sevs  (Bio-Rad)

Journal: Materials Today Bio

Article Title: Rational design of FAP-targeted sEVs delivered by microneedles for precision treatment of hypertrophic scars via ferroptosis in hypertrophic scar fibroblasts

doi: 10.1016/j.mtbio.2026.103117

Figure Lengend Snippet: Construction and characterization of sEVs ErF . (A) Schematic of sEVs ErF preparation from ADSC-derived sEVs, including erastin loading and UAMC1110 conjugation. (B, C) UV-Vis spectra of erastin and calibration curve at 280 nm. (D) Erastin loading content and loading efficiency at increasing incubation concentrations. (E) Synthetic route of SUC-Lys (Ac)-PEG 3 -UAMC1110. (F) Nano-flow cytometry of ligand-conjugated sEVs. (G) Fluorescence images of HSFs and HDFs incubated with SUC-Lys (FITC)-PEG 3 -UAMC1110-labeled sEVs ErF (FITC, green; phalloidin, red; DAPI, blue). Scale bar, 10 μm. (H) Image-based quantification of cellular fluorescence. (I, J) Uptake-positive cells (%) and representative flow-cytometry histograms for HSFs (left) and HDFs (right). (K) NTA size distributions of sEVs, sEVs Er , and sEVs ErF . (L) Western blot analysis of sEV-positive markers (CD9, CD63, TSG101) and the negative marker Calnexin in parental ADSC lysates and the indicated sEV formulations (sEVs, sEVs Er , sEVs ErF ). (M) TEM images showing cup-shaped morphology of the three vesicle types. Scale bar, 100 nm. Data are mean ± SD; ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: HSFs were treated with sEVs ErF (erastin-equivalent concentration: 15 μM) in the presence or absence of Ferrostatin-1 (Fer-1, 30 nM; MedChemExpress, USA).

Techniques: Derivative Assay, Conjugation Assay, Incubation, Flow Cytometry, Fluorescence, Labeling, Western Blot, Marker

Journal: Materials Today Bio

Article Title: Rational design of FAP-targeted sEVs delivered by microneedles for precision treatment of hypertrophic scars via ferroptosis in hypertrophic scar fibroblasts

doi: 10.1016/j.mtbio.2026.103117

Figure Lengend Snippet: In vitro antifibrotic effects of sEVs ErF on HSFs. (A, B) EdU staining and quantification of proliferating HSFs treated with Con, Er, sEVs Er , or sEVs ErF . Scale bar, 100 μm. (C) CCK-8 assay showing dose-dependent viability changes. (D, E) Transwell migration images and quantification of migrated HSFs. Scale bar, 100 μm. (F, G) Wound healing assay images and migration area (%) over time. Scale bar, 400 μm. (H-K) Western blot and densitometric analysis of COL I, COL III, and α-SMA; GAPDH, loading control. (L, M) α-SMA immunofluorescence and quantitative fluorescence intensity in HSFs. Scale bar, 50 μm. (N, O) Calcein-AM/PI flow cytometry and quantification of PI-positive cells. Data are mean ± SD; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: HSFs were treated with sEVs ErF (erastin-equivalent concentration: 15 μM) in the presence or absence of Ferrostatin-1 (Fer-1, 30 nM; MedChemExpress, USA).

Techniques: In Vitro, Staining, CCK-8 Assay, Migration, Wound Healing Assay, Western Blot, Control, Immunofluorescence, Fluorescence, Flow Cytometry

Journal: Materials Today Bio

Article Title: Rational design of FAP-targeted sEVs delivered by microneedles for precision treatment of hypertrophic scars via ferroptosis in hypertrophic scar fibroblasts

doi: 10.1016/j.mtbio.2026.103117

Figure Lengend Snippet: sEVs ErF induces ferroptosis in HSFs. (A) TEM images of mitochondria in HSFs after the indicated treatments, showing ferroptosis-like damage in the sEVs ErF group (reduced volume, dense membranes, loss of cristae). Scale bars, 1 μm (left) and 500 nm (right). (B, C) Representative flow-cytometry plots of JC-1 and percentage of Δψm-low cells. (D, E) Cellular MDA and GSH levels. (F) Labile Fe 2+ content measured by FerroOrange. (G-I) Western blots of ACSL4 and GPX4 and corresponding densitometric analysis. (J, K) Total ROS assessed by DCFH-DA flow cytometry and quantitative fluorescence. (L, M) Lipid ROS measured by C11-BODIPY 581/591 and corresponding quantification. Data are mean ± SD; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: HSFs were treated with sEVs ErF (erastin-equivalent concentration: 15 μM) in the presence or absence of Ferrostatin-1 (Fer-1, 30 nM; MedChemExpress, USA).

