Journal: Bioactive Materials

Article Title: Mesenchymal stromal cells-loaded 3D radially aligned composite scaffold with potentiated paracrine signaling for sequential bone regeneration

doi: 10.1016/j.bioactmat.2026.02.059

Figure Lengend Snippet: Temporal analysis of the BMSC paracrine profile on different scaffolds. (A) Confocal microscopy images from Live/Dead fluorescence staining of BMSCs encapsulated within the PCL/HAp-GelMA/BMSCs scaffold after 1, 3, 5, and 14 d of 3D culture (live cells, green; dead cells, red). (B) The concentrations of key paracrine factors (TGF-β, PGE2, VEGF, HGF, and BMP-2) from BMSCs cultured in different scaffolds, quantified from culture supernatants at day 3 and day 7. (C) Corresponding relative mRNA expression levels of TGFB1, PTGS2, VEGFA, HGF, and BMP-2 in BMSCs at day 3 and day 7, as determined by qPCR analysis. Data are presented as mean ± SD (n = 3) *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns: not significant.

Article Snippet: ELISA kits for PGE2 (Cat. No. E-EL-0034), TGF-β (Cat. No. E-EL-0162), VEGF (Cat. No. E-EL-R2603), and HGF (Cat. No. E-EL-R0496) were purchased from Elabscience (Wuhan, China).

Techniques: Confocal Microscopy, Fluorescence, Staining, Cell Culture, Expressing

Journal: Bioactive Materials

Article Title: Chronic inflammation-responsive hydrogel restores myeloid-T cell crosstalk to reinvigorate antitumor immunity against metastatic colorectal cancer

doi: 10.1016/j.bioactmat.2026.03.012

Figure Lengend Snippet: PGE2 blockade modulates immune cell phenotypes in antitumor resp onses. (A) Inflammatory gene expression across cancer types (GEPIA2 database). (B) Gene expression of Il1b , Cxcl8 , and Lif in colon adenocarcinoma (COAD) tumor tissue and normal tissue (GEPIA2 database). (C and D) Correlation between Ptgs2 and inflammatory genes in various cancers (C) and COAD (D) (TIMER 2.0). (E) Schematic of immune cells co-incubated with CXB treated tumor conditional medium (TCM) (Source material from BioRender). (F and G) Cell viability (F) and Cell cycle arrest (G) detection of CT26 tumor cells treated with gradient concentrations of CXB; n = 3. (H) PGE2 concentration in CT26 cell supernatants; n = 3. (I) The proportion of CD103 + DC within BMDCs after CXB treatments in vitro ; n = 3. (J and K) Maturation (J) and Antigen processing capability (K) on BMDCs; n = 3. (L – N) Flow charts of CD86 or CD206 expression on Raw 264.7 cells (L). Quantification of CD86 (M) and CD206 (N) expression on Raw 264.7 cells; n = 3. (O and P) Flow charts (O) and Quantification (P) of CD69 and CD137 expression on splenic T cells exposed to CXB-pretreated TCM; n = 3. (Q) IFN-γ secretion by T cells co-cultured with CXB-pretreated TCM; n = 3. Data are presented as mean ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Significance was calculated using One-way ANOVA.

Article Snippet: ELISA kits for mouse IL-1β, IL-6, IL-12p70, TNFα (BioLegend, USA), PGE2 (Cayman Chemical Company, USA), IL-2 (Dakewe Biotech, Shenzhen, China), IFN-γ, and TGF-β (Invitrogen, Thermo Fisher Scientific, USA) were used for cytokine quantification.

Techniques: Gene Expression, Incubation, Concentration Assay, In Vitro, Expressing, Cell Culture

Journal: Bioactive Materials

Article Title: Chronic inflammation-responsive hydrogel restores myeloid-T cell crosstalk to reinvigorate antitumor immunity against metastatic colorectal cancer

