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Fluorimetric ex vivo quantification of protease debriding efficacy on natural substrate
Author(s) -
Vootukuri Reddy Sreekanth,
Philpott Mike P.,
Trigiante Giuseppe
Publication year - 2020
Publication title -
wound repair and regeneration
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.847
H-Index - 109
eISSN - 1524-475X
pISSN - 1067-1927
DOI - 10.1111/wrr.12864
Subject(s) - eschar , ex vivo , debridement (dental) , in vivo , collagenase , proteases , wound healing , chemistry , papain , fibrin , in vitro , biology , biochemistry , pathology , medicine , surgery , immunology , microbiology and biotechnology , enzyme
Debridement is the process of removal of necrotic and infected tissue to clean a wound or burn and expedite healing. Proteases such as papain, bromelain, and collagenase that promote debridement by degrading proteins in the dead tissue are in use today. However, the only method to measure debriding efficacy in vitro is the fluorescent monitoring of the digestion of an Artificial Wound Eschar (AWE) substrate. This AWE substrate contains a pellet of only three eschar matrix proteins collagen, elastin, and fibrin which do not account for the complexity and the composition of necrotic tissue. Here, we describe an ex vivo method using dry necrotic full thickness human skin and ortho‐phthalaldehyde (OPA), a molecule commonly used for sensitive fluorimetric protein detection to monitor debridement activity. We advocate this simple yet sensitive approach to detect debridement efficacy that can readily be used commercially to benchmark products prior to in vivo testing.