Journal: Materials Today Bio

Article Title: Human iPSC-EV-loaded nanofiber stent coatings accelerate vascular repair by enhancing EGFR/HIF-1α signaling and suppressing ROCK1-mediated remodeling

doi: 10.1016/j.mtbio.2026.103564

Figure Lengend Snippet: In vivo evaluation of re-endothelialization and inhibition of intimal hyperplasia following stent implantation. (A–E) SEM images of the luminal surface treated with EV-loaded (A, B) or DPBS-loaded (D, E) nanofibrous membranes. The EV-treated group exhibited a continuous, smooth, and intact endothelial layer (A, B), whereas the DPBS group displayed incomplete coverage with substantial endothelial disruption (D, E). (C, F) H&E-stained cross-sections revealed the extent of neointimal formation above the stent struts (black arrows). The EV group (C) demonstrated significantly reduced intimal thickening compared with the pronounced hyperplasia observed in the DPBS group (F). (G–H) Statistical analysis confirmed that EV treatment significantly increased endothelial coverage (G) and suppressed intimal hyperplasia thickness (H). (I–K) EV treatment significantly upregulated the expression of the angiogenic factor VEGF (I), the endothelial marker CD31 (J), and the proliferation marker PCNA (K), confirming accelerated vascular repair. (L–N) EV treatment prevented phenotypic switching and matrix accumulation, as evidenced by reduced levels of the contractile/hyperplasia marker α-SMA (L). Furthermore, the EV group showed decreased expression of matrix metalloproteinases MMP-2 and MMP-9 (M) and beneficial modulation of collagen composition, with increased Collagen I and the Collagen I/III ratio (N). (O–P) The EV group exhibited significantly downregulated levels of the profibrotic factor TGF-β1 (O) and the inflammatory cytokine IL-6 (P). N = 5 (G and H), and n = 3 (I-P). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.000.

Article Snippet: Smooth muscle and fibrosis markers: α-SMA (1:1000, A17910, ABclonal), SM22α (1:1000, ab14106, Abcam), Calponin-1 (1:1000, ab46794, Abcam), Osteopontin (1:1000, ab8448, Abcam), Collagen I (1:1000, A5786, ABclonal), and Collagen III (1:1000, A3795, ABclonal).

Techniques: In Vivo, Inhibition, Disruption, Staining, Expressing, Marker

Journal: Aging Cell

Article Title: IGF ‐1/ SLC25A33 Maintains Blood‐Labyrinth Barrier Integrity by Suppressing TGF ‐β/Smad2‐Mediated Pericyte Activation in Age‐Related Hearing Loss

doi: 10.1111/acel.70712

Figure Lengend Snippet: SLC25A33 deficiency induces pericyte differentiation via mitochondrial ROS accumulation, which were regulated by the TGF‐β/Smad2 signaling pathway. (A) Flow cytometry analysis using MitoSOX Red shows increased mtROS production in SLC25A33‐suppressed HBVPs compared to control and SLC25A33‐overexpressing groups. (B) Quantification of mtROS fluorescence intensity confirms this increase. (C, D) qPCR analysis reveals upregulated α‐Sma and downregulated E‐cadherin mRNA levels in Slc25a33 ‐knockdown HBVPs. (E) Representative Western blot images of α‐SMA and E‐cadherin protein expression. (F, G) Densitometric quantification shows increased α‐SMA and decreased E‐cadherin protein levels. (H) qPCR analysis shows upregulated Tgfb1 mRNA in SLC25A33‐deficient HBVPs. (I–K) No significant changes are observed in Smad2 , Smad3 , or Smad4 mRNA levels among groups. (L) Representative western blots of TGF‐β, p‐Smad2, Smad2, p‐Smad3, Smad3, Smad4, and GAPDH. (M) TGF‐β protein levels are significantly elevated in the knockdown group. (N) Phosphorylated Smad2 (p‐Smad2) is markedly increased, indicating pathway activation. (O–R) No significant differences are detected in p‐Smad3, total Smad2, Smad3, or Smad4 protein expression. Control: HBVPs without treatment; SLC25A33 (−): HBVPs treated with LV‐ Slc25a33 ‐shRNA; SLC25A33 (+): HBVPs treated with LV‐ Slc25a33 ‐OE. Data are mean ± SEM; n = 3.

Article Snippet: Membranes were blocked in 5% non‐fat milk for 1 h and incubated overnight at 4°C with primary antibodies against: E‐cadherin (Bioworld, BS80070, 1:500), α‐SMA (Bioss, bs‐10196R, 1:2000), TGFβ1 (Bioworld, BS80030, 1:500), Smad2 (Bioworld, BS1425, 1:500), Smad3 (Bioworld, BS1426, 1:500), Smad4 (Bioworld, BS45204, 1:1000), p‐Smad2 (Bioworld, MB11895, 1:500), p‐Smad3 (Bioworld, BZ16381, 1:500), and GAPDH (Cell Signaling Technology, 1:5000).

