Table 2.
Summary of technical details and main results of studies on MSC-EVs in neonatal brain diseases.
| Reference | MSC source | MSC product | Isolation | Disease model | Route | Dose/ frequency | Main result/action↑↓ | Pathway/active factor |
|---|---|---|---|---|---|---|---|---|
| Ophelders et al. (2016) | BM-MSCs (human) | EVs | MSC-CM filtration, PEG, low-speed centrifugation | In vivo: HIE transient UCO in the preterm ovine fetus |
In utero IV | 2 Doses 2 × 107 cell equivalents 1 h following UCO and 4 days after the insult |
↓ Total number and duration of seizures Preserved baroreceptor reflex sensitivity ↓ hypomyelination |
|
| Drommelschmidt et. al. (2017) | BM-MSCs (human) | EVs | PEG, UC | In vivo: LPS induced perinatal brain injury |
IP | 2 doses 1 × 108 cell equivalents/kg 3 h prior to and 24 h after IP injection of the vehicle or LPS |
↓Neuronal degeneration ↓ microgliosis ↓ reactive astrogliosis Prevented myelination deficits and white matter microstructural abnormalities ↑ cognitive function |
|
| Joerger-Messerli et al. (2018) | WJMSC (human) | EVs | Serial centrifugation | In vitro: OGD in the mouse neuroblastoma cell line neuro2a (N2a) |
1 Dose 0.1 mg/mL or 1 mg/mL of EVs either 24 h or 1 h before, or 6 h after OGD induction |
↓ DNA fragmentation and Casp3 expression | Delivery of let-7-5p-miR targeting proapoptotic genes | |
| C. Sisa et al. (2019) | BM-MSCs (human) | EVs | UC | In vivo: HIE modified Rice–Vannucci model |
IN | 1 Dose 6 µL of EVs 1.25 × 109 particles/dose |
↓ Microglia activation ↓ apoptosis ↓ brain tissue volume loss ↑ behavioral outcomes |
|
| R. Gussenhoven et al. (2019) | BM-MSCs (human) | EVs | PEG, low-speed centrifugation and UC | In vivo: HIE model ovine fetus UCO model in vitro: OGD in primary fetal endothelial cells |
IV | 2 Doses 2 × 107 cell equivalents at 1 h and 4 days after injury |
In vivo: ↓ BBB leakage in vitro: Restored endothelial barrier integrity |
Annexin A1 (ANXA1) in MSC-EVs targets the formyl peptide receptor (FPR) activation. |
| Thomi et al. (2019a) | WJ-MSC (human) |
Exosome | Serial centrifugation and UC | In vivo: LPS induced perinatal brain injury & modified Rice–Vannucci model in vitro: LPS stimulation of BV-2 microglia and primary mixed glial cells. |
IN | 1 Dose of 50 mg/kg. | In vivo: ↓ neuroinflammation ↓ pro-inflammatory cytokine production ↓ microgliosis in vitro: ↓inflammatory gene expression |
interfered with the TLR-4 signaling pathway, ↓degradation of IkBa and ↓phosphorylation of MAP kinase family molecules. |
| G.Thomi et al. (2019b) | WJ-MSC (human) |
Exosomes | Serial centrifugation and UC | In vivo: LPS induced perinatal brain injury & modified Rice–Vannucci model |
IN | 1 Dose of 50 mg/ kg | ↑ Animal survival ↓neuronal cell death Preserved: myelination, mature oligodendroglia and neuron cell counts ↑functional recovery ↑ the learning ability of treated animals. |
|
| Xin et al. (2019) | BM-MSCs (mouse) |
EVs | Centrifugation, filtration, ultrafiltration and EVs isolation kit (qEV, iZonScience) | In vivo: HIE modified Rice–Vannucci model |
ICV | 1 Dose of 100μg/ml 24 h after HI |
Neuroprotective effect ↓neuronal apoptosis and neuroinflammation skewed microglia and brain monocyte/macrophage toward a more anti-inflammatory phenotype. |
MSC-EVs transfer of miR-21a-5q to neurons which targets Timp3 gene. |
| Kaminski et al. (2019) | BM-MSCs (human) | EVs | Sequential centrifugation, PEG and UC | In vivo: HIE modified Rice–Vannucci model |
IP | 3 Doses (day 1, 3, and 5 after injury) 1 × 105 cell equivalents/g |
