Journal: Bioactive Materials

Article Title: Immunomodulatory supramolecular hydrogel for rheumatoid arthritis management via adenosine A2A receptor-mediated macrophage remodeling

doi: 10.1016/j.bioactmat.2025.11.031

Figure Lengend Snippet: The design of an inflammatory macrophages-targeted, acid-sensitive PNSH for delivery MTX and its role in promoting A2AR activation on M2 macrophages for arthritis therapy. (A)The structure of acid-sensitive polymer-nanomedicine supramolecular hydrogels, composed of drug-loaded MTX NPs. (B) Schematic illustration of arthritis therapy targeting inflammatory joint network via mPECN NPs-mediated release of MTX. This approach leverages MTX targeting the A2AR and repolarization of macrophages from the pro-inflammatory M1 phenotype to the anti-inflammatory M2 phenotype. Activation of A2AR and macrophage repolarization by mPECN-MTX NPs synergistically enhanced the anti-inflammatory effect of PNSH-mediated nanomedicine in an arthritis rat model.

Article Snippet: The selective adenosine A2A receptor (A2AR) antagonist SCH58261 (HY-19533) was purchased from MedChemExpress (Shanghai, China).

Techniques: Activation Assay, Polymer

Journal: Bioactive Materials

Article Title: Immunomodulatory supramolecular hydrogel for rheumatoid arthritis management via adenosine A2A receptor-mediated macrophage remodeling

doi: 10.1016/j.bioactmat.2025.11.031

Figure Lengend Snippet: MTX@PNSH promotes anti-inflammatory M2 macrophages through the A2AR signaling pathway. (A) The intracellular cAMP levels in Raw 264.7 cells were determined by homogeneous time-resolved fluorescence (HTRF) following different treatments. (B) qRT-PCR analysis of relative mRNA of A2ar with different treatments. (C) Immunostaining of A2AR expression in Raw 264.7 cells with different treatments. (D) Flow cytometry analysis of A2AR + cells in Raw 264.7 cells with different treatments and the percentage of A2AR + cells in Raw 264.7 cells with different treatments. (E) qRT-PCR analysis of relative mRNA expression of Pdl1 without LPS treatment. (F) qRT-PCR analysis of relative mRNA expression of Ido1 without LPS treatment. (G) qRT-PCR analysis of relative mRNA expression of Pdl1 with LPS treatment. (H) qRT-PCR analysis of relative mRNA expression of Ido1 with LPS treatments. (I) Western blotting analysis of A2AR protein level in macrophages after 24 h of different treatment with or without A2AR inhibitor. (J) Quantitative analysis of A2AR protein level in different treatments with or without A2AR inhibitor. (K) Flow cytometry analysis of CD11b and CD39 in Raw 264.7 cells with different treatments and the percentage of CD11b + CD39 + macrophages. (L) Flow cytometry analysis of CD11b and CD73 in Raw 264.7 cells with different treatments and the percentage of CD11b + CD73 + macrophages. (M) Flow cytometry analysis of CD11b and PD-L1 in Raw 264.7 cells after 24 h of different treatments and the percentage of CD11b + PD-L1 + macrophages in Raw 264.7 cells after 24 h of different treatments. Data are presented as mean ± SD, n = 3. Statistical significance was determined using one-way ANOVA, followed by Dunnett's post hoc test for comparisons between groups.

Article Snippet: The selective adenosine A2A receptor (A2AR) antagonist SCH58261 (HY-19533) was purchased from MedChemExpress (Shanghai, China).

Techniques: Fluorescence, Quantitative RT-PCR, Immunostaining, Expressing, Flow Cytometry, Western Blot

Journal: bioRxiv

Article Title: MG53 Protects Against Intestinal Inflammation by Inhibiting NLRP3 Inflammasome Activation

doi: 10.64898/2026.02.18.706669

Figure Lengend Snippet: A . Immunoblot of IL-1β p17 and caspase-1 p20 in supernatants (Sup.) from lipopolysaccharide (LPS)-primed mouse bone marrow–derived macrophages (BMDMs) (100 ng/ml, 3 h) treated with rhMG53 for 1 h and stimulated with nigericin for 1 h. Pro–IL-1β, pro–caspase-1, NOD-like receptor family pyrin domain containing 3 (NLRP3), and MG53 were analyzed in cell lysates. B . IL-1β in culture supernatant was measured by enzyme-linked immunosorbent assay (ELISA). Treatments as in ( A ). n=3. C . Immunoblot of IL-1β p17 and caspase-1 p20 in supernatants from LPS-primed BMDMs treated with rhMG53 (10 μg/ml, 3 h) and stimulated with nigericin (10 μg/ml, 1 h), adenosine 5′-triphosphate disodium salt (ATP) (5 mM, 30 min), or monosodium urate crystals (MSU) (150 μg/ml, 5 h). Pro–IL-1β, pro– caspase-1, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) and MG53 were assessed in lysates. D . ELISA of IL-1β release in culture supernatants. Treatments as in ( C ). n=3. E. BMDMs were treated with rhMG53 (1, 5, 10 μg/ml) for 1h before or after LPS stimulation, and NLRP3 inflammasome-related proteins were examined by immunoblotting. F . Immunoblot of IL-1β p17 and caspase-1 p20 in culture supernatants of LPS-primed WT or MG53-overexpressing transgenic mice (TPA) BMDMs treated for 4 h and stimulated with nigericin (10 μg/ml, 1 h) or ATP (5 mM, 30 min). MG53, NLRP3, ASC, pro-IL-1β, and pro-caspase-1 were analyzed in cell lysates. G . ELISA of IL-1β release in supernatants from LPS-primed WT or TPA BMDMs treated for 3 h and stimulated with nigericin (10 μg/ml,1h) or ATP (5 mM, 30 min). n=5. H . THP-1 cells differentiated with PMA were primed with LPS (0.5 μg/ml, 4 h), treated with rhMG53 at the indicated concentrations for 1 h, and stimulated with nigericin (1 μg/ml, 2 h). Supernatants and cell lysates were analyzed by immunoblotting. IL-1β p17 and caspase-1 p20 were detected in supernatants, while pro-caspase-1, pro-IL-1β, NLRP3, ASC, and MG53 were detected in lysates. I . ELISA measurement of IL-1β release in culture supernatants. n=3. Data are mean ± SD. Statistical significance was determined by one-way ANOVA ( B and I ) and unpaired Student’s t-test ( D and G ), *** p < 0.005, ** p < 0.01, * p < 0.05.

Article Snippet: Lipopolysaccharide (LPS) from Escherichia coli 0111: B4 (tlrl-eblps), Nigericin sodium salt (tlrl-nig), adenosine 5′-triphosphate disodium salt (ATP, tlrl-atpl) and monosodium urate crystals (MSU, tlrl-msu) were from InvivoGen.

Techniques: Western Blot, Derivative Assay, Enzyme-linked Immunosorbent Assay, Transgenic Assay