doi: 10.1016/j.bioactmat.2026.03.012

Figure Lengend Snippet: Sustained PGE2 blockade prompts immune activ ation. (A) Structure of hydrogel matrix and scheme of Gel-CXB preparation (Source material from BioRender). (B) Microstructure of the hydrogel. (C) Rheological evaluation of Gel-CXB. (D) CXB release from Gel-CXB in PBS or PBS containing 0.5 mM H 2 O 2 ; n = 3. (E and F) Flow chart (E) and Quantification (F) of CD103 + DC within BMDCs; n = 3. (G and H) Flow chart (G) and Heatmap (H) of costimulatory molecular expression on CD103 - DC, CD103 + DC, or total DC with different treatments; n = 3. (I and J) CXCL9 (I) and Costimulatory molecular expression (J) on cDC1; n = 3. (K – M) CD86 and CD206 expression (K), MHC-II expression (L), and Antigen processing capability (M) of BMDMs incubated with different TCM; n = 3. (N and O) CD69 (N) and CD137 (O) expression on CD8 + T cells co-incubated with different TCM; n = 3. (P) Scheme of Gel-CXB-regulated CT26 TME at different time points in vivo . (Q) Changes of several immune cells within TME at Day 1, 5, and 9; n = 3. (R) Tumor volume of mice treated with CXB alone or Gel-CXB in vivo ; n = 5. (S) CD137 expression on CD8 + T cells in vivo ; n = 3. Data are presented as mean ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Significance was calculated using One-way ANOVA.

Article Snippet: ELISA kits for mouse IL-1β, IL-6, IL-12p70, TNFα (BioLegend, USA), PGE2 (Cayman Chemical Company, USA), IL-2 (Dakewe Biotech, Shenzhen, China), IFN-γ, and TGF-β (Invitrogen, Thermo Fisher Scientific, USA) were used for cytokine quantification.

Techniques: Expressing, Incubation, In Vivo

Journal: Bioactive Materials

Article Title: Sulfated polysaccharide prevents senescent adipocyte-driven osteonecrosis by stem cell fate reprogramming

doi: 10.1016/j.bioactmat.2025.11.039

Figure Lengend Snippet: SCS reprograms the lineage commitment of MSCs after GC treatment and inhibits the generation of primary senescent adipocytes. ( A ) Schematic illustration of the in vitro investigation of SCS targeting the prostaglandin/PPARγ/INK positive feedback loop in MPS-induced primary senescent adipocytes. ( B ) Representative flow cytometry plot showing p16 + senescent cells in adipocytes derived from bone marrow after 14 days of in vivo MPS induction and subsequently treated with SCS in vitro . ( C ) qPCR analysis of 12 senescence-associated markers in primary senescent adipocytes after in vitro SCS treatment. n = 3 biological replicates. ( D ) ELISA analysis of IL-1β levels in adipocyte supernatant following in vitro SCS treatment. n = 6 biological replicates. ( E ) ELISA analysis of secreted prostaglandins PGD2 and PGE2 in adipocytes under different treatment conditions. D-PBS: bone marrow adipocytes isolated from mice treated in vivo with the solvent control DMSO, followed by in vitro treatment with PBS; M-PBS: bone marrow adipocytes isolated from mice treated in vivo with MPS, followed by in vitro treatment with PBS. M-SCS: bone marrow adipocytes isolated from mice treated in vivo with MPS, followed by in vitro treatment with SCS. ( F ) Western blot analysis of intracellular COX-2 protein levels in adipocytes across the three treatment conditions. ( G ) Schematic illustration of competitive osteogenic–adipogenic differentiation of CD45 − Ter119 − CD31 − LepR + MSCs after 7 days of in vivo SCS and MPS co-treatment. ( H ) qPCR analysis of pan-adipocyte markers ( Fabp4 , Adipoq , Plin1 , Cd36 , and Lep ) in CD45 − Ter119 − CD31 − LepR + MSCs after 14 days of in vitro competitive lineage differentiation. n = 3 biological replicates. ( I and J ) Representative immunofluorescence images (I) and quantification (J) of perilipin + adipocytes and osteopontin + mature osteoblasts derived from lineage-committed MSCs. n = 6 biological replicates. (Scale bars, 30 μm, 15 μm and 15 μm). ( K ) Western blot analysis of adipogenesis-related markers C/EBPα, PPARγ, and C/EBPβ in the lineage-mixed cells after in vitro competitive differentiation of CD45 − Ter119 − CD31 − LepR + MSCs. ( L ) qPCR analysis of lipogenesis-related markers Fasn , Scd1 , Srebf1 , Acaca , and Acacb . n = 3 biological replicates. ( M and N ) Representative H&E staining images (M) of the femurs at day 14 following SCS and MPS co-treatment. Yellow arrows indicate bone marrow adipocytes. Magnified images show hypertrophic adipocyte morphology, with quantification of adipocyte diameter (N). n = 19 biological replicates. (Scale bars, 200 μm, 50 μm and 20 μm). Data are presented as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; ns, not significant. Statistical significance was determined using an unpaired two-tailed Student's t -test ( C, D, H, J, L and N ), or one-way ANOVA with Tukey's post hoc test ( E ).

