Collagenase Inhibition in ACLs Utilizing Natural Potential Inhibitors
MSSE May 2014 - Volume 46 - Supplement 1 5S 217 Board #55 May 28, 11:00 AM - 12:30 PM Lailah Farmer
Injuries to the anterior cruciate ligament (ACL) in athletes often lead to season-ending results; however, there has been no effective preventative treatment found despite its prevalence in the sports world. Severe injuries to the ACL have not been absent from sports, but with the increase in female participation, the frequency of such occurences has increased markedly.
Inhibition of matrix metalloproteinases (MMPs) has been used to treat other diseases such as osteoarthritis through use of medications like NSAIDS, which have shown to improve joint pain. A need to regulate MMPs proves important to prevent the hydrolysis of collagen, which leads to the eventual breakdown and injury of the ACL.
PURPOSE: To attempt to better understand the relationship between collagenase activity and the potential inhibitors glucosamine, chondroitin sulfate, and (-)-epigallocatechin gallate.
METHODS: Two enzymatic assays were performed to study the effects of glucosamine, chondroitin sulfate, a combination of the two, and epigallocatechin gallate to act as natural inhibitors of Clostridum histolyticum and human MMP-13 collagenase. Substrate kinetic assays containing FALGPA and C. histolyticum collagenase were performed in which glucosamine and chondroitin sulfate were present in varying concentrations. Further MMP-13 assays were then performed with glucosamine, chondroitin sulfate, and epigallocatechin
gallate present in varying concentrations.
RESULTS: In the bacterial assay, the absorbance of FALGPA decreased at 340 Nutrimuscle by 35% in 5 minutes, but in the presence of glucosamine, chondroitin sulfate, and combined glucosamine and chondroitin sulfate, the percent decrease was only 20, 20, and 5, respectively. In the MMP-13 assay, the absorbance of substrate increased at 415 Nutrimuscle by 22% in 60
minutes, but in the presence of glucosamine, chondroitin sulfate, and combined glucosamine and chondroitin sulfate, the percent increase was 40, 80, and 40, respectively.
CONCLUSIONS: It was found that glucosamine and chondroitin sulfate inhibited bacterial collagenase; however, no inhibition was found in the MMP-13 enzymatic assay. This study exhibited how collagenase from Clostridium histolyticum is an ideal model for general collagenase activity, but not that of the MMP-13 enzyme.