Total fraction of mononuclear cell or stromal vascular fraction transplant associated with cellulosic membrane in experimental cutaneous wounds in rabbits

The skin presents various important functions to the organism. The maintenance of its integrity is fundamental, among others, to prevent penetration of microorganisms and exit of liquids essential to life maintenance. Due to its constant environment exposure, the skin is highly susceptible to trauma which can result solutions of continuity. The healing of wounded skin should be fast and many alternatives are searched for, aiming to reduce repair time and to guarantee a functional and esthetically acceptable scar. One option to repair injured tissues which is ever more viable is cellular therapy with adult stem cells. The aims of this study consisted in evaluating the transplant of total mononuclear cell fraction (TMCF) from bone marrow (BM) or stromal vascular fraction (SVF) from adipose tissue (AT), associated with the use of cellulose membrane. To carry out this experiment, 20 rabbits were randomly divided in four groups with five animals each. After induction of the experimental cutaneous wound, Group A did not receive any treatment; group B received only cellulose membrane; and groups C and D, in addition to receiving the membrane, were submitted to autologous transplant of TMCF, with total cell value between 6,92 x 10(6) and 4,91 x 10(7) and a viability of 82 to 97% or SVF, with total cell value between 9,6 x 10(5) and 6,5 x 10(6) and a viability of 66 a 87%, respectively. At the end of the evaluation period, the three treated groups presented significant statistical difference of wound area in relation to the control group, and the group which received SVF from adipose tissue presented the shortest wound healing time.A pele apresenta diversas funções importantes para o organismo. A manutenção de sua integridade é fundamental, entre outras, para impedir a penetração de microrganismos e a perda de líquidos essenciais à manutenção da vida. Por estar constantemente exposta ao meio ambiente, a pele é altamente susceptível a traumas que podem acarretar soluções de continuidade. A cicatrização da pele ferida deve ser rápida e diversas alternativas são buscadas, visando à redução do tempo de reparo a fim de garantir cicatriz funcional e esteticamente aceitável. Uma opção cada vez mais viável para reparar tecidos danificados é a terapia celular com células-tronco adultas. Os objetivos deste trabalho consistiram em avaliar o transplante da fração total de células mononucleares (FTCM) da medula óssea (MO) e da fração vascular estromal (FVE) do tecido adiposo (TA), associado ao uso da membrana celulósica. Para realização deste experimento, 20 coelhos foram divididos aleatoriamente em quatro grupos com cinco animais cada. Após a indução da ferida cutânea experimental, o grupo A, não recebeu nenhum tratamento, o grupo B, recebeu apenas membrana celulósica e os grupos C e D, além de receberem a membrana, foram submetidos a transplante autólogo da FTCM, com valor total de células entre 6,92 x 10(6) a 4,91 x 10(7) e uma viabilidade de 82 a 97% ou da FVE, com valor total de células entre 9,6 x 10(5) e 6,5 x 10(6) e uma viabilidade de 66 a 87%, respectivamente. Ao final do período de avaliação, os três grupos tratados apresentaram diferença estatística significativa da área da ferida em relação ao grupo controle e o grupo que recebeu a FVE do tecido adiposo apresentou o menor tempo de cicatrização da ferida

Remoção da cartilagem articular associada ou não a implante homógeno ou enxerto autógeno de osso esponjoso em cães submetidos à artrodese atlantoaxial Joint cartilage removal associated or not to homologous implant or autologous cancellous bone graft in dogs submitted to atlantoaxial arthrodesis

