elas foram associadas com a idade do animal, massa corporal e grau de luxação. Foi avaliada também a porcentagem de casos com recidiva. As lesões patelares observadas foram erosão da cartilagem articular, exposição óssea subcondral, superfície patelar achatada ou côncava e entesófitos. As lesões extra patelares incluíram sinovite, osteófitos, ausência do sulco troclear, erosão das bordas condilares, espessamento da cápsula, lesão do tendão do músculo extensor digital, ruptura do ligamento cruzado cranial e prolapso de menisco. As lesões foram encontradas com maior frequência em animais com luxação de Grau II ou III e idade de 24 meses ou mais, sendo mais graves em cães com massa corporal superior a 15 kg. A luxação patelar ocasiona alterações que favorecem a degeneração articular e devem ser tratadas cirurgicamente. O tratamento conservativo alivia a dor, mas não corrige as alterações teciduais.(AU)

Evaluation of the presence of Leishmania infantum in bone stored in glycerol or sterilized by autoclave for cortical bone grafting / Avaliação da presença de Leishmania infantum em ossos armazenados em glycerol ou esterilizados em autoclave para enxertia óssea cortical

This study was performed to evaluate the effects of autoclaving and storage in 85% glycerol on cortical bone from dogs infected with Leishmania sp. We used 42 cadavers with leishmaniasis. The dogs were evaluated for the presence of Leishmania sp. in culture of bone marrow and by culturing cortical bone. From the infected animals, we harvested 42 diaphysis of the right femur for culture of cortical bone before and after autoclaving or storage in glycerol. There was no significant difference in growth of the parasite in culture of bone marrow or cortical bone. There was no growth of Leishmania sp. in culture of samples after autoclaving or storage in glycerol. Both treatments were effective in preventing the growth of the parasite in vitro, so it was considered viable for grafting.(AU)

Este estudo teve por objetivo avaliar os efeitos da esterilização em autoclave e do armazenamento em glicerol a 85% no osso cortical de cães infectados por Leishmania sp. Foram utilizados 42 cadáveres de cães com leishmaniose. Os cães foram avaliados para a presença de Leishmania sp. em cultura de medula óssea e de osso cortical. Foram coletadas 42 diáfises do fêmur direito para cultura do osso cortical antes e após a esterilização e o armazenamento em glicerol. Não houve diferença significativa no crescimento do parasito em cultura utilizando-se medula óssea ou osso cortical. Não houve crescimento de Leishmania sp. em cultura de amostras de osso cortical após a autoclavagem ou o armazenamento em glicerol. Ambos os tratamentos foram eficazes na prevenção do crescimento do parasita in vitro, de modo que foram considerados viáveis para enxerto.(AU)

Biocerâmica de fosfato de cálcio nanoestruturada micro-macroporosa em grânulos de absorção rápida no preenchimento de defeito crítico em rádio de coelhos (Oryctolagus cuniculus) / Bioceramics of calcium phosphate nano-structured micro-macro porous granules rapidly absorbed in filling critical radial defect in rabbits (Oryctolagus cuniculus)

O objetivo do presente trabalho foi avaliar, por radiografia, histologia e densitometria óssea, o efeito da HA/ßTCP em grânulos de absorção rápida em defeito ósseo crítico em rádio de coelhos. Foram utilizados 35 coelhos machos, da raça Nova Zelândia, e realizou-se um defeito crítico nos rádios direito e esquerdo. Os animais foram distribuídos em GI, enxerto autólogo e GII, HA/ßTCP em grânulos de absorção rápida. Avaliações radiográficas foram feitas antes da cirurgia, após, aos oito, 15, 30, 45 e 60 dias e avaliações histológicas e de densitometria. Verificou-se diferença significativa ao se comparar a densidade mineral óssea obtida ao longo do tempo de estudo. Observou-se formação de rede vascular entre os poros da biocerâmica desde o primeiro tempo de avaliação, (oito dias). Foram observados tecido ósseo primário e trabéculas em tecido ósseo preexistente a partir de 30 dias da implantação. Aos 60 dias, constatou-se presença de matriz óssea em segmentos ósseos preexistentes, caracterizando a formação óssea centrípeta. A biocerâmica HA/ßTCP nanoestruturada micro-macroporosa em grânulos de absorção rápida não causa alterações microscópicas indicativas de rejeição, permite a invasão e a multiplicação celular, bem como propicia a regeneração óssea, constituindo um implante apropriado para preenchimento de falhas ósseas críticas.(AU)

