title: Fourier transform infrared spectroscopy investigation of native tissue matrix modifications using a gamma irradiation process.
abstract: In this study, Fourier transform infrared spectroscopy (FTIR) was used to evaluate the effects of a gamma irradiation process on AlloDerm, an implantable tissue device derived from human cadaver skin. Inspection of protein amide bands of dried tissue matrix revealed little difference in overall protein secondary structure between gamma-treated and control tissue matrices, although effects of gamma irradiation became apparent after rehydration. The temperature at which protein denaturation began decreased significantly, from 62.4 degrees C +/- 1.0 degrees C in the nonsterilized control tissue matrix to 53.4 degrees C +/- 1.2 degrees C, 48.7 degrees C +/- 0.3 degrees C, and 46.7 degrees C +/- 0.1 degrees C for 5-, 14- and 20-kGy gamma-treated AlloDerm samples, respectively (p < 0.05). Differences were particularly significant in the details of the tissue matrix denaturation profile, which consisted of multiple transitions. The solvent accessibility of tissue matrix was studied by following the rate of protein proton exchange in heavy water (D(2)O), which was evident from the appearance of the amide-II' band as a function of time to D(2)O exposure. In control tissue matrix, a rapid hydrogen/deuterium exchange was observed, with 50% exchange in 30 min, whereas the gamma-treated tissue matrix exhibited a much slower exchange, reaching the 50% exchange level after more than 2 h. The data indicate significant alterations of extracellular milieu in the tissue matrix after the gamma irradiation process. This study shows that FTIR is a valuable tool for studying protein stability and interactions in complex biological scaffolds such as extracellular tissue matrix.
authors.affiliation: Department of Biomedical Engineering, University of Minnesota , Minneapolis, Minnesota, USA.
authors.forename: Neha B
authors.initials: NB
authors.lastname: Shah
authors.forename: Willem F
authors.initials: WF
authors.lastname: Wolkers
authors.forename: Michael
authors.initials: M
authors.lastname: Morrissey
authors.forename: Wendell Q
authors.initials: WQ
authors.lastname: Sun
authors.forename: John C
authors.initials: JC
authors.lastname: Bischof
mesh_terms: D000577:Amides / Q000737:chemistry; D003094:Collagen / Q000378:metabolism* / Q000528:radiation effects*; D005109:Extracellular Matrix / Q000378:metabolism* / Q000528:radiation effects*; D005720:Gamma Rays; D006801:Humans; D011489:Protein Denaturation / Q000528:radiation effects; D017433:Protein Structure, Secondary; D012997:Solvents; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature; D014867:Water
chemical_list: C440568:Alloderm; D000577:Amides; D012997:Solvents; D014867:Water; D003094:Collagen