Figure 2.

Methods to facilitate the release of EVs. ① Acute Hypoxia: Catabolism of HIF-1α is inhibited by acute hypoxia, which stabilizes the P53 gene and activates the P21 gene, leading to apoptosis and promoting the release of EVs (91–93). ② UV: After UV irradiation, a large number of free radicals are generated to attack nucleic acids and proteins, causing apoptosis and increasing the release of EVs (94). ③ Photodynamic Treatment: Laser irradiation at a specific wavelength excites the tissue-absorbing photosensitizer, and the excited state of the photosensitizer transmits energy to the surrounding oxygen, generating strongly reactive monomorphic oxygen, which may reacts oxidatively with the surrounding neighboring biomolecules, resulting in a cytotoxic effect that causes apoptosis and also promoting the release of EVs (95). ④ Complement Proteins: The membrane attack complex (MAC) is composed of complement proteins (C5b, C6, C7, C8 and C9). MAC is cleared from the cell surface by cytosolic or cytocytic action to help release EVs (96, 97). ⑤ Chemotherapy: The use of chemotherapeutic agents (e.g., doxorubicin, methotrexate, and cisplatin) causes cellular damage and EVs release (95, 98). ⑥ Toxic Lipids: Toxic lipids activates the DR5 pro-apoptotic signaling cascade, which in turn activates ROCK1 and promotes the release of EVs from hepatocytes (90). ⑦ Nutritional Deficiency: Activation of Caspase 3, ROCK1 signaling pathway and promotion the release of EVs (99). ⑧ Infection factors (100) and ⑨ focused ultrasound (101) can also promote the release of EVs.