Journal: Cardiovascular Therapeutics

Article Title: CircIdh2 Attenuates Endothelial‐to‐Mesenchymal Transition and Myocardial Fibrosis and Exhibits Translational Potential

doi: 10.1155/cdr/2005876

Figure Lengend Snippet: AAV‐mediated delivery of circIdh2 or circIdh2‐aa attenuated myocardial fibrosis and EndMT‐related marker changes in vivo. (A) HE staining of myocardial infarction tissues from the sham, model, circIdh2‐AAV, and circIdh2‐aa‐AAV groups. (B) Masson staining of MI tissues. (C) IF detection and quantification of fibrotic markers (Collagen I and TGF‐ β ) in myocardial tissues. Scale bars, 100 μ m. (D) Western blot analysis of EndMT‐related proteins (VE‐cadherin, vimentin, and N‐cadherin). Data are mean ± SD; n = 3 rats per group; one‐way ANOVA; ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: The sections were then incubated overnight at 4°C with primary antibodies against Collagen I (1:200, #66761‐1‐Ig, Proteintech) and TGF‐ β 1 (1:200, #81746‐2‐RR, Proteintech).

Techniques: Marker, In Vivo, H&E Stain, Staining, Western Blot, Analysis

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Renal Tubular Epithelial Cells Senescence Induced by 1‐Aminopyrene Activates Fibroblast via BRD4 Driven Senescence‐Associated Secretory Phenotype Signaling

doi: 10.1002/jbt.71124

Figure Lengend Snippet: 1 ‐ AP induces tubular epithelial senescence and promotes paracrine fibroblast activation via a SASP‐mediated mechanism. (A) Representative images and quantification of SA‐β‐gal positive RTECs following exposure to 1‐AP after 72 h. Scale bars, 5 µm. (B) DAPI‐stained nuclei (blue) and quantification showing a significant reduction in cell number following exposure to 1‐AP after 72 h. Scale bars, 2 µm. (C) Representative images and quantification of immunofluorescence analysis of Ki67 in RTECs following 1‐AP treatment after 72 h. Scale bars, 5 µm. (D) RT‐qPCR analysis of p16, p21, and p53 mRNA expression showing upregulation of senescence‐associated cell cycle regulators in RTECs following exposure to 1‐AP after 48 h. (E) Representative images and quantification of immunofluorescence analysis of p21 in RTECs following 1‐AP treatment after 72 h. Scale bars, 5 µm. (F) Heatmap for RT‐qPCR analysis of senescence‐associated secretory phenotype (SASP) factors (IL‐1β, IL‐6, IL‐8, MCP‐1, TGF‐β1, and CXCL1) in RTECs following 1‐AP treatment after 48 h. (G) Schematic illustration of the experimental design showing 1‐AP induced senescence in RTECs and paracrine activation of renal fibroblasts via conditioned media may leads to TIF or renal fibrosis. (H) Heatmap for RT‐qPCR analysis of fibrotic markers ( α‐SMA, POSTN, Col1a2, TIMP1 , and CTGF ) in NRK52F renal fibroblasts exposed to conditioned media from 1‐AP‐treated TECs for 48 h. (I–K) Representative images and quantification of immunofluorescence analysis of α‐SMA (I), POSTN (J), and COL1A1 (K) in NRK52F fibroblasts, demonstrating significant fibroblast activation and extracellular matrix induction following exposure to TEC‐derived conditioned media after 48 h. Scale bars, 20 µm. Data are presented as mean ± SD, n = 3. Statistical analysis was performed using two‐tailed unpaired Student's t‐test. Exact p ‐values are indicated in the graphs; ns, not significant. DAPI used as nuclear counterstain.