O objetivo deste estudo foi avaliar o grau de fusão articular e formação óssea na articulação atlantoaxial de cães submetidos à artrodese após a remoção da cartilagem articular associada ou não ao implante homógeno ou enxerto autógeno de osso esponjoso. Foram utilizados 12 cães, adultos, distribuídos aleatoriamente em três grupos iguais. Grupo I (GI): realizada apenas a remoção da cartilagem articular e imobilização articular com pinos e resina acrílica. Grupo II (GII): feita a remoção da cartilagem articular e imobilização da articulação, seguida da colocação e modelagem do implante ósseo esponjoso homógeno entre as superfícies articulares. Grupo III (GIII): foi realizado o mesmo procedimento do GII, mais o enxerto ósseo esponjoso autógeno no local determinado. Realizaram-se exames radiográficos em todos os animais aos 30, 60 e 90 dias de pós-operatório (PO). Aos 90 dias de PO foi feita a eutanásia para o emprego do teste de palpação manual, avaliação tomográfica e histopatológica. Para análise estatística da associação entre o grau de fusão articular, aplicou-se o Teste Qui-quadrado de independência. Os resultados dos testes foram avaliados pela significância exata e considerados significantes a 5% (P<0,05). Pelo teste de palpação manual e pelas imagens tomográficas, não houve diferença entre os grupos aos 90 dias de PO. A análise radiográfica da articulação atlantoaxial mostrou que o grau de fusão era semelhante entre os tratamentos entre os tempos de evolução. Com relação ao estudo histopatológico da articulação atlantoaxial dos cães, decorridos 90 dias de PO, foi verificado que a proliferação óssea no grupo I apresentou grau de intensidade proporcional, no grupo II, 75% leve e 25% acentuada e, no grupo III, 25% moderada e 75% acentuada. Não existe diferença quanto ao grau de fusão articular e formação óssea quando a técnica de artrodese atlantoaxial em cães for associada ao enxerto de osso esponjoso autógeno ou implante homógeno. A remoção da cartilagem articular isoladamente é uma alternativa eficiente e apresenta resultados semelhantes, quando associada ao enxerto autógeno ou implante homógeno.<br>The aim of this study was to evaluate the degree of joint fusion and bone formation in dogs undergoing atlantoaxial arthrodesis after removal of articular cartilage associated or not to implant homogenous or autogenous cancellous bone. Twelve dogs, weighing between 8 and 12kg were randomly divided into three groups. Group I (GI) performed only the removal of joint cartilage and joint immobilization with acrylic resin and pins. Group II (GII): after removel of joint cartilage and articular immobilization was performed modeling and placement of homogenous cancellous bone at the given location. The volume of homograft placed in the joint was measured using a precision balance and all animals received the same amount. Group III (GIII): this was the same procedure as GI, however, was used autogenous cancellous bone graft at the given location. Radiographic examinations were performed on all animals at 30, 60 and 90 days postoperatively. At 90 days of PO was carried out euthanasia of animals for testing of manual palpation and CT evaluation and histological. Samples were kept in solution in 10% buffered formaldehyde for fixation for a minimum of 72 hours and after, decalcified in formic acid and sodium citrate for subsequent processing and included in paraffin. Sections were stained with hematoxylin-eosin for light microscopy evaluation. Statistical analysis of the association between the degree of joint fusion assessed by manual palpation, the digitized radiographic images and CT scans and treatment groups, we applied the Chi-square test of independence. The test results were evaluated by the exact significance and considered statistically significant at 5% significance (P<0.05). By manual palpation test and by the CT images can be seen that there was no statistically significant difference between groups at 90 days postoperatively. Radiographic analysis of the atlantoaxial joint showed that the degree of fusion was similar among treatments, no statistical difference at 30, 60 and 90 days postoperatively. Regarding the histopathological study of atlantoaxial joint of dogs after 90 days postoperatively was found that bone formation in group I had 25% of each intensity (absent, mild, moderate, severe) in group II, 75% lighter and 25% sharp and in group III, 25% moderate and 75% severe. It can be concluded that the use of graft homogenous preserved in 98% glycerin in dogs undergoing atlantoaxial fusion method is a viable alternative for treatment of atlantoaxial instability. There is no difference in the degree of joint fusion and bone formation when the technique of atlantoaxial arthrodesis in dogs is associated with autogenous cancellous bone graft or implant homogenous. The removal of articular cartilage alone is an effective and shows similar results to the use of autograft or homogenous implant

Minimally Invasive Approach to Renal Artery Using Different Types of Catheter by Conventional Radiology

