Journal: Biomaterials Research

Article Title: Anti-senescence Effects of Nanovesicles Derived from Cluster of Differentiation-146-Positive Tonsil Mesenchymal Stem Cells via Modulation of the Tumor Protein 53 Pathway

doi: 10.34133/bmr.0371

Figure Lengend Snippet: In vitro analysis of CD146 + TMSC-NVs in a passage-associated cellular senescence model. (A) Morphological changes of human dermal fibroblasts (HDFs) after treatment with CD146 + TMSC-NVs (scale bar = 200 μm). (B) Senescence-associated β-galactosidase (SA-β-gal) assay after treatment with CD146 + TMSC-NVs (scale bar = 200 μm). (C) Cell proliferation test using Cell Counting Kit-8 (CCK-8) assay during treatment with CD146 + TMSC-NVs. (D) Quantitative data following colorization with the SA-β-gal-positive cells. (E to G) Quantitative data of the polymerase chain reaction for the mRNA expression of elastin, COL1, and HMOX1. Statistical significance is shown as the mean ± standard deviation (* P < 0.05, *** P < 0.001, **** P < 0.0001; ns, not significant).

Article Snippet: The senescence-associated β-galactosidase (SA-β-gal) assay was conducted using a cellular senescence staining kit (Cell Biolabs, San Diego, CA, USA).

Techniques: In Vitro, Cell Counting, CCK-8 Assay, Polymerase Chain Reaction, Expressing, Standard Deviation

Journal: Biomaterials Research

Article Title: Anti-senescence Effects of Nanovesicles Derived from Cluster of Differentiation-146-Positive Tonsil Mesenchymal Stem Cells via Modulation of the Tumor Protein 53 Pathway

doi: 10.34133/bmr.0371

Figure Lengend Snippet: Schematic of the in vivo analysis of CD146 + TMSC-NVs using an intrinsic aging mice model. PBS, phosphate-buffered saline.

Article Snippet: The senescence-associated β-galactosidase (SA-β-gal) assay was conducted using a cellular senescence staining kit (Cell Biolabs, San Diego, CA, USA).

Techniques: In Vivo, Saline

Journal: Biomaterials Research

Article Title: Anti-senescence Effects of Nanovesicles Derived from Cluster of Differentiation-146-Positive Tonsil Mesenchymal Stem Cells via Modulation of the Tumor Protein 53 Pathway

doi: 10.34133/bmr.0371

Figure Lengend Snippet: In vivo analysis of the skin in the intrinsic aging mice model after treatment with CD146 + TMSC-NVs. (A) Hematoxylin and eosin (H&E) and Masson’s trichrome (MT) staining (scale bar = 100 μm). (B) Immunohistochemistry staining to confirm the expression of COL1 and elastin in the intrinsic aging model following treatment with CD146 + TMSC-NVs (scale bar = 100 μm). (C and D) Quantitative data from the immunohistochemistry staining in the skin of the intrinsic aging mice following treatment with CD146 + TMSC-NVs. Statistical significance is shown as the mean ± standard deviation (* P < 0.05, ** P < 0.01, **** P < 0.0001; ns, not significant).

Article Snippet: The senescence-associated β-galactosidase (SA-β-gal) assay was conducted using a cellular senescence staining kit (Cell Biolabs, San Diego, CA, USA).

Techniques: In Vivo, Staining, Immunohistochemistry, Expressing, Standard Deviation

Journal: Biomaterials Research

Article Title: Anti-senescence Effects of Nanovesicles Derived from Cluster of Differentiation-146-Positive Tonsil Mesenchymal Stem Cells via Modulation of the Tumor Protein 53 Pathway

doi: 10.34133/bmr.0371

Figure Lengend Snippet: In vivo analysis of the brain in the intrinsic aging mice model after treatment with CD146 + TMSC-NVs. (A) Immunohistochemistry staining to confirm the expression of phospho-Tau (p-Tau) and amyloid beta (Aβ) in the intrinsic aging model after treatment with CD146 + TMSC-NVs (scale bar = 100 μm). (B and C) Quantitative data from the immunohistochemistry staining. (D) Western blotting of p-Tau, Aβ, and β-actin from the brains of the intrinsic aging mice model after treatment with CD146 + TMSC-NVs. Statistical significance is shown as the mean ± standard deviation (** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant).

