Ординатура / Хирургия / Библиотека им академика М.И. Перельмана / Книга_5645_Библиотеки_им_академика_М_И_Перельмана

Electroporation (electric pulses to produce transitory pores in the lipid bilayer) facilitates the incorporation of therapeutic cargos such as drugs, nucleic acids, or proteins in exosomes. The electric pulses create temporary openings in the exo­some membrane, enabling the efficient loading of therapeutic molecules into the exosomes. The therapeutic-loaded exosomes generated by electroporation methods have shown promise in various therapeutic applications. For targeted drug delivery applications, the loaded exosomes are engineered to specifically deliver therapeutic cargos to desired cell types or tissues. Additionally, these exosomes can be utilized in gene therapy, where exosomes with loaded nucleic acids can deliver genetic material to target cells for therapeutic purposes. Greco et al., [21] revealed that Mesenchymal stromal cells (MSC) and Human Embryonic Kidney 293 (HEK293) exosomes were suspended in an electroporation buffer containing several forms of siRNA at a specific concentration. Following that, the exosome-siRNA mixture was put into a cuvette and electroporated with a Bio-Rad® Gene Pulse XCell electroporation equip­ment. The authors were able to measure the exosome loading efficiency using siRNA that was allophycocyanin-labeled. The UMUC3 bladder cancer cells were incubated with the exosomes for 6hours. They observed that the fluorescence intensity was more than 28-fold higher in the siRNA-loaded exosome-containing cell population. The enhanced fluorescence intensity demonstrated that electroporation was effec­tive in transferring siRNA into exosomes, and consequently into the bladder cancer cell. A modified calcium chloride transfection technique is capable of introducing miRNA into exosomes [22]. This technique offers a versatile platform for loading a wide range of therapeutics. However, there is a chance that this approach will result in excessive aggregation or that the exosomes’ membrane integrity could be dam­aged. Electroporation also requires specific equipment to use, such as the Neon® Transfection System [22, 23]. It is important to note that further research is required to optimize electroporation methods for therapeutic loading in exosomes. Factors