The objective of this study was to evaluate the effect of HA/ ßTCP on rapid absorption granules in rabbit radiography, histology, and bone densitometry. Thirty - five male rabbits of the New Zealand breed were used and a critical defect was performed on the right and left radios. The animals were distributed in GI, autologous graft and GII HA / ßTCP in rapidly absorbed granules. Radiographic, histological, and densitometry evaluations were performed before surgery, then after eight, 15, 30, 45 and 60 days. A significant difference was found when comparing the bone mineral density obtained over the study time. Formation of vascular network between the bioceramic pores was observed by the first evaluation time, (eight days). Primary bone tissue and trabeculae were observed from preexisting bone tissue after 30 days of implantation. At 60 days, the presence of bone matrix was observed from the preexisting bone segments, characterizing the centripetal bone formation. The micro-macroporous nanocomposite HA / ßTCP of rapidly absorbing granules do not cause microscopic changes indicative of rejection, allows invasion, cell multiplication, and promotes bone regeneration, constituting an appropriate implant for filling of critical bone failures.(AU)

Avaliação histológica e por microscopia eletrônica de varredura da biocerâmica de fosfato de cálcio nano-estruturada micromacro porosa em grânulos em defeito crítico de rádio de coelhos / Histological and nano-structured calcium phosphate bioceramic micro-macro porous granules in critical defect rabbits radius scanning electron microscopy evaluation

Objetivou-se avaliar, histologicamente e por microscopia eletrônica de varredura (MEV), a evolução de defeitos críticos experimentais em rádio de coelhos preenchidos ou não com biocerâmica fosfocálcica nanoestruturada micromacro porosa em grânulos. Utilizaram-se 70 coelhos, Nova Zelândia, adultos jovens, machos, e realizou-se um defeito crítico nos rádios. Os membros constituíram os grupos: GI, biocerâmica lenta, GII, biocerâmica moderada e GIII, controle negativo. Após cada período experimental, os animais foram sacrificados, e os rádios coletados. As avaliações histológicas foram realizadas aos oito, 15, 30, 45, 60, 90 e 120 dias, e as análises de MEV aos 60, 90 e 120 dias. Histologicamente, observou-se processo de reparação óssea mais adiantado nos grupos GI e GII comparando-se ao GIII. Na MEV, constatou-se maior proporção de osso maduro e presença de ósteons secundários nos GI e GII, sendo mais evidente no GII, confirmando os achados histológicos. As cerâmicas promoveram preenchimento completo do defeito, enquanto no grupo controle houve preenchimento no centro do defeito, permanecendo espaços vazios nas laterais. Conclui-se que o emprego das biocerâmicas de absorção lenta e moderada favorece a regeneração óssea completa em defeitos críticos sendo indicadas como substituto ósseo. A maturação óssea ocorre mais precocemente quando se emprega a cerâmica de absorção moderada.(AU)

The objective was to evaluate, through histologic exam and by scanning electron microscopy (SEM), the evolution of experimental critical defects in radio or not filled with nano-structured calcium phosphate micro-macro porous bioceramic granules rabbits. We used 70 rabbits, New Zealand, young adults, males, there was a critical defect in radio. The members were the groups: GI, slow bioceramic, GII, GIII and bioceramic moderate, negative control. After each experimental period, the animals were sacrificed and the radios harvested. Histological evaluations were performed at eight, 15, 30, 45, 60, 90 and 120 days and SEM analyzes at 60, 90 and 120 days. Histologically there was bone healing process earlier in GI and GII compared to GIII. In SEM we observed a higher proportion of mature bone and presence of secondary osteons in GI and GII, being more evident in the GII, confirming the histological findings. Ceramic promoted complete filling of the defect, while the control group was filling in the center of the defect, with empty spaces remaining on the sides. In conclusion, the use of bioceramics, slow and moderate absorption favor complete bone regeneration in critical defects being indicated as a bone substitute. The maturation occurs earlier when employing the ceramic moderate absorption.(AU)

Antioxidant activity and chemical composition of oleoresin from leaves and flowers of Brunfelsia uniflora.

