Anti-inflammatory Effect of Self-assembling Glycol-Split Glycosaminoglycan-Stearylamine Conjugates in Lipopolysaccharide-Stimulated Macrophages

Abstract

Glycosaminoglycans (GAGs) play important roles in various biological processes such as cell adhesion and signal transduction, as well as promote anti-inflammatory activity. We previously revealed that glycolsplit heparin (HP)-aliphatic amine conjugates form self-assembled nanoparticles and suppress the production of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β in lipopolysaccharide (LPS)-stimulated macrophages much more strongly than native HP (J. Control. Release, 194, 2014, Babazada et al.). Considering that HP is not the only GAG to have anti-inflammatory activity, the present study was initiated to examine whether conjugation of GAGs with aliphatic amines is generally effective in their activity augmentation against LPS-stimulated macrophages. We newly synthesized the stearylamine conjugates of chondroitin sulfate (CS), hyaluronic acid (HA), and low-molecular-weight heparin (LH), and investigated the effect of the position and degree of sulfation and molecular weight of GAGs on their anti-inflammatory activity. All of the conjugates formed self-assembled nanoparticles in aqueous solution. The IC 50 value for suppression of TNF-α production from the macrophages was the smallest with the derivative of LH, followed by HP, CS, and HA. The degree of sulfation appeared to be important in determining their anti-inflammatory activity, which would correspond to previous results using the derivatives of siteselectively desulfated HP. Comparison of HP and LH derivatives revealed that fractionated smaller heparin has greater anti-inflammatory activity.Key words glycosaminoglycan; self-assembled nanoparticle; anti-inflammatory activity; Toll-like receptor 4; macrophage; structure-activity relationship Glycosaminoglycans (GAGs) are linear polysaccharides that play important roles in processes such as cell adhesion and signal transduction. 1) Heparin (HP) is one of the highly sulfated GAGs that consist of repeats of alternating uronic acid and D-glucosamine.2) HP is a well-known anticoagulant that activates antithrombin, leading to anticoagulation 3) ; moreover, it has a variety of additional biological effects such as antiangiogenesis and anti-inflammation. Chondroitin sulfate (CS), consisting of repeats of alternating D-glucuronic acid and N-acetyl-D-galactosamine, 2,4) is used clinically to reduce pain and improve articular function in patients with osteoporosis.5,6) Hyaluronic acid (HA) consists of repeats of alternating D-glucuronic acid and N-acetyl-D-glucosamine, but characteristically lacks sulfo groups. 2,4) HA has been used to protect synovial membranes and heal injuries. [7][8][9] Although the biological activity varies for each GAG, a common feature is the presence of an anti-inflammatory effect. Several in vitro [10][11][12][13][14] and in vivo 15,16) studies have shown that HP, CS, and HA suppress the lipopolysaccharide (LPS)-induced production of inflammatory cytokines by inhibiting the activation of MyD88, p38 mitogen-activated protein kinase (MAPK), an...

Capturing Cytokines with Advanced Materials: A Potential Strategy to Tackle COVID‐19 Cytokine Storm

“…Furthermore, investigations of the relationship between anti-inflammatory activity and conjugate structure suggested a critical role of the sulfation levels in regulating activity of such antibody-nanoparticle conjugates. [80] In another study, heparin-embedded HA hydrogel was used to salvage brain tissue after the onset of stroke. [81,82] The loading of vascular endothelial growth factor (VEGF) in the HA hydrogel reduced levels of TNF-α in damaged areas of the brain; meanwhile heparin nanoparticles captured cytokines, reduced scar formation, and eventually facilitated tissue repair after the stroke.…”

Learning from BMPs and their biophysical extracellular matrix microenvironment for biomaterial design

“…In their recent review, Hachim et al [192] described how GAGs-based biomaterials can also be used to reduce the dose of BMPs, given their ability to bind and slowly release BMPs, to their natural composition recognized by remodeling ECM-enzymes and to their anti-inflammatory effect [193]. Heparin has been proposed as carriers for BMPs in several types of biomaterials: microparticles [194][195][196], hydrogels [197] and scaffolds [198,199].…”

Potential nanoparticle applications for prevention, diagnosis, and treatment of COVID‐19

“…For example, a series of NPs with heparin and GAG derivatives have been shown to suppress the production of inflammatory cytokines, such as IL-6, TNF-α, and IL-1β, in lipopolysaccharides-stimulated MΦs. 65,66 Another treatment that could potentially reduce the severity of the CRS systemic effects are cell-membranecoated NPs. MΦ-membrane-coated NPs possess a similar antigenic morphology to MΦs that can capture and neutralize pro-inflammatory cytokines, including IL-6 and TNF-α.…”

“…Such dynamic binding interactions have been integrated into NP formulations for cytokine neutralization. For example, a series of NPs with heparin and GAG derivatives have been shown to suppress the production of inflammatory cytokines, such as IL‐6, TNF‐α, and IL‐1β, in lipopolysaccharides‐stimulated MΦs 65,66 …”