Article Snippet: To further investigate the anti-senescent effects of SCS on primary senescent adipocytes, ELISA was performed on adipocyte supernatant to quantify senescence-associated factors including IL-1β (Neobioscience, EMC001b.96), IL-18 (Neobioscience, EMC011.96), TNF-α (Neobioscience, EMC102a.96), and prostaglandins PGJ2 (NOVUS, NBP2-61285), PGD2 (Cayman Chemical, 500151), and PGE2 (R&D Systems, KGE004B).

Techniques: In Vitro, Flow Cytometry, Derivative Assay, In Vivo, Enzyme-linked Immunosorbent Assay, Isolation, Solvent, Control, Western Blot, Immunofluorescence, Staining, Two Tailed Test

Journal: Redox Biology

Article Title: Stromal cell-derived itaconate promotes endometriosis via macrophage NRF2 and lysosomal pH modulation

doi: 10.1016/j.redox.2026.104101

Figure Lengend Snippet: Activation of the itaconate metabolic pathway in a mouse model of endometriosis (A) Schematic diagram of the mouse endometriosis model. (B) Representative intra-abdominal images. Top: normal uterus from control mice. Middle and bottom: recipient mice with endometriotic lesions (red circles); lower panels show magnified views. (C) Bubble plot of quantitative enrichment analysis showing altered metabolic pathways among ectopic lesions, eutopic endometrium, and normal endometrium in mice. (D) Heat map of selected metabolites across normal, eutopic, and ectopic tissues from the mouse endometriosis model. Row-wise z-scored intensities (no log transform); colors reflect relative abundance per metabolite, centered at 0 (≈[−1, 1]). (E) Quantification of itaconate in ectopic lesions (n = 10), eutopic endometrium (n = 5), and normal endometrium (n = 5). (F, G) Western blot analysis (F) and quantification (G) of ACOD1 protein in lesion tissues from endometriosis (EM) and control mice (n = 3/group). (H, I) Western blot analysis (H) and quantification (I) of ACOD1 in stromal cells from normal (nor-ESC), eutopic (eu-ESC), and ectopic (ec-ESC) endometrium (n = 3/group). (J) qRT-PCR analysis of Acod1 mRNA in nor-ESC, eu-ESC, and ec-ESC (n = 3/group); LPS-stimulated macrophages from control mice serve as positive control. (Data are presented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ns: not significant.) E2, estradiol benzoate; EM, endometriosis; NC, non-EM control; LPS, lipopolysaccharide.

Article Snippet: Estrous cycles were synchronized with subcutaneous estradiol benzoate (E2) (3 μg/mouse) (MCE; Nanjing, China).

Techniques: Activation Assay, Control, Western Blot, Quantitative RT-PCR, Positive Control

Journal: Redox Biology

Article Title: Stromal cell-derived itaconate promotes endometriosis via macrophage NRF2 and lysosomal pH modulation

doi: 10.1016/j.redox.2026.104101

Figure Lengend Snippet: Itaconate Pathway Intervention Alters Lesion Burden and Fibrosis in Endometriosis Mice (A)Schematic of IRG1-IN-1 (0.5 mg/kg) or PBS administration in the mouse endometriosis (EM) model. (B) Representative images of endometriotic lesions in PBS- and IRG1-IN-1-treated mice. (C) Gross morphology of excised lesions from each group. (D) Quantification of lesion weight (n = 6/group). (E) Schematic of the experimental design for si-Irg1-LNP administration in EM mice. (F, G) Representative images (F) and gross morphology (G) of lesions in si-ctrl-LNP and si-Irg1-LNP groups. (H) Quantification of lesion weight (n = 6/group). (I) Western blot analysis of ACOD1 expression in lesions from si-ctrl-LNP and si-Irg1-LNP groups.(J) Schematic of the experimental design for 4-OI intervention in EM mice. (K) Representative images of endometriotic lesions after treatment with PBS or 4-OI (red arrows). (L) Gross morphology of lesions from each group. (M) Quantification of lesion weight (n = 6/group). (N) Masson's trichrome staining of lesions with quantification of fibrotic area (n = 6/group). (O) Measurement of itaconate content in peritoneal macrophages from EM and non-EM mice (n = 8/group). (P) Western blot analysis of ACOD1 in peritoneal macrophages from EM and control mice E2, estradiol benzoate; EM, endometriosis; LNP, lipid nanoparticle; LPS, lipopolysaccharide; 4-OI, 4-octyl itaconate; PBS, phosphate-buffered saline; si- Irg1 , small interfering RNA targeting Irg1.

