Journal: Biomaterials Research

Article Title: Multiaction Antimicrobial, Anti-inflammatory, and Prohealing Hydrogel as a Novel Strategy for Preventing Postoperative Pancreatic Fistula

doi: 10.34133/bmr.0194

Figure Lengend Snippet: In vitro compatibility of hydrogels. (A) Appearance of hydrogel in vitro hemolysis test (digital camera view) ( n = 3). (B) Statistical results of hemolysis rate of hydrogels in vitro. (C) Statistical results of cell viability of hydrogel samples with different compositions. (D) Dual staining of live and dead cells under the microscope for hydrogel samples with different composition ( n = 3). (E) TUNEL staining under the microscope for PC-OHAD@MP hydrogel ( n = 3). (F) Apoptosis flow cytometry results for PC-OHAD@MP hydrogel ( n = 3). (G) Statistical results of apoptosis flow cytometry for PC-OHAD@MP hydrogel.

Article Snippet: TUNEL (terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end labeling) assay kit, live-dead double staining kit, annexin V-APC/7-AAD apoptosis kit, and cell cycle flow cytometry kit were obtained from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

Techniques: In Vitro, Staining, Microscopy, TUNEL Assay, Flow Cytometry

Journal: Biomaterials Research

Article Title: Multiaction Antimicrobial, Anti-inflammatory, and Prohealing Hydrogel as a Novel Strategy for Preventing Postoperative Pancreatic Fistula

doi: 10.34133/bmr.0194

Figure Lengend Snippet: Hydrogel enhances cell proliferation and migration. (A) Scratch assay under the microscope for hydrogel samples with different compositions ( n = 3). (B) Statistical results of scratch assay for hydrogel samples with different compositions. (C) Cell cycle flow cytometry results for PC-OHAD@MP hydrogel ( n = 3). (D) Statistical results of cell cycle apoptosis for PC-OHAD@MP hydrogel. (E) Relative mRNA expression levels of key regulatory genes in the cell cycle for PC-OHAD@MP hydrogel ( n = 3). (F) Quantitative analysis results of protein expression levels of key regulatory genes in the cell cycle for PC-OHAD@MP hydrogel ( n = 3). (G) Statistical results of protein expression levels of key regulatory genes in the cell cycle for PC-OHAD@MP hydrogel.

Article Snippet: TUNEL (terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end labeling) assay kit, live-dead double staining kit, annexin V-APC/7-AAD apoptosis kit, and cell cycle flow cytometry kit were obtained from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

Techniques: Migration, Wound Healing Assay, Microscopy, Flow Cytometry, Expressing

Journal: iScience

Article Title: CircSOBP suppresses the progression of glioma by disrupting glycolysis and promoting the MDA5-mediated immune response

doi: 10.1016/j.isci.2023.107897

Figure Lengend Snippet: Increased expression of circSOBP inhibits the proliferation, invasion, and migration of glioma cells (A) The spliced sequence of circSOBP was cloned into the pEGFP-C1 vector, with the upstream and downstream containing the identified active ALU elements. RT-qPCR analysis of circSOBP overexpression in U87 and U251 cells. (B) CCK-8 analysis of the effect of circSOBP overexpression on the proliferation of U87 and U251 cells. (C) Cell viability was detected at 0 h, 24 h, 48 h, and 72 h after transfection with pEGFP-C1 or pEGFP-C1-circSOBP plasmids in U87 and U251 cells. (D) Edu assay to analyze the effect on cell replication after overexpression of circSOBP in U87 and U251 cells. Scale bars, 50 μm. (E) TUNEL analysis of the effect of circSOBP overexpression on apoptosis of U87 and U251 cells. Scale bars, 50 μm. (F) Transwell assay to assess the invasive ability of glioma cells after overexpression of circSOBP. (G) Wound healing assay to detect the migration level of glioma cells after overexpression of circSOBP. All statistics of error bars, S.E.M. from three independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001 by two-tailed Student’s t test. Normalized by the value of the control group.

Article Snippet: Next, TUNEL assay was performed using the One-Step TUNEL Apoptosis Kit (RiboBio, China) according to the manufacturer’s instructions.

Techniques: Expressing, Migration, Sequencing, Clone Assay, Plasmid Preparation, Quantitative RT-PCR, Over Expression, CCK-8 Assay, Transfection, EdU Assay, TUNEL Assay, Transwell Assay, Wound Healing Assay, Two Tailed Test, Control

Journal: iScience

Article Title: CircSOBP suppresses the progression of glioma by disrupting glycolysis and promoting the MDA5-mediated immune response

doi: 10.1016/j.isci.2023.107897

Figure Lengend Snippet:

Article Snippet: Next, TUNEL assay was performed using the One-Step TUNEL Apoptosis Kit (RiboBio, China) according to the manufacturer’s instructions.

