Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: Epigenetic metabolite lipid nanoparticles alleviate venous thrombosis via bone marrow reprogramming

doi: 10.1016/j.bvth.2025.100086

Figure Lengend Snippet: Immunomodulatory effects of ITA-LNP in vitro and in vivo. (A) Immunoblot probing for IL-1β in BMDMs treated with 100 ng/mL LPS in the presence of various doses of ITA-LNPs. (B) Schematic of the experiments in C57BL/6 mice subjected to LPS-induced endotoxemia. (C) Kaplan-Meier survival analysis after LPS-induced (15 mg/kg) endotoxemia in C57BL/6 mice pretreated with Ctrl/ITA-LNPs or anti–IL-1β mAb. (D) Rectal temperature measured in the mice from panel C before and 6 hours after LPS injections. (E-F) The levels of IL-1β and IL-6 in the serum from mice of a separate cohort injected as in panel B but with a sublethal dose of 8 mg/kg LPS; n = 10 to 15 mice per group. (G) NETs in primary human neutrophils incubated for 5 hours in vitro with 50 μM LNPs as indicated and in the presence of 100 nM PMA followed by immunofluorescence analysis; n = 6 for Ctrl-LNP and ITA-LNP without PMA; n = 8 for Ctrl-LNP with PMA; n = 9 for ITA-LNP with PMA; and n = 4 for PBS groups with or without PMA. (H) Cytokine analysis in supernatants from experiments in panel G. Statistical analysis is through pairwise t test with Holm post hoc test . N.d., not detected; PBS, phosphate-buffered saline; ROI, region of interest; T, temperature.

Article Snippet: C57BL/6 male mice (10 weeks old, n = 15 per group) were IV injected with 50 mg/kg ITA- or control (Ctrl)-LNPs or 50 mg/kg anti-mouse IL-1β monoclonal antibody (clone 7E3, InvivoGen mil1b-mab9-10).

Techniques: In Vitro, In Vivo, Western Blot, Injection, Incubation, Immunofluorescence, Saline

Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: Epigenetic metabolite lipid nanoparticles alleviate venous thrombosis via bone marrow reprogramming

doi: 10.1016/j.bvth.2025.100086

Figure Lengend Snippet: ITA-LNPs alleviate venous thrombosis in IVC ligation model. (A) Schematic of the experiments. WT mice IV pretreated (50 mg/kg, bolus) with ITA- or Ctrl-LNPs were subjected to IVC ligation surgery followed by the isolation of the blood clots; n = 6 for Ctrl-LNP and n = 8 for ITA-LNP. (B) Gross pathology with clot weight quantification. (C) Hematoxylin and eosin staining of blood clot sections from these experiments. (D) Immunofluorescence staining of the same sections using Ly6G/C mAb (Red) and nuclei (blue). (E) MPO (top, alkaline phosphatase immunohistochemistry) and IL-1β (bottom, immunofluorescence) staining in the same clots. (F) Quantification of the staining in panel E. For each biological replicate (n = 6 for Ctrl-LNP and n = 8 for ITA = LNP), 3 to 4 histopathological locations at different depths (100 μm apart) were analyzed. (G) NET staining in the same sections using triple-positive identification through colocalization of the signals from MPO (green), H4Cit3 (red), and DNA (DAPI; blue). (H) Quantification of the staining in panel G. Quantification was accomplished using 2 to 3 different histopathological locations and the same aforementioned biological replicates. Statistical analysis is through a 2-tailed t test. DAPI, 4′,6-diamidino-2-phenylindole; H4Cit3, citrullinated histone H4; MPO, myeloperoxidase; WT, wild type.

Article Snippet: C57BL/6 male mice (10 weeks old, n = 15 per group) were IV injected with 50 mg/kg ITA- or control (Ctrl)-LNPs or 50 mg/kg anti-mouse IL-1β monoclonal antibody (clone 7E3, InvivoGen mil1b-mab9-10).

Techniques: Ligation, Isolation, Staining, Immunofluorescence, Immunohistochemistry