Background: Several routes of administration have been used for cell and gene therapy delivery in kidney disease in the last years. Moreover, a more homogenous distribution and a higher response was observed when administered intra-arterially. The main objective of the present study was to describe and to compare the use of Fogarty thru-lumen embolectomy catheter with angiographic catheter through the femoral artery by using conventional radiology as a guide.Materials, Methods & Results: Seven healthy dogs were used in these experiments. First, a Fogarty thru-lumen embolectomy catheter was introduced followed by an angiographic catheter. The primary outcome was the number of attempts for the placement of the introducer and the number of radiographic studies to carry out nephrography. Preceding the surgical procedure, ultrasonographic evaluation was performed in all animals in order to measure the diameter of the left and right femoral arteries, left and right renal and abdominal aorta at the renal level. After trichotomy and antisepsis of the medial face of the two pelvic limbs, the 6F valve introducer was placed based on the modified Seldinger technique. Thereafter, aortography was performed, showing its collateral arteries, with contrast injection, and it was estimated how much of the Fogarty 5,5F catheter had to be introduced. Subsequently, a guidewire was inserted through the catheter followed by the Fogarty catheter. Another x-ray was taken to show the position of the catheter. It was observed whether the catheter was positioned correctly and the cuff was filled, and then another contrasting radiograph was performed. If the contrast was not going to the kidneys, the cuff of the catheter was deflated, the catheter was repositioned and a new contrast radiography was performed. For the angiographic catheters technique, it was estimated how much of the catheter needed to be introduced, and then the curvature of the catheter was directed to the left kidney, followed by placement of the guidewire. The guidewire was then removed and X-rays were taken to confirm the position as many times as necessary for the correct positioning of the catheter. After selective catheterization of the left kidney, the catheter was removed and its tip was repositioned for catheterization of the right kidney, repeating the same procedure. After the radiographic evaluation, the 6F valve introducer was removed followed by the hemostasis of the femoral artery. In five out of seven animals it was possible to place the introducer tube 6F. Angiographic catheter technique needed significantly more radiographic films compared to Fogarty catheter when both arteries were evaluated. No significant difference was observed when compared to only one renal artery. The only complication observed was hematoma in three animals.Discussion: The access of the renal artery for drug delivery and cell therapy is possible using both Fogarty thru-lumen embolectomy catheter and angiographic catheter with conventional radiology as a guide in dogs over 10 kg. However, we considered the Fogarty catheter easier to be applied, since only one procedure is needed to access both renal arteries, thus making it less expensive and faster to run. If access to only one renal artery is required, the angiographic catheter is recommended, since cell therapy would be administered more selectively in the target kidney

Minimally invasive approach to renal artery using different types of catheter by conventional radiology

