Fig 2: Illustration of how electric microenvironment promotes osteogenesis in diabetes mellitus by modulating macrophage polarization. After corona poling treatment, the surface of the ferroelectric nanocomposite membrane displays induced charges. Within the electrical microenvironment, macrophages exhibit less M1 polarization and more M2 polarization. The polarized nanocomposite membranes downregulate AKT2-IRF5 signaling that induces macrophages to the M2 phenotype, which indicates a favorable osteo-immunomodulatory environment. Hence mimicking the endogeneous electrical microenvironment promotes osteogenic differentiation of BM-MSCs mediated by macrophages, thereby improving bone regeneration. The purple arrow denotes the osteogenesis of BM-MSCs. The grey arrows denote the suppressed M1 phenotype of macrophages. The red arrows denote the downregulated expression of AKT2 and IRF5, and improved M2 phenotype of macrophages.

Fig 3: Persistent exposure of HUVECs to HG increases miR-6832-5p, miR-6842-5p and miR-8056 expression by downregulating MIR181A2HG. (A) MTS assay detected the viability of HUVECs treated with different doses of D-glucose for the indicated times. **P<0.01 and ***P<0.001 vs. day 0. (B) Heatmap representation of differential expression of lncRNAs between HUVECs treated with 5.5 mM (L5.5) and 30 mM D-glucose (H30) for 24 h. (C) RT-qPCR was used to detect the relative expression of MIR181A2HG in HUVECs cultured in medium containing 30 mM D-glucose for the indicated times. *P<0.05 and ***P<0.001 vs. day 0. (D) FISH was used to detect the location and expression of MIR181A2HG in HUVECs exposed to 5.5 or 30 mM D-glucose for the indicated times. (E) Predicted binding sites of the miRNAs on MIR181A2HG. (F) Luciferase reporter assay was used to verify the direct binding between MIR181A2HG and miRNAs. *P<0.05 and **P<0.01 vs. miR-Ctl. (G) RT-qPCR was used to detect the relative expression of the indicated miRNAs in HUVECs cultured in medium containing 30 mM D-glucose for 96 h. *P<0.05 and **P<0.01 vs. Ctl. (H) RNA pulldown assay was performed to detect the direct binding between MIR181A2HG and the 3 miRNAs. **P<0.01 vs. Ctl. (I and J) RT-qPCR was used to detect the relative expression of (I) MIR181A2HG and (J) miRNAs in HASMCs cultured in medium containing 5.5 or 30 mM D-glucose. (K and L) RT-qPCR and western blot analysis detected the relative expression of AKT2 mRNA and protein in HASMCs transfected with miR-181-3p mimics, miR-181-5p mimics or their corresponding control. HUVECs, human umbilical vein endothelial cells; HASMCs, human aortic smooth muscle cells.

Fig 4: Electrical microenvironment mediates the phenotypic changes of macrophages by the AKT2-IRF5 signaling pathway. (a) Pathway Enrichment analysis of normalized gene expression data in each group by microarray analysis. (b) The mRNA expression levels of pathway genes in each group. (c) The AKT2 and phosphorylatd AKT2 were activated under high glucose conditions and inhibited in an electrical microenviroenvirnment. (d, e) The expression of IRF5 and HIF-1a in cells treated with or without AKT inhibitor. (f, g) Flow cytometry analysis of cell surface markers CD11c and CCR7 under different conditions. (h) The schematic representation of molecular signaling events that mediates electrical microenvironment-induced macrophage polarization. *denotes significantly different from the UP-NG group, #denotes significantly different from the UP-HG group (n = 3).

Fig 5: miR-758-3p promoted the tumorigenesis of melanoma cells via targeting TCEAL7. a, b In vivo tumor formation assay was carried out to detect the tumorigenesis in A375 cells from mimic-NC + Lenti-NC, mimics + Lenti-NC and mimics + Lenti-TCEAL7 groups. c RT-PCR analysis of the expression levels of miR-758-3p in tumor tissues. d, e RT-PCR and western blotting assays were applied to detect the mRNA and protein levels of TCEAL7, AKT1, AKT2 and c-Myc in tumor tissues. (*P < 0.05 and **P < 0.01, compared with mimic-NC + Lenti-NC group; #P < 0.05, compared with mimics + Lenti-NC group)