title: Identification and Characterization of Novel Matrix-Derived Bioactive Peptides: A Role for Collagenase from Santyl® Ointment in Post-Debridement Wound Healing?
abstract: Debridement, the removal of diseased, nonviable tissue, is critical for clinicians to readily assess wound status and prepare the wound bed for advanced therapeutics or downstream active healing. Removing necrotic slough and eschar through surgical or mechanical methods is less specific and may be painful for patients. Enzymatic debridement agents, such as Clostridial collagenase, selectively and painlessly degrade devitalized tissue. In addition to its debriding activities, highly-purified Clostridial collagenase actively promotes healing, and our past studies reveal that extracellular matrices digested with this enzyme yield peptides that activate cellular migratory, proliferative and angiogenic responses to injury in vitro, and promote wound closure in vivo. Intriguingly, while collagenase Santyl® ointment, a sterile preparation containing Clostridial collagenases and other non-specific proteases, is a well-accepted enzymatic debridement agent, its role as an active healing entity has never been established. Based on our previous studies of pure Clostridial collagenase, we now ask whether the mixture of enzymes contained within Santyl® produces matrix-derived peptides that promote cellular injury responses in vitro and stimulate wound closure in vivo. Here, we identify novel collagen fragments, along with collagen-associated peptides derived from thrombospondin-1, multimerin-1, fibronectin, TGFβ-induced protein ig-h3 and tenascin-C, generated from Santyl® collagenase-digested human dermal capillary endothelial and fibroblastic matrices, which increase cell proliferation and angiogenic remodeling in vitro by 50-100% over controls. Using an established model of impaired healing, we further demonstrate a specific dose of collagenase from Santyl® ointment, as well as the newly-identified and chemically-synthesized ECM-derived peptides significantly increase wound re-epithelialization by 60-100% over saline-treated controls. These results not only confirm and extend our earlier studies using purified collagenase- and matrix-derived peptides to stimulate healing in vitro and in vivo, but these Santyl®-generated, matrix-derived peptides may also represent exciting new opportunities for creating advanced wound healing therapies that are enabled by enzymatic debridement and potentially go beyond debridement.
authors.affiliation: Graduate Program in Cellular & Molecular Physiology, The Sackler School of Graduate Biomedical Sciences, Tufts University, 136 Harrison Ave, Boston, MA, 02111, United States of America.|Department of Developmental, Molecular and Chemical Biology, School of Medicine, Tufts University, 136 Harrison Ave, Boston, MA, 02111, United States of America.|The Center for Innovations in Wound Healing Research, School of Medicine, Tufts University, 136 Harrison Ave, Boston, MA, 02111, United States of America.
authors.forename: Anthony R
authors.identifier: http://orcid.org/0000-0002-8641-636X
authors.initials: AR
authors.lastname: Sheets
authors.affiliation: The Center for Innovations in Wound Healing Research, School of Medicine, Tufts University, 136 Harrison Ave, Boston, MA, 02111, United States of America.
authors.forename: Tatiana N
authors.initials: TN
authors.lastname: Demidova-Rice
authors.affiliation: Smith & Nephew PLC, 3909 Hulen St., Fort Worth, TX, 76107, United States of America.
authors.forename: Lei
authors.initials: L
authors.lastname: Shi
authors.affiliation: University of North Texas Health Science Center, College of Pharmacy, 3500 Camp Bowie Blvd., Fort Worth, TX, 76107, United States of America.
authors.forename: Vincent
authors.initials: V
authors.lastname: Ronfard
authors.affiliation: Smith & Nephew PLC, 3909 Hulen St., Fort Worth, TX, 76107, United States of America.
authors.forename: Komel V
authors.initials: KV
authors.lastname: Grover
authors.affiliation: Graduate Program in Cellular & Molecular Physiology, The Sackler School of Graduate Biomedical Sciences, Tufts University, 136 Harrison Ave, Boston, MA, 02111, United States of America.|Graduate Program in Cell, Molecular and Developmental Biology, The Sackler School of Graduate Biomedical Sciences, Tufts University, 136 Harrison Ave, Boston, MA, 02111, United States of America.|Department of Developmental, Molecular and Chemical Biology, School of Medicine, Tufts University, 136 Harrison Ave, Boston, MA, 02111, United States of America.|The Center for Innovations in Wound Healing Research, School of Medicine, Tufts University, 136 Harrison Ave, Boston, MA, 02111, United States of America.
authors.forename: Ira M
authors.initials: IM
authors.lastname: Herman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017364:Collagenases / Q000378:metabolism*; D003646:Debridement; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D051379:Mice; D003012:Microbial Collagenase / Q000378:metabolism; D023421:Models, Animal; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D059748:Proteolysis; D012038:Regeneration; D014945:Wound Healing
chemical_list: D010455:Peptides; D017364:Collagenases; D003012:Microbial Collagenase