Techniques: Virus, Recombinant, Staining, Transfection, Software, Fractionation, Labeling, Northern Blot, Cell Viability Assay, TUNEL Assay, Silver Staining, Luciferase, Reporter Gene Assay, ATP Assay, Enzyme-linked Immunosorbent Assay, Purification, Titration, Real-time Polymerase Chain Reaction

Journal: Bone Research

Article Title: Melanoma bone metastasis-induced osteocyte ferroptosis via the HIF1α-HMOX1 axis

doi: 10.1038/s41413-024-00384-y

Figure Lengend Snippet: B16F10 Cell-Induced Osteocyte Ferroptosis in Bone Metastasis. a H&E staining of cortical bone from control and B16F10-injected mice ( n ≥ 6). White arrows indicate normal osteocytes; green arrows, dying osteocytes; and red arrows, dead osteocytes. Scale bars: 20 μm. b TUNEL staining of tibial bone sections from control and B16F10-injected mice ( n = 5). White arrows mark TUNEL-positive cells. Scale bars: 20 μm. c mRNA expression of Dmp1, Dkk1, Phex, Sclerostin, Col1a1 , and Runx2 in long bone (without bone marrow) from control and B16F10-injected mice ( n ≥ 6). d PCA plot showing sample patterns of individual samples in the control and B16F10 groups. e KEGG pathway analysis indicating “Ferroptosis” as the significantly altered pathway. f Heatmap of differentially expressed genes in ferroptosis pathways. Statistical significance was determined by a 2-tailed Student’s t-test (A, B, C)

Article Snippet: To quantify osteocyte death percentages, both in vivo and in vitro, the One-step TUNEL In Situ Apoptosis Kit (biomol, Cat# E-CK-A320) protocol was followed as per the manufacturer’s instructions.

Techniques: Staining, Control, Injection, TUNEL Assay, Expressing

Journal: Bone Research

Article Title: Melanoma bone metastasis-induced osteocyte ferroptosis via the HIF1α-HMOX1 axis

doi: 10.1038/s41413-024-00384-y

Figure Lengend Snippet: Znpp Rescues B16F10-Induced Autophagy-Dependent Ferroptosis In Vivo. a H&E staining of cortical bones from mice injected with B16F10 and treated with DMSO, 1 mg/kg Fer-1, or 10 mg/kg Znpp (i.p.). Quantification of filled, dying, and empty lacunae ( n ≥ 8 per group). White arrows: normal osteocytes; green arrows: dying osteocytes; red arrows: dead osteocytes. Scale bars: 20 μm. b µCT analysis of trabecular bone from mice described in (A), including bone volume per total volume (Tb.BV/TV), separation (Tb.Sp), number (Tb.N), thickness (Tb.Th), and connective density (Conn.D) in tibial bones ( n = 4 per group). c µCT analysis of cortical bone mice described in (A), including bone volume per total volume (Ct.BV/TV), separation (Ct.Sp), and thickness (Ct.Th) in tibial bones ( n = 4 per group). d TUNEL staining and quantification of TUNEL-positive cells in cortical bones ( n ≥ 4 per group) mice described in (A). White arrows indicate TUNEL-positive cells. Statistical significance was determined by a 2-tailed Student’s t-test

Article Snippet: To quantify osteocyte death percentages, both in vivo and in vitro, the One-step TUNEL In Situ Apoptosis Kit (biomol, Cat# E-CK-A320) protocol was followed as per the manufacturer’s instructions.

Techniques: In Vivo, Staining, Injection, TUNEL Assay

Journal: Bone Research

Article Title: Melanoma bone metastasis-induced osteocyte ferroptosis via the HIF1α-HMOX1 axis

doi: 10.1038/s41413-024-00384-y

Figure Lengend Snippet: Roxadustat Enhances B16F10-Induced Ferroptosis in Osteocytes. a HIF1α immunofluorescence staining and quantification of HIF1α-positive cells in proximal tibia from B16F10-injected mice treated with DMSO or 10 mg/kg Roxadustat (i.p.) daily. Scale bars: 20 μm. b H&E staining of cortical bone from mice described in (A). Quantification of filled, dying, and empty lacunae ( n ≥ 8 per group). White arrows: normal osteocytes; green arrows: dying osteocytes; red arrows: dead osteocytes. Scale bars: 20 μm. c µCT analysis of trabecular bone from mice described in (A), showing trabecular bone volume (Tb.BV/TV), separation (Tb.Sp), number (Tb.N), thickness (Tb.Th), and connectivity density. d µCT analysis of cortical bone from mice described in (A), showing cortical bone volume (Ct.BV/TV), separation (Ct.Sp), and thickness (Ct.Th). e TUNEL staining and quantification of TUNEL-positive cells in cortical bone from mice described in (A). Scale bars: 20 μm. f mRNA expression of Dmp1, Dkk1, Sclerostin, and Phex in long bone tissue from mice described in (A). Statistical significance was determined by a 2-tailed Student’s t-test

Article Snippet: To quantify osteocyte death percentages, both in vivo and in vitro, the One-step TUNEL In Situ Apoptosis Kit (biomol, Cat# E-CK-A320) protocol was followed as per the manufacturer’s instructions.

