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Degradation of extracellular matrix components by defined proteinases from the greenbottle larva Lucilia sericata used for the clinical debridement of non-healing wounds.
Br J Dermatol. 2003 Jan; 148(1):14-23.BJ

Abstract

BACKGROUND

Larvae of the greenbottle fly Lucilia sericata are used routinely for the clinical treatment of difficult necrotic and infected wounds. Degradation by proteinases contained in larval excretory/secretory (ES) products is thought to contribute to wound debridement by removal of dead tissue. However, proteinase activity may also affect host tissue remodelling processes.

OBJECTIVES

To identify proteolytic enzymes derived from L. sericata ES products with activities against fibrin and extracellular matrix (ECM) components.

METHODS

Larval proteinase activities were assayed in vitro using class-specific substrates and inhibitors. Their action against fibrin and ECM components was examined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis.

RESULTS

Three classes of proteolytic enzyme were detected in the secretions using fluorescein isothiocyanate-labelled casein as a model substrate. The predominant activity belonged to serine proteinases (pH optima 8-9) of two different subclasses (trypsin-like and chymotrypsin-like), with a weaker aspartyl proteinase (pH 5) and a metalloproteinase (pH 9) with exopeptidase characteristics also present. Using skin-relevant ECM components as substrates L. sericata ES products solubilized fibrin clots and degraded fibronectin, laminin and acid-solubilized collagen types I and III. Hydrolysis of ECM macromolecules was inhibited by preincubating ES products with phenylmethylsulphonyl fluoride but not 4-amidinophenylmethylsulphonyl fluoride, indicating that degradation was due to the 'chymotrypsin-like' serine proteinase.

CONCLUSIONS

These data suggest that a combination of L. sericata ES proteinases involving chymotrypsin-like and trypsin-like activities could potentially influence wound healing events when maggots are introduced into necrotic and infected wounds, with the chymotrypsin-like activity involved in the remodelling of ECM components.

Authors+Show Affiliations

Chambers L
Immune Modulation Group, School of Pharmaceutical Sciences, Boots Science Building, University of Nottingham, UK.
Woodrow S
No affiliation info available
Brown AP
No affiliation info available
Harris PD
No affiliation info available
Phillips D
No affiliation info available
Hall M
No affiliation info available
Church JC
No affiliation info available
Pritchard DI
No affiliation info available

MeSH

AnimalsChymotrypsinCollagenDipteraElectrophoresis, Polyacrylamide GelEndopeptidasesExtracellular MatrixFibrinFibronectinsHydrogen-Ion ConcentrationLamininLarvaSerine EndopeptidasesTrypsinWound Healing

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12534589

Citation

Chambers, L, et al. "Degradation of Extracellular Matrix Components By Defined Proteinases From the Greenbottle Larva Lucilia Sericata Used for the Clinical Debridement of Non-healing Wounds." The British Journal of Dermatology, vol. 148, no. 1, 2003, pp. 14-23.
Chambers L, Woodrow S, Brown AP, et al. Degradation of extracellular matrix components by defined proteinases from the greenbottle larva Lucilia sericata used for the clinical debridement of non-healing wounds. Br J Dermatol. 2003;148(1):14-23.
Chambers, L., Woodrow, S., Brown, A. P., Harris, P. D., Phillips, D., Hall, M., Church, J. C., & Pritchard, D. I. (2003). Degradation of extracellular matrix components by defined proteinases from the greenbottle larva Lucilia sericata used for the clinical debridement of non-healing wounds. The British Journal of Dermatology, 148(1), 14-23.
Chambers L, et al. Degradation of Extracellular Matrix Components By Defined Proteinases From the Greenbottle Larva Lucilia Sericata Used for the Clinical Debridement of Non-healing Wounds. Br J Dermatol. 2003;148(1):14-23. PubMed PMID: 12534589.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Degradation of extracellular matrix components by defined proteinases from the greenbottle larva Lucilia sericata used for the clinical debridement of non-healing wounds. AU - Chambers,L, AU - Woodrow,S, AU - Brown,A P, AU - Harris,P D, AU - Phillips,D, AU - Hall,M, AU - Church,J C T, AU - Pritchard,D I, PY - 2003/1/22/pubmed PY - 2003/3/20/medline PY - 2003/1/22/entrez SP - 14 EP - 23 JF - The British journal of dermatology JO - Br J Dermatol VL - 148 IS - 1 N2 - BACKGROUND: Larvae of the greenbottle fly Lucilia sericata are used routinely for the clinical treatment of difficult necrotic and infected wounds. Degradation by proteinases contained in larval excretory/secretory (ES) products is thought to contribute to wound debridement by removal of dead tissue. However, proteinase activity may also affect host tissue remodelling processes. OBJECTIVES: To identify proteolytic enzymes derived from L. sericata ES products with activities against fibrin and extracellular matrix (ECM) components. METHODS: Larval proteinase activities were assayed in vitro using class-specific substrates and inhibitors. Their action against fibrin and ECM components was examined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis. RESULTS: Three classes of proteolytic enzyme were detected in the secretions using fluorescein isothiocyanate-labelled casein as a model substrate. The predominant activity belonged to serine proteinases (pH optima 8-9) of two different subclasses (trypsin-like and chymotrypsin-like), with a weaker aspartyl proteinase (pH 5) and a metalloproteinase (pH 9) with exopeptidase characteristics also present. Using skin-relevant ECM components as substrates L. sericata ES products solubilized fibrin clots and degraded fibronectin, laminin and acid-solubilized collagen types I and III. Hydrolysis of ECM macromolecules was inhibited by preincubating ES products with phenylmethylsulphonyl fluoride but not 4-amidinophenylmethylsulphonyl fluoride, indicating that degradation was due to the 'chymotrypsin-like' serine proteinase. CONCLUSIONS: These data suggest that a combination of L. sericata ES proteinases involving chymotrypsin-like and trypsin-like activities could potentially influence wound healing events when maggots are introduced into necrotic and infected wounds, with the chymotrypsin-like activity involved in the remodelling of ECM components. SN - 0007-0963 UR - https://www.unboundmedicine.com/medline/citation/12534589/Degradation_of_extracellular_matrix_components_by_defined_proteinases_from_the_greenbottle_larva_Lucilia_sericata_used_for_the_clinical_debridement_of_non_healing_wounds_ DB - PRIME DP - Unbound Medicine ER -