Date of Graduation

12-2011

Document Type

Thesis

Degree Name

Master of Science in Chemistry (MS)

Degree Level

Graduate

Department

Chemistry & Biochemistry

Advisor

Julie Stenken

Committee Member

Narayan C. Rath

Second Committee Member

Suresh Kumar Thallapuranam

Third Committee Member

Paul D. Adams

Keywords

Pure sciences; Anal.chem; Chem

Abstract

Biomaterial implantation induces the foreign body response (FBR). Development of longer-term implants relies on the thorough understanding of the FBR. The progression of the FBR is regulated by a number of biomolecules including cytokines, chemokines, and matrix metalloproteinases (MMPs). The nature of the FBR requires the spatial and temporal regulation of these mediators. MMPs are an extremely large and diverse group of enzymes that play key roles in regulating the FBR. Precise spatiotemporal regulation of MMPs defines their proteolytic activities. The aim of this project is to develop a new bioanalytical method to visualize the localization of active MMPs at the implant site in ex vivo tissue samples. The localization of active MMP-2 and MMP-9 surrounding the implanted biomaterial in the ex vivo tissue sample was determined using MALDI imaging mass spectrometry. The MALDI imaging scheme employed a synthetic FRET substrate specific for MMP-2 and MMP-9. The intensity of the m/z value corresponding to the product of active MMP-2 and MMP-9 digestion in each spectrum was extracted to produce a two-dimensional image showing the localization of active MMP-2 and MMP-9 in the ex vivo tissue sample. Fluorescence imaging was used as a complementary technique.

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