Table 1.
Overview of studies investigating the therapeutic potential of EVs in neonatal diseases.
| References | Sources of EVs | Model | Protocol of Administration | Dosing | Therapeutic Effects | Elucidated Mechanisms |
|---|---|---|---|---|---|---|
| HYPOXIC–ISCHEMIC ENCEPHALOPATHY | ||||||
| [245] 2016 |
human bone marrow MSCs |
the Rice–Vannucci model | total dose—2.4 × 1010 EVs route of administration—intravenous administrations per day—2 days—1 |
1 unit * |
↓ total number and duration of seizures ↓ pathological fluctuations of blood pressure |
↑ baroreflex-mediated heart rate response |
| [156] 2019 |
human bone marrow MSCs |
the Rice–Vannucci model | total dose—2.4 × 1010 EVs route of administration—intravenous administrations per day—2 days—1 |
1 unit |
↓ permeability of blood–brain barrier | ↑ Annexin A1/FPR in neonatal brain endothelial cells and microglia |
| [151] 2019 |
human bone marrow MSCs |
the Rice–Vannucci model | total dose—1.25 × 109 EVs route of administration—intranasal administrations per day—1 days—1 |
0.1 units |
↓ of tissue loss ↓ % of cell death ↓ microglial activation ↑ behavioral outcomes (negative geotaxis test) |
|
| [93] 2019 |
human Wharton’s jelly MSCs |
the Rice–Vannucci model + intraperitoneal injection of LPS | total dose—325 µg of EV protein per animal route of administration—intranasal administrations per day—1 days—1 |
0.3 units |
↓ microgliosis ↓ neuroinflammation |
↓ LPS/TLR4 signaling in microglia |
| [246] 2020 |
rat bone marrow MSCs (H2S preconditioning) |
the Rice–Vannucci model | total dose—1.5 × 108 EVs route of administration—intracardial injection administrations per day—1 days—1 |
0.06 units |
↓ water content and infarct volume of the brain ↓ % of cell apoptosis ↑polarization toward the anti-inflammatory M2 phenotype ↑ memory function |
↑ miR-7b-5p ↓ FOS → ↓ Iba1+ in microglia |
| [152] 2020 |
human bone marrow MSCs |
the Rice–Vannucci model | total dose—2.7 × 108 EVs route of administration—intraperitoneal administrations per day—1 days—3 (1,3,5 post HI) |
0.03 units |
↓ striatal tissue loss ↓ M1 micro- and A1 astroglia activation ↑ neurogenensis and angiogenesis ↑ myelination |
|
| [247] 2021 |
mice bone marrow MSCs |
the Rice–Vannucci model | total dose—100 µg of EV protein route of administration—intracardial injection administrations per day—1 days—1 |
0.1 units |
↓ HI-induced edema, infarction, infiltrating monocytes ↓ phagocytosis of viable neurons ↑ synaptic densities |
↓ p-NF-κB → ↓OPN → ↓ Iba1 in M1 microglia |
| [248] 2021 |
mice bone marrow MSCs |
the Rice–Vannucci model | total dose—5 µg of EV protein route of administration—intranasal administrations per day—1 days—1 |
0.005 units |
↓ injury volumes ↓ microglial activation ↓ neuroinflammation |
↓ Iba1 → ↓ Casp3 in microglia |
| [249] 2021 |
rat primary astrocytes (P1) |
the Rice–Vannucci model | total dose—2.5 µg of EVs protein route of administration—intraperitoneal administrations per day—1 days—1 |
0.0024 units | ↓ the area of cerebral infarction ↓ HIBD-induced neuronal apoptosis ↓ oxidative stress ↓ neuroinflammation ↑ body weight ↑ cognitive functions(grip test, negative geotaxis test |
↑ miR-17-5p → ↓ BNIP → ↓ Bax in brain tissue |
| [250] 2022 |
brain tissues of neonatal mice (P9) after HI |
the Rice–Vannucci model | total dose—8 × 109 EVs route of administration—intranasal administrations per day—2 days—1 |
0.066 units |
↓ infarct size ↓ Casp3 expression |
↑ miR-342-3p and miR-330-3p in brain tissue |
| [251] 2022 |
mice bone marrow MSCs |
the Rice–Vannucci model | total dose—2 × 109 EVs route of administration—intranasal administrations per day—1 days—1 |
0.2 units |
↑ animal survival ↓ infarct volume of brain ↓ % of apoptosis cells ↓ neuroinflammation ↑ proprioceptive function |
↑ miR-93 → ↓ JMJD3 → ↑ KLF2 → ↓ Casp3,Bax in neurons |
| [252] 2023 |
immortalized human bone marrow MSCs | the Rice–Vannucci model | total dose—2.7 × 108 EVs route of administration—intranasal administrations per day—1 days—3 (1,3,5 post HI) |
0.03 units |
↑ neurogenesis and angiogenesis ↓ monocyte infiltration ↓ astrogliosis and microgliosis |