Article Snippet: Primary antibodies for AhR antibody (182705‐10 G, Meck), Phospho‐Histone H2A.X (Ser139) Antibody (20E3, cell signaling), BRD4 antibody (ER1901‐02, Thermo‐Fischer), Ki‐67 antibody (27309‐1‐AP, proteintech), p21 (2947, cell signaling), α‐SMA antibody (14395‐1‐AP, proteintech), periostin (19899‐1‐AP, proteintech), COL1A1 antibody (55131‐1‐AP, proteintech), Histone H3K9/14ac (Acetyl H3K9/14) Antibody (A‐4021‐025, epigentek), Histone H3K27me3 (H3K27 Trimethyl) Antibody (A‐4039‐025, epigentek), Histone H3K9me3 (H3K9 Trimethyl) Antibody (A‐4036‐025, epigentek), IL‐6 (21865‐1‐AP, proteintech), TGF‐beta1 (21898‐1‐AP, proteintech) and Secondary antibody anti‐Mouse Alexa‐Fluro 555 ( A31570 , Thermo‐Fischer) and anti‐Rabbit Alexa‐Fluro 488 (A11008, Thermo‐Fischer), TUNEL Assay Kit (25879, cell signaling) and Senescence β‐Galactosidase (SA‐β‐gal) Staining Kit (9860, cell signaling) were purchased.

Techniques: Activation Assay, Staining, Immunofluorescence, Analysis, Quantitative RT-PCR, Expressing, Derivative Assay, Two Tailed Test

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Renal Tubular Epithelial Cells Senescence Induced by 1‐Aminopyrene Activates Fibroblast via BRD4 Driven Senescence‐Associated Secretory Phenotype Signaling

doi: 10.1002/jbt.71124

Figure Lengend Snippet: AhR signaling is activated in RTECs by 1‐AP but is insufficient for SASP‐mediated fibroblast activation. (A) Representative images and quantification of immunofluorescence analysis of AHR expression in RTECs following 1‐AP treatment ± AhR inhibitor (10 μM) after 48 h. Scale bars, 20 µm. (B) Heatmap for RT‐qPCR analysis of AhR target genes ( CYP1A1, CYP1A2 ) and senescence markers ( p16, p21, p53 ) following 1‐AP treatment ± AhR inhibitor after 48 h. (C) Heatmap for RT‐qPCR analysis of SASP genes ( IL‐1β, IL‐6, IL‐8, MCP‐1, TGF‐β1, CXCL1 ) following 1‐AP treatment ± AhR inhibitor after 48 h. Conditioned media (CM) transfer from NRK‐52E/RTECs treated with 1‐AP ± AhR inhibitor to NRK‐52F fibroblasts after 48 h showing (D) Heatmap for RT‐qPCR analysis of fibroblast activation and extracellular matrix genes ( α‐SMA, POSTN, Col1a2, TIMP‐1, CTGF ), (E–G) Quantification of immunofluorescence analysis of α‐SMA, POSTN and COL1A1 in NRK52F fibroblasts. Data are presented as mean ± SD, n = 3. Statistical analysis was performed using two‐tailed unpaired Student's t‐test. Exact p ‐values are indicated in the graphs; ns, not significant. DAPI used as nuclear counterstain.

Article Snippet: Primary antibodies for AhR antibody (182705‐10 G, Meck), Phospho‐Histone H2A.X (Ser139) Antibody (20E3, cell signaling), BRD4 antibody (ER1901‐02, Thermo‐Fischer), Ki‐67 antibody (27309‐1‐AP, proteintech), p21 (2947, cell signaling), α‐SMA antibody (14395‐1‐AP, proteintech), periostin (19899‐1‐AP, proteintech), COL1A1 antibody (55131‐1‐AP, proteintech), Histone H3K9/14ac (Acetyl H3K9/14) Antibody (A‐4021‐025, epigentek), Histone H3K27me3 (H3K27 Trimethyl) Antibody (A‐4039‐025, epigentek), Histone H3K9me3 (H3K9 Trimethyl) Antibody (A‐4036‐025, epigentek), IL‐6 (21865‐1‐AP, proteintech), TGF‐beta1 (21898‐1‐AP, proteintech) and Secondary antibody anti‐Mouse Alexa‐Fluro 555 ( A31570 , Thermo‐Fischer) and anti‐Rabbit Alexa‐Fluro 488 (A11008, Thermo‐Fischer), TUNEL Assay Kit (25879, cell signaling) and Senescence β‐Galactosidase (SA‐β‐gal) Staining Kit (9860, cell signaling) were purchased.

