Correlating Changes in Collagen Secondary Structure with Aging and Defective Type II Collagen by Raman Spectroscopy

Abstract

A novel application of Raman spectroscopy for monitoring damage to ocular collagen in wild-type mice and Del1 (+/–) transgenic mice, a murine animal model of osteoarthritis, is described. In order to understand the progression of diseases of collagen, it is necessary to use methods that can recognize alterations in affected tissue due to chemical and/or genetic modifications. The heterozygous Del1 (+/–) transgenic mouse is established as a model for early-onset osteoarthritis caused by modifications to the type II collagen gene (COL2A1) that result in a truncated collagen fiber. We expect that abnormal type II collagen is expressed in articular cartilage and eye tissue of the Del1 (+/–) mouse. Eyes excised from a subset of specimens from another study using Del1 (+/–) mice were examined by Raman spectroscopy for evidence of defective collagen. Spectral contributions from the collagen protein were readily observed. The amide III envelope (1220–1280 cm−1) was used to characterize changes in collagen secondary structure. Raman spectra of the sclera component of eyes taken from transgenic and older wild-type mice show an increased collagen disorder, as expected. These preliminary results suggest that Raman is capable of recognizing and measuring abnormality in eye collagen and may have potential as a diagnostic tool for ocular collagen damage.