Journal: Materials Today Bio

Article Title: Senolytic-loaded asymmetric wound dressing for targeted senescent cell clearance in diabetic wound healing

doi: 10.1016/j.mtbio.2025.102741

Figure Lengend Snippet: Screening and evaluation of senolytic agents in vehicle-treated and etoposide (ETO)-treated L929 cells. (A) Cell viability assays were performed on vehicle-treated (CON, n = 3) and ETO-treated (n = 3) L929 cells exposed to varying concentrations of ABT-263, Fisetin, A1155463, Dasatinib, Quercetin, and D + Q for 48 h. Cell viability was assessed using CCK-8 and expressed as a percentage relative to untreated controls. Dose-response curves and IC 50 values were calculated for each compound. The selectivity index (SI) was determined by dividing the IC 50 in proliferating cells by that in senescent cells. (B) Cytotoxicity of each senolytic agent (n = 3 for each agent), shown with representative images, at its optimal concentration after 48-h treatment of vehicle-treated L929 cells, assessed via cell count quantification. (C) Representative images of SA-β-Gal staining of ETO-treated senescent L929 cells treated with each senolytic agent (n = 3 for each agent) at its optimal concentration for 48 h, with quantification of SA-β-Gal-positive cell percentages. Values represent mean ± SEM. Statistical analysis was performed using Student's t-test or one-/two-way ANOVA. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: Cellular senescence was induced by co-cultivating L929 and HUVECs in a cell culture medium supplemented with etoposide (ETO, HY-13629, MCE, USA) at a concentration of 2.5 μM or Advanced glycation end products-modified bovine serum albumin (AGEs, BGT-CMP-100, Biogradetech) 200 μg/mL for a duration of 48 h.

Techniques: CCK-8 Assay, Concentration Assay, Cell Characterization, Staining

Journal: Materials Today Bio

Article Title: Senolytic-loaded asymmetric wound dressing for targeted senescent cell clearance in diabetic wound healing

doi: 10.1016/j.mtbio.2025.102741

Figure Lengend Snippet: Evaluation of cell viability and apoptosis following treatment with ABT-263-CGH in senescent L929 cells. (A) Representative images of Calcein-AM/PI staining used to evaluate cell viability and apoptosis in etoposide (ETO)-induced senescent L929 cells after 48-h treatment with unloaded CGH (n = 3) or ABT-263-CGH (n = 3), compared to untreated controls (ETO, n = 3). Live cells were stained green (Calcein-AM), and dead cells were stained red (Propidium Iodide, PI); the live/dead cell ratio was quantified. (B) Quantitative assessment of cell viability in senescent L929 cells treated with unloaded CGH (n = 3) or ABT-263-CGH (n = 3) for 48 h, compared to untreated controls (ETO, n = 3), measured using the CCK-8 assay over 7 days. (C) Western blot analysis of p21 protein expression in senescent L929 cells after 48-h treatment with unloaded CGH (n = 3) or ABT-263-CGH (n = 3), compared to untreated controls (ETO, n = 3), with densitometric quantification. (D) RT-qPCR analysis of mRNA levels of SASP factors ( Il-1 , Il-1b , Cxcl1 , and Mmp10 ) in senescent L929 cells after treatment (n = 3). (E) Representative images of SA-β-Gal staining in senescent L929 cells treated with unloaded CGH (n = 3) or ABT-263-CGH (n = 3) for 48 h, compared to untreated controls (ETO, n = 3), with quantification of SA-β-Gal-positive cells. Values are expressed as mean ± SEM. Statistical analysis was performed using Student's t-test or one-/two-way ANOVA. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Cellular senescence was induced by co-cultivating L929 and HUVECs in a cell culture medium supplemented with etoposide (ETO, HY-13629, MCE, USA) at a concentration of 2.5 μM or Advanced glycation end products-modified bovine serum albumin (AGEs, BGT-CMP-100, Biogradetech) 200 μg/mL for a duration of 48 h.

Techniques: Staining, CCK-8 Assay, Western Blot, Expressing, Quantitative RT-PCR

Journal: Aging Cell

Article Title: A Novel Human Cellular System for Studying Normal Aging and for Anti‐Aging Discovery

doi: 10.1111/acel.70352

Figure Lengend Snippet: The hTSC‐STB system enables simple, fast, and quantitative evaluation of known anti‐aging molecules. (A) Western blot analysis of hCG levels in hTSCs and STBs on D2, D4, and D6 treated with different concentrations of Rapamycin (RAPA), INK128, and Fisetin. These known anti‐aging molecules are able to prevent, to some extent, hTSCs from developing into fully mature STBs; (B) Western blotting analysis showed significantly decreased γH2AX in Rapamycin, INK128, and Fisetin treated STB cells, whereas the treated STBs express higher HP1γ, Lamin B1 and Ki67 than non‐treated STBs; (C) Schematic graph of the CGA‐T2A‐H2B‐EGFP reporter allele where the T2A‐H2B‐EGFP cassette is inserted into the CGA locus for fusion peptide production and subsequent cleavage into CGA and H2B‐EGFP; (D) The CGA‐EGFP hTSC reporter cells (left) efficiently differentiate into STBs (right) with particularly high EGFP signals in the nucleus (live cells); (E) Sum intensity of EGFP gradually increases in CGA‐EGFP hTSC reporter cells to STBs. EGFP signals appear to have reached a plateau on D4 as the signals are similar in D4 and D6 cells; (F) Representative immunostaining images of CGA‐EGFP hTSCs and CGA‐EGFP STB cells of D2, D4, D6 showing EGFP signals co‐stained with DAPI; (G, H) Using the CGA‐EGFP reporter hTSCs to quantitively evaluate known anti‐aging molecules. The treatments start on day 0 of hTSCs to STBs development; Normalized sum GFP intensity refers to DAPI signals. The graphs in (G) are mean ± SEM; (I) An example of a screened 96‐well library plate. Rapamycin and INK128 were used as the positive controls. Magenta rectangle marked the potential candidates (Anti‐aging and Pro‐aging); (J) Quantification of EGFP signals of the 96‐well plate in Figure . The sum intensity was normalized referring to DAPI signals; (K) β‐gal staining of STBs (D6) in the presence of candidate molecules (Anisomycin and Nitidine) in its development from hTSCs to STBs; (L) Quantification of β‐gal staining cell in (K); (M) Representative β‐gal staining images of HFF‐1 cells treated with DMSO, VP‐16, VP‐16 + INK128 25 nM, VP‐16 + Anisomycin 25 nM, and VP‐16 + Nitidine 25 nM; (N) Quantification of β‐gal staining cell in (M). One‐way ANOVA with Dunnett's test was used in statistical analysis. Ns, p > 0.05; * p ≤ 0.05, ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001. All experiments have been independently repeated three times. Scale bars, 10 μm.

Article Snippet: Etoposide (VP‐16) , MedChemExpress , HY‐13629.

Techniques: Western Blot, Immunostaining, Staining