0000000000367989
AUTHOR
Francisco Orbis
A surgical model for isolating the pig liver in vivo for gene therapy.
Several studies report results that suggest the need of vascularization blocking for efficient gene transfer to the liver, especially in nonviral gene therapy. In this study, we describe a surgical strategy for in vivo isolation of the pig liver, resulting in a vascular watertight organ that allows the evaluation of several gene injection conditions. The hepatic artery and portal, suprahepatic and infrahepatic cava veins were dissected. Then, liver vascularization was excluded for 5-7 min. In that time, we first injected 200 ml saline solution containing the p3c-eGFP plasmid (20 µg/ml) simultaneously through two different catheters placed in the portal and cava veins, respectively. Vital co…
Efficacy of interleukin 10 gene hydrofection in pig liver vascular isolated ‘in vivo’ by surgical procedure with interest in liver transplantation
AIM Liver transplantation is the only curative strategy for final stage liver diseases. Despite the great advances achieved during the last 20 years, the recipient immune response after transplantation is not entirely controlled. This results in high rates of acute cell rejection and, approximately, 10% of early mortality. Therapeutic treatment could be improved by efficiently transfecting genes that encode natural immunosuppressant proteins, employing safe procedures that could be transferred to clinical setting. In this sense, interleukin 10 plays a central role in immune tolerance response by acting at different levels. METHODS hIL10 gene was hydrofected by retrograde hydrodynamic inject…
Surgical anatomy applied to transperitoneal approaches of the abdominal aorta and visceral trunks. Dynamic article.
Access to the abdominal aorta and its visceral trunks is possible through several approaches. Dissections of five cadavers performed during three National Surgical Anatomy courses applied to Aorta, Hepatobiliopancreatic and Digestive Surgery. Videos and pictures were taken throughout the dissections and showed different abdominal aorta approaches. Abdominal aorta and visceral trunks approaches: longitudinal inframesocolic access, supraceliac clamping, celiac trunk dissection, superior mesenteric artery approaches (retroperitoneal after Kocher menoeuvre, supramesocolic or inframesocolic), Cattell-Braasch manoeuvre and mattox manoeuvre: retrorenal and prerenal. Correct knowledge of the intraa…
Anatomía quirúrgica aplicada a abordajes transperitoneales de la aorta abdominal y los troncos viscerales. Artículo dinámico
Resumen Los cirujanos cardiovasculares y del aparato digestivo deberian estar al corriente de las multiples alternativas de abordaje de la aorta abdominal y sus troncos viscerales. Articulo narrativo, ilustrado y dinamico de las diferentes maniobras quirurgicas descritas con este objetivo. Diseccion de 5 cadaveres realizadas durante tres cursos nacionales de Anatomia Quirurgica aplicada a aorta integral, Cirugia hepatobiliopancreatica y Cirugia abdominal digestiva. Maniobras quirurgicas descritas: abordaje aortico inframesocolico longitudinal, abordaje aortico supraceliaco, abordaje del tronco celiaco, tres tipos de abordaje de la arteria mesenterica superior: retroperitoneal tras maniobra …
Low RNA translation activity limits the efficacy of hydrodynamic gene transfer to pig liver “in vivo”
Background Hydrodynamic gene delivery has proved an efficient strategy for nonviral gene therapy in the murine liver but it has been less efficient in pigs. The reason for such inefficiency remains unclear. The present study used a surgical strategy to seal the whole pig liver in vivo. Methods A solution of enhanced green fluorescent protein (eGFP) DNA was injected under two different venous injection conditions (anterograde and retrograde), employing flow rates of 10 and 20 ml/s in each case, with the aim of identifying the best gene transfer conditions. The gene delivery and information decoding steps were evaluated by measuring the eGFP DNA, mRNA and protein copy number 24 h after transf…