Article Snippet: The senescence-associated β-galactosidase (SA-β-gal) assay was conducted using a cellular senescence staining kit (Cell Biolabs, San Diego, CA, USA).

Techniques: In Vivo, Immunohistochemistry, Staining, Expressing, Western Blot, Standard Deviation

Journal: Biomaterials Research

Article Title: Anti-senescence Effects of Nanovesicles Derived from Cluster of Differentiation-146-Positive Tonsil Mesenchymal Stem Cells via Modulation of the Tumor Protein 53 Pathway

doi: 10.34133/bmr.0371

Figure Lengend Snippet: In vivo analysis of the liver in the intrinsic aging mice model after treatment with CD146 + TMSC-NVs. (A) Immunohistochemistry staining to confirm the expression of alpha-smooth muscle actin (α-SMA) and perilipin-1 in the liver from the intrinsic aging model after treatment with CD146 + TMSC-NVs (scale bar = 100 μm). (B and C) Quantitative data from the immunohistochemistry staining. Statistical significance is shown as the mean ± standard deviation (* P < 0.05, ** P < 0.01, **** P < 0.0001; ns, not significant).

Article Snippet: The senescence-associated β-galactosidase (SA-β-gal) assay was conducted using a cellular senescence staining kit (Cell Biolabs, San Diego, CA, USA).

Techniques: In Vivo, Immunohistochemistry, Staining, Expressing, Standard Deviation

Journal: Biomaterials Research

Article Title: Anti-senescence Effects of Nanovesicles Derived from Cluster of Differentiation-146-Positive Tonsil Mesenchymal Stem Cells via Modulation of the Tumor Protein 53 Pathway

doi: 10.34133/bmr.0371

Figure Lengend Snippet: Anti-senescence properties of CD146 + TMSC-NVs via modulation of the p53 signaling pathway. (A) Immunofluorescence image to confirm the expression of p-p53 and p21 after treatment with CD146 + TMSC-NVs. p-p53 (green, top row) and p21 (red, bottom row) were co-stained with Hoechst 33342 (blue) to visualize the nuclei (scale bars = 50 μm). (B) Quantitative data of p-p53-positive cells. (C) Quantitative data of p21-positive cells. (D) Western blotting of p53 and caspase-3. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an endogenous control. (E) Quantitative data of western blotting of p53. (F) Quantitative data of western blotting of caspase-3. Statistical significance is shown as the mean ± standard deviation (* P < 0.05, *** P < 0.001, **** P < 0.0001; ns, not significant).

Article Snippet: The senescence-associated β-galactosidase (SA-β-gal) assay was conducted using a cellular senescence staining kit (Cell Biolabs, San Diego, CA, USA).

Techniques: Immunofluorescence, Expressing, Staining, Western Blot, Control, Standard Deviation

Journal: Biomaterials Research

Article Title: Anti-senescence Effects of Nanovesicles Derived from Cluster of Differentiation-146-Positive Tonsil Mesenchymal Stem Cells via Modulation of the Tumor Protein 53 Pathway

doi: 10.34133/bmr.0371

Figure Lengend Snippet: miRNA profiling. Heatmap showing selected miRNAs enriched in CD146 + TMSC-NVs. miRNAs were categorized based on their functional relevance to extracellular matrix remodeling, oxidative stress, and cellular senescence, along with highly enriched miRNAs representing CD146 + TMSC-NVs. Expression values were normalized as log2(TPM + 1).

Article Snippet: The senescence-associated β-galactosidase (SA-β-gal) assay was conducted using a cellular senescence staining kit (Cell Biolabs, San Diego, CA, USA).

