Usefulness of Kidney Donor Profile Index (KDPI) to predict graft survival in a South Brazilian Cohort / Utilidade do Índice de Perfil de Doadores de Rins (KDPI) para prever a sobrevida do enxerto em uma coorte do Sul do Brasil

Abstract Introduction: Kidney Donor Profile Index (KDPI) has been incorporated in the United States to improve the kidney transplant allocation system. Objectives: To evaluate deceased kidney donors' profile using KDPI and compare to the previous United Network for Organ Sharing (UNOS) definition of expanded criteria donors (ECD) and assess the KDPI applicability to predict five-year graft survival and renal function in our sample. Methods: Retrospective cohort of 589 kidney transplants from deceased donors performed from January 2009 to May 2013 with follow-up until May 2018. Results: In 589 kidney transplants, 36.6% of donors were classified as ECD and 28.8% had KDPI ≥ 85%. Mean KDPI was 63.1 (95%CI: 60.8-65.3). There was an overlap of standard and ECD in KDPI between 60 and 95 and a significantly lower death-censored graft survival in KDPI ≥ 85% (78.6%); KDPI 0-20: 89.8%, KDPI 21-59: 91.6%, and KDPI 60-84: 83.0%; p = 0.006. The AUC-ROC was 0.577 (95%CI: 0.514-0.641; p = 0.027). Renal function at 5 years was significantly lower according to the incremental KDPI (p < 0.002). KDPI (HR 1.011; 95%CI 1.001-1.020; p = 0.008), donor-specific antibodies (HR 2.77; 95%CI 1.69-4.54; p < 0.001), acute rejection episode (HR 1.73; 95%CI 1.04-2.86; p = 0.034) were independent and significant risk factors for death-censored graft loss at 5 years. Conclusion: In our study, 36.6% were classified as ECD and 28.8% had KDPI ≥ 85%. KDPI score showed a moderate power to predict graft survival at 5 years. Renal function was significantly lower in patients with higher KDPI.

Resumo Introdução: O Índice de Perfil de Doadores de Rins (KDPI) foi adotado nos Estados Unidos para melhorar o sistema de alocação de transplantes renais. Objetivos: avaliar o perfil dos doadores de rim falecidos usando o KDPI e comparar com a definição anterior do United Network for Organ Sharing (UNOS) de doadores de critérios expandidos (DCE) e avaliar a aplicabilidade do KDPI para prever a sobrevida do enxerto em cinco anos e a função renal em nossa amostra. Métodos: Coorte retrospectiva de 589 transplantes renais de doadores falecidos, realizada de janeiro de 2009 a maio de 2013, com acompanhamento até maio de 2018. Resultados: Em 589 transplantes renais, 36,6% dos doadores foram classificados como DCE e 28,8% apresentaram KDPI ≥ 85%. O KDPI médio foi de 63,1 (IC 95%: 60,8-65,3). Houve uma sobreposição de padrão e DCE no KDPI entre 60 e 95 e uma sobrevida do enxerto censurada por óbito significativamente menor no KDPI ≥ 85% (78,6%); KDPI 0-20: 89,8%, KDPI 21-59: 91,6% e KDPI 60-84: 83,0%; p = 0,006. A ASC-ROC foi de 0,577 (IC 95%: 0,514-0,641; p = 0,027). A função renal aos 5 anos foi significativamente menor de acordo com o aumento do KDPI (p <0,002). KDPI (HR 1.011; 95% CI 1.001-1.020; p = 0.008), anticorpos específicos contra doadores (HR 2,77; 95% CI 1,69-4,54; p <0,001), episódio de rejeição aguda (HR 1,73; 95% CI 1,04-2,86; p = 0,034) foram fatores de risco independentes e significativos para perda do enxerto censurada por óbito em 5 anos. Conclusão: Em nosso estudo, 36,6% foram classificados como DCE e 28,8% apresentaram KDPI ≥ 85%. O escore KDPI mostrou potencial moderado para prever a sobrevida do enxerto em 5 anos. A função renal foi significativamente menor nos pacientes com maior KDPI.

Kidney transplantation with donors in severe acute kidney injury. Should we use these organs? Retrospective Case Series / Transplante renal com doadores em lesão renal aguda severa. Devemos utilizar esses órgãos? Série de casos retrospectiva

ABSTRACT Introduction: The number of incident and prevalent patients on dialysis has increased, as well as the number of candidates for renal transplantation in Brazil, without a proportional increase in the number of organ donors. The use of expanded kidneys, as to renal function, may be an alternative to increase the supply of organs. Objective: to discuss the feasibility of using expanded kidneys for renal function, which are in severe acute renal injury. Methods: All cases of renal transplantation of deceased donors performed at the Hospital das Clínicas de Botucatu of UNESP, from January 2010 to June 2018, totaling 732 cases were evaluated. Cases with final donor creatinine greater than 6 mg/dL were selected. Results: four patients were selected, of whom all donors were in severe acute kidney injury (AKI). These donors presented rhabdomyolysis as a probable cause of severe AKI, were young, with no comorbidities and had decreased urinary volume in the last 24 hours. The clinical evolution of all the recipients was satisfactory, with a glomerular filtration rate after transplantation ranging from 48 to 98 mL/min/1.73 m2. Conclusion: this series of cases shows the possibility of using renal donors in severe AKI, provided the following are respected: donor age, rhabdomyolysis as the cause of AKI, and implantation-favorable biopsy findings. Additional studies with better designs, larger numbers of patients and longer follow-up times are needed.