In this study, the temperature and pressure of supercritical CO2 extraction were evaluated to obtain oleoresin of Brunfelsia uniflora leaves and flowers. The oleoresin compounds were identified by gas chromatography-mass spectrometry. The antioxidant activity was evaluated by three different methods. The highest oleoresin yields were 3.32% at 40°C and 200 bar for the leaves, and 1.03% at 60°C and 200 bar for the flowers. The main extracted compounds from leaves were phytol varying from 11.95 to 36.42% and α-tocopherol from 15.53 to 43.10%, and from flowers were geranyl linalool from 11.05 to 21.42% and α-amyrin from 9.66 to 22.12%. Oleoresin obtained at 60°C and 150 bar from leaves presented high antioxidant activity by DPPH (IC50 1.90 mg/mL) and by FRAP (1.8 µmol Fe2+/mg). ß-carotene/linoleic acid co-oxidation oleoresin from leaves at 0.25 mg/mL presented higher antioxidant activity than Trolox. The total phenolic content of the oleoresin from leaves ranged from 66.20 to 83.33 µg/mg and from flowers it was just up to 12.46 µg/mg. The extraction conditions affected yield, chemical composition, and antioxidant activity of oleoresin from leaves and flowers. This is the first report on the antioxidant activity of B. uniflora oleoresin from leaves and flowers and provides subsidies for potential applications in chemical, pharmaceutical, and food industries.

Morgan line and its relationship with distraction index, angle of inclination and degenerative joint disease in the diagnosis of canine hip dysplasia / Linha Morgan e sua relação com índice de distração, ângulo de inclinação e doença articular degenerativa no diagnóstico da displasia coxofemoral canina

We evaluated 160 hip joint radiographs of 40 dogs of different large breeds (25 females and 15 males) from the metropolitan area of Belo Horizonte, Minas Gerais, Brazil. The radiographs of each dog were obtained at two different stages: stage 1 (mean 7.23 months) and stage 2 (mean 14.25). The conventional radiographic method (CRM) and the radiographic distraction method (RDM) were used, carried out in both stages. CRM measured the Norberg angle (NA), the angle of inclination (AI) and evaluated the presence of degenerative joint disease (DJD). The MRD was performed to establish the distraction index (DI). The aims were to evaluate the presence of the Morgan line and other signs of DJD and correlate them with the degree of canine hip dysplasia (CHD) and also check if the DI greater than 0.3 (first stage) was associated with the presence of ML (second stage). It was found that DI, AI and changes of femoral neck and the formation of osteophytes were associated with the presence of ML. It was observed that if the DI is greater than 0.3 at the first stage, the chance of a positive outcome of ML in the second stage increases by 7.2 times. Thus, 49 joints showed DI > 0.3 at the first stage, in which 31 (63.3 %) presented ML at the second stage. Of the 31 animals that showed DI ≤ 0.3 at first, six (19.4%) had LM at the second stage. There has been a significant association between the presence of ML and the degree of CHD. The more severe the CHD, the higher the percentage of positive ML results. Thus, among the 24 (60 %) animals that showed ML, 11 (45.83 %) were classified as severe dysplastics, 5 (20.83%) as moderate and 8 (33.33 %) as mild. None of the animals classified as normal or borderline presented ML. Among the 8 animals classified as mild dysplastics, 5 showed only ML as DJD.(AU)

Avaliaram-se 160 radiografias das articulações coxofemorais de 40 cães de diferentes raças de grande porte, provenientes de Belo Horizonte, Brasil. As radiografias de cada animal foram obtidas em dois momentos: um (média de 7,23 meses de idade) e dois (média de 14,25 meses). Utilizaram-se o método radiográfico convencional (MRC) e o método radiográfico em distração (MRD), ambos realizados nos dois momentos. No MRC, mensurou-se o ângulo de Norberg (AN), o ângulo de inclinação (AI) e avaliou-se a presença de doença articular degenerativa (DAD). O MRD foi realizado para se estabelecer o índice de distração (ID). Os objetivos foram avaliar a presença da linha Morgan (LM) e de outros sinais de DAD, correlacionando-os com o grau de displasia coxofemoral (DCF); além de se verificar se o ID maior que 0,3 (primeiro momento) possui associação com a presença de LM (segundo momento). Verificou-se que ID, AI e encurtamento e/ou espessamento do colo femoral e formação de osteófitos foram associados à presença da LM. Observou-se que, quando o ID era maior que 0,3 no primeiro momento, a chance de um resultado positivo da LM no segundo momento aumentava em 7,2 vezes. No primeiro momento, 49 articulações apresentaram ID > 0,3. Destas, 31 (63,3%) apresentaram LM no segundo momento. Dos 31 animais que apresentaram ID ≤ 0,3 no primeiro momento, seis (19,4%) desenvolveram LM no segundo momento. Houve associação significativa entre presença de LM com o grau de DCF, e quanto mais grave a DCF, maior foi o percentual de resultados positivos da LM. Dos 24 (60%) animais que apresentaram LM, 11 (45,83%) foram classificados como displásicos graves, cinco (20,83%) como médios e oito (33,33%) como leves. Nenhum dos animais considerados suspeitos ou normais apresentou LM. Dos oito animais classificados como displásicos leves, cinco apresentaram apenas LM como sinal de DAD.(AU)