Background: Several routes of administration have been used for cell and gene therapy delivery in kidney disease in the last years. Moreover, a more homogenous distribution and a higher response was observed when administered intra-arterially. The main objective of the present study was to describe and to compare the use of Fogarty thru-lumen embolectomy catheter with angiographic catheter through the femoral artery by using conventional radiology as a guide. Materials, Methods & Results: Seven healthy dogs were used in these experiments. First, a Fogarty thru-lumen embolectomy catheter was introduced followed by an angiographic catheter. The primary outcome was the number of attempts for the placement of the introducer and the number of radiographic studies to carry out nephrography. Preceding the surgical procedure, ultrasonographic evaluation was performed in all animals in order to measure the diameter of the left and right femoral arteries, left and right renal and abdominal aorta at the renal level. After trichotomy and antisepsis of the medial face of the two pelvic limbs, the 6F valve introducer was placed based on the modified Seldinger technique Thereafter, aortography was performed, showing its collateral arteries, with contrast injection, and it was estimated how much of the Fogarty 5,5F catheter had to be introduced. Subsequently, a guidewire was inserted through the catheter followed by the Fogarty catheter. Another x-ray was taken to show the position of the catheter. It was observed whether the catheter was positioned correctly and the cuff was filled, and then another contrasting radiograph was performed. If the contrast was not going to the kidneys, the cuff of the catheter was deflated, the catheter was repositioned and a new contrast radiography was performed. For the angiographic catheters technique, it was estimated how much of the catheter needed to be introduced, and then the curvature of the catheter was directed to the left kidney, followed by placement of the guidewire. The guidewire was then removed and X-rays were taken to confirm the position as many times as necessary for the correct positioning of the catheter. After selective catheterization of the left kidney, the catheter was removed and its tip was repositioned for catheterization of the right kidney, repeating the same procedure After the radiographic evaluation, the 6F valve introducer was removed followed by the hemostasis of the femoral artery. In five out of seven animals it was possible to place the introducer tube 6F. Angiographic catheter technique needed significantly more radiographic films compared to Fogarty catheter when both arteries were evaluated. No significant difference was observed when compared to only one renal artery. The only complication observed was hematoma in three animals. Discussion: The access of the renal artery for drug delivery and cell therapy is possible using both Fogarty thru-lumen embolectomy catheter and angiographic catheter with conventional radiology as a guide in dogs over 10 kg. However, we considered the Fogarty catheter easier to be applied, since only one procedure is needed to access both renal arteries, thus making it less expensive and faster to run. If access to only one renal artery is required, the angiographic catheter is recommended, since cell therapy would be administered more selectively in the target kidney

Minimally invasive approach to renal artery using different types of catheter by conventional radiology

Background: Several routes of administration have been used for cell and gene therapy delivery in kidney disease in the last years. Moreover, a more homogenous distribution and a higher response was observed when administered intra-arterially. The main objective of the present study was to describe and to compare the use of Fogarty thru-lumen embolectomy catheter with angiographic catheter through the femoral artery by using conventional radiology as a guide. Materials, Methods & Results: Seven healthy dogs were used in these experiments. First, a Fogarty thru-lumen embolectomy catheter was introduced followed by an angiographic catheter. The primary outcome was the number of attempts for the placement of the introducer and the number of radiographic studies to carry out nephrography. Preceding the surgical procedure, ultrasonographic evaluation was performed in all animals in order to measure the diameter of the left and right femoral arteries, left and right renal and abdominal aorta at the renal level. After trichotomy and antisepsis of the medial face of the two pelvic limbs, the 6F valve introducer was placed based on the modified Seldinger technique Thereafter, aortography was performed, showing its collateral arteries, with contrast injection, and it was estimated how much of the Fogarty 5,5F catheter had to be introduced. Subsequently, a guidewire was inserted through the catheter followed by the Fogarty catheter. Another x-ray was taken to show the position of the catheter. It was observed whether the catheter was positioned correctly and the cuff was filled, and then another contrasting radiograph was performed. If the contrast was not going to the kidneys, the cuff of the catheter was deflated, the catheter was repositioned and a new contrast radiography was performed. For the angiographic catheters technique, it was estimated how much of the catheter needed to be introduced, and then the curvature of the catheter was directed to the left kidney, followed by placement of the guidewire. The guidewire was then removed and X-rays were taken to confirm the position as many times as necessary for the correct positioning of the catheter. After selective catheterization of the left kidney, the catheter was removed and its tip was repositioned for catheterization of the right kidney, repeating the same procedure After the radiographic evaluation, the 6F valve introducer was removed followed by the hemostasis of the femoral artery. In five out of seven animals it was possible to place the introducer tube 6F. Angiographic catheter technique needed significantly more radiographic films compared to Fogarty catheter when both arteries were evaluated. No significant difference was observed when compared to only one renal artery. The only complication observed was hematoma in three animals. Discussion: The access of the renal artery for drug delivery and cell therapy is possible using both Fogarty thru-lumen embolectomy catheter and angiographic catheter with conventional radiology as a guide in dogs over 10 kg. However, we considered the Fogarty catheter easier to be applied, since only one procedure is needed to access both renal arteries, thus making it less expensive and faster to run. If access to only one renal artery is required, the angiographic catheter is recommended, since cell therapy would be administered more selectively in the target kidney