Techniques: Functional Assay, Expressing

Journal: Redox Biology

Article Title: ERRα-NOS2-mediated citrulline metabolism attenuates tubular epithelial cell senescence in diabetic kidney disease

doi: 10.1016/j.redox.2026.104065

Figure Lengend Snippet: Downregulation of ERRα in TECs and its association with senescence (A) Representative images and quantification of GLB1 (red), LTL (green) and DAPI (blue)-immunofluorescent stained kidney sections from control (n = 12), MCD (n = 12) and DKD (n = 24) patients. (B) Representative images and quantification of p16 (red), LTL (green) and DAPI (blue)-immunofluorescent stained kidney sections from control (n = 12), MCD (n = 12) and DKD (n = 24) patients (C) Representative images and quantification of p21 (red), LTL (green) and DAPI (blue)-immunofluorescent stained kidney sections from control (n = 12), MCD (n = 12) and DKD (n = 24) patients (D) Representative images and quantification of ERRα (red), LTL (green) and DAPI (blue)-immunofluorescent stained kidney sections from control (n = 12), MCD (n = 12) and DKD (n = 24) patients (E) Correlation analysis of ERRα expression and GLB1 expression in DKD (n = 24) patients (F) Representative images and quantification of ERRα (red), LTL (green) and DAPI (blue)-immunofluorescent stained kidney sections from Vehicle (n = 6) and STZ/HFD (n = 6) groups of mice (G) Representative images and band density quantification of Western blotting using isolated renal tubules from Vehicle (n = 6) and STZ/HFD (n = 6) groups of mice to quantify protein expression of ERRα normalized to β-actin. (H) Representative images and band density quantification of Western blotting using primary TECs from each mouse group to quantify protein expression of ERRα normalized to β-actin (n = 3) n = 6 independent group mice. Data are expressed as mean ± SD. Statistical differences between two groups were assessed using t -test. For comparisons among three or more groups, one-way ANOVA followed by Tukey's post hoc test was used. ∗∗∗ p < 0.001.

Article Snippet: To assess cellular senescence, β-galactosidase (β-gal) staining was performed using a commercial kit (Cell Signaling Technology).

Techniques: Staining, Control, Expressing, Western Blot, Isolation

Journal: Redox Biology

Article Title: ERRα-NOS2-mediated citrulline metabolism attenuates tubular epithelial cell senescence in diabetic kidney disease

doi: 10.1016/j.redox.2026.104065

Figure Lengend Snippet: TECs-specific knockout of ERRα exacerbates TECs injury and senescence in diabetic mice (A) Schematic diagram of intraperitoneal injection of Streptozotocin (STZ) and 12 weeks treatment of high-fat diet (HFD) or normal saline (Vehicle) to ERRα podKO and ERRα ctrl mice (n = 6) (B – D) Blood urea nitrogen (BUN), serum creatinine (Cr) and urine albumin-to-creatinine ratio (ACR) levels in each mouse group (n = 6) (E – F) Representative HE, Masson ' s trichrome staining and β-gal staining images, quantification of tubular interstitial damage, Masson staining positive area and immunofluorescent staining of GLB1 (red), LTL (green), and DAPI (blue) in kidney sections from each mouse group (n = 6) (G – H) Representative TEM images, DHE staining and quantification of proportion of damaged mitochondria, DHE staining positive area in kidney sections from each mouse group (n = 6). (I – J) Representative images and band density quantification of Western blotting using isolated renal tubules from each mouse group (n = 6) to quantify protein expression of ERRα, p16 and p21 normalized to β-actin (n = 6) (K – L) IL-1β and IL-6 contents in isolated renal tubules from each mouse group (n = 6). n = 6 independent group mice. Data are expressed as mean ± SD. Statistical differences among three or more groups were evaluated using one-way ANOVA followed by Tukey's post hoc test. ns: not significant ( p > 0.05), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: To assess cellular senescence, β-galactosidase (β-gal) staining was performed using a commercial kit (Cell Signaling Technology).