Techniques: Flow Cytometry, Western Blot, Fluorescence

Journal: Materials Today Bio

Article Title: Rational design of FAP-targeted sEVs delivered by microneedles for precision treatment of hypertrophic scars via ferroptosis in hypertrophic scar fibroblasts

doi: 10.1016/j.mtbio.2026.103117

Figure Lengend Snippet: Characterization and performance of sEVs ErF -loaded dissolvable microneedle patches (sEVs ErF -DMNPs). (A) Schematic of GelMA-PVA bilayer microneedle fabrication and loading of sEVs ErF into needle tips. (B) Photograph of the microneedle array. (C) SEM images showing microneedle geometry from different views. Scale bar, 500 μm. (D) Bending test of the PVA backing demonstrating flexibility. (E) Confocal 3D reconstruction of DiO-labeled sEVs ErF (green) localized in GelMA tips and red-labeled PVA backing. (F) Compression test showing mechanical strength of the patch. (G, H) In vivo fluorescence imaging of DiI-labeled sEVs ErF delivered by DMNP or intradermal injection and corresponding signal decay over 7 days. Scale bar, 1 cm. (I) Trypan-blue staining of rabbit ear skin showing microchannels created by microneedles. Insertion success rate: 88.3 ± 2.9% (n = 3 patches, 300 needles per patch). Scale bar, 1 mm. (J) Closure of puncture sites within 20 min after patch removal. Scale bar, 1 mm. (K, L) Time-dependent reduction in microneedle height and representative images during dissolution. Scale bar, 100 μm. Data are mean ± SD.

Article Snippet: HSFs were treated with sEVs ErF (erastin-equivalent concentration: 15 μM) in the presence or absence of Ferrostatin-1 (Fer-1, 30 nM; MedChemExpress, USA).

Techniques: Labeling, In Vivo, Fluorescence, Imaging, Injection, Staining, Dissolution

Journal: Materials Today Bio

Article Title: Rational design of FAP-targeted sEVs delivered by microneedles for precision treatment of hypertrophic scars via ferroptosis in hypertrophic scar fibroblasts

doi: 10.1016/j.mtbio.2026.103117

Figure Lengend Snippet: In vivo anti-scar efficacy of sEVs ErF -DMNPs in a rabbit HS model. (A) Experimental scheme of rabbit ear HS induction and treatment schedule. (B) Macroscopic images of scars in each group at day 0, 28, 35, 42, and 49, showing flatter and lighter scars in the sEVs ErF -DMNPs group. Scale bar, 2 mm. (C) Day-49 histology of scar sites: H&E, Masson's trichrome, and Sirius Red (bright-field and polarized light), demonstrating reduced dense collagen bundles and a shift toward type III collagen in the sEVs ErF -DMNPs group. Scale bars, 400 μm. (D) Scar elevation index (SEI) at day 28 and day 49. (E, F) MDA and GSH levels in scar tissues of the seven groups. (G) Serum IL-6 levels in the seven treatment groups at day 49 (n = 3). Data are mean ± SD; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 vs NS; #p < 0.05, ##p < 0.01, ###p < 0.001, ####p < 0.0001 vs HS. n = 3 rabbits per group.

Article Snippet: HSFs were treated with sEVs ErF (erastin-equivalent concentration: 15 μM) in the presence or absence of Ferrostatin-1 (Fer-1, 30 nM; MedChemExpress, USA).

Techniques: In Vivo

Journal: Materials Today Bio

Article Title: Rational design of FAP-targeted sEVs delivered by microneedles for precision treatment of hypertrophic scars via ferroptosis in hypertrophic scar fibroblasts

doi: 10.1016/j.mtbio.2026.103117

Figure Lengend Snippet: In vivo molecular validation of antifibrotic and ferroptosis-related changes. (A) Western blots of COL I, COL III, α-SMA, GPX4, and ACSL4 in scar tissues from NS, HS, Er, sEVs Er , sEVs ErF , sEVs ErF -DMNPs, and TA groups; GAPDH, loading control. (B-D) Densitometric analysis of COL I, COL III, and α-SMA relative to HS. (E, F) Densitometric analysis of GPX4 and ACSL4 relative to HS. (G) Volcano plot of DEGs between HS and sEVs ErF -DMNPs tissues (RNA-seq). (H) KEGG enrichment bubble plot showing fibrosis- and ferroptosis-related pathways. (I) GO enrichment (BP, CC, MF) highlighting ECM remodeling, glycosaminoglycan metabolism, and stress signaling. (J) Heatmap of fibrosis-related genes (e.g., CCN2, COL1A1, FKBP10, TGFBR2, MMP1, MMP3). (K) Heatmap of ferroptosis-related genes, including ACSL4, STEAP3, TFRC, SLC11A2, GPX4, GSS, FTH1, SLC40A1, NFE2L2, GCLM, GCLC, HMOX1, and SLC7A11/xCT, in scar tissues from HS and sEVs ErF -DMNPs groups. Data are mean ± SD; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 vs NS; #p < 0.05, ##p < 0.01, ###p < 0.001, ####p < 0.0001 vs HS.