Techniques: Immunofluorescence, Staining, Injection, TUNEL Assay, Expressing

Journal: Frontiers in Pharmacology

Article Title: Bone marrow mesenchymal stem cell-derived small extracellular vesicles promote liver regeneration via miR-20a-5p/PTEN

doi: 10.3389/fphar.2023.1168545

Figure Lengend Snippet: BMSC-sEV infusion enhances liver regeneration in LPS/D-GalN-induced acute liver failure. (A) Representative images of PCNA (dark brown nuclei) staining in liver tissue. Scale bar: 100 μm. (B) PCNA-reactive hepatocyte nuclei were quantified by digital image analysis. (C) Representative images of TUNEL staining in liver tissue at 12 h. Scale bar: 100 μm. (D) Quantification of TUNEL-positive cells. (E) Distribution of DiR-labeled BMSC-sEVs in organs at 3/6 h after intravenous injection in mice. Data are presented as mean ± SEM. n = 6. For each animal, the value was calculated from the average of four randomly chosen fields of view under the microscope. Statistical analysis was performed using two-way ANOVA, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: At 48 h after ALF induction, gene expression was measured, cell proliferation was detected by EdU staining (RiboBio Co. Ltd., #C10310), and apoptosis was detected by TUNEL staining (RiboBio Co. Ltd., #C11012), according to the manufacturer’s instructions.

Techniques: Staining, TUNEL Assay, Labeling, Injection, Microscopy

Journal: Frontiers in Pharmacology

Article Title: Bone marrow mesenchymal stem cell-derived small extracellular vesicles promote liver regeneration via miR-20a-5p/PTEN

doi: 10.3389/fphar.2023.1168545

Figure Lengend Snippet: miR-20a-5p promotes hepatocyte proliferation and inhibits apoptosis via the PTEN/AKT pathway. (A) The viability of L-02 cells transfected with miR-20a-5p mimic/inhibitor was tested using the CCK-8 assay under normal conditions or after hydrogen peroxide-induced damage. (B-C) Western blotting was used to detect protein levels at 48 h after hydrogen peroxide-induced damage in L-02 cells transfected with miR-20a-5p mimic/inhibitor. (D-E) EdU staining was used to detect proliferation at 48 h after hydrogen peroxide-induced damage in L-02 cells transfected with miR-20a-5p mimic. (F-G) TUNEL staining was used to detect apoptosis. Scale bar: 200 μm. The positive rate of cells was calculated based on the average of four random fields per well under the microscope. (H) qPCR was used to detect the relative expression levels of PTEN, CCND1, BCL2 , and BAX . Data are presented as mean ± SEM. Statistical analysis was performed using Student’s t- test, * p < 0.05, ** p < 0.01, *** p < 0.001. n = 3.

Article Snippet: At 48 h after ALF induction, gene expression was measured, cell proliferation was detected by EdU staining (RiboBio Co. Ltd., #C10310), and apoptosis was detected by TUNEL staining (RiboBio Co. Ltd., #C11012), according to the manufacturer’s instructions.

Techniques: Transfection, CCK-8 Assay, Western Blot, Staining, TUNEL Assay, Microscopy, Expressing

Journal: Frontiers in Pharmacology

Article Title: Bone marrow mesenchymal stem cell-derived small extracellular vesicles promote liver regeneration via miR-20a-5p/PTEN

doi: 10.3389/fphar.2023.1168545

Figure Lengend Snippet: BMSC-sEVs inhibit PTEN expression and promote proliferation through miR-20a-5p but inhibit apoptosis independently of miR-20a-5p. L-02 cells transfected with miR-20a-5p inhibitors were rescued with BMSC-sEVs after hydrogen peroxide-induced injury. (A-B) EdU staining was used to detect cell proliferation. (C-D) Detection of apoptosis by TUNEL staining. Scale bar: 200 μm. The positive rate of cells was calculated based on the average of four random fields per well under the microscope. n = 3. (E-F) Protein levels of PTEN and cleaved caspase-3. n = 3. (G) Levels of miR-20a precursors in liver tissues of mice with LPS/D-GalN-induced ALF after BMSC-sEV treatment. n = 6. (H) The intracellular miR-20a precursor levels were detected at 24 h after adding BMSC-sEVs to hydrogen peroxide-damaged L-02 cells. n = 3. Data are presented as mean ± SEM. Statistical analysis was performed using Student’s t- test, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: At 48 h after ALF induction, gene expression was measured, cell proliferation was detected by EdU staining (RiboBio Co. Ltd., #C10310), and apoptosis was detected by TUNEL staining (RiboBio Co. Ltd., #C11012), according to the manufacturer’s instructions.

Techniques: Expressing, Transfection, Staining, TUNEL Assay, Microscopy