Journal: Biomaterials Research

Article Title: Bioactive self-healing hydrogel based on tannic acid modified gold nano-crosslinker as an injectable brain implant for treating Parkinson’s disease

doi: 10.1186/s40824-023-00347-0

Figure Lengend Snippet: Biocompatibility evaluation of the hydrogels for treating PD rats. A In vivo immunofluorescent images of CD86 positive (CD86+) M1 macrophages (red) and CD163 positive (CD163+) M2 macrophages (green) for the explanted tissue after implantation in the brain for 14 days. Quantification for ( B ) the fluorescent intensity of M1 and M2 macrophages and ( C ) the ratios of M2/M1 in vivo. * p < 0.05, ** p < 0.01, and **** p < 0.0001 between the indicated groups

Article Snippet: The primary antibodies used in this study involved Anti Iba1 (Rabbit, for immunocytochemistry, 019–19,741, 1:500, Wako Pure Chemical Industries, Ltd., Japan), ABclonal rabbit polyclonal anti-CD86 antibody (A1199, 1:200, ABclonal Inc., USA), ABclonal rabbit polyclonal anti-CD163 antibody (A8383, 1:200, ABclonal Inc., USA), recombinant anti- tyrosine hydroxylase (TH) antibody (EP1533Y/ab75875, 1:50, Abcam, UK), and mouse monoclonal anti-GFAP antibody (14–9892-82, 1:100, Invitrogen, USA).

Techniques: In Vivo

Journal: Materials today. Bio

Article Title: Curcumin-loaded polydopamine nanoparticles-based antioxidant scaffold promote spinal cord repair though dural-regulation of macrophage polarization.

doi: 10.1016/j.mtbio.2025.101895

Figure Lengend Snippet: Fig. 2. In vitro culture reveals the macrophage polarization regulation by curcumin release. NC:cell culture without LPS, Ctrl:cell culture with LPS, Gel:cell culture on gel coated plate with LPS. PDA@Gel: cell culture on PDA included gel with LPS, Cur-PDA@Gel:cell culture on Cur-PDA micellel included gel with LPS. A: Immune staining of ROS positive cells in different conditions. ROS positive cells (green), Dapi (blue). B: Ratio of ROS positive cells quantified by flow cytometry. Scale bar indicate 30 μm. C: Quantification of M1 (CD86 positive) polarized macrophage and M2 (CD206 positive) polarized macrophage RAW 264.7 through flow cytometry. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Rabbit anti-Stat6 polyclonal antibod, rabbit anti-p-Stat6 polyclonal antibody, rab-p65 polyclonal antibody, rabbit anti-p-p65 polyclonal antibod were purchased from Abmart® (Shanghai, China).APC anti-CD206 antibody and PE anti-CD86 antibody were obtained from Elabscience (Wuhan, China).

Techniques: In Vitro, Cell Culture, Staining, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: Huanshaodan regulates microglial glucose metabolism reprogramming to alleviate neuroinflammation in AD mice through mTOR/HIF-1α signaling pathway

doi: 10.3389/fphar.2024.1434568

Figure Lengend Snippet: Expression levels of CD86, Arg1 and IL-1β protein in hippocampus and cortex of SAMP8 mice detected by Western-blot, the expression levels of IL-6 and TNF-α in hippocampus and cortex of SAMP8 mice by ELISA. (A,E) Blotting of CD86, Arg1 and IL-1β protein in hippocampus and cortex of mice in each group; (B–D) Expression of CD86, Arg1 and IL-1β protein in hippocampus of mice in each group; (F–H) Expression of CD86, Arg1 and IL-1β protein in cortex of mice in each group; (I–L) Expression of IL-6 and TNF-α protein in hippocampus and cortex of mice in each group. Model group compared with the control group * p < 0.05, ** p < 0.01, *** p < 0.001, Donepezil group compared with the model group # p < 0.05, ## p < 0.01, ### p < 0.001, HSD group compared with the model group $ p < 0.05, $$ p < 0.01, $$$ p < 0.001.