Article Snippet: Estrous cycles were synchronized with subcutaneous estradiol benzoate (E2) (3 μg/mouse) (MCE; Nanjing, China).

Techniques: Western Blot, Expressing, Staining, Control, Saline, Small Interfering RNA

Journal: Redox Biology

Article Title: Stromal cell-derived itaconate promotes endometriosis via macrophage NRF2 and lysosomal pH modulation

doi: 10.1016/j.redox.2026.104101

Figure Lengend Snippet: Itaconate-mediated modulation of macrophage activity alters endometriotic lesion p rogression (A) mRNA expression of IL1B , IL6 , TNFA , and iNOS in peritoneal macrophages from endometriosis (EM) and non-EM patients, as well as non-EM macrophages co-cultured with either nor-ESC or ectopic ec-ESC for 12 h (n = 3/group).(B) Migration of si- Irg1 or NC-treated ectopic ESCs was assessed after co-culture with PBMCs for 48 h. Quantification of migrated cells in five random fields per group are shown (n = 3/group). (C-D) PKH67-labeled human ectopic ESCs pretreated with si- Irg1 or NC were co-cultured with PBMCs for 8 h. Phagocytosis of ESCs by macrophages was analyzed by flow cytometry with representative gating (C), PKH67 signal and quantification of PKH67-positive macrophages (D) (n = 6/group).(E,G) Flow cytometry analysis and quantification of iNOS + peritoneal macrophages from mouse model in E (E), A (G). (n = 6/group). (F,H) mRNA levels of Il1b , Il6 , Nos2 , and Tnf in peritoneal macrophages from mouse model in E (E), A (G) (n = 6/group).(I)Quantification of itaconate in endometriotic lesion tissue by LC–MS from mice treated with IRG1-IN-1 or vehicle. (n = 3/group).(J) Flow cytometry analysis and quantification of iNOS + peritoneal macrophages from mouse model in J (n = 6/group).(K,L) Migration of mESCs induced by peritoneal macrophages from PBS- or 4-OI-treated mice was assessed by transwell assay; representative images and quantification of migrated cells are shown (n = 5/group). (M) Flow cytometry analysis and quantification of phagocytosis of PKH67-labeled mESCs by peritoneal macrophages (n = 6/group).(N) Schematic of the experimental design for clodronate liposome-mediated macrophage depletion and 4-OI intervention in EM mice. (O, P) Representative images (O) and gross morphology (P) of endometriotic lesions in control, clodronate, PBS, and 4-OI groups. (Q) Quantification of lesion weight in mice treated with clodronate liposomes or control liposomes, with or without 4-OI (n = 6/group). (Data are presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.) E2, estradiol benzoate; EM, endometriosis; LNP, lipid nanoparticle; LPS, lipopolysaccharide; 4-OI, 4-octyl itaconate; Clod, clodronate liposome; PBS, phosphate-buffered saline; si- Irg1 , small interfering RNA targeting Irg1.

Article Snippet: Estrous cycles were synchronized with subcutaneous estradiol benzoate (E2) (3 μg/mouse) (MCE; Nanjing, China).

Techniques: Activity Assay, Expressing, Cell Culture, Migration, Co-Culture Assay, Labeling, Flow Cytometry, Liquid Chromatography with Mass Spectroscopy, Transwell Assay, Control, Liposomes, Saline, Small Interfering RNA

Journal: Redox Biology

Article Title: Stromal cell-derived itaconate promotes endometriosis via macrophage NRF2 and lysosomal pH modulation

