Biological activities of cytokine-neutralizing hyaluronic acid-antibody conjugates.
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.
Sun
Liang Tso
LT
Department of Biomedical Engineering, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA.
Bencherif
Sidi A
SA
Gilbert
Thomas W
TW
Farkas
Adam M
AM
Lotze
Michael T
MT
Washburn
Newell R
NR
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*
D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D018796:Immunoconjugates; D053583:Interleukin-1beta; D014409:Tumor Necrosis Factor-alpha; D006820:Hyaluronic Acid