Table 5.
Secondary outcomes related to microRNA/circRNA profile and signaling pathway/gene expression analysis.
| microRNA/circRNA Profile | Functions/Signaling Pathways | Reference |
|---|---|---|
| Exo-D3 versus Exo-P6: one miRNA differentially expressed, which enriches three pathways related to osteogenic differentiation. Exo-D21 versus Exo-P6: nine differentially expressed miRNAs that enrich twenty pathways related to osteogenic differentiation. Exo-D21 versus Exo-D3: sixteen differentially expressed miRNAs that enrich twenty pathways related to osteogenic differentiation. |
Exo-D3 versus Exo-P6: Hippo signaling pathway, adherens junction, ECM–receptor interaction. Exo-D21 versus Exo-P6: Wnt, Hippo, MAPK, cAMP, PI3K-Akt, TGF-beta, TNF, VEGF, insulin, and AMPK signaling pathways, among others. Exo-D21 versus Exo-D3: Wnt, Hippo, MAPK, cAMP, PI3K-Akt, TGF-β, TNF, HIF-1, insulin, and AMPK signaling pathways, among others. |
[36] |
| 11 circRNAs were upregulated in Exo7. ↑ circLPAR1 and ↓ hsa-miR-31 in Exo7-treated DPSCs |
circLPAR1 was predicted to bind to hsa-miR-31, a miRNA that showed an inhibitory effect against osteogenic differentiation. circLPAR1 would be the target of hsa-miR-31. Both downregulation of hsa-miR-31 and upregulation of circLPAR1 promoted osteogenic differentiation of DPSCs. | [31] |
| ↑ of 72 miRNAs and ↓ of 35 miRNAs in exosomes derived from osteogenically differentiated PDLSCs. ↑of miR-122-5p, miR-142-5p, miR-25-3p, miR-192-5p. ↓ of miR-125b-5p, let-7b-5p, and miR-100-5p. |
Predicted functions: catalytic activity, protein binding, metabolic process, transport, and phosphate-containing compound metabolic process. Processes related to target genes: 2-oxocarboxylic acid metabolism, adipocytokine signaling pathway, AMPK signaling pathway, insulin signaling pathway, and MAPK signaling pathway. |
[33] |
| Upregulation of 160, 166, 193, and 136 miRNAs and downregulation of 130, 139, 150, and 191 miRNAs were were observed in the Exo-D4, Exo-D10, Exo-D15, and Exo-D20 exosomes, respectively. ↑ expression of osteogenic miRNAs (Hsa-miR-146a-5p, Hsa-miR-503-5p, Hsa-miR-483-3p, and Hsa-miR-129-5p) and ↓ expression of anti-osteogenic miRNAs (Hsa-miR-32-5p, Hsa-miR-133a-3p, and Hsa-miR-204-5p) in 10- and 15-day osteogenically induced exosomes. |
Predicted signaling pathways: PI3K/Akt and MAPK. | [40] |
| ↑ of 201 miRNAs and ↓ of 33 miRNAs in exosomes derived from osteogenically differentiated ASCs. ↑ of five miRNAs (miR-130a-3p, miR-30b-5p, miR-34a-5p, miR-324-5p, and miR-378f) and ↓ of miR-513b-5p. |
Predicted processes affected: axon guidance, MAPK signaling, Wnt signaling, endocytosis, regulation of actin cytoskeleton, and TGF-β signaling pathway. Functions affected: enzyme binding, cell projection, transcription factor activity, regulation of gene expression, and cell metabolism. Mir-130a03p had the highest differential expression and was predicted to bind to SIRT7. The downregulation of SIRT7 may enhance the osteogenic differentiation of BMSCs. |
[34] |
| NA | Phosphorylation of STAT6, GSK-3α/β, STAT5b, and STAT5a/b increased following stimulation with 0d-Exo, whereas phosphorylation levels of FAK, PRAS40, and WNK1 were downregulated. SMAD 4 and BMPR2 were upregulated in 0d-Exo treatment. SMAD1/8, BMPR1A, and BMPRIB showed no difference between treatments. |
[35] |