RESUMO Introdução: O número de pacientes incidentes e prevalentes em diálise tem aumentado, assim como o número de candidatos ao transplante renal no Brasil, sem um aumento proporcional do número de doadores de órgãos. O uso de rins expandidos, quanto à função renal, pode ser uma alternativa para aumentar a oferta de órgãos. Objetivo: discutir a viabilidade do uso de rins expandidos quanto à função renal, que estejam em lesão renal aguda severa. Métodos: foram avaliados todos os casos de transplante renal de doador falecido realizados no Hospital das Clínicas de Botucatu da UNESP, de janeiro de 2010 a junho de 2018, totalizando 732 casos. Selecionou-se os casos com creatinina final do doador maior do que 6 mg/dL. Resultados: quatro pacientes foram selecionados, dos quais todos os doadores estavam em lesão renal aguda (LRA) severa. Esses doadores apresentavam rabdomiólise como provável causa de LRA severa, eram jovens, sem comorbidades e apresentavam diminuição de volume urinário nas últimas 24 horas. A evolução clínica de todos os receptores foi satisfatória, com taxa de filtração glomerular após o transplante variando entre 48 a 98 mL/min/1,73m2. Conclusão: essa série de casos mostra a possibilidade de utilização de doadores renais em LRA severa, desde que respeitadas as condições seguintes: idade do doador, rabdomiólise como causa de LRA e achados de biópsia favoráveis à implantação. Estudos adicionais com melhores desenhos, maior número de pacientes e maiores tempos de seguimento são necessários.

Is the combination of fat grafts and platelet rich plasma effective in rats?

ABSTRACT PURPOSE: To investigate if the association of fat grafts and platelet-rich plasma (PRP) improves graft viability in female rats. METHODS: This is an experimental, randomized and blinded study, which involved 47 rats. Fat was harvested from the inguinal region and grafted to the cranial region. The experimental group consisted of PRP-enriched fat grafts (n=22) whilst the control group consisted of fat graft only (n=25). After a 100-day period, the animals were euthanised and the fat grafts were analyzed using scores from 0 (absent) to 4 (abundant), in optical microscopy by two independent and blinded pathologists. RESULTS: Regarding fat graft cell viability, the PRP group scored moderate/abundant in 63% of cases and the fat graft only group scored absent/slight in 72% of cases (p=0.03). The PRP group also presented lower fat necrosis scores when compared to the fat graft only group (p=0.03). Tumors (dermoid cysts) within the fat grafts were observed in three animals in which the grafts were mixed with PRP. CONCLUSION: Platelet-rich plasma improves the viability and integration of fat grafts in rats, but more studies are needed to fully understand the exact mechanisms that lead to this improvement and assess the safety of the method for use in humans.

Expanding the pool of kidney donors: use of kidneys with acute renal dysfunction / Ampliando o pool de doadores de rim: utilização de órgãos com disfunção renal aguda

ABSTRACT Given the shortage of organs transplantation, some strategies have been adopted by the transplant community to increase the supply of organs. One strategy is the use of expanded criteria for donors, that is, donors aged >60 years or 50 and 59 years, and meeting two or more of the following criteria: history of hypertension, terminal serum creatinine >1.5mg/dL, and stroke as the donor´s cause of death. In this review, emphasis was placed on the use of donors with acute renal failure, a condition considered by many as a contraindication for organ acceptance and therefore one of the main causes for kidney discard. Since these are well-selected donors and with no chronic diseases, such as hypertension, renal disease, or diabetes, many studies showed that the use of donors with acute renal failure should be encouraged, because, in general, acute renal dysfunction is reversible. Although most studies demonstrated these grafts have more delayed function, the results of graft and patient survival after transplant are very similar to those with the use of standard donors. Clinical and morphological findings of donors, the use of machine perfusion, and analysis of its parameters, especially intrarenal resistance, are important tools to support decision-making when considering the supply of organs with renal dysfunction.

RESUMO Diante da escassez de órgãos para transplante, algumas estratégias têm sido adotadas pela comunidade transplantadora, no sentido de ampliar a oferta de órgãos. Uma delas é a utilização de rins de doadores com critérios expandidos, ou seja, doadores com idade >60 anos ou entre 50 e 59 anos, e que atendem a dois ou mais dos seguintes critérios: história de hipertensão, creatinina sérica terminal >1,5mg/dL e acidente vascular cerebral como causa de morte do doador. Nesta revisão, foi dada ênfase à utilização de doadores com disfunção renal aguda, condição considerada por muitos uma contraindicação para a aceitação de órgãos e, portanto, uma das principais causas de descarte de órgãos. Desde que sejam doadores bem selecionados e que não tenham doença renal crônica, como hipertensão ou diabetes, muitos trabalhos mostraram que o uso de doadores com disfunção renal aguda deve ser encorajado, pois, em geral, a disfunção renal aguda é de caráter reversível. Embora, a maioria dos estudos tenha demonstrado que há uma maior taxa de função retardada do enxerto com a utilização desses órgãos, os resultados de sobrevida do enxerto e do paciente após o transplante são muito semelhantes aos resultados obtidos da utilização de doadores padrão. Os achados clínicos e morfológicos do doador, a utilização da máquina de perfusão e a análise de seus parâmetros, principalmente a resistência intrarrenal, são importantes ferramentas de apoio para tomada de decisão no momento da oferta de órgãos com disfunção renal.

