Journal: Bioactive Materials

Article Title: Mechanically sensitized hydrogel microspheres trigger membrane receptor switch for cartilage repair

doi: 10.1016/j.bioactmat.2026.03.017

Figure Lengend Snippet: OBNC microspheres activate integrin receptors and mechanosensitive calcium channels. A) Mechanistic diagram of integrin activation verified using fluorophores. B-C) Fluorescence microscopy images of MSCs loaded on HAMA or OBNC hydrogel. D) Fluorescence intensity of single cell in each group was quantified. E) Fluorescence microscopy of MSCs loaded on OBNC hydrogel after different treatments. F) Fluorescence intensity in the whole field of view for each group. G) Fluorescence intensity in the single cell for each group. H) Schematic representation of patch clamp experiments. I) Electrical signals generated by MSCs in response to mechanical stimulation. J) Statistical analysis of poking currents (n = 6). K) The concentration of calcium ions in stem cells of different groups as detected by flow cytometry (siRNA1: targeting the TRPM4 gene, siRNA2: targeting the PIEZO1 gene). L) Quantitative analysis of flow cytometric results (∗ symbol represents comparison with HAMA group, # symbol represents comparison with OBNC group). (ns: non-significant, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ## P < 0.01, ### P < 0.001).

Article Snippet: TRPC1 inhibitor (0.3 nM, Pico145, CAS No. 1628287-16-0), TRPM7 inhibitor (1.0 μM, VPC4, CAS No. 945604-76-2), TRPV2 inhibitor (5.0 μM, compound IV2-1, CAS No. 2242724-49-6), TRPM4 inhibitor (1.5 μM, CBA, CAS No. 351424-20-9), PIEZO1 inhibitor (2.5 μM, GsMTx4, CAS No. 1209500-46-8), integrin αvβ5 inhibitor (8.0 nM, Compound 12, CAS No.: 2615912-33-7), integrin αvβ1 inhibitor (0.3 nM, Compound C8, CAS No. 1689540-62-2), integrin α5β1 inhibitor (10 μM, ATN-161, 904763-27-5), and CDK5 inhibitor (5 nM, CDK5-IN-1, 2,639,540-19-3) were purchased from MCE Biotechnology Co., LTD. After the MSCs were treated, the cRGD solution was added at a concentration of 1:200 and incubated in the dark for 15 min, and the results were observed by fluorescence microscopy.

Techniques: Activation Assay, Fluorescence, Microscopy, Single Cell, Patch Clamp, Generated, Concentration Assay, Flow Cytometry, Comparison

Journal: Advanced Science

Article Title: Nanovesicles With Mechanically Induced Adjuvanticity for Robust Melanoma Vaccination Toward Tumor‐Associated Macrophages

doi: 10.1002/advs.76773

Figure Lengend Snippet: Activation of YAP/TAZ and Piezo1 by P‐P m . (A) Immunofluorescence confocal images of YAP/TAZ in RAW264.7 cells exposed to PEAMP, P‐P 20 , P‐P 30 , and P‐P 50 for 4 h, the intensity of gray values along the white line from left to right in the corresponding image was delineated, blue for the nucleus and red for YAP/TAZ, scale bar: 10 µm. (B) Mean fluorescence intensity of YAP/TAZ in RAW264.7 cells exposed to PEAMP, P‐P 20 , P‐P 30 , and P‐P 50 for 4 h ( n = 3). (C) Quantification of nuclear to cytoplasmic ratios of YAP/TAZ in RAW264.7 cells exposed to PEAMP, P‐P 20 , P‐P 30 , and P‐P 50 for 4 h ( n = 3). (D) WB analysis of YAP and pYAP, GAPDH proteins expression in RAW264.7 cells treated with PEAMP, P‐P 20 , P‐P 30 , and P‐P 50 for 4 h. (E) Time trace of Flou‐4 fluorescence intensity and relative peak amplitude detected by CLSM in RAW264.7 cells treated with PBS (as control), PEAMP, P‐P 20 , P‐P 30 , P‐P 50 , and Yoda1 (5 µM) with or without GsMTx4 (3 µ m , 0.5 h) pre‐treatment ( n = 6). Real‐time imaging was conducted for capturing images every 15 s for a 10 min period. ΔF representing the change of Flou‐4 fluorescence intensity. The inserted photos represent the fluorescence intensity of macrophages in 0 or 600 s. (F) Relative mRNA expression of Piezo1 in RAW264.7 cells exposed to PEAMP, P‐P 20 , P‐P 30 , and P‐P 50 for 24 h ( n = 6). (G) Flou‐4 fluorescence intensity of RAW264.7 cells treated with PBS (as control), PEAMP, P‐P 20 , P‐P 30 , P‐P 50 , and Yoda 1 (5 µ m ) for 2 h with or without GsMTx4 (3 µ m , 0.5 h) pre‐treatment ( n = 3). (H) WB analysis of NF‐κB p65 and GAPDH proteins expression in RAW264.7 cells treated with PEAMP, P‐P 20 , P‐P 30 , and P‐P 50 for 24 h with or without GsMTx4 (3 µ m ) pre‐treatment. Data are presented as mean ± s.d. Statistical significance was determined by one‐way ANOVA with Tukey's multiple‐comparisons test. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; n.s., not significant.

Article Snippet: Verteporfin and GsMTx4 was purchased from MedChemExpress LLC.

Techniques: Activation Assay, Immunofluorescence, Fluorescence, Expressing, Control, Imaging