Techniques: Knock-Out, Injection, Saline, Staining, Western Blot, Isolation, Expressing

Journal: Redox Biology

Article Title: ERRα-NOS2-mediated citrulline metabolism attenuates tubular epithelial cell senescence in diabetic kidney disease

doi: 10.1016/j.redox.2026.104065

Figure Lengend Snippet: TECs-specific knock-in of ERRα exacerbates TECs injury and senescence in diabetic mice (A) Schematic diagram of intraperitoneal injection of Streptozotocin (STZ) and 12 weeks treatment of high-fat diet (HFD) or normal saline (Vehicle) to ERRα podKI and ERRα ctrl mice (n = 6). (B – D) Blood urea nitrogen (BUN), serum creatinine (Cr) and urine albumin-to-creatinine ratio (ACR) levels in each mouse group (n = 6). (E – F) Representative HE and β-gal staining images, quantification of tubular interstitial damage, and immunofluorescent staining of GLB1 (red), LTL (green), and DAPI (blue) in kidney sections from each mouse group (n = 6). (G – H) Representative TEM images, DHE staining and quantification of proportion of damaged mitochondria, DHE staining positive area in kidney sections from each mouse group (n = 6) (I – J) Representative images and band density quantification of Western blotting using isolated renal tubules from each mouse group (n = 6) to quantify protein expression of ERRα, p16 and p21 normalized to β-actin (K – L) IL-1β and IL-6 contents in isolated renal tubules from each mouse group (n = 6). n = 6 independent group mice. Data are expressed as mean ± SD. Statistical differences among three or more groups were evaluated using one-way ANOVA followed by Tukey's post hoc test. ns: not significant ( p > 0.05), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: To assess cellular senescence, β-galactosidase (β-gal) staining was performed using a commercial kit (Cell Signaling Technology).

Techniques: Knock-In, Injection, Saline, Staining, Western Blot, Isolation, Expressing

Journal: Redox Biology

Article Title: ERRα-NOS2-mediated citrulline metabolism attenuates tubular epithelial cell senescence in diabetic kidney disease

doi: 10.1016/j.redox.2026.104065

Figure Lengend Snippet: ERRα expression is associated with high glucose-induced senescence in HK-2 cells (A) Representative β-gal staining images quantification of positive area from each HK-2 cell group (n = 3). (B–C) IL-1β and IL-6 contents from each HK-2 cell group (n = 3). (D) Representative images and band density quantification of Western blotting from each HK-2 cell group (n = 3) to quantify protein expression of p16 and p21 normalized to β-actin. (E) Representative β-gal staining images quantification of positive area from each HK-2 cell group (n = 3) (F – G) IL-1β and IL-6 contents from each HK-2 cell group (n = 3). (H) Representative images and band density quantification of Western blotting from each HK-2 cell group (n = 3) to quantify protein expression of p16 and p21 normalized to β-actin n = 3 cultures per group. HG, High gluose (40 mM, 24h); ERRα-KO, Knockout of ERRα in HK-2 cells using CRISPR/Cas9 technology. OE-ERRα, overexpression ERRα group. Data are expressed as mean ± SD. Statistical differences among three or more groups were evaluated using one-way ANOVA followed by Tukey's post hoc test. ns: not significant ( p > 0.05), ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: To assess cellular senescence, β-galactosidase (β-gal) staining was performed using a commercial kit (Cell Signaling Technology).

Techniques: Expressing, Staining, Western Blot, Knock-Out, CRISPR, Over Expression

Journal: Redox Biology

Article Title: ERRα-NOS2-mediated citrulline metabolism attenuates tubular epithelial cell senescence in diabetic kidney disease

doi: 10.1016/j.redox.2026.104065

Figure Lengend Snippet: NOS2 is downregulated in DKD and associated with TECs senescence (A) NOS2 expression in the kidney of different age groups of the population using ADEIP database. (B) Representative images and quantification of NOS2 (red), LTL (green) and DAPI (blue)-immunofluorescent stained kidney sections from control (n = 12), MCD (n = 12) and DKD (n = 24) patients. (C) Correlation analysis of NOS2 expression and GLB1 expression in DKD (n = 24) patients (D-E) Representative images and band density quantification of Western blotting using isolated renal tubules from each mouse group (n = 6) to quantify protein expression of NOS2 normalized to β-actin. (F) Representative images and quantification of NOS2 (red), LTL (green) and DAPI (blue)-immunofluorescent stained kidney sections from each mouse group (n = 6). (G) Representative images and band density quantification of Western blotting from each HK-2 cells group to quantify protein expression of NOS2 normalized to β-actin (n = 3) (H) Representative images and band density quantification of Western blotting from each HK-2 cells group to quantify protein expression of NOS2 normalized to β-actin (n = 3) (I) Representative β-gal staining images quantification of positive area from each HK-2 cell group (n = 3) n = 3 cultures per group. HG, High gluose (40 mM, 24h); ERRα-KO, knockout of ERRα in HK-2 cells using CRISPR/Cas9 technology. si-ERRα, knockdown ERRα group. OE-NOS2, overexpression NOS2 group. Data are expressed as mean ± SD. Statistical differences among three or more groups were evaluated using one-way ANOVA followed by Tukey's post hoc test. ns: not significant ( p > 0.05), ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: To assess cellular senescence, β-galactosidase (β-gal) staining was performed using a commercial kit (Cell Signaling Technology).

