Table 2.

Summary of the therapeutic roles of Mφ-EVs in diseases.

Disease models	EVs source and Isolation method	EV subtypes (diameter)	Therapeutic molecules	EV doses and routes	Therapeutic outcomes	References
Tissue repair
Myocardial I/R injury	- Rat peritonea macrophages - Ultracentrifugation	Exos (~100 nm)	miR-148a	- 2 h before I/R procedure - 2–3 μg per rat by single caudal vein	- Reduced the dysregulation of cardiac enzymes and Ca2+ overload - Reduced apoptosis and the number of broken cardiomyocytes	112
Diabetic skin defects	- RAW 264.7 cell line - Ultracentrifugation	Exos (~95 nm)	Not studied	- The day when wounds were produced - 0.1 or 1 mg EVs in 1 ml PBS per rat by single subcutaneously	- Elevated angiogenesis, migration and proliferation ability of high glucose treated HUVECs by anti-inflammation - Accelerated wound contraction and reduced wound length - Therapeutic effects with dose-dependent	113
Hair loss	- RAW 264.7 cell line - Ultracentrifugation	EVs (~128.8 nm)	Wnt3a/Wnt7b	- 2 days after the hair removed - 0.1 or 1 mg EVs per mouse by intradermally three times weekly for 4 weeks	- Increased proliferation and migration of dermal papilla cells by activating Wnt/β-Catenin pathway - Increased hair follicle number, elongation of hair and thickness of dermis with time- and dose-dependent - Therapeutic effects with dose-dependent	114
Radiation-induced gastrointestinal syndrome	- Mice BMDM - Ultracentrifugation and precipitation	EVs (undefined)	Functional Wnt ligands	Unclear	Elevated survival of mice with wild type BMDM cell medium	115
Inflammatory bowel disease	- Mice BMDM with IL-13/IL-10/IL-1β - Precipitation	Exos (30–150 nm)	CCL1	- The day when administration with DSS - 50 mg/mouse by intraperitoneally once a day for 8 days	- Reduced length and inflammatory damage of colon - Increased number of Treg cells in spleen - Alleviated weight loss, diarrhea and bleeding in mice with colitis	116
Cutaneous Wound	- Mice BMDM - Ultracentrifugation	Exos (~69 nm)	not specific	- 100 μg/mouse by subcutaneously injection - Once a day at day 1 and 4	- Increased expression of Arginase and decreased expression of iNOS in M1-like Mφ - Enhanced fibroblast migration and EC tube formation - Increased wound dermal cellularity and collagen production	117
Pathogen control
Systemic candidiasis	- THP-1 cell line with Candida albican - Ultracentrifugation	EVs (~30–369 nm)	Proteins	Unclear	- Elevated proinflammatory cytokines by activating ERK2 and p38 - Decreased candidacidal activity	122
Tuberculosis	- THP-1 cell line with mycobacteria - Sucrose gradient centrifugation	Exos (50–100 nm)	Not studied	- 20 μg EVs per mouse by intranasally - Single dose	- Elevated inflammatory response	118
	- Mouse BMDM with M.tb - Ultracentrifugation	EVs (undefined)	M.tb-RNA	- 3 weeks after infection - 5 μg/mouse EVs intratracheally at 4 weeks post‐infection	- Decreased replication and survival of M.tb by increased IFN-γ - Elevated autophagy by activating LC-3-associated phagocytosis pathway	119
	- RAW 264.7 cell line with M.tb culture filtrate protein - Ultracentrifugation	Exos (50–150 nm)[112] Exos (30–100 nm)[113]	Immunized purified EVs	- Six weeks before M.tb infection - 40 μg/mouse EVs three times by intranasally at an interval of 2 weeks	- Elevated Th1 response and limited Th2 response - Reduced mycobacterial numbers in lung	120,121
Dengue	- U937 Mφ with DENV - Ultracentrifugation	EVs (~100 nm)	NS3 and miRs	Unclear	Elevated production of proinflammatory cytokines in ECs	123
HCV	- Human monocytes-derived Mφ with HIV RNA - Precipitation	Exos (50–100 nm)	miR-29	Unclear	Decreased HCV replication by inducing the expression of antiviral genes such as IFN-α, ISGs, OAS-1	124
	- THP-1 cell line with IFN - Ultracentrifugation	EVs (50–400 nm)	Not studied	Unclear	Profound inhibition of HCV RNA replication after 72 h or 96 h exposure	126
Drug delivery
Pulmonary metastases	- RAW 264.7 cell line for vitro; mice BMDM for vivo - Precipitation	Exos (~110 nm)	EVs loaded with PTX	- 40 h after i.v. injecting tumor cells - 4 × 1011 particles per mouse by i.v. at the days 1, 4, and 7	- Higher drug loading and targeting capacity towards cancer cells - Superior antineoplastic efficacy and prolonged lifespan	132
Breast cancer	- RAW 264.7 cell line - Ultracentrifugation	EVs (~110.8 nm)	EVs loaded with DOX or PTX	- Tumor volume reached ~50 mm3 - EVs-PTX (0.5 mg/kg); EVs-DOX (2.5 mg/kg) by i.v. once every 3 days	- Improved loading efficiency of DOX when pH close to PI, of PTX when dissolved in ethanol - Higher affinity towards tumor sites, more robust inhibitory potency on tumor growth and prolonged lifespan of both	133
	- RAW 264.7 cell line - Ultracentrifugation	Exos (~97.3 nm)	EVs loaded with DOX	- Tumor volumes reached 60 mm3 - At DOX dose of 5 mg/kg by i.v. every 3 days for total 18 days	- Prolonged circulation time of DOX and better accumulation of DOX at tumor tissue - Highest tumor inhibition efficacy - Decreased other tissue lesions	134
	- J774A.1 cell line - Membrane filter extrusion	EVs (~139 nm)	Hybrid EVs with liposomes and loaded with DOX	Unclear	- Higher drug release in acidic microenvironment - Higher biocompatibility as a safe delivery system with lower cytotoxicity - Decreased K7M2, 4T1, and NIH/3T3 cell viability	135
Vaccine adjuvant
Melanoma	- RAW264.7 cell line with γ-IFN - Ultracentrifugation	Exos (~50 nm)	cytokines such as IL-6, IL-12, and γ-IFN	- Tumor volume reached ~50 mm3 - 10 μg EVs by single subcutaneously at 24 h after injection of vaccine	- Elevated apoptosis of melanoma cancer cells - Decreased celluar scar tissues as well as many infiltrating immune cells - Significant inhibitory effects on tumor growth	131