MONITORING OF WOUNDS BY MEASUREMENT OF PROTEASE AND PROTEASE INHIBITOR LEVELS IN WOUND FLUIDS
A diagnostic test apparatus for determining a ratio of: (a) at least one endogenous protease enzyme inhibitor, to (b) at least one endogenous protease enzyme, in a sample of a wound fluid. Suitably the protease enzyme is a neutrophil elastase and the protease enzyme inhibitor is alpha-1-antitrypsin. Also provided are wound treatment systems comprising an apparatus according to the invention and a wound dressing comprising an oxidized cellulose. Also provided are methods of diagnosis, prognosis and treatment of wounds using the apparatus and systems of the invention.
1 . A diagnostic test apparatus for determining a ratio of: (a) at least one endogenous protease enzyme inhibitor, to (b) at least one endogenous protease enzyme, in a sample of a wound fluid, the diagnostic test apparatus comprising:
an indicator moiety that is immobilized or inhibited by a chemical moiety, wherein said chemical moiety comprises an exogenous peptide substrate for a protease enzyme, and said exogenous peptide substrate is cleavable by the protease enzyme to release or activate said indicator moiety.
2 . The diagnostic test apparatus according to claim 1 , wherein said step of determining comprises determining whether said ratio falls within a predetermined range.
3 . The diagnostic test apparatus according to claim 1 , wherein the said at least one protease enzyme comprises one or more proteases selected from the group consisting of neutrophil elastase, matrix metalloproteinases ( e.g. MMP-9, MMP-8, MMP-1, MMP-12), proteinase 3, plasmin, low molecular weight gelatinases and latent or active elastases, interleukin converting enzymes and tumor necrosis factor (TNFa) converting enzymes.
4 . The diagnostic test apparatus according to claim 1 , wherein the said at least one protease comprises one or more proteases selected from the group consisting of neutrophil elastase and matrix metalloproteinases.
5 . The diagnostic test apparatus according to claim 1 , wherein the said at least one protease enzyme inhibitor is selected from the group consisting of elastinil, elafin, secretory leukocyte proteinase inhibitor, alpha-1-macroglobulin, alpha-1-antitrypsin (AAT), and mixtures thereof.
6 . The diagnostic test apparatus according to claim 1 , wherein the said at least one protease enzyme is a neutrophil elastase, and the said at least one protease enzyme inhibitor is AAT.
7 . The diagnostic test apparatus according to claim 1 , wherein said apparatus comprises a single diagnostic device specifically adapted for detecting both said at least one endogenous protease enzyme and said at least one endogenous protease inhibitor in said sample.
8 . The diagnostic test apparatus according to claim 1 , further comprising a dip-stick or swab for sampling of said wound fluid.
9 . A wound treatment system comprising the apparatus according to claim 1 , and a wound dressing comprising oxidized cellulose.
10 . The wound treatment system of claim 9 , wherein the wound dressing comprises a combination of oxidized regenerated cellulose with collagen and/or chitosan in the dry weight ratio of from about 10:1 to about 1:10.
11 . A diagnostic test apparatus for determining a ratio of: (a) at least one endogenous protease enzyme inhibitor, to (b) at least one endogenous protease enzyme, in a sample of a wound fluid, the diagnostic test apparatus comprising:
a solid support material having an immunological binding partner for an analyte moiety covalently linked thereto.
12 . The diagnostic test apparatus according to claim 11 , wherein said step of determining comprises determining whether said ratio falls within a predetermined range.
13 . The diagnostic test apparatus according to claim 11 , wherein the said at least one protease enzyme comprises one or more proteases selected from the group consisting of neutrophil elastase, matrix metalloproteinases (e.g. MMP-9, MMP-8, MMP-1, MMP-12), proteinase 3, plasmin, low molecular weight gelatinases and latent or active elastases, interleukin converting enzymes and tumor necrosis factor (TNFa) converting enzymes.
14 . The diagnostic test apparatus according to claim 11 , wherein the said at least one protease comprises one or more proteases selected from the group consisting of neutrophil elastase and matrix metalloproteinases.
15 . The diagnostic test apparatus according to claim 11 , wherein the said at least one protease enzyme inhibitor is selected from the group consisting of elastinil, elafin, secretory leukocyte proteinase inhibitor, alpha-1-macroglobulin, alpha-1-antitrypsin (AAT), and mixtures thereof.
16 . The diagnostic test apparatus according to claim 11 , wherein the said at least one protease enzyme is a neutrophil elastase, and the said at least one protease enzyme inhibitor is AAT.
17 . The diagnostic test apparatus according to claim 11 , wherein said apparatus comprises a single diagnostic device specifically adapted for detecting both said at least one endogenous protease enzyme and said at least one endogenous protease inhibitor in said sample.
18 . The diagnostic test apparatus according to claim 11 , further comprising a dip-stick or swab for sampling of said wound fluid.
19 . A wound treatment system comprising the apparatus according to claim 11 , and a wound dressing comprising oxidized cellulose.
20 . The wound treatment system of claim 19 , wherein the wound dressing comprises a combination of oxidized regenerated cellulose with collagen and/or chitosan in the dry weight ratio of from about 10:1 to about 1:10.