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. 2022 Aug 4;10:965006. doi: 10.3389/fbioe.2022.965006

TABLE 3.

Functional tests on Perinatal cell-derived small extracellular vesicles (sEV) alone or compared to perinatal cell-conditioned medium (CM).

Perinatal cell-derived small extracellular vesicles (sEV)
PnD Functional in vitro tests Outcome Reference
hUC-MSC-sEV (i) Cell differentiation assay (i) hUC-MSC-sEV inhibited α-SMA and collagen I and III expression in HDF cultivated at high cell density. (ii) hUC-MSC-sEV restrict HaCaT and HDF proliferation at high cell densities, but promote cell proliferation at low densities. (Zhang et al., 2016)
(ii) Cell proliferation assay
(i) Cell proliferation assay hUC-MSC-sEV promoted the (i, ii) proliferation, (iii, v) migration, and (iv) tube formation of a HUVEC-derived cell line in a dose-dependent manner. (Zhang et al., 2015)
(ii) Cytotoxicity assay
(iii) Scratch assay
(iv) Tube formation assay
(v) Chemotaxis assay
(i) Cell proliferation assay hUC-MSC-sEV promoted (i) the proliferation, (ii) migration and (iii) tube-formation of HUVEC. hUC-MSC-sEV contained Ang-2, and treatment with hUC-MSC-sEV enhanced the expression of the Ang-2 in HUVEC through exosome-mediated Ang-2 transfer. (Liu et al., 2021)
(ii) Chemotaxis assay
(iii) Tube Formation Assay
Perinatal cell-derived small extracellular vesicles (sEV) compared to perinatal cell-CM
PnD Functional in vitro tests Outcome Reference
a) hUC-MSC-sEV (i) Cell apoptosis assay hUC-MSC-CM and hUC-MSC-sEV (i) decreased H2O2-induced cell apoptosis of HaCaT by inhibiting AIF and upregulating PARP-1 and poly ADP-ribose, (ii) increased HaCaT proliferation, in contrast to hUC-MSC-sEV-dp. (iii) hUC-MSC-CM improved the viability of HaCaT. (iv, vi) hUC-MSC-sEV and hUC-MSC-CM promoted cell migration relative to the hUC-MSC-sEV-dp. (vi) (v) ROS intensity in the hUC-MSC-sEV group and hUC-MSC-CM group was lower than in the control group. (Zhao et al., 2020)
b) hUC-MSC-CM (ii) Cell proliferation assay
(iii) Cell viability assay
(iv) Chemotaxis assay
(v) ROS generation assay
(vi) Scratch wound assay
(i) Cell proliferation assay (i) hUC-MSC-sEV/Pluronic F-127 hydrogel promoted HUVEC proliferation better than hUC-MSC-sEV and hUC-MSC-CM. (ii) hUC-MSC-sEV and CM groups showed greater cell migration than the Pluronic F-127 hydrogel and control group. The hUC-MSC-sEV/Pluronic F-127 hydrogel group exhibited the best performance. (Yang et al., 2020)
(ii) Scratch wound assay
a) hDMSC-sEV Cell cycle assay Cell differentiation assay Cell proliferation assay Scratch wound assay Senescence assay ROS generation assay (i) hDMSC-sEV enhanced proliferation of HG aged HDF, (iv) increased their migration rate, (ii) promoted differentiation of HG-aged HDF into myofibroblasts (increased α-SMA and collagen I protein expression), (v) inhibited senescence associated β-galactosidase expression and (vi) inhibited ROS generation in HG aged HDF. hDMSC-CM improved (iii) proliferation and (iv) migration of HDF. The sEV blocker GW4869 reduced both effects, indicating that hDMSC-sEV in hDMSC-CM probably enhance the proliferation and migration abilities of HDF. (Bian et al., 2020)
b) hDMSC-CM

Abbreviations; Ang-2: angiopoietin-2, AIF: apoptosis-inducing factor, α-SMA: alpha-smooth muscle actin, CM: conditioned medium derived from hDMSC, hUC-MSC, HaCAT: immortalized human keratinocytes, HDF: human dermal fibroblast, hDMSC: human decidua mesenchymal stromal cells, HG: high glucose, hUC-MSC: human umbilical cord mesenchymal stromal cells, PARP-1: poly ADP, ribose polymerase 1, ROS: reactive oxygen species, sEV: small extracellular vesicles derived from hDMSC, hUC-MSC, sEV-dp: conditioned medium depleted from small extracellular vesicle. Perinatal cell-derived small extracellular vesicles (sEV).