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Thirty\u2010four (34) female athymic rats were inoculated with HeLa (<jats:italic>N<\/jats:italic>\u2009=\u200917) or Ace\u20101 (<jats:italic>N<\/jats:italic>\u2009=\u200917) cancer cells lines producing osteolytic or mixed (osteolytic and osteoblastic) metastases, respectively. A maximum of 21 days was allowed between inoculation and rat sacrifice for vertebrae extraction. High performance liquid chromatography (HPLC) was utilized to determine modifications in collagen\u2010I parameters such as proline hydroxylation and the formation of specific enzymatic and non\u2010enzymatic (pentosidine) cross\u2010links. Raman spectroscopy was used to determine relative changes in mineral crystallinity, mineral carbonation, mineral\/collagen matrix ratio, collagen quality ratio, and proline hydroxylation. HPLC results showed significant increase in the formation of pentosidine and decrease in the formation of the enzymatic cross\u2010link deoxy\u2010pryridinoline within osteolytic bone compared to mixed bone. Raman results showed decreased crystallinity, increased carbonation, and collagen quality (aka 1660\/1690 sub\u2010band) ratio with osteolytic bone compared to mixed bone and healthy controls along with an observed increase in proline hydroxylation with metastatic involvement. The mineral\/matrix ratio decreased in both osteolytic and mixed bone compared to healthy controls. Quantifying modifications within the intrinsic characteristics of bone tissue will provide a foundation to assess the impact of current therapies on the material behavior of bone tissue in the metastatic spine and highlight targets for the development of new therapeutics and approaches for treatment. \u00a9 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. 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