Techniques: Flow Cytometry, Analysis, Control, Fluorescence, Knockdown, Western Blot, Expressing, Activation Assay, shRNA

Journal: Aging Cell

Article Title: IGF ‐1/ SLC25A33 Maintains Blood‐Labyrinth Barrier Integrity by Suppressing TGF ‐β/Smad2‐Mediated Pericyte Activation in Age‐Related Hearing Loss

doi: 10.1111/acel.70712

Figure Lengend Snippet: SLC25A33 modulates α‐SMA and E‐cadherin expression through the TGF‐β/Smad2 pathway in vivo. (A) Immunofluorescence staining shows α‐SMA (red) and E‐cadherin (green) expression in the stria vascularis. Nuclei are counterstained with DAPI (blue). (B, C) Quantification reveals increased α‐SMA and decreased E‐cadherin in the Slc25a33 ‐knockdown group ( n = 5, five typical sections from five individual mice were analyzed per group). (D, E) qPCR confirms upregulation of α‐Sma and downregulation of E‐cadherin mRNA levels in the Slc25a33 ‐knockdown group ( n = 3 mice). (F, G) TGF‐β and (H, I) p‐Smad2 immunoreactivity are enhanced in the knockdown group and reduced in the overexpression group ( n = 5, five typical sections from five individual mice were analyzed per group). (J) qPCR confirms upregulation of Tgf‐β mRNA levels in the knockdown group and downregulation levels in the overexpression group ( n = 3 mice). Control: wild‐type C57BL/6J mice without treatment; SLC25A33 (−): mice treated with AAV‐ie‐sh Slc25a33 ; SLC25A33 (+): mice treated with AAV‐ie‐ Slc25a33 ‐OE. Data are mean ± SEM.

Article Snippet: Membranes were blocked in 5% non‐fat milk for 1 h and incubated overnight at 4°C with primary antibodies against: E‐cadherin (Bioworld, BS80070, 1:500), α‐SMA (Bioss, bs‐10196R, 1:2000), TGFβ1 (Bioworld, BS80030, 1:500), Smad2 (Bioworld, BS1425, 1:500), Smad3 (Bioworld, BS1426, 1:500), Smad4 (Bioworld, BS45204, 1:1000), p‐Smad2 (Bioworld, MB11895, 1:500), p‐Smad3 (Bioworld, BZ16381, 1:500), and GAPDH (Cell Signaling Technology, 1:5000).

Techniques: Expressing, In Vivo, Immunofluorescence, Staining, Knockdown, Over Expression, Control

Journal: Aging Cell

Article Title: IGF ‐1/ SLC25A33 Maintains Blood‐Labyrinth Barrier Integrity by Suppressing TGF ‐β/Smad2‐Mediated Pericyte Activation in Age‐Related Hearing Loss

doi: 10.1111/acel.70712

Figure Lengend Snippet: Exogenous IGF‐1 rescues blood‐labyrinth barrier integrity and improves hearing function in aged mice. (A) TEM images reveal ultrastructural changes in the stria vascularis. In the IGF‐1‐inhibited group, pericytes exhibit abnormal morphology, loose association with endothelial cells, and widened tight junctions (yellow arrows). These defects are ameliorated by exogenous IGF‐1 administration. (B) Click‐evoked ABR thresholds are significantly elevated in IGF‐1‐suppressed mice but partially rescued by IGF‐1 supplementation. (C–E) At 12 kHz, ABR thresholds are significantly lower in the IGF‐1‐treated group compared to control and suppressed groups, indicating improved auditory sensitivity. (F) Proposed model of IGF‐1/SLC25A33‐mediated regulation of pericyte phenotype and BLB integrity in age‐related hearing loss. Under physiological conditions, pericytes maintain a quiescent state characterized by low α‐SMA and high E‐cadherin expression, contributing to BLB stability. With aging, declining IGF‐1 levels lead to reduced SLC25A33 expression, resulting in mtROS accumulation and activation of the TGF‐β/Smad2 signaling pathway. This drives pericyte transition toward a contractile, pro‐fibrotic phenotype (high α‐SMA, low E‐cadherin), disrupting BLB integrity and promoting cochlear dysfunction. Targeting this axis may offer therapeutic potential for ARHL. Control: wild‐type C57BL/6J mice without treatment; IGF‐1 (−): mice treated with NVP‐AEW541; IGF‐1 (+): mice treated with recombinant IGF‐1. Data are mean ± SEM; n = 6 mice.

Article Snippet: Membranes were blocked in 5% non‐fat milk for 1 h and incubated overnight at 4°C with primary antibodies against: E‐cadherin (Bioworld, BS80070, 1:500), α‐SMA (Bioss, bs‐10196R, 1:2000), TGFβ1 (Bioworld, BS80030, 1:500), Smad2 (Bioworld, BS1425, 1:500), Smad3 (Bioworld, BS1426, 1:500), Smad4 (Bioworld, BS45204, 1:1000), p‐Smad2 (Bioworld, MB11895, 1:500), p‐Smad3 (Bioworld, BZ16381, 1:500), and GAPDH (Cell Signaling Technology, 1:5000).

Techniques: Morphology, Control, Sensitivity, Expressing, Stability, Activation Assay, Recombinant