doi: 10.1016/j.redox.2026.104101

Figure Lengend Snippet: Inhibition of NRF2 Signaling Reverses the Anti-inflammatory and Anti-endometriotic Effects of Itaconate in Macrophages and a Mouse Model of End ometriosis (A,B) Western blot analysis (A) and quantification (B) of NRF2 protein levels in bone marrow-derived macrophages (BMDMs) pretreated with LPS (100 ng/mL), 4-octyl itaconate (4-OI, 250 μM), or both for 12 h (n = 3/group). (C,D) Western blot analysis (C) and quantification (D) of NRF2 in BMDMs treated with LPS, 4-OI, and the NRF2 inhibitor ML385 (2.5 μM) for 12 h (n = 3/group). (E) Flow cytometry analysis and quantification of iNOS + BMDMs after indicated treatments (n = 3/group). (F) mRNA expression of pro-inflammatory genes ( Il1b , Il6 , Nos2 , Tnf ) in BMDMs under different conditions (n = 3/group). (G) Flow cytometry analysis and quantification of iNOS + BMDMs following NRF2 knockdown (si Nrf2 ) with or without 4-OI, compared to negative control (NC) (n = 3/group).(H) Schematic of the experimental design for ML385 administration in a mouse model of endometriosis. (I) Representative images of endometriotic lesions in PBS- and ML385-treated mice (lesions marked by red circles). (J) Gross morphology of lesions in both groups(n = 6/group). (K) Quantification of lesion weight (n = 6/group). (L) Flow cytometry analysis and quantification of iNOS + peritoneal macrophages from EM mice treated with PBS or ML385 (n = 6/group). (M) mRNA expression of Il1b , Il6 , Nos2 , and Tnf in peritoneal macrophages from each group (n = 6/group). (Data are presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.) E2, estradiol benzoate; EM, endometriosis; LPS, lipopolysaccharide; 4-OI, 4-octyl itaconate; ML385, NRF2 inhibitor.

Article Snippet: Estrous cycles were synchronized with subcutaneous estradiol benzoate (E2) (3 μg/mouse) (MCE; Nanjing, China).

Techniques: Inhibition, Western Blot, Derivative Assay, Flow Cytometry, Expressing, Knockdown, Negative Control

Journal: Redox Biology

Article Title: Stromal cell-derived itaconate promotes endometriosis via macrophage NRF2 and lysosomal pH modulation

doi: 10.1016/j.redox.2026.104101

Figure Lengend Snippet: Itaconate Suppresses NOX2-Derived ROS to Regulate Macrophage Function and Lesion Progression in End ometriosis (A-B) qPCR analysis of NOX2 mRNA in peritoneal macrophages (PMs) from human (A) and mouse (B) NC and EM groups (n = 3/group). (C-D) Western blot and quantification of NOX2 protein in PBMC-derived macrophages treated with LPS or LPS + 4-OI (n = 5/group). (E) NOX2 enzyme activity in BMDMs (n = 4/group). (F, G) Flow cytometry and quantification of ROS production in BMDMs after LPS or LPS + 4-OI (n = 3/group). (H, I) Flow cytometry and quantification of iNOS + BMDMs after LPS, LPS + 4-OI, or LPS + 4-OI + DPI treatment (n = 4/group). (J) Intracellular Ca 2+ dynamics in BMDMs measured by Fluo-4 after LPS, 4-OI, or DPI treatment. (K) Schematic of 4-OI and DPI intervention in the mouse endometriosis model. (L, M) Representative images (L) and gross morphology (M) of endometriotic lesions after PBS, 4-OI, or DPI treatment. (N) Quantification of lesion weight (n = 6/group). (O, P) Flow cytometry and quantification of iNOS + peritoneal macrophages (percentage and MFI) in peritoneal lavage (n = 6/group). (Q) mRNA levels of Il1b , Il6 , Nos2 , and Tnf in peritoneal macrophages after treatments (n = 6/group). (Data are presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001, ns: not significant.) E2, estradiol benzoate; EM, endometriosis; NC, non-EM control; LPS, lipopolysaccharide; 4-OI, 4-octyl itaconate; DPI, NOX2 inhibitor.

Article Snippet: Estrous cycles were synchronized with subcutaneous estradiol benzoate (E2) (3 μg/mouse) (MCE; Nanjing, China).

Techniques: Derivative Assay, Western Blot, Activity Assay, Flow Cytometry, Control

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Interleukin-1β-induced arthritis involves chondrocyte oxiapoptophagy

doi: 10.4196/kjpp.25.279

Figure Lengend Snippet: (A, B) IL-1β increases ROS production in chondrocytes. Scale bar = 100 μm. (C–E) The expression of inflammatory mediators, including inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nitric oxide (NO), and prostaglandin E 2 (PGE 2 ), are upregulated in chondrocytes stimulated with IL-1β. Values are presented as mean ± SD. DCF-DA, dichlorfluorescein-diacetate.

Article Snippet: The production of PGE 2 in the chondrocytes stimulated with IL-1β was measured using a PGE 2 Parameter Assay Kit (R&D System), according to the manufacturer’s instruction.

Techniques: Expressing