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.