Enzyme Immobilization on Chitosan-Glutaraldehyde Films by Covalent Attachment: Application in wound healing

Matrix Metalloproteinases (MMPs) are zinc endopeptidases which present in both acute and chronic wounds disturbance of the balance between synthesis and degradation of extracellular matrix (ECM) lead to abnormal wound healing. MMPs regulate extracellular structural proteins and consequent tissue remodeling. Controlling levels of inflammation and protease expression is a critical part of normal wound healing, elevated and prolonged expression of proteases produced during the inflammatory phase of healing can lead to excessive extracellular matrix ECM degradation associated with impaired healing.There are various investigations about the roles of proteases within both acute and chronic wounds and how the manipulation of protease activity may aid healing. Indeed, numerous wound dressings have been developed that target such proteases in an attempt to promote wound healing. The wound healing dressings based on modified cellulose, chitosan and various immobilized therapeutic agents are commonly used. This study disclosed the preparation of aldehyde functionalized chitosan film and immobilization of enzyme on chitosan-glutaraldehyde films. The immobilization capacity of the supports and specific activity of the immobilized derivatives were investigated. Response surface methodology (RSM) and a central composite rotatable design (CCRD) was used to study the effects of four factors, glutaraldehyde concentration, enzyme loading, immobilization pH, and glutaraldehyde functionalizing time on the specific activity of immobilized preparations. Under optimized experimental conditions (2.5% glutaraldehyde, 7.5 U mL-1 enzyme loading, pH 6.0, and 30 min treatment with glutaraldehyde) the specific activity was about 4.8 U/mg. This wound dressing could effectively accelerate the wound healing process at BALB/c mouse skin wounds. These results indicate that enrichment of wound dressings with enzyme can significantly enhance their potential efficacy as wound dressing material.

کلیدواژه ها:

Chitosan ، Enzyme ، Wound dressing