alpha-chymotrypsin and Varicose-Ulcer

Venous leg ulcer slough is unpleasant to the patient and difficult to manage clinically. It harbours infection, also preventing wound management materials and dressings from supporting the underlying viable tissues. In other words, slough has significant nuisance value in the tissue viability clinic. In this study, we have sought to increase our knowledge of slough by building upon a previous but limited analysis of this necrotic tissue. In particular, slough has been probed using Western blotting for the presence of proteins with the capacity to engage microbial surface components recognising adhesive matrix macromolecules. Although the samples were difficult to resolve, we detected fibrinogen, fibronectin, IgG, collagen, human serum albumin and matrix metalloproteinase-9. Furthermore, the effect of a maggot-derived debridement enzyme, chymotrypsin 1 on macromolecules in slough was confirmed across seven patient samples. The effect of chymotrypsin 1 on slough confirms our thesis that this potential debridement enzyme could be effective in removing slough along with its associated bacteria, given its observed resistance to intrinsic gelatinase activity. In summary, we believe that the data provide scientists and clinicians with further insights into the potential molecular interactions between bacteria, wound tissue and Lucilia sericata in a clinically problematic yet scientifically interesting wound ecosystem.

Topics: Animals; Chymotrypsin; Collagen; Debridement; Diptera; Fibronectins; Gelatinases; Humans; Matrix Metalloproteinase 9; Skin; Tissue Survival; Varicose Ulcer; Wound Healing

Larvae of the greenbottle Lucilia sericata are used to debride nonhealing wounds and stimulate the production of fresh granulation tissue. Previous publications have shown that secretions from L. sericata contain a number of proteolytic activities including a chymotrypsin that degrades a number of extracellular matrix components such as fibronectin, laminin and collagen.. To produce a recombinant L. sericata chymotrypsin (chymotrypsin I) and determine its effects on the degradation of patient wound eschar.. An active recombinant chymotrypsin I from L. sericata was cloned and expressed in Sf9 cells and its subsequent effects ex vivo on eschar from venous leg ulcers were determined by two-dimensional electrophoresis.. The recombinant enzyme had the attributes of a chymotrypsin, possessing sequence homology with other chymotrypsins and demonstrating attributes of the native enzyme including cleavage of the chymotrypsin substrate succinyl-alanyl-alanyl-prolyl-phenylalanyl-7-amino-4-methyl coumarin, inhibition by phenylmethylsulphonyl fluoride and lack of inhibition by amidinophenylmethylsulphonyl fluoride. Importantly, the recombinant chymotrypsin cleaved the majority of proteins from slough/eschar from venous leg ulcers in a superior manner to chymotrypsins from human and bovine sources.. The ex vivo degradation of eschar from venous leg ulcers indicates the potential value of recombinant chymotrypsin I as a novel, stand-alone debridement agent.

Topics: Animals; Blotting, Western; Cattle; Chymotrypsin; Diptera; Electrophoresis, Gel, Two-Dimensional; Humans; Larva; Proteomics; Recombinant Proteins; Sequence Homology, Amino Acid; Varicose Ulcer; Wound Healing

Topics: Anti-Bacterial Agents; Arteriosclerosis; Burns; Chymotrypsin; Debridement; Deoxyribonucleases; Endarteritis; Enzyme Therapy; Female; Humans; Male; Nervous System Diseases; Skin Transplantation; Streptodornase and Streptokinase; Streptokinase; Transplantation, Autologous; Trypsin; Ulcer; Varicose Ulcer; Veins; Wounds and Injuries

Topics: Adult; Chymotrypsin; Female; Humans; Leg; Male; Middle Aged; Streptodornase and Streptokinase; Thrombophlebitis; Trypsin; Varicose Ulcer

Topics: Aged; Anti-Bacterial Agents; Chymotrypsin; Cicatrix; Female; Framycetin; Humans; Varicose Ulcer

Topics: Chymotrypsin; Esculin; Flavonoids; Heparin; Humans; Leg; Nutritional Status; Varicose Ulcer; Veins