Table 1.
Synopsis of technical details and main results of studies on MSC-EVs in BPD.
| Reference | MSC source | MSC product | Isolation | Disease model | Route | Dose/ frequency | Main result/ action↑↓ | Pathway/active factor |
|---|---|---|---|---|---|---|---|---|
| Lee et al. (2012) | BM-MSCs (mouse) and WJMSCs (human) | CM and Exosomes | Ultrafiltration, PEG, size exclusion chromatography, UC | In vivo: HPH |
IV | 1 Dose of CM: 50 μL of 5 μg protein 1 or 2 doses of exosomes: 0.1 μg or 10 μg protein |
Inhibited vascular remodeling and HPH ↓ Influx of macrophages ↓ Pro-inflammatory and pro-proliferative mediators |
↓ STAT3activation and ↑ miR-17 superfamily and lung levels of miR-204. |
| Willis et al. (2018) | BM-MSCs (human) and WJMSCs (human) | EVs angiogenesis | Differential centrifugation, TFF, OptiPrepTM cushion density flotation | In vivo: BPD (hyperoxia 75% O2) In vitro: mouse BMDM Or alveolar macrophages |
IV | 1 dose 50 μL of EVs ≈ 0.5 × 106 cell equivalents |
Prevented lung fibrosis ↑ Alveolarization, ↑ Pulmonary function and modulated macrophage phenotype |
Modulate macrophage Phenotype: ↓ pro-inflammatory and ↑ Anti-inflammatory state |
| Ahn et al. (2018) | hUC MSCs | Cells and EVs | UC | In vivo: BPD hyperoxia (90%O2) in vitro: rat lung epithelial cell line challenged with H2O2 |
IT | 1 Dose onPN5 50 µL of: MSCs: 1 × 105 cells OR EVs: 20 µg of protein |
MSCs and EVs equally: ↑alveolarization,↑vascularization and ↓inflammatory response |
Transfer of VEGF |
| Chaubey et al. (2018) | GA hUC-MSCs | CM and Exosomes | 10-Fold concentrated CM &UC | In vivo: BPD hyperoxia (95% O2) in vitro: lung epithelial cell line challenged with H2O2 |
IP | 2 Doses PN2 & PN4 CM:10 μg protein. Exosomes: 0.7 × 106 cell equivalents |
↓ Pulmonary inflammation ↓ Alveolar-capillary leakage ↓alveolar simplification, ↓PH and RVH ↓ Cell death in brain and ↑Myelination |
TSG-6 |
| Braun et al. (2018) | BM-MSCs (Rat) | Exosomes | UC | In vivo: BPD hyperoxia (85%O2) in vitro: HUVEC tube formation assay |
IP | Daily injection of 15 mg protein ≈ 3.4 × 109 exosomes | In vivo: ↓alveolar simplification, ↑Small vessel number and inhibited RVH In vitro: ↑ tube-like formation by HUVEC |
VEGF mediated mechanism |
| Porzionato et al. (2018) | hUC MSCs | Cells and EVs | TFF | In vivo: BPD hyperoxia (60%O2) |
IT | 3 Doses (PN3, PN7, PN10) MSCs: 6 × 106 cells EVs: 0.64 × 1010 particles/ml |
Both EVs and MSCs: ↓hyperoxia-induced damage. EVs: better alveolarization and vascularization |
|
| Porzionato et al. (2020) | hUC MSCs | EVs | TFF | In vivo: BPD hyperoxia (60%O2) |
IT | 4 Times (PN3, PN7, PN10, and PN21) EVs: 0.64 x1010 particles/ml |
↑ Alveolarization ↓ PA remodeling MSC-EV preserved: –The interstitial, alveolar and perivascular CD163+ macrophages –↑ Cell proliferation |
M2 macrophage polarization could play a role through anti-inflammatory and proliferative mechanisms. |
| Willis et al. (2020) | WJMSCs (human) | EVs | Differential centrifugation, TFF, OptiPrepTM cushion density flotation | in vivo: BPD (hyperoxia 75% O2) In vitro: mouse BMDM |
IV | For early intervention: at PN4 1 dose of 0.5 × 106 cell equivalents For late intervention: at PN18 1 dose of 1 × 106 cell equivalents For serial late intervention: 4 doses (PN18-PN39) of 1 × 106 cell equivalents |
Early intervention: See previous publication Late intervention: –1 dose:partially restores alveolar simplification. –serial doses: ↑ alveolarization,↓ fibrosis, ↓ vascular muscularization and ↓microvascular loss Early and late MEx interventions ↓ RVH and ↑ functional exercise capacity |
|
| You et al. (2020) | hUC MSCs | EVs | Serial centrifugation, UC | In vivo: BPD (85% O2) In vitro: HUVEC tube formation assay and cell survival of MLE-2 under hyperoxic conditions |
