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Radiomarcaje y estudios de biodistribucion de nanopartículas polimericas como adyuvantes para la vacunacion oftalmica frente a la brucelosis

, : Radiomarcaje y estudios de biodistribucion de nanopartículas polimericas como adyuvantes para la vacunacion oftalmica frente a la brucelosis.

Optimizar el radiomarcaje con 99mTc de nanopart culas de Gantrez manosiladas y cargadas con el ant geno de Brucella ovis (Man-NP-HS) y llevar a cabo estudios de biodistribuci n en un rat n tras la administraci n de las nanopart culas por v a ocular. Las Man-NP-HS se obtuvieron por el m todo de desplazamiento del disolvente. Se purificaron, liofilizaron y caracterizaron. A continuaci n, se marcaron con 74MBq de 99mTcO4 ? previamente reducido con una disoluci n cida de cloruro de esta o, trabajando en ausencia de ox geno y con un pH final de 4. El rendimiento del marcaje se evalu mediante TLC. Los estudios de biodistribuci n se llevaron a cabo en ratones tras la administraci n oft lmica de la formulaci n y de un control de 99mTcO4 ? libre. Para ello, se sacrificaron los animales a las 2 y a las 24h tras la administraci n ocular y se contaron los rganos en un contador gamma. Se obtuvo un rendimiento de marcaje superior al 90%. Los estudios de biodistribuci n de 99mTc-Man-NP-HS permitieron detectar la actividad concentrada en la mucosa nasal y ocular y el tracto gastrointestinal tanto a las 2 como a las 24h frente a la biodistribuci n de 99mTcO4 ? libre que permaneci concentrado en la piel alrededor del ojo y en el tracto gastrointestinal. Los estudios de biodistribuci n de 99mTc-Man-NP-HS tras la administraci n oft lmica han permitido demostrar su biodistribuci n en las mucosas y el tracto gastrointestinal, caracter stica indispensable como sistema de liberaci n de ant genos a trav s de la mucosa ocular. Esto, junto con su elevada respuesta inmune, efectiva protecci n y no virulencia, convierte a estas nanopart culas en una vacuna ideal antibrucelosis. To optimize radiolabeling with 99mTc of mannosylated Gantrez nanoparticles loaded with the Brucella Ovis antigen (Man-NP-HS) and to carry out biodistribution studies in mice after ocular administration of the nanoparticles. Man-NP-HS nanoparticles were prepared by the solvent displacement method. They were purified, lyophilized and characterized. Following this, they were radiolabeled with 74 MBq of 99mTcO4 ? previously reduced with an acidic stannous chloride solution, working in absence of oxygen and at a final pH of 4. Radiolabeling yield was evaluated by TLC. Biodistribution studies were carried out in mice after ocular administration of the formulation and control of free 99mTcO4 ?. To do so, the animals were humanely killed at 2 and 24hours after the ocular administration and activity in organs was measured in a Gamma counter. Radiolabeling yield obtained was greater than 90%. Biodistribution studies of 99mTc-Man-NP-HS showed radioactivity accumulated at 2 and 24hours in nasal and ocular mucosa and gastrointestinal tract, in contrast to biodistribution of free 99mTcO4 ? that remained concentrated in the skin around the eye and gastrointestinal tract. Biodistribution studies of 99mTc-Man-NP-HS after ocular instillation have made it possible to demonstrate its biodistribution in nasal mucosa and gastrointestinal tract. This characteristic is essential as an antigenic delivery system throughout the ocular mucosa. This, together with its elevated immune response, effective protection and intrinsic avirulence make them a suitable anti-Brucella vaccine candidate.


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