| D21 versus D0 (do not specify the type of cell): ↑ of miR-186, miR-210, miR-181c-5p, and miR-146a-5p and ↓ of miR-133 and miR-485. | Potential signaling pathways: TGF-beta signaling pathway, Hippo signaling pathway, Map kinase, and Wnt signaling pathway, among others. | [32] |
| NA | ↓ PTEN in DMOG-MSC-Exos. The deficiency of PTEN is related to increased migration and invasion of HUVECs. The downstream target of PTEN, AKT/mTOR, was blocked, and DMOG-MSC-Exos lost their superior pro-angiogenic abilities. | [20] |
| ↑ of miR-146a in BMSCs by Sr-CS extracts | The miR-146a inhibition led to the downregulation of miR-146a in both BMSCs and BMSC-Exos. Treatment with Sr-CS + 146I-Exo (derived from miR-146a inhibition) diminished its angiogenic ability in HUVECs. Prediction targets showed that miR-146a directly targets Smad4 and NF2 (reported to inhibit angiogenesis). |
[41] |
| ↓ of IGFBP3 associated with miR-375 overexpression. ↑ cell differentiation in IGFBP3-deficient cells cultured in OM. Furthermore, the increase in osteogenic differentiation induced by Exo (miR-375) treatment was reversed by IGFBP3 recombinant treatment. | [38] | |
| NA | BMP2 FEEs were able to trigger SMAD 1/5/8 phosphorylation, and control EVs had no effect beyond the resting cell state. | [21] |
| ↑ of 94 miRNAs and ↓ of 39 miRNAs in the Hypo-Exos group when compared to the Exos group. ↑ of miR-126, miR-855–5p, miR-146b, miR-223, and miR-451. |
Knockdown of miR-126 inhibited Hypo-Exos-mediated proliferation, migration, and angiogenesis in vitro and in vivo. SPRED1 is a target of miR-126 and was increased in miR-126 knockdown Hypo-Exos-treated cells. Silencing of SPRED1 promoted cell proliferation, migration, and tube formation during treatments with miR-126 knockdown Hypo-Exos. Hypo-Exos suppressed SPRED1 by activating the Ras/Erk pathway. |
[17] |
| 565 miRNAs were differentially expressed in SM-Exo. ↑ of 16 miRNAs and ↓ of 9 miRNAs in SM-Exo compared to Exos. ↑ of mir-10a-5p and mir-10b-5p and ↓ of mir-212-3p in SM-Exo. |
NA | [39] |
ALP, alkaline phosphatase; AMPK, AMP-activated protein kinase; ARS, alizarin red staining; ASCs, adipose stem cells; BMSCs, bone marrow mesenchymal stem cells; BMP2, bone morphogenetic protein 2; BMPR2, bone morphogenetic protein receptor type 2; circRNA, circular RNA; DMOG, dimethyloxalylglycine; ECM, extracellular matrix; EVH1 domain-containing protein 1; Exo, exosome; FAK, focal adhesion kinase; FEEs, functionally engineered extracellular vesicles; GSK, glycogen synthase kinase; HUVECs, human umbilical vein endothelial cells; IGFBP3, insulin-like growth-factor-binding protein 3; IRS1, insulin receptor substrate 1; JAK-STAT, Janus kinase (JAK) signal transducer and activator of transcription; LPAR1, lysophosphatidic acid receptor 1; miRNA, microRNA; MAPK, mitogen-activated protein kinase; NA, not applicable; NF2, neurofibromatosis type 2; OM, osteogenic medium; PDLSCs, periodontal ligament stem cells; PDSCs, dental pulp stem cells; PLK2, Polo-like kinase 2; PRAS40, proline-rich AKT substrate of 40 kDa; PTEN, phosphatase and tensin homolog; SIRT7, sirtuin 7; SM-Exo, strain-microenvironment-derived exosome; SPRED1, Sprouty-related; Sr-CS, strontium-substituted calcium silicate; STAT, signal transducer and activator of transcription; TGF-β, transforming growth factor beta; TNF, tumor necrosis factor; VEGF, vascular endothelial growth factor; WNK1, lysine deficient protein kinase 1; ↑, upregulated; ↓, downregulated).