| Autor: |
Sebastian V; Instituto de Nanociencia y Materiales de Aragón (INMA), CSIC-Universidad de Zaragoza, Zaragoza, Spain. victorse@unizar.es.; Department of Chemical Engineering and Environmental Technologies, University of Zaragoza, Zaragoza, Spain. victorse@unizar.es.; Networking Research Center on Bioengineering Biomaterials and Nanomedicine (CIBER-BBN), Madrid, Spain. victorse@unizar.es., Sancho-Albero M; Instituto de Nanociencia y Materiales de Aragón (INMA), CSIC-Universidad de Zaragoza, Zaragoza, Spain.; Department of Chemical Engineering and Environmental Technologies, University of Zaragoza, Zaragoza, Spain.; Networking Research Center on Bioengineering Biomaterials and Nanomedicine (CIBER-BBN), Madrid, Spain., Arruebo M; Instituto de Nanociencia y Materiales de Aragón (INMA), CSIC-Universidad de Zaragoza, Zaragoza, Spain.; Department of Chemical Engineering and Environmental Technologies, University of Zaragoza, Zaragoza, Spain.; Networking Research Center on Bioengineering Biomaterials and Nanomedicine (CIBER-BBN), Madrid, Spain., Pérez-López AM; Cancer Research UK Edinburgh Centre, MRC Institute of Genetics & Molecular Medicine, University of Edinburgh, Edinburgh, UK.; Institut für Biotechnologie, Technische Universität Berlin, Berlin, Germany., Rubio-Ruiz B; Cancer Research UK Edinburgh Centre, MRC Institute of Genetics & Molecular Medicine, University of Edinburgh, Edinburgh, UK.; Pfizer-University of Granada-Andalusian Regional Government Centre for Genomics and Oncological Research (GENYO) and Department of Medicinal and Organic Chemistry, Faculty of Pharmacy, University of Granada, Granada, Spain., Martin-Duque P; Networking Research Center on Bioengineering Biomaterials and Nanomedicine (CIBER-BBN), Madrid, Spain.; Instituto Aragonés de Ciencias de la Salud-Fundación Araid/IIS Aragón, Centro de Investigaciones Biomédicas de Aragón, Universidad San Jorge, Zaragoza, Spain., Unciti-Broceta A; Cancer Research UK Edinburgh Centre, MRC Institute of Genetics & Molecular Medicine, University of Edinburgh, Edinburgh, UK., Santamaría J; Instituto de Nanociencia y Materiales de Aragón (INMA), CSIC-Universidad de Zaragoza, Zaragoza, Spain.; Department of Chemical Engineering and Environmental Technologies, University of Zaragoza, Zaragoza, Spain.; Networking Research Center on Bioengineering Biomaterials and Nanomedicine (CIBER-BBN), Madrid, Spain. |
| Abstrakt: |
The use of exosomes as selective delivery vehicles of therapeutic agents, such as drugs or hyperthermia-capable nanoparticles, is being intensely investigated on account of their preferential tropism toward their parental cells. However, the methods used to introduce a therapeutic load inside exosomes often involve disruption of their membrane, which may jeopardize their targeting capabilities, attributed to their surface integrins. On the other hand, in recent years bio-orthogonal catalysis has emerged as a new tool with a myriad of potential applications in medicine. These bio-orthogonal processes, often based on Pd-catalyzed chemistry, would benefit from systems capable of delivering the catalyst to target cells. It is therefore highly attractive to combine the targeting capabilities of exosomes and the bio-orthogonal potential of Pd nanoparticles to create new therapeutic vectors. In this protocol, we provide detailed information on an efficient procedure to achieve a high load of catalytically active Pd nanosheets inside exosomes, without disrupting their membranes. The protocol involves a multistage process in which exosomes are first harvested, subjected to impregnation with a Pd salt precursor followed by a mild reduction process using gas-phase CO, which acts as both a reducing and growth-directing agent to produce the desired nanosheets. The technology is scalable, and the protocol can be conducted by any researcher having basic biology and chemistry skills in ~3 d. |
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