Multiplexed analysis of matrix metalloproteinases in leg ulcer tissue of patients with chronic venous insufficiency before and after compression therapy

Abstract

Elevated matrix metalloproteinases (MMP) levels have been implicated in the pathogenesis of chronic venous insufficiency ulcers. Quantitative measurements of a broad range of MMP proteins in human tissue treated with compression bandaging have not been reported. The goal of this study was to determine the expression of a wide range of proteases in untreated venous leg ulcer tissue and the changes in these levels after 4 weeks of high-strength compression therapy. Twenty-nine limbs with new or untreated chronic venous insufficiency and leg ulceration received therapy for 4 weeks with sustained high compression bandaging. Biopsies were obtained from healthy tissue and from ulcerated tissue before and after therapy. A novel multiplexed protein assay was used to measure multiple MMPs in a single sample. MMP protein activity, TIMP protein levels, and gene expression levels were also addressed. MMP1, 2, 3, 8, 9, 12, and 13 protein levels were elevated in ulcer tissue compared with healthy tissue. MMP8 and 9 were highly expressed in ulcer tissue. MMP3, 8, and 9 significantly decreased following treatment. Reduction in the levels of MMP1, 2, and 3 was associated with significantly higher rates of ulcer healing at 4 weeks. We conclude that compression therapy results in a reduction of the pro-inflammatory environment characterizing chronic venous ulcers, and ulcer healing is associated with resolution of specific elevated levels of protease expression.

Wound Fluid Diagnostics in Diabetic Foot Ulcers

“…For instance, Grimstad et al measured the concentration of 27 different cytokines in wound fluid derived from acute wounds using a bead-based system (Grimstad et al 2011). Beidler et al applied this technology to measure the concentrations of matrix metalloproteinases and cytokines in wound tissue lysate (Beidler et al 2008(Beidler et al , 2009). In combination with appropriate wound sampling procedures, the complex protein-composition of wound exudate together with limitations in the anticipated sample volume renders protein array technology -be it planar or bead-based-as an interesting tool for the analysis of wound-associated biomarkers (see section 6.1).…”

“…Due to the frequently found low exudate levels in chronic wounds along with the inherent complexity of wound fluid, appropriate technologies are needed to accurately and reliably determine multiple parameters with www.intechopen.com minimal sample consumption. In this context, protein microarrays are important tools for the simultaneous detection and quantification of multiple parameters in complex biological mixtures, such as plasma (Cassatella et al 1997) or tissue lysates (Beidler et al 2008). This technology has been shown to exhibit high sensitivity and optimal signal to noise ratios even in the case of low target concentrations (Ekins 1989(Ekins , 1998.…”

Enzymatic activity analysis of MMP-2 and 9 collected by swab from lower limb venous ulcers

“…10 Furthermore, the course of the healing process in chronic wounds is associated with reduced expression of certain MMPs. 14 In view of this, development of a simple technique for collection of biological material for analysis of MMPs makes it possible to conduct serial evaluations of the process of ulcer healing and opens a wide range of applications for clinical research.…”

“…[7][8][9]14 Among the different MMPs, MMP-9 appears to be a prognostic marker of healing of chronic ulcers. 10 Furthermore, the course of the healing process in chronic wounds is associated with reduced expression of certain MMPs.…”

Tracking Healing Process of Experimental Liver Injuries Treated with Different Sealants and Adhesives Biomaterials. Matrix Metalloproteinase Evaluation

“…[25] Prolonged MMP1 activity in chronic skin ulcers can have a critical effect on tissue re-epithelialization. [26] Gelatinases (MMP2 and MMP9) play an important role in the formation and maturation of granulation tissue during wound healing. [27] Both MMP2 and MMP9 have been reported to act synergistically with collagenases.…”