Parâmetros radiográficos de displasia coxofemoral na raça Rottweiler / Radiographic parameters of hip dysplasia in Rottweilers

In this study, 139 radiographs of the Rottweiller breed dog, 44 males and 95 females were evaluated. Radiographs were classified by the degree of hip dysplasia, and we measured the angle of inclination, Norberg angle, the cortical index and percentage of coverage of the femoral head on both sides. The aim of this study was to evaluate that parameters based on gender and age of animals correlating them with the incidence and severity of hip dysplasia. In dysplastic dogs the mean Norberg angle was 100.09 ± 7.86 °, the mean percent coverage of the femoral head was 41.50 ± 8.59%, the mean femoral cortical index was 0.33 ± 0.04, and the femoral angle mean was 153.06 ± 9.81 °. Higher inclination angles were associated with smaller angles of Norberg and lowest percentage of coverage of the femoral head. In this study gender, cortical index and angle of inclination did not show any association with the DCF.(AU)

Matriz porosa do BV60S no tratamento de defeitos ósseos críticosem rádios de cães / Porous matrix of BG60S in the treatment of critical bone defects in the radius of dogs

O objetivo deste estudo foi avaliar o efeito da matriz porosa do biovidro de composição molar 60% SiO2 - 36% CaO - 4% P2O5 (BV60S) no tratamento de defeitos ósseos críticos de cães. Foram utilizados 20 cães, machos, sem raça definida, com dois anos e massa corporal média de 25kg. Foram constituídos três grupos experimentais: defeitos ósseos preenchidos com BV60S (BV), com osso autógeno (C+) e defeitos não preenchidos (C-). A regeneração óssea foi avaliada por meio de exames radiográficos, densitométricos e histomorfométricos ao longo de 90 dias. A matriz do BV60S mostrou rápida reabsorção com redução média de 12,62% a cada 15 dias. A regeneração foi completa no grupo C+ e incompleta nos grupos BV e C-, aos 90 dias. A área de neoformação óssea foi semelhante entre os grupos BV e C-, em todos os tempos estudados. Conclui-se que a matriz porosa do BV60S possui rápida reabsorção, não sendo eficiente no tratamento de defeitos ósseos críticos em rádios de cães.

The aim of this study was to evaluate the effect of the porous matrix of bioglass with molar composition of 60% SiO2, 36% CaO, 4% P2O5 (BV60S) in the treatment of critical bone defects in dogs. 20 male mongrel dogs at two years of age and a mean weight of 25 kg were used. Three experimental groups were formed: bone defects filled with BV60S (BV), with autogenous bone (C+) and unfilled defects (C-). Bone regeneration was evaluated by radiography, bone densitometry and histoforfometry over 90 days. The matrix of BV60S showed rapid absorption with an average reduction of 12.62% every 15 days. Regeneration was complete in group C+ and incomplete in the BV-group and C- group at 90 days. The area of new bone formation was similar between BV and C- groups at all time points. It was concluded that the porous matrix BV60S has fast resorption not being effective in the treatment of critical bone defects in the radius of dogs.