|
| BRONCHOPULMONARY DYSPLASIA | ||||||
| [167] 2018 |
human Wharton’s jelly MSCs |
hyperoxia (HYRX)-induced BPD mice model (P1–P7 75% O2) |
total dose—0.9 µg of EV protein route of administration—intravenous administrations per day—1 days—1 |
0.001 units |
↑ lung architecture ↓ lung fibrosis ↓ peripheral pulmonary arterial remodeling |
|
| [175] 2018 |
preterm human Wharton’s jelly MSCs | HYRX-induced BPD mice model (c P1–P4 95% O2) | total dose—4.5 × 108 EVs route of administration—intraperitoneal administrations per day—1 days—2 (P2 and P4) |
0.038 units |
↑ lung architecture ↓ infiltration of neutrophils ↓ pulmonary hypertension ↓ alveolar-capillary leak |
↑ TSG-6 signaling in lung tissue |
| [253] 2018 |
human Wharton’s jelly MSCs | HYRX-induced BPD rat model (P1–P14 60% O2) |
total dose—0.213 × 1010 EVs route of administration—intratracheal administrations per day—1 days—3 (P3, P7 and P10) |
0.27 units |
↑ alveolar development ↓ pulmonary vascular remodeling |
|
| [176] 2018 |
rat bone marrow MSCs | HYRX-induced BPD rat model (P0–P14 85% O2) | total dose—3.4 × 109 EVs route of administration—intraperitoneal administrations per day—1 days—14 (P1–P15) |
1.96 units |
↑ alveolar growth ↑ lung blood vessel density ↓ pulmonary hypertension |
↑ VEGF signaling in lung tissue |
| [177] 2018 |
human umbilical cord blood MSCs |
HYRX-induced BPD rat model (P1–P14 90% O2) | total dose—20 µg of EV protein route of administration—intratracheal administrations per day—1 Days—1 (P5) |
0.019 units |
↑ alveolarization and angiogenesis | ↑ VEGF signaling in lung tissue |
| [254] 2020 |
human Wharton’s jelly MSCs | HYRX-induced BPD mice model (P0–P14 75% O2) | total dose—6 × 108 EVs route of administration—intravenous administrations per day—1 days—1 (PN4) |
0.025 units |
↓ alveolar simplification ↓ septal collagen disposition ↑ blood vessel count ↓ pulmonary hypertension ↑ functional exercise capacity |
|
| [255] 2021 |
human bone marrow MSCs. | In utero induced BPD rat model (antenatal injection of E. coli endotoxin e20) | total dose—0.25 × 106 EVs route of administration— intra-amniotic administrations per day—10 per pregnant rat days—1 (e20) |
0.17 units |
↓ lung simplification ↑ vascularization ↓ pulmonary hypertension ↑ lung mechanical function |
|
| [174] 2021 |
human Wharton’s jelly MSCs | HYRX-induced BPD mice model (P–P7 75% O2) | total dose—6 × 108 EVs route of administration—intravenous administrations per day—1 days—1 (PN4) |
0.025 units |
↑ thymic development ↑ proportion of CD4+FoxP3+ regulatory T cells ↓ alveolar simplification |
|
| [253] 2021 |
human umbilical cord blood MSCs | HYRX-induced BPD rat model (P1–P14 60% O2) | total dose—0.64 × 1010 EVs route of administration—intratracheal administrations per day—1 days—4 (P3, P7, P10 and P21) |
0.27 units |
↑ alveolar development ↓ deposition of fibrous tissue ↑ density of M2 macrophages ↓ pulmonary hypertension |
|
| [111] 2021 |
amniotic fluid-derived EVs (full-term cesarean sections) | HYRX-induced BPD rat model (P1–P14 85% O2) | total dose—1 × 1010 EVs route of administration—intratracheal administrations per day—1 days—1 (P3) |
0.42 units |
↑ alveolar development ↓ pulmonary hypertension |
|
| [256] 2022 |
human breast milk-derived EVs | HYRX-induced BPD rat model (P1–P7 85% O2) | total dose—140 µg of EV protein route of administration—intragastric administrations per day—1 days—1 (PN7) |
0.133 units |
↓ lung tissue collapse ↓ cleaved caspase 3 |
↓ IL-17/↓ FADD in Type II alveolar epitheliocytes |
| [171] 2022 |
human Wharton’s jelly MSCs | HYRX-induced BPD rat model (P1–P14 85% O2) | total dose—96 × 108 EVs route of administration—intratracheal administrations per day—1 days—1 (PN3) |
0.04 units |
↑ lung vascular density and alveolar structure ↓ lung inflammation ↓ pulmonary hypertension |
↑ VEGF/eNOS in lung tissue |
| [182] 2022 |
human amniotic epithelial cells (term birth after caesarean sections) |
in utero induced BPD mice model (injection of LPS e16) + (P3.5–P28 65% O2) |
total dose—10 µg of EV protein route of administration—intravenous administrations per day—1 days—1 (PN4) |