Article Snippet: HSFs were treated with sEVs ErF (erastin-equivalent concentration: 15 μM) in the presence or absence of Ferrostatin-1 (Fer-1, 30 nM; MedChemExpress, USA).

Techniques: In Vivo, Biomarker Discovery, Western Blot, Control, RNA Sequencing

Journal: Bioactive Materials

Article Title: Bioengineered extracellular vesicles escape lysosomal degradation and deliver Tet-PKM2 for macrophage immunometabolic reprogramming and periodontitis treatment

doi: 10.1016/j.bioactmat.2026.01.002

Figure Lengend Snippet: Generation of bioengineered LEVs (LEVs@TA) through in situ TA modifications. ( A ) Schematic illustration showing the interaction of TA with the phospholipid bilayer of LEVs via hydrogen bonding and the uptake of LEVs@TA by macrophages. ( B ) The percentages of CY5-TA-modified cells following incubation with gradient concentrations of CY5-TA (0, 0.1, 1, 5, and 10 μM) for 24 h (flow cytometry assay). ( C ) Colocalization of CY5-TA on HEK293T cells following incubation in 10 μM CY5-TA for 24 h (fluorescence microscopy). The nuclei were stained with DAPI (blue). ( D ) The percentages of CY5-TA-modified LEVs following incubation with gradient concentrations of CY5-TA (0, 10, 20, 50, and 100 μM) for 24 h (flow cytometry assay). ( E ) Colocalization of CY5-TA and PKH67-labeled LEVs (green) following incubation with 100 μM CY5-TA for 24 h (fluorescence microscopy). ( F ) Snapshots of CGMD simulations depicting the uptake of LEVs and LEVs@TA by macrophages at 0, 5, 10, 15, and 20 ns. ( G ) Representative in vivo fluorescence images showing good stability of DIO-labeled-LEVs@CY5-TA in vivo .

Article Snippet: PKH67-labeled SEVs and LEVs resuspended in complete medium were used to treat RAW 264.7 cells for 24 h. The cells were then fixed with 4 % PFA (Coolaber), permeabilized, and stained for cytoskeletal visualization using fluorescein phalloidin (1:1000 dilution; MCE) for 30 min.

Techniques: In Situ, Modification, Incubation, Flow Cytometry, Fluorescence, Microscopy, Staining, Labeling, In Vivo

Journal: Bioactive Materials

Article Title: Bioengineered extracellular vesicles escape lysosomal degradation and deliver Tet-PKM2 for macrophage immunometabolic reprogramming and periodontitis treatment

doi: 10.1016/j.bioactmat.2026.01.002

Figure Lengend Snippet: Endo/lysosomal escape capacity of bioengineered LEVs@TA following uptake by macrophages. ( A ) Schematic illustration showing the endo/lysosomal escape process of LEVs@TA within the cytoplasm of macrophages. After uptake by macrophages, LEVs@TA were entrapped within endo/lysosomes, and then TA underwent protonation and disassembled from LEVs in an acidic environment, leading to rupture of the endo/lysosomal structure. ( B ) Snapshots of CGMD simulations showing the disassembly of TA and LEVs in the lysosomal environment. ( C ) Colocalization of LysoTracker-labeled endo/lysosomes (violet) and PKH67-labeled LEVs or LEVs@TA (green) (fluorescence microscopy). The nuclei were stained with Hoechst (blue). ( D ) Quantification of the colocalization of endo/lysosomes and LEVs or LEVs@TA using the Pearson correlation coefficient ( n = 12). ( E ) Schematic illustration showing the leakage of calcein into the cytosol when TA diffused from LEVs@TA and destabilized the endo/lysosomal membranes. ( F ) The distribution of calcein (green) in macrophages treated with PBS, LEVs, and LEVs@TA (fluorescence microscopy). (G) Representative TEM images of macrophages showing the structure of lysosomes in macrophages treated with LEVs and LEVs@TA. The data are expressed as the mean ± SEM. Statistical analysis was performed with Student's t -test ( D ). ∗∗∗ p < 0.001 indicates significant differences between the indicated columns.

Article Snippet: PKH67-labeled SEVs and LEVs resuspended in complete medium were used to treat RAW 264.7 cells for 24 h. The cells were then fixed with 4 % PFA (Coolaber), permeabilized, and stained for cytoskeletal visualization using fluorescein phalloidin (1:1000 dilution; MCE) for 30 min.

Techniques: Labeling, Fluorescence, Microscopy, Staining