Efeito do produto iônico do biovidro 60S na diferenciação osteogênica de células-tronco mesenquimais do tecido adiposo de cães / Effect of ionic product of bioglass 60S in osteogenic differentiation of mesenchymal stem cells from the adipose tissue of dogs

O objetivo do presente trabalho foi avaliar o produto iônico do biovidro 60S (BV60S) na diferenciação osteogênica de células-tronco mesenquimais de origem adiposa (CTM-AD) de cães. As CTM-AD foram diferenciadas sem OSTe com o produto iônico (PI OST) por sete, 14 e 21 dias. Avaliou-se o MTT, a fosfatase alcalina (FA), o colágeno, mineralização e as expressões de osterix (OSX), sialoproteína óssea (BSP), osteonectina (ON) e osteocalcina (OC). O grupo PI OSTmostrou menor conversão de MTT aos sete dias e maior conversão aos 21 dias. A atividade de FA foi maior no grupo OST, aos 14 e 21 dias. A síntese de colágeno foi maior no grupo OST aos sete e 21 dias. Verificou-se maior área mineralizada no grupo PI OSTem todos os tempos. Não houve diferenças nas expressões de OSX e OC entre os grupos. Observou-se maior expressão de BSP no grupo PI OST, aos 14 e 21 dias. A expressão de ON foi maior no grupo OST aos 14 dias. Concluiu-se que o produto iônico do BV60S favorece a osteogênese in vitro de CTM-AD de cães.

The aim was to evaluate the ionic product of 60S bioglass (BV60S) in osteogenic differentiation of mesenchymal stem cells from adipose tissue (ADMSCs) in dogs. ADMSCs were differentiated without the ionic product (OST) and with the ionic product (PI-OST) for 7, 14 and 21 days. We evaluated the MTT, alkaline phosphatase (ALP), collagen mineralization and expressions of osterix (OSX), bone sialoprotein (BSP), osteonectin (ON) and osteocalcin (OC). The PI-OST group had a lower MTT conversion to 7days and higher conversion at 21 days. The ALP activity was higher in the OST group at 14 and 21 days. Collagen synthesis was higher in the OST group at 7 and 21 days. A higher mineralized area in the PI-OST group was observed at all times. There were no differences in expressions of OSX and OC between groups. We observed increased expression of BSP in the PI-OST group at 14 and 21 days. The expression of ON was higher in the OST group at 14 days. It was concluded that the ionic product of BV60S promotes in vitro osteogenesis of MSC-AD from dogs.

Microwave-assisted extraction of green coffee oil and quantification of diterpenes by HPLC.

The microwave-assisted extraction (MAE) of 13 different green coffee beans (Coffea arabica L.) was compared to Soxhlet extraction for oil obtention. The full factorial design applied to the microwave-assisted extraction (MAE), related to time and temperature parameters, allowed to develop a powerful fast and smooth methodology (10 min at 45°C) compared to a 4h Soxhlet extraction. The quantification of cafestol and kahweol diterpenes present in the coffee oil was monitored by HPLC/UV and showed satisfactory linearity (R(2)=0.9979), precision (CV 3.7%), recovery (<93%), limit of detection (0.0130 mg/mL), and limit of quantification (0.0406 mg/mL). The space-time yield calculated on the diterpenes content for sample AT1 (Arabica green coffee) showed a six times higher value compared to the traditional Soxhlet method.

Preliminary crystallographic studies of a Schistosoma mansoni antigen (Sm21.7) dynein light-chain (DLC) domain.

Schistosomiasis is an inflammatory chronic disease that represents a major health problem in tropical and subtropical countries. The drug of choice for treatment, praziquantel, is effective in killing adult worms but fails to kill immature forms and prevent reinfection. One prominent antigen candidate for an anti-schistosomiasis vaccine is the protein Sm21.7 (184 amino-acid residues) from Schistosoma mansoni, a tegumental protein capable of reducing the worm burden in a murine immunization model. In the present work, the Sm21.7 gene was cloned and expressed in Escherichia coli and the full-length protein was purified to homogeneity. Crystals of recombinant Sm21.7 suitable for X-ray diffraction were obtained using PEG monomethyl ether 2000 as a precipitant. X-ray diffraction images of a native crystal (at 2.05 Šresolution) and a quick-cryosoaked NaI derivative (at 1.95 Šresolution) were collected on the W01B-MX2 beamline at the Laboratório Nacional de Luz Síncrotron (LNLS, Brazilian Synchrotron Light Laboratory/MCT). Both crystals belonged to the hexagonal space group P6122, with similar unit-cell parameters a=b=108.5, c=55.8 Å. SIRAS-derived phases were used to generate the first electron-density map, from which a partial three-dimensional model of Sm21.7 (from Gln89 to Asn184) was automatically constructed. Anaysis of dissolved crystals by SDS-PAGE confirmed that the protein was cleaved in the crystallization drop and only the Sm21.7 C-terminal domain was crystallized. The structure of the Sm21.7 C-terminal domain will help in the localization of the epitopes responsible for its protective immune responses, constituting important progress in the development of an anti-schistosomiasis vaccine.

