Nondestructive production of exosomes loaded with ultrathin palladium nanosheets for targeted bio-orthogonal catalysis.
Victor Sebastian CabezaMaria Sancho-AlberoManuel ArrueboAna M Perez-LopezBelén Rubio-RuizPilar Martin-DuqueAsier Unciti-BrocetaJesús SantamaríaPublished in: Nature protocols (2020)
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.
Keyphrases
- mesenchymal stem cells
- stem cells
- reduced graphene oxide
- induced apoptosis
- randomized controlled trial
- cancer therapy
- metal organic framework
- cell cycle arrest
- highly efficient
- visible light
- drug delivery
- quantum dots
- healthcare
- bone marrow
- oxidative stress
- room temperature
- minimally invasive
- signaling pathway
- risk assessment
- cell proliferation
- cell death
- mass spectrometry
- human health
- high resolution
- pi k akt
- climate change