Techniques: Activation Assay, Immunofluorescence, Analysis, Expressing, Quantitative RT-PCR, Two Tailed Test

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Renal Tubular Epithelial Cells Senescence Induced by 1‐Aminopyrene Activates Fibroblast via BRD4 Driven Senescence‐Associated Secretory Phenotype Signaling

doi: 10.1002/jbt.71124

Figure Lengend Snippet: BRD4 recruitment in RTECs sustains SASP transcription and paracrine fibroblast activation. (A) Representative images and quantification of immunofluorescence analysis of BRD4 expression in RTECs following 1‐AP treatment ± JQ1 (0.5 μM) after 48 h. Scale bars, 20 µm. (B) Heatmap for RT‐qPCR analysis of senescence regulators ( p16, p21, p53 ) and SASP genes ( IL‐1β, IL‐6, IL‐8, MCP‐1, TGF‐β1, CXCL1 ) in RTECs following 1‐AP treatment ± JQ1 after 48 h. Conditioned media (CM) transfer from NRK‐52E/RTECs treated with 1‐AP ± JQ1 to NRK‐52F fibroblasts after 48 h showing (C) Heatmap for RT‐qPCR analysis of fibroblast activation and extracellular matrix genes ( α‐SMA, POSTN, Col1a2, TIMP‐1, CTGF ), (D–F) Quantification of immunofluorescence analysis of α‐SMA, POSTN and COL1A1 in NRK52F fibroblasts. Data are presented as mean ± SD, n = 3. Statistical analysis was performed using two‐tailed unpaired Student's t‐test. Exact p ‐values are indicated in the graphs; ns, not significant. DAPI used as nuclear counterstain.

Article Snippet: Primary antibodies for AhR antibody (182705‐10 G, Meck), Phospho‐Histone H2A.X (Ser139) Antibody (20E3, cell signaling), BRD4 antibody (ER1901‐02, Thermo‐Fischer), Ki‐67 antibody (27309‐1‐AP, proteintech), p21 (2947, cell signaling), α‐SMA antibody (14395‐1‐AP, proteintech), periostin (19899‐1‐AP, proteintech), COL1A1 antibody (55131‐1‐AP, proteintech), Histone H3K9/14ac (Acetyl H3K9/14) Antibody (A‐4021‐025, epigentek), Histone H3K27me3 (H3K27 Trimethyl) Antibody (A‐4039‐025, epigentek), Histone H3K9me3 (H3K9 Trimethyl) Antibody (A‐4036‐025, epigentek), IL‐6 (21865‐1‐AP, proteintech), TGF‐beta1 (21898‐1‐AP, proteintech) and Secondary antibody anti‐Mouse Alexa‐Fluro 555 ( A31570 , Thermo‐Fischer) and anti‐Rabbit Alexa‐Fluro 488 (A11008, Thermo‐Fischer), TUNEL Assay Kit (25879, cell signaling) and Senescence β‐Galactosidase (SA‐β‐gal) Staining Kit (9860, cell signaling) were purchased.

Techniques: Activation Assay, Immunofluorescence, Analysis, Expressing, Quantitative RT-PCR, Two Tailed Test

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: pH-responsive Zn 2+ -coordinated gellan gum/γ-polyglutamic acid hydrogel microspheres as functional biomaterials for intra-articular delivery: physicochemical design, smart release, and chondroprotective performance in osteoarthritis

doi: 10.3389/fbioe.2026.1924086

Figure Lengend Snippet: qPCR primer information.

Article Snippet: The sections were subsequently reacted with primary antibodies targeting COL2A1 (Proteintech, 28459-1-AP), MMP13 (Proteintech, 18165-1-AP), p-PI3K (HUABIO, HA721672), and p-AKT (Proteintech, 66444-1-Ig) for 16 h at 4 °C.

