Nucleic acid scavenging microfiber mesh inhibits trauma-induced inflammation and thrombosis

Abstract

Trauma patients produce a host of danger signals and high levels of damage-associated molecular patterns (DAMPs) after cellular injury and tissue damage. These DAMPs are directly and indirectly involved in the pathogenesis of various inflammatory and thrombotic complications in patients with severe injuries. No effective therapeutic agents for the removal of DAMPs from blood or tissue fluid have been developed. Herein, we demonstrated that nucleic acid binding polymers, e.g., polyethylenimine (PEI) and polyamidoamine dendrimers, immobilized onto electrospun microfiber mesh can effectively capture various DAMPs, such as extracellular DNAs and high mobility group box 1 (HMGB1). Furthermore, treatment with PEI-immobilized microfiber mesh abrogated the ability of DAMPs, released from dead and dying cells in culture or found in patients following traumatic injury, to activate innate immune responses and coagulation in vitro and in vivo. Nucleic acid scavenging microfiber meshes represent an effective strategy to combat inflammation and thrombosis in trauma.

Damage‐Associated Molecular Patterns Induce Inflammatory Injury During Machine Preservation of the Liver: Potential Targets to Enhance a Promising Technology

“…To assess the biological activity of DAMPs released into the perfusate, an in vitro assay was performed using commercially available TLR reporter cell lines. In this assay, TLR stimulation produces activation of NF‐κB, which is reflected by secretion of alkaline phosphatase in the cell media and detected by a colorimetric assay . Perfusate samples were incubated with TLR3, TLR4, and TLR9 reporter cell lines, which are activated by extracellular RNA, HMGB1, and exDNA, respectively …”

“…In this assay, TLR stimulation produces activation of NF-κB, which is reflected by secretion of alkaline phosphatase in the cell media and detected by a colorimetric assay. (23) Perfusate samples were incubated with TLR3, TLR4, and TLR9 reporter cell lines, which are activated by extracellular RNA, HMGB1, and exDNA, respectively. (24,25) Perfusate samples from the end of MP demonstrated differential capacity for TLR stimulation with significantly increased activation of TLR3 and TLR9 by perfusate from the MP37 group (Fig.…”

Cationic nanoparticle as an inhibitor of cell-free DNA-induced inflammation

“…Using these nucleic acid-binding polymers (NABP) to remove the DAMP molecules, we had demonstrated that the scavenging approach works in animal models of CpG-induced or RNA-induced acute liver injury 19 , thrombosis 29 , and SLE 20 , 30 . To reduce toxicity, we had immobilized the NABP on fibrous membrane to remove the DAMP molecules in a local 31 or ex vivo manner 32 . Motivated by the high NA-binding capacity of the immobilized configuration of NABP in these recent studies, we propose to use cNP as a DAMP scavenger, which has not been studied before, for RA treatment.…”

Damage- and pathogen-associated molecular patterns play differential roles in late mortality after critical illness

“…Upon binding to their cognate ligands, TLR signaling activates NF-κB that leads to expression and release SEAP from the TLR reporter cells. The level of NF-κB activity was measured by quantification of SEAP in culture media using a colorimetric assay, as described previously (39). To stimulate mouse macrophages, RAW264.7 cells were incubated overnight with DAMPs or PAMPs.…”

“…Plasma coagulation assay was performed by described previously (39). Briefly, 50 μL normal pooled mouse plasma in sodium citrate (C57BL/6) (Biochemed Services) or 50 μL normal pooled human plasma in sodium citrate (George King Bio-Medical Inc.) was incubated for 5 minutes with 5 μL DAMPs or PAMPs at 37°C, followed by the addition of CaCl 2 (25 mM).…”