¿Influye el medio de preservación del injerto en los resultados y costos del trasplante hepático de donante fallecido?: Comparación de dos soluciones de preservación / Outcomes using two preservation solutions (UW/HTK) in liver transplantation from brain death donors

Background: Preservation solutions are critical for organ transplantation. In liver transplant (LT), the solution developed by the University Of Wisconsin (UW) is the gold-standard to perfuse deceased brain death donor (DBD) grafts. Histidine-Tryptophan-Ketoglutarate (HTK), formerly a cardioplegic infusion, has been also used in solid organ transplantation. Aim: To compare the outcomes of LT in our center using either HTK or UW solution. Patients and Methods: Retrospective study including 93 LT DBD liver grafts in 89 patients transplanted between March 1994 and July 2010. Forty-eight grafts were preserved with UW and 45 with HTK. Donor and recipient demographics, total infused volume, cold ischemia time, post-reperfusion biopsy, liver function tests, incidence of biliary complications, acute rejection and 12-month graft and patient survival were assessed. Preservation solution costs per liver graft were also recorded. Results: Donor and recipient demographics were similar. When comparing UW and HTK, no differences were observed in cold ischemia time (9.6 ± 3 and 8.7 ± 2 h respectively, p = 0.23), biliary complications, the incidence of acute rejection, primary or delayed graft dysfunction. Histology on post-reperfusion biopsies revealed no differences between groups. The infused volume was significantly higher with HTK than with UW (9 (5-16) and 6 (3-11) l, p < 0.001). The cost per procurement was remarkably lower using HTK. Conclusions: Perfusion of DBD liver grafts with HTK is clinically equivalent to UW, with a significant cost reduction.

Comparación de resultados entre el retrasplante y el trasplante renal primario en el Hospital Hermanos Ameijeiras de 1984 a 2012 / Comparison of results between retransplantation and primary kidney transplantation on Hermanos Ameijeiras Hospital from 1984 to 2012

Introducción: el retrasplante constituye la mejor opción terapéutica para los enfermos que pierden un primer trasplante renal y vuelven a diálisis, existen disímiles criterios en cuanto a sus resultados al compararlos con los trasplantes renales primarios. Objetivo: analizar el porcentaje de retrasplantes, revisar la supervivencia del injerto y del enfermo, el comportamiento de variables que pueden incidir en los resultados y compararlos con los de los enfermos que reciben un primer trasplante renal. Métodos: se realizó un estudio analítico, descriptivo, retrospectivo, de los trasplante renales realizados en el Hospital Hermanos Ameijeiras desde 1984 hasta diciembre de 2012; quedaron excluidos, los terceros trasplante, dobles (2 riñones a un mismo receptor), combinados (páncreas-riñón e hígado-riñón) y aquellos en los que no fue posible obtener la información requerida para la investigación. Se compararon (entre los grupos retrasplantes y primeros trasplantes) variables de índole general: edad de los receptores y donantes, sexo del receptor, enfermedad que ocasionó la insuficiencia renal, porcentaje de reactividad ante un panel de linfocito (PRA), compatibilidades HLA, tipo de donante (vivo o cadáver), tiempos de isquemia, presencia y duración de necrosis tubular aguda (donante cadáver), rechazo y supervivencia del injerto y el paciente. Resultados: los retrasplantes constituyeron el 5,4 por ciento de la muestra. No existieron diferencias entre edades, sexo, PRA, compatibilidades ni tipo de donante entre los segundos y primeros injertos. Los enfermos que llegaron a la insuficiencia renal por riñones poliquísticos nunca han recibido en nuestro centro un segundo trasplante. Resultó significativamente estadístico el uso de terapia cuádruple secuencial como inmunosupresión de inducción en los retrasplantes (55,9 por ciento vs. 9,7 por ciento de los primarios...

Introduction: retransplant constitutes the best therapeutic choice for patients who lose a first renal transplant and return to dialysis, existing dissimilar criteria as to its results when ranking them with renal primary transplant. Objective: to analyze the percentage of retransplantation, to revise graft and patient survival, to review the behavior of variables that can affect the results and to compare them with patients receiving a first renal transplant. Methods: an analytic, descriptive, retrospective study was accomplished, including all renal transplant performed at the Hermanos Ameijeiras Hospital from 1984 to December of 2012. Third transplants, double transplants (two kidneys to the same receptor), combined transplants (pancreas-kidney and liver-kidney) and those where it was not possible to obtain the information required for this research were excluded. Variables of general nature were compared between retransplantation groups and first transplants, such as: age of recipient and donor, sex of the recipient, a disease that caused kidney failure, percentage of reactivity to a lymphocyte panel (PRA), HLA compatibility, donor type (living or dead), ischemia time, presence and duration of acute tubular necrosis (dead donor), rejection and graft and patient survival. Results:rRetransplant constituted only 5.4 percent of the sample (34 patients). There were no differences in age, sex, PRA, donor type or compatibilities between the second and first grafts. Patients who reached the renal failure due to polycystic kidneys have never had a second transplant in our institution. The use of sequential quadruple therapy as induction immunosuppression, retransplantation (55.9 percent vs. 9.7 percent of primary) was statistically significant...

