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: Metabolic reprogramming and enhanced mitochondrial function in LPS-activated macrophages in response to LEV Tet−PKM2 @TA treatment. The macrophages were pretreated with 100 ng/mL LPS for 24 h and then treated with PBS (Control), 100 μg/mL LEVs PKM2 , LEVs Tet−PKM2 , or LEVs Tet−PKM2 @TA for another 24 h. ( A ) Heatmap representing differentially detected metabolites involved in glycolysis and the TCA cycle in the Control, LEVs PKM2 , LEVs Tet−PKM2 , or LEVs Tet−PKM2 @TA groups ( n = 4). ( B ) Concentrations of key glycolysis and TCA metabolites in Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups ( n = 4). ( C ) Schematic illustration revealing changes in key glycolysis and TCA metabolites in the LEVs Tet−PKM2 @TA group versus the Control group. The up (down) arrows indicate increased (decreased) levels of metabolites in macrophages. ( D ) Kinetic profile of the ECAR in LPS-activated macrophages in response to sequential injections of glucose, oligomycin, and 2-DG in various groups (Seahorse XF test) ( n = 4). ( E ) Quantification of glycolysis, glycolytic capacity and glycolytic reserve in the Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups ( n = 4). ( F ) Kinetic profile of the OCR in LPS-activated macrophages in response to sequential injections of oligomycin, FCCP, and Rot/AA in various groups (Seahorse XF test) ( n = 4). ( G ) Quantification of basal respiration, ATP production, and maximal respiration in the Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups ( n = 4). ( H ) JC-1 aggregation (red fluorescence) in healthy mitochondria and cytosolic JC-1 monomers in compromised mitochondria (green fluorescence) (immunofluorescence assays). ( I ) Quantitative analysis of MMP levels determined by the relative ratio of red/green fluorescence intensity in the Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups ( n = 4). ( J ) Intracellular ATP levels of LPS-activated macrophages in the Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups ( n = 3). ( K-M ) The macrophages were pretreated with 100 ng/mL LPS for 24 h and then treated with PBS (Control), 10 μM UK-5099, 100 μg/mL LEVs Tet−PKM2 @TA, or 10 μM UK-5099 plus 100 μg/mL LEVs Tet−PKM2 @TA for another 24 h. ( K ) Schematic illustration revealing mechanism of LEVs Tet−PKM2 @TA promotes macrophage metabolic reprogramming depending on pyruvate influx into the TCA cycle. ( L ) Kinetic profile of the OCR in LPS-activated macrophages in response to sequential injections of oligomycin, FCCP, and Rot/AA in various groups (Seahorse XF test) ( n = 3). ( M ) Quantification of basal respiration, ATP production, and maximal respiration in the Control, UK-5099, LEVs Tet−PKM2 @TA, and UK-5099 + LEVs Tet−PKM2 @TA groups ( n = 3). The data are expressed as the mean ± SEM. Statistical analysis was performed with one-way ANOVA ( B , E , G, I, J , and M ). ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 indicate significant differences between the indicated columns.

Article Snippet: The MMP of the macrophages was assessed using a JC-1 MMP Assay Kit (MCE).

Techniques: Control, Fluorescence, Immunofluorescence

Journal: International Journal of Oncology

Article Title: Diverse roles of SERPINE1 in regulating cellular proliferation and invasion

doi: 10.3892/ijo.2026.5871

Figure Lengend Snippet: SERPINE1 regulates MMP activity. (A) MMP and TIMP levels in the supernatants of shSE1 and shc cells after 24 h of incubation and 10-fold concentration. Statistical significance was determined using a two-sided Student's t test. The data are presented as the means ± SDs. n=4. (B) Quantification of active MMP-1 and MMP-13 levels in cell lysates using fluorescence ELISA. Statistical significance was determined using two-way ANOVA followed by Šídák's multiple comparisons test. The data are presented as the means ± SDs of three independent experiments. (C) Western blotting showing the levels of the indicated proteins in H4-shSE1 cells at 72 h after transfection with the si-HSP90AA1 or siNC. The numbers beneath the bands represent the ratios of the expression levels of the indicated proteins and represent mean ± SDs of three independent experiments. (D) Representative images and quantification of Transwell invasion assays through Matrigel and collagen type I in H4-shSE1 cells transfected with the si-HSP90AA1 or siNC (scale bar, 100 μ m). Statistical significance was determined using a two-sided Student's t-test. The data are presented as the means ± SDs of three independent experiments. (E) Diagram showing the mechanisms underlying the effects of SERPINE1 on cancer proliferation and invasion. SERPINE1 knockdown influences cell proliferation and invasion through distinct signaling pathways. With respect to proliferation, SERPINE1 knockdown reduces TGF-β levels, and this reduction alters the activity of SMAD3, p53, and MCM3 to promote cell cycle progression. SERPINE1 knockdown interferes with the uPAR-mediated balance of the ERK/p38 ratio; it may also affect this ratio by modulating HSP90α expression and p38 activity, which suppress cell proliferation. In terms of invasion, SERPINE1 downregulation increases MMP-1 levels via the HSP90α-p38 pathway, thereby promoting cellular invasion. *** P<0.001, ** P<0.01, ns, not significant. SERPINE1, serine protease inhibitor clade e member 1; MMP, matrix metalloproteinases; TIMP, tissue inhibitors of metal proteases; shRNA, short hairpin RNA; shSE1, shRNA targeting SERPINE1; si, short interfering RNA; siHSP90AA1, short interfering heat shock protein 90 alpha family class a member 1; NC, negative control; uPAR, urokinase-type plasminogen activator receptor; p-, phosphorylated; MCM3, minichromosome maintenance complex component 3; HSP90α, heat shock protein 90-alpha.

