title: Identification of Pyridinoline Trivalent Collagen Cross-Links by Raman Microspectroscopy.
abstract: Intermolecular cross-linking of bone collagen is intimately related to the way collagen molecules are arranged in a fibril, imparts certain mechanical properties to the fibril, and may be involved in the initiation of mineralization. Raman microspectroscopy allows the analysis of minimally processed bone blocks and provides simultaneous information on both the mineral and organic matrix (mainly type I collagen) components, with a spatial resolution of ~1 μm. The aim of the present study was to validate Raman spectroscopic parameters describing one of the major mineralizing type I trivalent cross-links, namely pyridinoline (PYD). To achieve this, a series of collagen cross-linked peptides with known PYD content (as determined by HPLC analysis), human bone, porcine skin, predentin and dentin animal model tissues were analyzed by Raman microspectroscopy. The results of the present study confirm that it is feasible to monitor PYD trivalent collagen cross-links by Raman spectroscopic analysis in mineralized tissues, exclusively through a Raman band ~1660 wavenumbers. This allows determination of the relative PYD content in undecalcified bone tissues with a spatial resolution of ~1 μm, thus enabling correlations with histologic and histomorphometric parameters.
authors.affiliation: Ludwig Boltzmann Institute of Osteology at Hanusch Hospital of WGKK and AUVA Trauma Centre Meidling, 1st Med. Dept. Hanusch Hospital, Vienna, Austria.
authors.forename: Sonja
authors.initials: S
authors.lastname: Gamsjaeger
authors.affiliation: Rowett Institute of Nutrition and Health, Bucksburn, Aberdeen, AB21 9SB, Scotland, UK.
authors.forename: Simon P
authors.initials: SP
authors.lastname: Robins
authors.affiliation: Division of Periodontology, College of Dentistry, The Ohio State University, Columbus, OH, USA.
authors.forename: Dimitris N
authors.initials: DN
authors.lastname: Tatakis
authors.affiliation: Ludwig Boltzmann Institute of Osteology at Hanusch Hospital of WGKK and AUVA Trauma Centre Meidling, 1st Med. Dept. Hanusch Hospital, Vienna, Austria.
authors.forename: Klaus
authors.initials: K
authors.lastname: Klaushofer
authors.affiliation: Ludwig Boltzmann Institute of Osteology at Hanusch Hospital of WGKK and AUVA Trauma Centre Meidling, 1st Med. Dept. Hanusch Hospital, Vienna, Austria. lpaschalis@gmx.net.|Ludwig Boltzmann Institute of Osteolgy, Hanusch Kh., Heinrich Collin Str. 30, 1140, Vienna, Austria. lpaschalis@gmx.net.
authors.forename: Eleftherios P
authors.initials: EP
authors.lastname: Paschalis
mesh_terms: D000596:Amino Acids / Q000378:metabolism*; D001842:Bone and Bones / Q000378:metabolism*; D003094:Collagen / Q000378:metabolism*; D003432:Cross-Linking Reagents; D006801:Humans; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods; D013059:Spectrum Analysis, Raman / Q000379:methods; D014070:Tooth / Q000473:pathology
chemical_list: D000596:Amino Acids; D003432:Cross-Linking Reagents; C015484:pyridinoline; D003094:Collagen
keywords: Chromatography; Collagen; Pyridinoline; Raman microspectroscopy; Trivalent collagen cross-links