Adipose-derived stem cells (ADSC) in the viability of random skin flap in rats

PURPOSE: To evaluate the effects of the adipose-derived stem cells (ADSC) in the viability of random skin flap in rats. METHODS: Thirty five adult male Wistar rats (weight 250-300 g) were used. ADSC were isolated from adult male rats (n=5). ADSC were separated, cultured and then analyzed. A dorsal skin flap measuring 10x4 cm was raised and a plastic barrier was placed between the flap and its bed. After the surgical procedure, the animals were randomized into two groups (n=15 each group), group control and group ADSC. In all groups the procedures were performed immediately after the surgery. The percentage of flap necrosis was measured on the seventh postoperative day. RESULTS: The ADSC were able to replicate in our culture conditions. We also induced their adipogenic, osteogenic and chondrogenic differentiation, verifying their mesenchymal stem cells potentiality in vitro. The results were statistically significant showing that the ADSC decreased the area of necrosis (p<0.05). CONCLUSION: The cells demonstrated adipogenic, osteogenic and chondrogenic differentiation potential in vitro. The administration of adipose-derived stem cells was effective to increase the viability of the random skin flaps in rats. .

meld score a reliable outcome predictor in living donor liver transplantation?

In patients undergoing orthotopic liver transplantation [OLT], some studies have shown that a higher preopcrative MELD score is associated with higher morbidity and mortality [Saab et aL 2003]. Other studies also suggest that higher MELD scores are associated with increased inlraoperativc risks [Xia et al 2008]. to evaluate the impact of preoperative MELD status on metabolic, electrolyte and haemodynamic changes during living donar liver transplantation, and consequently on outcome and for right choice of candidate who will get greatest benefit from transplantation. After approval iron- ethical committee and a written consent from every patient a prospective observational study including 40 patients scheduled for living donor liver transplantation in Wady El nile hospital. Patients classified according to MELD score into group 1 [Gl] with score <25 and group2 [G2] > 25. Haemodynamic, metabolic and electrolytes data collected during the three phases: pro anhepatic, anhepatic and post reperfusion phase and follow up for 1[st] post operative week then two years mortality. G2 associated with significant intraoperative haemodynamic disturbances in all stages compared to Gl in the form of lower mean arterial blood pressure, lower systemic vascular resistance, higher pulmonary vascular resistance, and higher cardiac output. Also there were more intraoperative metabolic and electrolyte disturbances as there were more acidosis, lower standard bicarbonate values, more base deficits, more negative anion gap and strong ion difference, higher serum potassium level, poor intraoperative glycemic control and much higher blood lactate level. Also there were higher early mortality in G2 8 cases [40%] while 3 cases [15%] in GL 2 years mortality in Gl 5 cases [25%] while 10 cases [50%] in G2,, time of ICU stay was 5.1 days in G1 and 7 days in G2 time of hospital stay was 21.6 days in Gl and 26.2 days in G2 High MELD score patients associated with more intraoperative haemodynamic, metabolic and electrolyte disturbances, longer postoperative ICU and hospital stay and higher mortality