Article Snippet: Following the blocking of nonspecific binding sites using 5% skimmed milk (cat. no. P0216; Beyotime Biotechnology) or bovine serum albumin (cat. no. NGP0028A; Beyotime Biotechnology) for 1 h at room temperature, the membranes were incubated with primary antibodies (incubation overnight at 4°C) against SERPINE1 (cat. no. 13801-1-AP; Proteintech Group, Inc), GAPDH (cat. no. sc-25778; Santa Cruz Biotechnology, Inc.), MCM3 (cat. no. PA5-79646; Thermo Fisher Scientific, Inc.), phosphorylated (p-)MCM3 (Ser112; cat. no. TA2362; Abmart Pharmaceutical Technology Co., Ltd.), ERK (cat. no. sc-514302; Santa Cruz Biotechnology, Inc.), uPAR (cat. no. ab10379; Abcam), Histone H3 (cat. no. 4499), p-p53 (Ser15; cat. no. 9284), p53 (cat. no. 2524), p-SMAD3 (Ser423/425; cat. no. 9520), SMAD3 (cat. no. 9523), p-Rb (Ser807/811; cat. no. 8516), Rb (cat. no. 9309), CyclinD1 (cat. no. 55506), CyclinE1 (cat. no. 20808), p21 (cat. no. 2947), p-p38 (cat. no. 4511), p38 (cat. no. 8690), p-ERK (cat. no. 4370), p-JNK (cat. no. 9255), JNK (cat. no. 9252), p-AKT (Ser473; cat. no. 4060), AKT (cat. no. 9272), HSP90α (cat. no. 4877) and MMP-1 (cat. no. 54376) were from Cell Signaling Technology, Inc.) and subsequently incubated with the corresponding secondary antibodies (dilution 1:4,000) for 2 h at room temperature.

Techniques: Activity Assay, Incubation, Concentration Assay, Fluorescence, Enzyme-linked Immunosorbent Assay, Western Blot, Transfection, Expressing, Knockdown, Protein-Protein interactions, Protease Inhibitor, shRNA, Small Interfering RNA, Negative Control

Journal: Materials Today Bio

Article Title: Fmoc-DDT@Fos hydrogel mitigates temporomandibular joint osteoarthritis through regulating macrophage reprogramming and ferroptosis

doi: 10.1016/j.mtbio.2026.102906

Figure Lengend Snippet: Impact of Fmoc-DDT@Fos on macrophage ferroptosis. (A) Fe 2+ in macrophages was marked using FerroOrange; (B) Total iron content in macrophages was measured by iron assay kit; (C) GSH/GSSG ratio in macrophages; (D-E) Visualization of ROS production in macrophages using DCFH-DA; (F) Measurement of MDA content in macrophages; (G) C11-BODIPY 581/591 staining for macrophages; qRT-PCR analysis of ferroptosis-related gene Gpx4 (H) and Slc7a11 (I); (J) TEM observation of mitochondrial morphology; (K) JC-1 assay for MMP in macrophages. ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: MMP was evaluated using the JC-1 MMP Assay Kit (40706ES60, Yeasen Biotechnology, China).

Techniques: Iron Assay, Staining, Quantitative RT-PCR

Journal: Non-coding RNA Research

Article Title: Hsa_circ_0101645 contributes to excessive autophagy and apoptosis in intervertebral disc degeneration by acting as a miR-1304-5p sponge modulating BNIP3 expression

doi: 10.1016/j.ncrna.2025.11.007

Figure Lengend Snippet: Hsa_circ_0101645 accelerating the IVDD process in vivo . A: Diagram of the animal procedure for this study. B: Grouping information for this section. C: Representative X-rays of each group of rats. Statistical graph demonstrating the disc height index (DHI) changes for L4/5 in each group of rats (N = 6) (One-way ANOVA test with Tukey's multiple comparisons test). D: HE staining exhibiting pathological changes of CEP, NP, and AP in IVD in each group of rats (N = 6). Scale bar: 500 μm. E: EdU staining was used to detect cell proliferation in IVD tissues of rats (N = 6) (One-way ANOVA test with Tukey's multiple comparisons test). Scale bar: 50 μm. F: TUNEL (white light) staining exhibiting TUNEL-positive cells in IVD tissues of rats (N = 6) (One-way ANOVA test with Tukey's multiple comparisons test). Scale bar: 50 μm. G: The effect of hsa_circ_0101645 on the protein levels of Collagen Ⅱ, Aggrecan, MMP-3 and MMP-13 in IVD was observed by IHC staining (N = 6) (One-way ANOVA test with Tukey's multiple comparisons test). Scale bar: 100 μm. H-I: The expression of hsa_circ_0101645 (H) and miR-1304-5p (I) in each group of IVD tissues (N = 6) (One-way ANOVA test with Tukey's multiple comparisons test). J-K: Changes in expression of apoptosis ( J; Caspase 3, Bcl-2 and Bax) and autophagy markers ( K; LC3B, Beclin and P62) in IVD tissues (N = 3) (One-way ANOVA test with Tukey's multiple comparisons test or Kruskal-Wallis test with Dunn's multiple comparisons test). ∗ indicates P < 0.05.

Article Snippet: Sections were then incubated with primary antibodies against Collagen II (28459-1-AP; 1:200; Proteintech, USA), Aggrecan (13880-1-AP; 1:100; Proteintech), MMP-3 (17873-1-AP; 1:200; Proteintech), and MMP-13 (18165-1-AP; 1:100; Proteintech), followed by the corresponding secondary antibodies.

Techniques: In Vivo, Staining, TUNEL Assay, Immunohistochemistry, Expressing