Modified cementless total coxofemoral prosthesis: development, implantation and clinical evaluation / Prótese coxofemoral não cimentada: desenvolvimento, implantação e avaliação clínica

The aim of this study was to modify canine coxofemoral prostheses and the clinical evaluation of the implantation. Fifteen canine hips and femora of cadavers were used in order to study the surface points of modification in prostheses and develop a perforation guide. Femoral stems and acetabular components were perforated and coated with biphasic calcium phosphate layer. Twelve young adult male mongrel dogs were implanted with coxofemoral prostheses. Six were operated upon and implanted with cemented canine modular hip prostheses, establishing the control group. The remaining six were implanted with a novel design of cementless porous tricalcic phosphate-hydroxyapatite coated hip prostheses. Clinical and orthopedic performance, complications, and thigh muscular hypotrophy were assessed up to the 120th post-operatory day. After 120 days, animals with cementless prostheses had similar clinical and orthopedic performance compared to the cemented group despite the increased pain thigh hypotrophy. Animals that underwent cementless hip prosthesis evidenced more pain, compared to animals with cemented hip prosthesis that required longer recuperation time. No luxations, two fractures and two isquiatic neurapraxies were identified in the course of the study. Using both the cemented and the bioactive coated cementless model were suitable to dogs, showing clinical satisfactory results. Osseointegration and biological fixation were observed in the animals with the modified cementless hip prosthesis.

O objetivo deste estudo foi modificar a prótese coxofemoral canina cimentada e avaliar os efeitos clínicos. Foram utilizadas 15 pelves e 15 fêmures de cadáveres de cães para modificações da haste, do componente acetabular e para a confecção de guia de perfuração. As hastes e componentes acetabulares foram perfurados e revestidos com uma camada de fosfato de cálcio bifásico. Doze cães machos, adultos jovens, sem raça definida, foram submetidos à cirurgia para o implante de prótese coxofemoral total. Seis cães receberam prótese modular cimentada, grupo controle, e seis, a prótese modificada não cimentada com revestimento de fosfato de cálcio bifásico. Foram avaliados a técnica de implantação, o desempenho clínico, o grau de hipotrofia muscular da coxa e as complicações durante 120 dias. Os animais com prótese não cimentada mostraram desempenho clínico similar aos animais com a prótese cimentada, porém mostraram maior hipotrofia muscular decorrente de dor e maior tempo de recuperação. Não foram observadas luxações. No entanto, duas fraturas e dois casos de neurapraxia isquiádica foram observados. A utilização de ambas as próteses coxofemorais, cimentada e não cimentada, com recobrimento bioativo são eficientes no cão, com resultados clínicos satisfatórios, mas a osteointegração e fixação biológica ocorreram na prótese com recobrimento de fosfato de cálcio bifásico, objetivo do tratamento que previne afrouxamento futuro.

Scintigraphy in postoperative follow-up of osteosynthesis of the femur with a bridge plate associated with an intramedullary pin in rabbits / Cintilografia no acompanhamento pós-operatório de osteossíntese do fêmur com placa em ponte associada ao pino intramedular em coelhos

The objective of this work was to monitor the consolidation of the femur after osteosynthesis with a bridge plate associated with the intramedullary pin using scintigraphy. We used seven New Zealand breed male rabbits, at 4 months of age, with a mean weight of 3.5 kg. We performed a three-phase bone scintigraphy with technetium-labeled methylene diphosphonate (99mTc-MDP) before and after surgery, and 20, 50 and 90 days postoperatively. The activity index (AI) was calculated by dividing the average number of uptake counts in the region of the osteotomy by the average number of counts in the corresponding region in the contralateral limb. Radiography was performed before surgery, after surgery, and 15, 30, 45, 60 and 90 days postoperatively. We found a direct relationship between the activity index and progress of bone scintigraphy in the evaluation sequence over the period of observation. Scintigraphy allows monitoring of bone metabolism and measurement of vascularization and/or bone or tissue perfusion. The images obtained in the blood pool and static phases are the most appropriate for assessing bone metabolism in the context of this study. The bridge plate associated with the intramedullary pin promotes osteosynthesis with sufficient stability to allow bone consolidation.