Management of traumatic upper limb arterial injuries: three years study

Upper limb vascular injury conditions requiring surgical intervention are multivarious and so are the surgical techniques employed. In the developing countries traumatic events predominate and the mechanism by which varies from society to another. Upper limb vascular injury must be approached promptly to avoid debilitating morbidity and mortality. is to assess the current management approach and to evaluate the outcome of patients with upper limb vascular trauma in Assiut University hospital. Forty two patients requiring operative intervention for upper limb vascular trauma from October 2009 - October 2012 were included. Patients were underwent full physical examination and resuscitation. The diagnosis of vascular injury is initially made by clinical examination as part of the full trauma assessment, plain x-ray, Doppler, Duplex and CT-angiography when indicated. Types of vascular repairs, orthopedic fixation, nerve repair, fasciotomy procedure and state of limb function after management were recorded. Forty two patients [37 males and 5 females] with mean age of 27H2.6 years [range 1.5 -70 ys] were included. The right upper limb was affected in 22 patients and the left was affected in 20 patients. Brachial artery was the most frequently affected 57.1%, followed by the axillary artery 31%, subclavian artery 9.5% and the ulnar artery 2.4%. The patients were presented 1-10 hours after injury [mean 6 +/- 2.1 hrs] with bleeding in 42.9%, ischemia 30.9%, ischemia 4 bleeding 9.5%, ischemia + hematoma 7.2% and expansile hematoma 9.5%. Penetrating trauma was in 90.4% and blunt trauma was in 9.6%. Injury by fire arm was in 40.4%, sharp objects 26%, motor car and train accidents 21%, iatrogenic 2.1% and fall from height 9.5%. Concomitant vein and/or nerve injuries were in 73%. Isolated nerve injury [brachial plexus or median nerve] was in 9.5%, isolated vein injury in 7.2% and both were injured in 57%. Orthopedic injuries occurred in 33% [open fracture 19%, closed fracture 12% and dislocation 2.4%]. Orthopaedic repair was done firstly before vascular repair except in delayed cases. Arterial repair in the form of end to end anastmosis in 42.8%, interposition vein graft 33.4%, simple repair 12%, ligation 4.7%, or thromboembolectomy in 4.7% were done. Heparin was applied both intralesional and systemic. Prophylactic fasciotomy with other vascular repair was also done in 30%. Primary nerve repair in non thermal [firearm] injuries was obtained while delayed repair was decided in thermal injuries. Small veins are ligated and large veins are managed by simple repair. The success rate of vascular repair was 39/42 [92.8%] assessed by the return of distal pulses at the end of the operation and limb salvage in follow up period. Three patients [7.2%], one of them [2.4%] died because of major head injury and above elbow amputation was done for the other two [4.8%]. All surviving patients were followed for one year for hand function and distal pulse. Clinical examination of the injured upper limb is a reliable diagnostic approach for evaluation of the vascular injury. Factors could help promoting good prognosis are; not wasting time with preoperative diagnostic tests, early intervention within 10 hours after trauma, performing orthopaedic repair firstly before vascular repair except in delayed cases, assessment and management of nerve injuries, use of intralesional and systemic anticoagulants, and prophylactic fasciotomy performance

Outcome of optical penetrating keratoplasties at a tertiary care eye institute in Western India.

Aim: To study the indications, risk factors, postoperative course, and long-term survival of corneal transplants done for optical purposes. Design: Retrospective case series. Materials and Methods: Data were obtained by reviewing the records of 181 patients operated at our institute (H.V. Desai Eye Hospital) between October 2005 and October 2007 for optical penetrating keratoplasty. Patients with less than one year of follow up, pediatric cases, therapeutic, tectonic, and lamellar keratoplasties were excluded. Kaplan Meier survival analysis was used to calculate median survival time of grafts and to see correlation between nine variables viz. age, gender, corneal vascularization, previous failed grafts, previous Herpes Simplex keratitis, post-perforation corneal scars, donor tissue quality, graft size, type of surgery and follow-up. These variables were also used for univariate and multivariate analysis using Cox Proportional Hazard Regression Modeling. Results: Median survival of the cohort was 27 months (95% confidence interval: 20.47-33.52). One- and two-year survival rates were 65% and 52.5%, respectively. Median survival was significantly lower in poor prognosis cases (14 months) than good prognosis cases (27 months, P = 0.0405). Graft survival was lower in vascularized corneas (18.55 months, P = 0.030) and in post-perforation corneal scars (17.96 months, P = 0.09, borderline significance). Multivariate analysis showed that the same factors were predictive of graft failure. Conclusion: Long-term survival of grafts at our center is different from centers in western world. More high-risk cases, paucity of excellent quality donor corneas, and differences in patient profile could be the contributory factors.

Efecto del plasma rico en plaquetas en la resistencia de un injerto tendíneo, en túnel óseo en modelo en animal / Effect of platelet-rich plasma in relation to mechanical strength of intra-tunel integration of tendon graft in animals

Introduction: Bone-tendon interface is one of the places for early failure in the ligament reconstruction surgery. In that way, osteintegration should be favored. Growth factors have been proposed to enhance tendon healing and tissue regeneration. Objective: To assess the effect of Platelet-Rich Plasma (PRP) in relation to the mechanical strength of intra-tunel integration of flexor tendon in a rabbit model. Methods: Experimental and control trial in animal model. Nine skeletally mature, New Zeland rabbits were used for this study (18 knees) performing a proximal tibia bone tunnel in one knee in which a flexor tendon graft was introduced, applying a dose of PRP in the bone tunnel. The other knee was used as control. Rabbits were sacrificed in three groups of three at de 2nd, 4th and 8th week. Biomechanical evaluation was performed, recording the force to achieve graft failure. Data analysis was done using Mann Whitney test. Results: The resistance obtained in the PRP group vs control group at 2nd week was 7.4 N vs 6.6 N (p = 0.51), at 4th week was 16 N vs 13.5 N (p = 0.27) and at 8th week was 21N vs 13.2 N (p = 0.51). Conclusion: Administration of local PRP in bone tunnels does not significantly modify mechanical strength of the graft in bone tunnel compared with controls.