Techniques: Expressing, Staining, Control, Western Blot, Isolation, Knock-Out, CRISPR, Knockdown, Over Expression

Journal: Redox Biology

Article Title: ERRα-NOS2-mediated citrulline metabolism attenuates tubular epithelial cell senescence in diabetic kidney disease

doi: 10.1016/j.redox.2026.104065

Figure Lengend Snippet: Overexpression of NOS2 Alleviates TECs Injury and Senescence in Diabetic Mice (A) Schematic diagram of overexpression of NOS2 in TECs using adeno-associated viruses (AAV). (B – D) Blood urea nitrogen (BUN), serum creatinine (Cr) and urine albumin-to-creatinine ratio (ACR) levels in each mouse group (n = 6). (E – F) Representative HE and β-gal staining images, quantification of tubular interstitial damage, and immunofluorescent staining of GLB1 (red), LTL (green), and DAPI (blue) in kidney sections from each mouse group (n = 6). (G – H) Representative TEM images, DHE staining and quantification of proportion of damaged mitochondria, DHE staining positive area in kidney sections from each mouse group (n = 6) (I – J) Representative images and band density quantification of Western blotting using isolated renal tubules from each mouse group (n = 6) to quantify protein expression of ERRα, NOS2, p16 and p21 normalized to β-actin (K – L) IL-1β and IL-6 contents in isolated renal tubules from each mice group (n = 6). n = 6 independent group mice. Data are expressed as mean ± SD. Statistical differences among three or more groups were evaluated using one-way ANOVA followed by Tukey's post hoc test. ns: not significant ( p > 0.05), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: To assess cellular senescence, β-galactosidase (β-gal) staining was performed using a commercial kit (Cell Signaling Technology).

Techniques: Over Expression, Staining, Western Blot, Isolation, Expressing

Journal: Redox Biology

Article Title: ERRα-NOS2-mediated citrulline metabolism attenuates tubular epithelial cell senescence in diabetic kidney disease

doi: 10.1016/j.redox.2026.104065

Figure Lengend Snippet: Citrulline Supplementation Alleviates TECs Injury and Senescence in Diabetic Mice (A) Schematic diagram of supplementation scheme of each group of mice with drinking water only or drinking waters containing citrulline (1 g/kg of body weight of mice) for 9 weeks. (B – D) Blood urea nitrogen (BUN), serum creatinine (Cr) and urine albumin-to-creatinine ratio (ACR) levels in each mouse group (n = 6). (E – F) Representative HE and β-gal staining images, quantification of tubular interstitial damage, and immunofluorescent staining of GLB1 (red), LTL (green), and DAPI (blue) in kidney sections from each mouse group (n = 6). (G – H) Representative TEM images, DHE staining and quantification of proportion of damaged mitochondria, DHE staining positive area in kidney sections from each mouse group (n = 6). (I – J) Representative images and band density quantification of Western blotting using isolated renal tubules from each mouse group (n = 6) to quantify protein expression of ERRα, NOS2, p16 and p21 normalized to β-actin. (K–K) IL-1β and IL-6 contents in isolated renal tubules from each mice group (n = 6). n = 6 independent group mice. Data are expressed as mean ± SD. Statistical differences among three or more groups were evaluated using one-way ANOVA followed by Tukey's post hoc test. ns: not significant ( p > 0.05), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: To assess cellular senescence, β-galactosidase (β-gal) staining was performed using a commercial kit (Cell Signaling Technology).

Techniques: Staining, Western Blot, Isolation, Expressing