A cintilografia foi empregada no acompanhamento da consolidação de osteotomia experimental do fêmur após osteossíntese com placa em ponte associada ao pino intramedular. Foram usados sete coelhos machos, raça Nova Zelândia, com massa corporal de 3,5kg e idade média de quatro meses. A cintilografia óssea trifásica com metilenodifosfonato marcado com tecnécio-99m (MDP-99mTc) foi obtida antes e após a cirurgia, e nos dias 20, 50 e 90 do pós-operatório.O índice de atividade (IA) foi calculado por meio do quociente da média do número de contagens na região da osteotomia pela média do número de contagens na região correspondente, no membro contralateral normal. Paralelamente ao estudo cintilográfico, foram realizadas radiografias antes dos procedimentos cirúrgicos e aos 15, 30, 45, 60 e 90 dias após, para acompanhamento do processo de cura óssea. Encontrou-se relação direta entre o índice de atividade e a evolução do processo de consolidação óssea na avaliação cintilográfica sequencial ao longo do período de observação. A cintilografia óssea trifásica permite acompanhar o metabolismo ósseo, avaliar e mensurar a vascularização e perfusão tecidual. As imagens obtidas na fase de pool sanguíneo e na fase óssea são as mais adequadas para avaliação do metabolismo ósseo. A placa em ponte associada com o pino intramedular promove osteossíntese com estabilidade suficiente para permitir a consolidação ossea.

Intra-articular injections of autologous platelet concentrates in dogs with surgical reparation of cranial cruciate ligament rupture: a pilot study.

OBJECTIVES: The objective of this study was to evaluate by clinical, radiographic, and force plate gait analyses the effect of post-surgical intra-articular injections of autologous platelet concentrates (PC) in a small group of dogs with cranial cruciate ligament (CCL) rupture. METHODS: The ten dogs used in this study were initially presented with CCL rupture and underwent ligament replacement surgery by fascia lata autograft guided by arthroscopy. Six dogs received three intra-articular injections of PC (PC group); one dose was injected immediately after surgery, and two additional doses were injected at two-week intervals. The remaining four dogs received only nutraceuticals (control group). All dogs were evaluated by clinical examination, serial radiography, and force plate gait analyses at monthly intervals up to 90 days. RESULTS: The clinical follow-up of the PC-treated group indicated a better outcome than the control group. Radiographic evaluation was not conclusive. Values of peak vertical reaction force and vertical impulse of the affected limbs were only significantly larger on the 90th postoperative day in the PC group compared to the control group. CLINICAL SIGNIFICANCE: Our results indicate that autologous PC might improve functional outcome after intra-articular cranial cruciate ligament repair. The effect of PC when using other repair procedures warrants additional studies.

Microwave-assisted methanolysis of green coffee oil.

Optimisation of a microwave-assisted methanolysis was performed to obtain cafestol and kahweol directly from green coffee oil (Coffea arabica). A two-factor (the methanolysis period and temperature), three-level, factorial experimental design (3(2)) was adopted. The methanolysis procedure was performed under microwave irradiation, using closed vessel and accurate fast responding internal fibre-optic temperature probe. The effects on the responses were measured by HPLC. After 3 min of microwave irradiation (hold time) at 100°C, with 500 mg of green coffee oil, a yield higher than 99% was obtained. The yield of this reaction is 26% after 2h when working under conventional heating. The methods described in the literature lead to long reaction times, poor yields and formation of side products. The microwave-assisted technique proved to be faster, avoided undesired side products and gave better conversion, when compared to conventional heating process.

Uso de plasma rico em plaquetas intra-articulares como tratamento pós-cirúrgico da ruptura do ligamento cruzado cranial num cão / The use of intra-articular platelet rich-plasma as a postsurgical treatment of cranial cruciate ligament rupture in a dog

Relata-se o caso de um cão que recebeu injeções intra-articulares de plasma rico em plaquetas (PRP) durante o pós-operatório do tratamento cirúrgico de ruptura do ligamento cruzado cranial (RLCCr). Os resultados clínicos e da avaliação da marcha mediante plataforma de força neste paciente sugerem a utilização de injeções intra-articulares de PRP como terapia pós-cirúrgica no tratamento da RLCCr.

We report the case of a dog that received intra-articular injections of platelet-rich plasma (PRP) during the postoperative period of surgical treatment of cranial cruciate ligament (CCL) rupture. Clinical, and gait analysis by the force plate in this patient might suggest the use of intra-articular injections of PRP as a post-surgical therapy in the treatment of CCL rupture.