Introducción: La interface tendón-hueso es uno de los sitios de falla precoz de la reconstrucción ligamentosa. Se han buscado distintas formas de aumentar la osteointegración del injerto. Existen estudios demostrando algún grado de utilidad del plasma rico en plaquetas (PRP) en relación a la cicatrización y regeneración de tejidos. Objetivo: Evaluar el efecto del plasma rico en plaquetas en la resistencia de la cicatrización intraósea de tendón flexor en un modelo de reconstrucción ligamentosa en conejos. Material y Método: Estudio experimental, controlado, en modelo animal. Nueve conejos de raza albina New Zeland, esqueléticamente maduros. Se estudiaron 9 conejos (18 rodillas), realizando un túnel óseo tibial proximal en una rodilla, donde se introdujo injerto de tendón flexor, administrándose una dosis autóloga de PRP en el túnel óseo. La otra rodilla se utilizó como control, los conejos se sacrificaron en grupos de 3 a la 2ª, 4ª y 8ª semana. Realizamos evaluación biomecánica, registrándose la fuerza necesaria para conseguir la falla del injerto. El análisis de datos se realizó mediante Test de Mann Whitney. Resultados: La resistencia obtenida en el grupo con PRP vs grupo control a la 2da semana fue 7,4 N vs 6,6 N (p = 0,51), a la 4ta semana fue 16 N vs 13,5 N (p = 0,27) y a la 8va semana fue 21 N vs 13,2 N (p = 0,51). Conclusiones: La administración local de PRP en los túneles óseos, no modifica en forma significativa la resistencia del injerto en el túnel óseo comparado con grupo control. Nivel de evidencia: II (ensayo clínico controlado no aleatorio).

Predicción de sobrevida del injerto a corto y largo plazo mediante el uso de índices gamagráficos en receptores de trasplante renal / Prediction of short and long term graft survival by 99mTc-MAG3 scintigraphy after renal transplantation

Background: 99Tcm-MAG3 is a non-invasive method for the evaluation of renal transplants, giving information about allograft function and short- and long-term allograft survival. However, few studies have validated this observation. Objetive: To determine the predictive power of different scintigraphic parameters on graft survival at 6 -12 months after renal transplantation. Methods: Between 1996-2007, receptors with varying degrees of kidney dysfunction were studied with 99mTc-MAG3 scintigraphy within 14 days after transplantation. Tubular injury score (TISS) ranging I-VI according to severity and renal extraction index (R20/3) were calculated. Survival analysis and Cox regression were used for analysis. Results: Three-hundred and four renograms were performed in 146 allografts (143 receptors, mean age 38.9 +/- 17 years, 81.5 percent from cadaveric donor). Mean follow-up time was 44 months. There was graft loss (GL) in 32 percent renal trasplants. According to severity range, graft survival at 6 and 12 months was: TISS I-II 85.23 percent and 81.17 percent; TISS III-IV 82.43 percent and 80.56 percent, and TISS V-VI 32 percent and 2.,43 percent respectively. TISS score of V-VI score was an independent predictor for GL (HR = 6.3 CI 95 percent 2.9–13; p<0.0001). R20/3 index was not a good predictor of GL. Conclusions: TISS score was an independent predictor of short- and long-term allograft survival using 99mTc-MAG3 scintigraphy performed early after kidney transplantation in patients with suspected patients renal dysfunction. TISS V-VI had a greater discriminatory power. R20/3 index individually did not present a good prognostic performance.

Study on the vaginal smear of rats submitted to autologous ovarian transplant: impact of remote ischemic preconditioning / Estudo do esfregaço vaginal de ratas submetidas a transplante autólogo de ovário: impacto do precondicionamento isquêmico remoto

PURPOSE: To evaluate the remote ischemic preconditioning (R-IPC) impact on the quality of the ovarian graft by means of vaginal smear of transplanted rats. METHODS: Sixty rats were used divided in six groups: Control; Fresh transplant (TxF); Cryopreserved transplant (TxC); R-IPC; R-IPC + fresh transplant (TxF+R-IPC); R-IPC + cryopreserved transplant (TxC+R-IPC). R-IPC was performed in the common iliac artery. Autologous ovarian tissue was implanted integrally in the retro peritoneum. On the first PO day, vaginal smear collection was daily initiated. After 30 days, the PO day when the estrous cycle was re-initiated was considered for analysis as well as the estrous days and the number of estrous cycles. RESULTS: R-IPC showed a tendency to an early estrus re-initiation (p>0.05) as well as increase the number of cycles in the fresh transplanted group while in the cryopreserved transplant the number of cycles was similar, regardless of the stimulus R-IPC (p>0.05). The animals which had undergone fresh grafts had a longer estrous period than the ones which had undergone cryopreserved grafts, with or without R-IPC (p<0.05). CONCLUSION: R-IPC promoted earlier re-initiation of ovarian activity in the PO and greater estrous frequency, with more consistent results in the fresh grafts than in the cryopreserved ones.

OBJETIVO: Avaliar o impacto do precondicionamento isquêmico remoto (PCI-R) na qualidade do enxerto ovariano através dos esfregaços vaginais dos animais transplantados. MÉTODOS: Foram utilizadas 60 ratas, distribuídas em seis grupos de estudo: Controle; Transplante fresco; Transplante criopreservado; PCI-R; PCI-R + Transplante fresco; PCI-R + Transplante criopreservado. O PCI-R foi realizado na artéria ilíaca comum. O tecido ovariano foi implantado íntegro no retroperiônio. No 1º dia de pós-operatório (PO) foram coletados esfregaços vaginais diariamente. Após 30 dias foram considerados para análise o dia de PO de retorno do estro, assim como o número de dias em estro e de ciclos estrais. RESULTADOS: PCI-R mostrou tendência em reinício mais precoce do cilo estral (p>0,05), assim como aumento no número de cilcos estrais nos grupos com transplante fresco enquanto que no criopreservado o número de ciclos foi semelhante, independente do PCI-R (p>0,05). Os animais que receberam enxertos frescos apresentaram mais dias na fase estro do que os criopreservados, com ou sem PCI-R. CONCLUSÃO: O PCI-R promoveu retorno mais precoce da atividade ovariana no PO e maior freqüência de estro, sendo os resultados mais consistentes nos enxertos frescos do que nos criopreservados.

