title: Biological activities of cytokine-neutralizing hyaluronic acid-antibody conjugates.
abstract: Wound healing represents a highly regulated, orchestrated response of cells recruited to sites of injury. High molecular weight hyaluronic acid was conjugated with monoclonal antibodies to the cytokine interleukin-1beta to create a matrix-forming polymer capable of modifying healing. Using gel electrophoresis and fluorescence immunosorbent assays, we determined a degree of antibody functionalization per hyaluronic acid chain of 13.6+/-1.6%. The biological activity of the conjugate in vitro, measured using a nuclear factor-kappaB translocation assay in activated THP-1 monocytes, was comparable in inhibiting cytokine signaling to a similar level as the unconjugated antibody. Incisional wound studies in Sprague-Dawley rats indicates that viscous hyaluronic acid solutions were immunologically active, but covalent functionalization with antibodies against tumor necrosis factor-alpha and interleukin-1beta resulted in significant reductions in the inflammatory response. Covalent attachment of cytokine-neutralizing antibodies to matrix-forming polymers could lead to the development of materials capable of locally regulating wound healing and inflammatory responses in the setting of tissue regeneration.
authors.affiliation: Department of Biomedical Engineering, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA.
authors.forename: Liang Tso
authors.initials: LT
authors.lastname: Sun
authors.forename: Sidi A
authors.initials: SA
authors.lastname: Bencherif
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Gilbert
authors.forename: Adam M
authors.initials: AM
authors.lastname: Farkas
authors.forename: Michael T
authors.initials: MT
authors.lastname: Lotze
authors.forename: Newell R
authors.initials: NR
authors.lastname: Washburn
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing / Q000276:immunology*; D045744:Cell Line, Tumor; D005109:Extracellular Matrix / Q000276:immunology; D006801:Humans; D006820:Hyaluronic Acid / Q000276:immunology*; D018796:Immunoconjugates; D066298:In Vitro Techniques; D007249:Inflammation / Q000276:immunology; D053583:Interleukin-1beta / Q000276:immunology*; D009000:Monocytes; D051381:Rats; D017207:Rats, Sprague-Dawley; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*; D014945:Wound Healing / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D018796:Immunoconjugates; D053583:Interleukin-1beta; D014409:Tumor Necrosis Factor-alpha; D006820:Hyaluronic Acid