Techniques: Sequencing

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: pH-responsive Zn 2+ -coordinated gellan gum/γ-polyglutamic acid hydrogel microspheres as functional biomaterials for intra-articular delivery: physicochemical design, smart release, and chondroprotective performance in osteoarthritis

doi: 10.3389/fbioe.2026.1924086

Figure Lengend Snippet: PGG-Zn@Cur restored matrix metabolic homeostasis in IL-1β-stimulated chondrocytes. (A) Representative immunofluorescence images of COL2A1 in chondrocytes after different treatments. COL2A1 was stained red, and nuclei were counterstained with DAPI. (B) Representative immunofluorescence images of MMP13 in chondrocytes after different treatments. MMP13 was stained green, and nuclei were counterstained with DAPI. (C) Quantitative analysis of relative fluorescence intensity for COL2A1 and MMP13 based on immunofluorescence staining. IL-1β stimulation decreased COL2A1 expression and increased MMP13 accumulation, whereas PGG-Zn@Cur treatment restored COL2A1 fluorescence intensity and suppressed MMP13-positive signals. (D–G) RT-qPCR analysis of cartilage matrix metabolism-related genes, including anabolic markers Col2a1 and Aggrecan, and catabolic markers Mmp13 and Adamts5, in chondrocytes after the indicated treatments. (H) Representative Western blot images of iNOS, COL2A1, MMP13, and ACAN protein expression, together with heatmap-based quantitative analysis of Western blot results across different treatment groups. β-Actin was used as the loading control. ( n = 3, Scale bar: 10 μm *: p < 0.05; **: p < 0.01; ***: p < 0.001; ****: p < 0.0001; ns: no significant).

Article Snippet: The sections were subsequently reacted with primary antibodies targeting COL2A1 (Proteintech, 28459-1-AP), MMP13 (Proteintech, 18165-1-AP), p-PI3K (HUABIO, HA721672), and p-AKT (Proteintech, 66444-1-Ig) for 16 h at 4 °C.

Techniques: Immunofluorescence, Staining, Analysis, Fluorescence, Expressing, Quantitative RT-PCR, Western Blot, Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: pH-responsive Zn 2+ -coordinated gellan gum/γ-polyglutamic acid hydrogel microspheres as functional biomaterials for intra-articular delivery: physicochemical design, smart release, and chondroprotective performance in osteoarthritis

doi: 10.3389/fbioe.2026.1924086

Figure Lengend Snippet: PGG-Zn@Cur attenuated PI3K-AKT activation and apoptosis-associated responses in IL-1β-stimulated chondrocytes. (A) Schematic workflow of RNA sequencing and subsequent experimental validation. (B) Heatmap of representative differentially expressed genes related to cartilage matrix metabolism, inflammation, apoptosis, and PI3K-AKT-associated signaling. (C) Volcano plot showing differentially expressed genes between the indicated groups, with representative genes related to cartilage homeostasis, apoptosis, and PI3K-AKT signaling labeled. (D) GO enrichment analysis of differentially expressed genes, including biological process, cellular component, and molecular function categories. (E) KEGG pathway enrichment analysis, with the PI3K-AKT signaling pathway highlighted as a candidate pathway. (F) Representative immunofluorescence images of p-PI3K, p-AKT, Cl-Caspase3, and COL2A1 in chondrocytes after different treatments. Nuclei were counterstained with DAPI. (G) Quantitative analysis of p-PI3K, p-AKT, Cl-Caspase3, and COL2A1 fluorescence intensity. (H) Representative Western blot images of PI3K-AKT pathway- and apoptosis-related proteins, including p-PI3K, PI3K, p-AKT, AKT, Cleaved-Caspase3, Caspase3, Bax, and BCL2. β-Actin was used as the loading control. (I) Heatmap-based quantitative analysis of Western blot results, summarizing changes in p-PI3K/PI3K, p-AKT/AKT, Cleaved-Caspase3/Caspase3, Bax/β-Actin, and BCL2/β-Actin across different groups. ( n = 3, Scale bar: 10 μm *: p < 0.05; **: p < 0.01; ***: p < 0.001; ****: p < 0.0001; ns: not significant).