Modified Acellularization for Successful Vascular Xenotransplantation

The purpose of this study was to estimate the possibilities of an acellular matrix using a modified acellularization protocol, which circumvents immunological, microbiological, and physiological barriers. We treated porcine subclavian arteries with various reagents to construct acellular grafts. Afterwards, these grafts were interposed in a mongrel dogs' abdominal aorta. Six dogs underwent interposition with fresh porcine grafts (control group), and seven had interposed acellular grafts (acellular group). The control and acellular group dogs were sacrificed at 1, 3, 5 (n=2 in each group) and 12 months (n=1 in acellular group) after the operation. Histopathological examinations were then performed, to assess the degree to which re-endothelialization, inflammation, thrombus formation, and calcification occurred. The entire acellular group, but none of the control group, exhibited re-endothelialization. The degrees to which inflammation, thrombosis, and calcification occurred were found to be lower in the acellular group. We also discovered many smooth muscle cells in the medial layer of the xenograft that had been implanted in the dog sacrificed 12 months after the operation. These results suggest that the construction of xenografts using our modified acellularization protocol may offer acceptable outcomes as a vascular xenograft.

Liver glutathione depletion after preservation and reperfusion in human liver transplantation

OBJETIVO: O estresse oxidativo é um importante mecanismo responsável pela disfunção dos enxertos após transplante de fígado (TF). Sabe-se que níveis baixos de Glutationa reduzida (GSH) deixam os enxertos vulneráveis aos danos de reperfusão. O objetivo deste estudo foi avaliar as concentrações de GSH e da Glutationa oxidada (GSSG), os danos hepatocelulares e a função em enxertos ótimos e subótimos após TF. MÉTODOS: Foram realizadas biópsias em 33 pacientes imediatamente antes do implante e duas horas após a reperfusão, permitindo a determinação do GSH, GSSG e o cálculo do índice de stress oxidativo (GSH/GSSG). Foram medidas as transaminases hepáticas e as atividades da Protrombina (TP) e do Fator V para avaliação dos danos hepatocelulares e da função do enxerto, respectivamente. O dano histopatológico foi avaliado através de um índice de cinco parâmetros. RESULTADOS: Houve uma diminuição nos níveis de GSH (p<0.01) 0.323 ± 0.062 ìmol/g to 0.095 ± 0.01 ìmol/g and 0.371 ± 0.052 ìmol/g to 0.183 ± 0.046 ìmol/g) e aumento nos níveis de GSSG (0.172 ± 0.038 ìmol/g to 0.278 ± 0.077 ìmol/g and 0.229 ± 0.048 ìmol/g to 0.356 ± 0.105 ìmol/g) (p<0.05). Houve diminuição do GSH/GSSG (2.23 ± 0.31 to 0.482 ± 0.042 and 2.47 ± 0.32 to 0.593 ± 0.068). Nenhuma diferença entre os grupos ótimo e subótimo foi vista após duas horas de reperfusão. Os escores de danos histopatológicos foram maiores no grupo subótimo (6.46 ± 0.4 vs. 5.39 ± 1.1) (p<0.05) e mostraram correlação com o TP e fator V no grupo Ótimo (p<0.05). A análise multivariada apontou a esteatose como um fator de risco independente para a ocorrência de danos histopatológicos (p<0.05). CONCLUSÃO: Houve uma significativa depleção de GSH e formação de GSSG após a preservação em solução devido a um intenso estresse oxidativo nos enxertos ótimos e subótimos, porém estes níveis não se correlacionaram com a viabilidade dos enxertos.

Effect of high frequency transcutaneous electrical nerve stimulation on viability of random skin flap in rats

OBJETIVO: Avaliar o efeito da Estimulação Elétrica Nervosa Transcutânea (TENS) em alta freqüência na viabilidade do retalho cutâneo randômico em ratos. MÉTODOS: Foram utilizados 75 ratos da linhagem Wistar. O retalho cutâneo apresentava 10 x 4 cm, sendo que entre o mesmo e a área doadora foi realizada a interposição de uma barreira plástica. Após o procedimento operatório todos os animais permaneceram anestesiados por mais uma hora com os eletrodos posicionados na base do retalho e submetidos ao tratamento de acordo com seus respectivos grupos. Este procedimento se repetiu nos outros dois dias subseqüentes. G1: simulação da TENS, G2: TENS (f = 80 Hz e I = 5 mA), G3: TENS (f = 80 Hz e I = 10 mA), G4: TENS (f = 80 Hz e I = 15 mA), G5: TENS (f = 80 Hz e I = 20 mA). RESULTADOS: A porcentagem média de área de necrose foi de 43,11, 34,65, 49,44, 23,52, 45,10 nos grupos 1, 2, 3, 4 e 5 respectivamente. CONCLUSÃO: Os animais estimulados com amplitude de 15 mA apresentaram menor área de necrose quando comparados ao grupo controle e a Estimulação Elétrica Nervosa Transcutânea foi eficiente no aumento de viabilidade do retalho cutâneo randômico em ratos.

