Journal: Molecular Metabolism

Article Title: Indoleamine 2,3-dioxygenase mediates the therapeutic effects of adipose-derived stromal/stem cells in experimental periodontitis by modulating macrophages through the kynurenine-AhR-NRF2 pathway

doi: 10.1016/j.molmet.2022.101617

Figure Lengend Snippet: ASC therapeutic effects on experimental periodontitis in rats. (A) The schematic diagram showed the induction of the experimental rat model of periodontitis for 21 days and ASC injection after the removal of ligatures. (B) Micro-CT imaging and bone parameters (CEJ-ABC distance, BV/TV, Tb. Th, and Tb. Sp) between healthy and experimental rats of periodontitis. Scale bar, 1 mm. (C) Micro-CT imaging displayed the alveolar bone of PBS-injected and ASC-injected rats on day 7, day 14, and day 21 Scale bar, 1 mm. (D) The CEJ-ABC distance and bone parameters (BV/TV, Tb. Th, and Tb. Sp) between the PBS-injected and ASC-injected rats. (E) Representative immunofluorescence staining of M1 phenotype macrophages (iNOS + ; Green) and M2 macrophages (CD206 + ; Red) in sagittal sections of maxillary molars in PBS-injected and ASC-injected rats. Scale bar, 20 μm. (F) The calculated ratio of iNOS + /CD206 + macrophages in PBS-injected and ASC-injected groups. CEJ-ABC, cementoenamel junction and alveolar bone crest. All data were expressed as mean ± SD; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: For tissues, the sections were incubated with a mouse anti-CD206 monoclonal antibody (1:100, 60143-1-lg, Proteintech), a rabbit anti-NRF2 monoclonal antibody (1:100, A0674, Abclonal), a rabbit anti-inducible nitric oxide synthase (iNOS) polyclonal antibody (1:100, 18985-1-AP, Proteintech) overnight at 4 °C.

Techniques: Injection, Micro-CT, Imaging, Immunofluorescence, Staining

Journal: Molecular Metabolism

Article Title: Indoleamine 2,3-dioxygenase mediates the therapeutic effects of adipose-derived stromal/stem cells in experimental periodontitis by modulating macrophages through the kynurenine-AhR-NRF2 pathway

doi: 10.1016/j.molmet.2022.101617

Figure Lengend Snippet: ASCs modulated macrophage polarization through NRF2. (A) Representative immunohistochemical staining of NRF2 in the PBS-injected and ASC-injected rats on day 7, day 14, and day 21. Scale bar, 50 μm. (B) Representative immunofluorescence staining of NRF2 (Green) and M2 macrophages (CD206 + ; Red) in sagittal sections of maxillary molars in PBS-injected and ASC-injected rats. Scale bar, 20 μm. (C) Schematic diagram of knocking down NRF2 in macrophages and macrophages cocultured with ASCs and LPS stimulation. (D, E) RT-qPCR and western blotting were used to measure the NRF2 siRNA or siNC in macrophages. (F, G) The mRNA expression and protein levels of M1 markers were increased, while those of M2 markers were decreased after silencing NRF2 in the cocultured groups. R, root; PDL, periodontal ligament; AB, alveolar bone; siRNA, small interfering RNA; NC, negative control. All data were expressed as mean ± SD; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: For tissues, the sections were incubated with a mouse anti-CD206 monoclonal antibody (1:100, 60143-1-lg, Proteintech), a rabbit anti-NRF2 monoclonal antibody (1:100, A0674, Abclonal), a rabbit anti-inducible nitric oxide synthase (iNOS) polyclonal antibody (1:100, 18985-1-AP, Proteintech) overnight at 4 °C.

Techniques: Immunohistochemical staining, Staining, Injection, Immunofluorescence, Quantitative RT-PCR, Western Blot, Expressing, Small Interfering RNA, Negative Control

Journal: Molecular Metabolism

Article Title: Indoleamine 2,3-dioxygenase mediates the therapeutic effects of adipose-derived stromal/stem cells in experimental periodontitis by modulating macrophages through the kynurenine-AhR-NRF2 pathway

doi: 10.1016/j.molmet.2022.101617

Figure Lengend Snippet: Inhibition of IDO activity reduced the therapeutic potential of ASCs in experimental periodontitis and downregulated NRF2 expression. (A) Three-dimensional reconstruction of maxillary alveolar bone in ASC-injected and 1-MT pretreated ASC-injected rats on day 7, day 14, and day 21. Scale bar, 1 mm. (B) Analysis of bone parameters. (C) Immunohistochemical staining of IL-1β, OCN, and NRF2 in the ASC-injected and 1-MT pretreated ASC-injected rats. Scale bar, 50 μm. (D) Representative immunofluorescence staining of M1 phenotype macrophages (iNOS + ; Green) and M2 macrophages (CD206 + ; Red) in sagittal sections of maxillary molars in ASC-injected and 1-MT pretreated ASC-injected rats. Scale bar, 20 μm. (E) The mean IOD of IL-1β, OCN, and NRF2 and the calculated ratio of iNOS + /CD206 + . IOD, integrated optical density.