Article Snippet: The sections were subsequently reacted with primary antibodies targeting COL2A1 (Proteintech, 28459-1-AP), MMP13 (Proteintech, 18165-1-AP), p-PI3K (HUABIO, HA721672), and p-AKT (Proteintech, 66444-1-Ig) for 16 h at 4 °C.

Techniques: Activation Assay, RNA Sequencing, Biomarker Discovery, Labeling, Analysis, Immunofluorescence, Fluorescence, Western Blot, Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: pH-responsive Zn 2+ -coordinated gellan gum/γ-polyglutamic acid hydrogel microspheres as functional biomaterials for intra-articular delivery: physicochemical design, smart release, and chondroprotective performance in osteoarthritis

doi: 10.3389/fbioe.2026.1924086

Figure Lengend Snippet: PGG-Zn@Cur attenuates OA-associated cartilage degeneration and preserves matrix homeostasis in vivo . (A) Representative Safranin O/Fast Green staining images of knee joint cartilage from the Sham, OA, OA+ PGG-Zn, OA+ Cur, and OA+ PGG-Zn@Cur groups. (B) Representative immunofluorescence images of COL2A1 and MMP13 in articular cartilage from different groups. Nuclei were counterstained with DAPI. (C, D) Quantitative analysis of cartilage degeneration using OARSI scores and modified Mankin’s scores. (E) Quantitative analysis of COL2A1 and MMP13 fluorescence intensity in articular cartilage. ( n = 5 mice per group, Scale bar: 50 μm *: p < 0.05; **: p < 0.01; ***: p < 0.001; ****: p < 0.0001; ns: not significant).

Article Snippet: The sections were subsequently reacted with primary antibodies targeting COL2A1 (Proteintech, 28459-1-AP), MMP13 (Proteintech, 18165-1-AP), p-PI3K (HUABIO, HA721672), and p-AKT (Proteintech, 66444-1-Ig) for 16 h at 4 °C.

Techniques: In Vivo, Staining, Immunofluorescence, Analysis, Modification, Fluorescence

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: pH-responsive Zn 2+ -coordinated gellan gum/γ-polyglutamic acid hydrogel microspheres as functional biomaterials for intra-articular delivery: physicochemical design, smart release, and chondroprotective performance in osteoarthritis

doi: 10.3389/fbioe.2026.1924086

Figure Lengend Snippet: PGG-Zn@Cur attenuates PI3K-AKT pathway activation and apoptosis-associated cartilage damage in vivo . (A) Representative immunofluorescence images of p-PI3K, p-AKT, and COL2A1 in articular cartilage from the Sham, OA, OA+ PGG-Zn, OA+ Cur, and OA+ PGG-Zn@Cur groups. Nuclei were counterstained with DAPI. (B) Representative TUNEL staining images of articular cartilage from different groups. (C) Quantitative analysis of p-PI3K, p-AKT, and COL2A1 fluorescence intensity in articular cartilage. (D) Quantitative analysis of TUNEL-positive chondrocytes in articular cartilage. ( n = 5 mice per group, Scale bar: 50 μm *: p < 0.05; **: p < 0.01; ***: p < 0.001; ****: p < 0.0001; ns: not significant).

Article Snippet: The sections were subsequently reacted with primary antibodies targeting COL2A1 (Proteintech, 28459-1-AP), MMP13 (Proteintech, 18165-1-AP), p-PI3K (HUABIO, HA721672), and p-AKT (Proteintech, 66444-1-Ig) for 16 h at 4 °C.

Techniques: Activation Assay, In Vivo, Immunofluorescence, TUNEL Assay, Staining, Analysis, Fluorescence