0.01 units |
↑ lung tissue-to-air space ratio ↓ lung inflammation ↑ type II alveolar epithelial cell ↓ pulmonary hypertension ↑ lung tissue elasticity |
|
| [179] 2022 |
human Wharton’s jelly MSCs | HYRX-induced BPD mice model (injection of LPS P7/P8) + 40% O2 P10 | total dose—1 × 106 EVs route of administration—intratracheal administrations per day—1 days—1 (PN9) |
0.0002 units |
↑ lung architecture ↑ blood vessel density ↑ mRNA of antiinflammatory cytokines in lung tissue |
|
| [257] 2023 |
human umbilical cord blood MSCs | HYRX-induced BPD mice model (P1–P14 85% O2) | total dose—15 × 105 EVs route of administration—intraperitoneal administrations per day—1 days—3 (P4–P6) |
0.000063 units | ↓ lung fibrosis ↑ vascular development |
↑ miR-185-5p →↓ CDK6 → ↑ angiogenesis in lung tissue |
| [169] 2023 |
human bone marrow MSCs | hypoxia—induced BPD rat model (10 min 40% O2 + 2 min 1% O2 12 times daily P1–P14) | total dose—2 × 105 EVs route of administration—intraperitoneal administrations per day—1 days—14 (P1–P14) |
0.00012 units | ↓ simplified alveolar structure ↓ pulmonary hypertension ↑ capillary distribution ↑ respiratory efficiency ↓ oxidative stress |
↑ PI3K/AKT → ↑ SOD in lung tisue |
| NECROTIZING ENTEROCOLITIS | ||||||
| [197] 2016 |
mice bone marrow MSCs | NEC—induced preterm mice model, Barlow et al. [258] (21e) + 90 s 100% N2 + 4 °C 10 min twice daily (P1–P4) | total dose—2.5 × 109 EVs route of administration—intraperitoneal administrations per day—1 days—1 (prior NEC) |
0.1 units |
↓ the overall incidence of NEC ↑ gut barrier function |
|
| [107] 2018 |
neonatal mice enteric neuronal stem cells | NEC—induced preterm rat model, Barlow et al. [258] (21e) + (90 s 100% N2 + 4 °C 10 min every 8 h + LPS every 4 h (P1–P4) | total dose—4 × 108 EVs route of administration—intraperitoneal administrations per day—1 days—1 (prior NEC) |
0.017 units |
↓ intestine villus destruction ↓ the overall incidence of NEC |
|
| [231] 2019 |
bovine milk-derived EVs | NEC—induced preterm mice model, Barlow et al. [258] (10 min 5% O2 3 times between P5–P9 + LPS 4 times between P6 and P7) | total dose—1.2 mg of EV protein route of administration—intragastric via gavage administrations per day—3 days—5 (P5–P9) |
1.14 units |
↑ intestine villus destruction ↑ number of goblet cells ↓ intestinal mucosal inflammation ↓ oxidative stress |
|
| [227] 2019 |
human breast milk-derived EVs | NEC—induced preterm rat model, Barlow et al. [258] (21e) 90 s 1.5% O2 + 4 °C 10 min 3 times daily P1–P4 + LPS 1 time P1 | total dose—2.4 × 1010 EVs route of administration—intragastric via gavage administrations per day—6 days—4 (P1–P4) |
1 unit |
↓ villus destruction ↓ the overall incidence of NEC |
|
| [194] 2020 |
rat amniotic fluid CD117 stem cells (e14.5) |
NEC—induced preterm mice model, Barlow et al. [258] (10 min 5% O2 3 times between P5–P9 + LPS 4 times between P6–P7) | total dose—3.5 × 108 EVs route of administration—intraperitoneal administrations per day—1 days—2 (P6–P7) |
0.015 units |
↑ gut epithelial regeneration ↓ intestinal inflammation |
↑ Wnt/β-catenin → increased intestinal epithelial proliferation |
| [259] 2022 |
human breast milk-derived EVs | NEC—induced preterm mice model, Barlow et al. [258] (1 min 100% N2 + 4 °C 5 min twice a day P6–P10) | total dose—30 µg of EV protein route of administration—intragastric via gavage administrations per day—3 days—3 (P8–P10) |
0.04 units |
↑ ileal crypts number ↓ inflammatory microenvironment |
|
| NEONATAL MENINGITIS | ||||||
| [190] 2022 |
human Wharton’s jelly-derived MSCs after thrombin preconditioning | Escherichia coli—induced meningitis in newborn rats (P11) | total dose—2.6 × 107 EVs route of administration—intraventricular administrations per day—1 days—1 (P11) |
0.001 units |
↓ neural cell death ↓ number of active microglia ↓ levels of inflammatory cytokines in brain tissues |
|
* the yield of EVs derived from supernatants of 4 × 107 MSCs that were cultured for 48 h was defined as 1 unit; 1 unit contains 2.4 × 1010 EVs or 1.05 mg of EV protein; ↓—downregulation, ↑—upregulation [14].