In order to closely mimic the actual mechanism of wound debridement by an enzymatic debriding agent, a synthetic wound eschar substrate (SWES) has been developed. The SWES is prepared by the enzymatic conversion of fibrinogen to fibrin in the presence of fibrous bovine collagen and elastin, to form an insoluble planar matrix composed of the three major proteins present in wound eschar. Each of the proteinaceous components is uniquely labeled prior to formation of SWES with different dye, and the debridement test is conducted using a Franz Diffusion System. The debriding agent is applied to the SWES mounted on the donor compartment of the cell, and the digested hydrolysate containing dye‐labeled peptide fragments is sampled from the receptor compartment. Accordingly, the progress of digestion is obtained in real time. Since the three proteins are labeled with spectrally distinct dyes, data on the hydrolysis of the substrates can be collected simultaneously. This method facilitates the testing of formulated debriding products in a manner that closely mimics application to a wound and provides a powerful tool for the evaluation of formulated enzymatic debriding agents.
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