Article Snippet: For tissues, the sections were incubated with a mouse anti-CD206 monoclonal antibody (1:100, 60143-1-lg, Proteintech), a rabbit anti-NRF2 monoclonal antibody (1:100, A0674, Abclonal), a rabbit anti-inducible nitric oxide synthase (iNOS) polyclonal antibody (1:100, 18985-1-AP, Proteintech) overnight at 4 °C.

Techniques: Inhibition, Activity Assay, Expressing, Injection, Immunohistochemical staining, Staining, Immunofluorescence

Journal: Journal of Neuroinflammation

Article Title: Presence and activation of pro-inflammatory macrophages are associated with CRYAB expression in vitro and after peripheral nerve injury

doi: 10.1186/s12974-021-02108-z

Figure Lengend Snippet: Quantification of the number of pro-inflammatory and immunosuppressive macrophages in WT and Cryab −/− crushed sciatic nerves. Representative images and quantification of the number of CD16/32 + Iba1 + DAPI + ( a , c ) and CD206 + Iba1 + DAPI + ( b , d ) profiles in the sciatic nerves of naïve (N), and 3, 5, 7, 14, 21, and 28 days post-crushed WT (white bars) and Cryab −/− (black bars) animals; representative of 2 experiments, n =3–4 animals/group, bar = 20 μm, data represent mean ± sem, * p < 0.05 independent t -test. e Graph displaying the ratio of CD16/32 + cells to CD206 + cells for WT and Cryab − / − nerves

Article Snippet: The tissues were incubated overnight at 4 °C with rabbit anti-Iba1 (Wako Chemicals, 019-1974, 1:200) or goat anti-Iba1 (Novus Biologicals, NB100-1028, 1:100), rat anti-mouse CD16/32 (BD Biosciences, 553842, 1:500), goat anti-mouse CD206 (Santa Cruz Biotechnologies, sc-34577, 1:500), chicken anti-P0 (Abcam, ab39375, 1:500), rabbit anti-CD68 (Cell Signaling, 97778, 1:100), rabbit anti-arginase 1 (Cell Signaling, 93668, 1:50), rabbit anti-iNOS (BD Transduction Laboratories, 1:100; gift from Dr. Samuel David, McGill University), and DAPI (Invitrogen, D3571, 1:2000).

Techniques:

Journal: Stem cell research & therapy

Article Title: Implantation and tracing of green fluorescent protein-expressing adipose-derived stem cells in peri-implant capsular fibrosis.

doi: 10.1186/s13287-023-03248-0

Figure Lengend Snippet: Fig. 2 A, B Representative images and quantification of α-smooth muscle actin (αSMA) in the control and green fluorescent protein-adipose-derived stem cell (GFP-ASC) groups. C, D Representative images and quantification of F4/80-positive macrophages in the control and GFP-ASC groups. E, F Representative images and quantification of iNOS-positive cells in F4/80-positive macrophages in the control and GFP-ASC groups. G, H Representative images and quantification of CD206-positive cells in F4/80-positive macrophages in the control and GFP-ASC groups. Values are presented as mean ± SD (n = 6). Scale bars = 100 μm. **p < 0.01. ***p < 0.001

Article Snippet: The cryosectioned slides were blocked with 5% bovine serum albumin diluted in PBS for 1 h at 37 °C, and then incubated with one of the following primary antibodies overnight at 4 °C: rabbit anti-mouse GFP antibody (AB3080; Millipore, Darmstadt, Germany), goat anti-mouse GFP antibody (ab5450; Abcam, Cambridge, UK), rat anti-mouse Ki67 antibody (14-5698-82; Invitrogen, Waltham, USA), rabbit anti-mouse PH3 antibody (9664; Cell signaling, Danvers, USA), rabbit anti-mouse Caspase3 antibody (53348; Cell signaling, Danvers, USA), mouse Cy3-conjugated anti-α-smooth muscle actin (αSMA) antibody (clone 1A4; Merck, Kenilworth, USA), rat anti-mouse F4/80 antibody (MAB5580; R&D Systems, Minneapolis, USA), mouse anti-mouse inducible nitric oxide synthase (iNOS) antibody (MAB9502; R&D Systems, Minneapolis, USA), goat anti-mouse CD206 antibody (AF2535; R&D Systems, Minneapolis, USA), or an Armenian hamster anti-mouse CD31 antibody (clone 2H8; Millipore, Darmstadt, Germany).

Techniques: Control, Derivative Assay

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