Article Snippet: The main materials that were used in this study were LPS (L8880, Solarbio), MHY1485 (MHY) (M9050-5MG, AbMole), RIPA buffer (high) (R0010, Solarbio), primary antibodies β-Actin and CD86 (42kDa, BM0627; 70kDa, BM4121, BOSTER), Arg1 and mTOR (40kDa, GB11285-100; 289kDa, GB111839-100, Servicebio), p-mTOR and HIF-1α (289kDa, 5536T; 120kDa, 48085S, Cell Signaling Technology), IL-1β (31kDa, 16806-1-AP, Proteintech), Horseradish peroxidase (HRP)-linked secondary antibodies against rabbit and mouse IgG of the primary antibodies (ZB-2301; ZB-2305, Beijing Zhong Shan-Golden Bridge Biological Technology), Glucose (GLU) Assay Kit, Lactic Acid (LD) assay kit, ATP assay kit, Lactate dehydrogenase (LDH) assay kit (A154-1-1; A019-2-1; A095-1-1; A020-2-2, Nanjing Jiancheng Bioengineering Institute), Pyruvate Dehydrogenase (PDH) Activity Assay Kit (E-BC-K650-M, Elabscience), BCA Protein Assay Kit (PC0020, Solarbio), ELISA Kit for Hexokinase 2 (HK2) activity based on enzyme standard of HK2, ELISA Kit for Tumor Necrosis Factor alpha (TNF-α) (MM-45418M1; MM-0132M1, MEIMIAN), ELISA Kit for Interleukin 6 (IL-6) (E-MSEL-M0001, Elabscience) and ELISA Kit for Pyruvate kinase isozymes M2 (PKM2) (SEA588Mu, Cloud Clone Corp), Cell Staining Buffer, PE Anti-Mouse F4/80 Antibody [CI:A3-1], PE/Cyanine7 Anti-Mouse CD86 Antibody [GL-1], Intracellular Fixation/Permeabilization Buffer Kit and APC Anti-Mouse CD206 Antibody (E-CK-A107; E-AB-F0995D; E-AB-F0994H; E-CK-A109; E-AB-F1135E, Elabscience).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: Frontiers in Pharmacology

Article Title: Huanshaodan regulates microglial glucose metabolism reprogramming to alleviate neuroinflammation in AD mice through mTOR/HIF-1α signaling pathway

doi: 10.3389/fphar.2024.1434568

Figure Lengend Snippet: The expression levels of p-mTOR/mTOR ratio, HIF-1α, and IL-1β in BV2 cells tested by western blot, the levels of GLU, LD, ATP, LDH and PDH quantified by biochemical test kit and the levels of polarization biomarkers of CD86 and CD206 detected by flow cytometry, the expression levels of IL-6 and TNF-α in BV2 cells tested by ELISA. (A) Blotting of p-mTOR, mTOR and HIF-1α protein in BV2 cells in each group; (B, C) Expression levels of p-mTOR/mTOR ratio and HIF-1α in BV2 cells in each group. (D) GLU content analysis chart; (E) LD content analysis chart; (F) ATP content analysis chart; (G) . LDH activity analysis graph; (H) PDH activity analysis graph. (I) Flow cytometry analysis charts; (J) Flow cytograms of BV2 cells in each group; (K) Blotting of IL-1β protein in BV2 cells in each group; (L) Expression level of IL-1β in BV2 cells in each group. (M, N) Expression level of IL-6 and TNF-α in BV2 cells in each group. Model group compared with control group * p < 0.05, ** p < 0.01, *** p < 0.001, HSD-containing serum-treated group compared with the model group $ p < 0.05, $$ p < 0.01, $$$ p < 0.001, Reverse validation group compared with the HSD-containing serum-treated group & p < 0.05, && p < 0.01, &&& p < 0.001.

Article Snippet: The main materials that were used in this study were LPS (L8880, Solarbio), MHY1485 (MHY) (M9050-5MG, AbMole), RIPA buffer (high) (R0010, Solarbio), primary antibodies β-Actin and CD86 (42kDa, BM0627; 70kDa, BM4121, BOSTER), Arg1 and mTOR (40kDa, GB11285-100; 289kDa, GB111839-100, Servicebio), p-mTOR and HIF-1α (289kDa, 5536T; 120kDa, 48085S, Cell Signaling Technology), IL-1β (31kDa, 16806-1-AP, Proteintech), Horseradish peroxidase (HRP)-linked secondary antibodies against rabbit and mouse IgG of the primary antibodies (ZB-2301; ZB-2305, Beijing Zhong Shan-Golden Bridge Biological Technology), Glucose (GLU) Assay Kit, Lactic Acid (LD) assay kit, ATP assay kit, Lactate dehydrogenase (LDH) assay kit (A154-1-1; A019-2-1; A095-1-1; A020-2-2, Nanjing Jiancheng Bioengineering Institute), Pyruvate Dehydrogenase (PDH) Activity Assay Kit (E-BC-K650-M, Elabscience), BCA Protein Assay Kit (PC0020, Solarbio), ELISA Kit for Hexokinase 2 (HK2) activity based on enzyme standard of HK2, ELISA Kit for Tumor Necrosis Factor alpha (TNF-α) (MM-45418M1; MM-0132M1, MEIMIAN), ELISA Kit for Interleukin 6 (IL-6) (E-MSEL-M0001, Elabscience) and ELISA Kit for Pyruvate kinase isozymes M2 (PKM2) (SEA588Mu, Cloud Clone Corp), Cell Staining Buffer, PE Anti-Mouse F4/80 Antibody [CI:A3-1], PE/Cyanine7 Anti-Mouse CD86 Antibody [GL-1], Intracellular Fixation/Permeabilization Buffer Kit and APC Anti-Mouse CD206 Antibody (E-CK-A107; E-AB-F0995D; E-AB-F0994H; E-CK-A109; E-AB-F1135E, Elabscience).

Techniques: Expressing, Western Blot, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Activity Assay, Control, Biomarker Discovery