Cotransplanted Bone Marrow Derived Mesenchymal Stem Cells (MSC) Enhanced Engraftment of Hematopoietic Stem Cells in a MSC-dose Dependent Manner in NOD/SCID Mice

Transplantation of marrow-derived mesenchymal stem cells (MSCs), expanded by culture in addition to whole bone marrow, has been shown to enhance engraftment of human hematopoietic stem cells (HSCs). Our hypothesis was that there might be an optimum ratio range that could enhance engraftment. We examined the percent donor chimerism according to the ratio of HSCs to MSCs in non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice. We tested a series of ratios of co-transplanted CD34+-selected bone marrow cells, and marrow-derived MSCs into sublethally irradiated NOD/SCID mice. In all experiments, 1 x 10(5) bone marrow derived human CD34+ cells were administered to each mouse and human MSCs from different donors were infused concomitantly. We repeated the procedure three times and evaluated engraftment with flow cytometry four weeks after each transplantation. Serial ratios of HSCs to MSCs were 1:0, 1:1, 1:2 and 1:4, in the first experiment, 1:0, 1:1, 1:2, 1:4 and 1:8 in the second and 1:0, 1:1, 1:4, 1:8 and 1:16 in the third. Cotransplantation of HSCs and MSCs enhanced engraftment as the dose of MSCs increased. Our results suggest that the optimal ratio of HSCs and MSCs for cotransplantation might be in the range of 1:8-1:16; whereas, an excessive dose of MSCs might decrease engraftment efficiency.

Cotransplanted Bone Marrow Derived Mesenchymal Stem Cells (MSC) Enhanced Engraftment of Hematopoietic Stem Cells in a MSC-dose Dependent Manner in NOD/SCID Mice

Transplantation of marrow-derived mesenchymal stem cells (MSCs), expanded by culture in addition to whole bone marrow, has been shown to enhance engraftment of human hematopoietic stem cells (HSCs). Our hypothesis was that there might be an optimum ratio range that could enhance engraftment. We examined the percent donor chimerism according to the ratio of HSCs to MSCs in non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice. We tested a series of ratios of co-transplanted CD34+-selected bone marrow cells, and marrow-derived MSCs into sublethally irradiated NOD/SCID mice. In all experiments, 1 x 10(5) bone marrow derived human CD34+ cells were administered to each mouse and human MSCs from different donors were infused concomitantly. We repeated the procedure three times and evaluated engraftment with flow cytometry four weeks after each transplantation. Serial ratios of HSCs to MSCs were 1:0, 1:1, 1:2 and 1:4, in the first experiment, 1:0, 1:1, 1:2, 1:4 and 1:8 in the second and 1:0, 1:1, 1:4, 1:8 and 1:16 in the third. Cotransplantation of HSCs and MSCs enhanced engraftment as the dose of MSCs increased. Our results suggest that the optimal ratio of HSCs and MSCs for cotransplantation might be in the range of 1:8-1:16; whereas, an excessive dose of MSCs might decrease engraftment efficiency.

Low endothelial cell count and clear corneal grafts.

Specular microscopic study on clear corneal grafts indicates that at times surprisingly low endothelial cell density can maintain the grafted cornea in a relatively dehydrated state. The critical limit of the endothelial cell count for corneal decompensation is thought to be 700 cells/mm2. This communication reports 13 cases of clear corneal graft with endothelial cell count below 700 cells/mm2.

Repair of lamellar scleral lesions in dogs with preserved equine renal capsule: short report

PURPOSE: To evaluate the use of equine renal capsule preserved in glycerin to repair lamellar scleral lesions in dogs. METHODS: Twelve healthy mongrel dogs, male and female, weighing 12 kg were used. The study was both clinical and morphological, and performed on the first, third, seventh, 15th 30th and 60th day after surgery. Temporal canthotomy was performed after standard preoperative and general anesthesia. Conjunctival and sclera square incisions of 0.5x0.5 cm were carried out in a one o' clock position, near the limbus. A fragment of hydrated biological prosthesis, of the saro shape, was sutured with 7-0 Vicryl® in an interrupted suture. RESULTS: The clinical evaluation showed blepharospasm/photophobia until the 7th day after surgery. Conjunctival edema appeared up to the 5th day after surgery. Mucoid ocular discharge was sustained until the 10th day after surgery. Hyperemia was observed until the end of the evaluation period. There were no signs of graft extrusion in all animals. The anterior and posterior segmente did not show clinical signs of inflammation. The optical microscopy mor phological evaluation showed an inflammatory exudation with acute aspects in the early and intermediate periods, and inflammatory exudation with chronic aspects in the late periods. There was incorporation of the implant by the recipient's sclera. CONCLUSION: These results suggest that the equine renal preserved capsule could be a useful alternative tissue to repair lamellar corneal lesions in dogs and humans.