Spectral fingerprinting of decellularized heart valve scaffolds.

Abstract

Decellularized heart valves hold promise for their use as bioscaffolds in cardiovascular surgery. Quality assessment of heart valves after decellularization processing and/or storage is time consuming and destructive. Fourier transform infrared spectroscopy (FTIR) allows rapid non-invasive assessment of biomolecular structures in tissues. In this study, IR-spectra taken from different layers of the pulmonary artery trunk and leaflet tissues of decellularized porcine heart valves were compared with those of pure collagen and elastin, the main protein components in these tissues. In addition, spectral changes associated with aging and oxidative damage were investigated. Infrared absorbance spectra of the arteria intima and media layer were found to be very similar, whereas distinct differences were observed when compared with spectra of the externa layer. In the latter, the shape of the CH-stretching vibration region (3050-2800 cm-1) resembled that of pure collagen. Also, pronounced νCOOH and amide-II bands and a relatively high content of α-helical structures in the externa layer indicated the presence of collagen in this layer. The externa layer of the artery appeared to be sensitive to collagenase treatment, whereas the media and intima layer were particularly affected by elastase and not by collagenase treatment. Protein conformational changes after treatment with collagenase were observed in all three layers. Collagenase treatment completely degraded the leaflet tissue sections. Spectra were also collected from scaffolds after 2 and 12 weeks storage at 37 °C, and after induced oxidative damage. Spectral changes related to aging and oxidative damage were particularly evident in the CH-stretching region, whereas the shape of the amide-I band, reflecting the overall protein secondary structure, remained unaltered.