IT | 1 Dose 20 mg of protein on PN7 |
In vivo: ↓ alveolar simplification and lung function,↓ PH, ↑ Ki-67+ and ↓ TUNEL+ lung cells, ↑ type II AECs, ↑ pulmonary vascular endothelial cells in vitro: ↑ tube formation of hyperoxic HUVECs ↑ proliferation and ↓apoptosis in MLE-12 |
PTEN/Akt signaling pathway: ↓ expression of PTEN & cleaved-caspase3, and ↑ expression of p-Akt and VEGF-a |
| Wu et al. (2021) | BM-MSCs (rat) | Exosomes | Exosome isolation reagent & centrifugation for 1 h at 12,000 g | In vivo: Hyperoxia lung injury (90% O2) in vitro: lung epithelial cell line (RLE-6TN) challenged with H2O2 |
IV | 1 Dose 800 μg of protein |
In vivo: ↑ alveolarization, ↓inflammatory influx in lung In vitro: ↓ oxidative damage on RLE-6TN, |
miR-425 in BMSCs-EVs activates the PI3K/AKT axis by targeting PTEN and thus inhibits hyperoxic injury |
| Willis GR et al. (2021) | WJMSCs and BM-MSCs (human) | MEx (small EVs) | Differential centrifugation, TFF, OptiPrepTM cushion density flotation | In vivo: BPD (hyperoxia 75% O2) in vitro: mouse BMDMy pretreated with MEx |
IV | 1 Dose 50 μL of EVs ≈ 0.5 × 106 cell equivalents on PN4 |
MSC-EVs: –co-localized with F4/80+, CD64+myeloid cells –preserved the lung CD45+ cells, especially AMφ and Ly6C low monocytes -↓ AMφ inflammatory activation Adoptive transfer of MSC-EV-educated-BMDMy prevented the hyperoxic injury similarly to MSC-EV treatment |
MSC-EV modulate myeloid cells into a Ly6C/G+, CX3CR1+, CCR2− phenotype, with immunosuppressive capacity, possibly through transcriptomic and epigenetic reprogramming |
| Reis M. et al. (2021) | WJMSCs and HDF (human dermal fibroblast) | MEx (small EVs) | Differential centrifugation, TFF, OptiPrepTM cushion density flotation | In vivo: BPD (hyperoxia 75% O2) in vitro: T cell autoreactivity assessment |
IV | 1 Dose 50 μL of EVs ≈ 0.5 × 106 cell equivalents on PN4 |
MSC-EVs: –prevented the development of BPD –preserved the thymic medullary architecture ↑development of regulatory T cells ↓ T cell autoreactivity ↑ genes related to maturation, antigen presentation and oxidative stress in DCs and mTECs |
Modulation of thymic antigen presenting cell populations (DCs and mTECs) |
| Fernandez-Gonzalez et al. (2021) | WJMSCs (human) & BMSCs(human) | MEx (small EVs) | Differential centrifugation, TFF, OptiPrepTM cushion density flotation | In vivo: BPD (hyperoxia 75% O2) |
IV | 1 Dose 50 μl of EVs ≈ 0.5x106 cell equivalents on PN4 |
Lung: ↑ Vascularization and ↑alveolarization Brain: ↑myelination ↓ astrogliosis ↓activation of microglial cells Retina: Preserved the retinal thickness, ↓gliosis and ↓microglial activation and invasion into the outer nuclear layer. |
Macrophage/microglia immunomodulation |
Abbreviations: AECs, alveolar epithelial cells; Amφ, alveolar macrophages; BMDM, bone marrow derived monocytes; BMDMy, bone marrow derived myeloid cells; BM-MSCs, bone marrow mesenchymal stem cells; BPD, bronchopulmonary dysplasia CM, conditioned media; DCs, dendritic cells; EVs, extracellular vesicles; GA hUC-MSCs, early gestational age mesenchymal stem cells; HDF, human dermal fibroblast; HPH, hypoxia-induced pulmonary hypertension; hUC MSCs, human umbilical cord blood mesenchymal stem cells; HUVEC, human endothelial cells; IP, intraperitoneally IT, intratracheally; IV, intravenously; MEx, mesenchymal stem cell derived small extracellular vesicles; miR, microRNA; mTEC, medullary thymic epithelial cells; PEG, polyethylene glycol; PH, pulmonary hypertension; PN, post-natal day; RVH, right ventricular hypertrophy; TFF, tangential flow filtration; TSG-6, tumor necrosis factor alpha-stimulated gene-6; UC, ultracentrifugation; VEGF, vascular endothelial growth factor; WJ-MSCs, umbilical cord Wharton’s jelly mesenchymal stem cells.