↓Striatal tissue loss ↓ microglia and astroglia activation In microglia: ↓ TNFa, ↑ YM-1 and TGFb In astroglia: ↓ C3,↑neural growth factors(BDNF, VEGF, EGF). ↑neuronal and vessel density ↑ cell proliferation in the neurogenic sub-ventricular zone juxtaposed to the striatum. Improved oligodendrocyte maturation and myelination |
Immunomodulation of microglia and astroglia phenotype (M1/M2 & A1/A2) |
| Chu et al. (2020) | BM-MSCs (mouse) |
EVs and H2S-EVs | Centrifugation, filtration, ultrafiltration and EVs isolation kit (qEV, iZonScience) | In vivo: HIEmodifiedRice–Vannucci model |
ICV | 1 Dose 100 μg EVs 1.5 × 108 particles 24 h following HI insult |
↑ Cognitive function MSC-EVs were found in both microglia and neurons 2h post-administration H2S-EVs were more potent at: ↓ brain tissue loss ↑ a more anti-inflammatory brain environment ↑ long-term cognitive and memory outcomes |
EV delivery of miR-7b-5p results in microglia and monocyte immunomodulation H2S MSC pre-treatment ↑ miR-7b-5p EV content miR-7b-5p delivery into the cells induces further miR-7b-5p expression |
| Han et al. (2021) | hUC-MSCs | EVs | Serial centrifugation & UC | In vivo: HIE modified Rice–Vannucci model in vitro: OGD to primary neurons |
IP | 4 Doses (prior and after the injury) 2 × 105 cell equivalents | In vitro: ↓ neuronal apoptosis in vivo: ↓ edema formationand infarction volume Ameliorated the neurological severity score |
EV delivery of miR-410 prevents neuronal apoptosis by an HDAC1-dependent EGR2/Bcl2 axis |
| Xin et al. (2021) | BM-MSCs(mouse) | EVs | Differential centrifugation & UC | In vivo: HIE modified Rice–Vannucci model |
ICV | 1 dose of 100 μg of EVs 24 h after HI |
↓ OPN expression induced by HI insult in microglia and macrophages restored synaptic reorganization ↑ synaptic protein expression ↓ edema and infarction volume |
EVs ↓OPN expression through NF-κB involvement |
| Ahn et al. (2020) | hUC-MSCs | Cells & EVs | UC | In vivo: IVH rodent model in vitro: rat cortical neuronal cells challenged with thrombin |
ICV | 1 × 105 MSCs or 20 μg of EVs at P6. |
In vitro: ↓ thrombin-induced neuronal cell death In vivo: ↓neuronal cell death, ↓ astrogliosis ↓ inflammatory responses ↑myelin basic protein and neurogenesis ↓ progression of post hemorrhagic hydrocephalus Ameliorated behavioral tests |
BDNF transfer via EVs |
| Pathipati et al. (2021) | BM-MSCs (mouse) |
EVs | Sequential centrifugation, filtration, ExoQuick TC Ultra |
In vivo: perinatal rodent stroke model in vitro: Microglia cells of HI mice |
ICV or IN | 1 Dose 1 μg/μL or 5 μg/μL |
↓ Edema and infarction volume MSC-sEV reside in microglia/macrophages of the injury site ↓ microglial morphological transformation ↓ cytokine/chemokine concentration ↓ caspase-3-dependent apoptotic cell death |
Modulate the microglia phenotype and cytokine production |
Abbreviations: BBB, blood–brain barrier; BDNF, brain derived neurotrophic factor; BM-MSCs, bone marrow mesenchymal stem cells; CM, conditioned media; EGF, epidermal growth factor; EVs, extracellular vesicles; H2S, hydrogen sulfide; H2S-EVs, hydrogen sulfide conditioned mesenchymal stem cell derived extracellular vesicles; HIE, hypoxic ischemic encephalopathy; hUC MSCs, human umbilical cord blood mesenchymal stem cells; ICV, intracerebroventricularly; IN, intranasally; IP, intraperitoneally; IT, intratracheally; IV, intravenously; LPS, lipopolysaccharide; MEx, mesenchymal stem cell derived small extracellular vesicles; miR, microRNA; OGD, oxygen/ glucose deprivation assay; OPN, osteopontin; PEG, polyethylene glycol; PN, post-natal day; TFF, tangential flow filtration; TNF-α, tumor necrosis factor alpha; UC, ultracentrifugation; UCO, umbilical cord occlusion; VEGF, vascular endothelial growth factor; WJ-MSCs, umbilical cord Wharton’s jelly mesenchymal stem cells; YM-1: CH3L3, chitinase 3 like protein 3.