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Meniscus samples (<jats:italic>n<\/jats:italic>\u2009=\u200924) were obtained from nonarthritic knees of human cadavers with no history of joint diseases. Specimens (<jats:italic>n<\/jats:italic>\u2009=\u200972) were obtained from three distinct sections of the meniscus, namely; anterior, center, posterior. Absorption spectra were acquired from each specimen in the VIS and NIR spectral range (400\u20131,100\u2009nm). Following spectroscopic probing, the specimens were subjected to biochemical analyses to determine the matrix composition, that is water, hydroxyproline, and uronic acid contents. Multivariate analytical techniques, including principal component analysis (PCA) and partial least squares (PLS) regression, were then used to investigate the correlation between the matrix composition and it spectral response. Our results indicate that the optical absorption of meniscus matrix is related to its composition, and this relationship is optimal in the NIR spectral range (750\u20131,100\u2009nm). High correlations (<jats:italic>R<\/jats:italic><jats:sup><jats:italic>2<\/jats:italic><\/jats:sup><jats:sub><jats:italic>uronic<\/jats:italic><\/jats:sub>\u2009=\u200986.9%, <jats:italic>R<\/jats:italic><jats:sup><jats:italic>2<\/jats:italic><\/jats:sup><jats:sub><jats:italic>water<\/jats:italic><\/jats:sub>\u2009=\u200983.8%, <jats:italic>R<\/jats:italic><jats:sup><jats:italic>2<\/jats:italic><\/jats:sup><jats:sub><jats:italic>hydroxyproline<\/jats:italic><\/jats:sub>\u2009=\u200981.7%, <jats:italic>p\u2009<\/jats:italic>&lt;\u20090.0001) were obtained between the spectral predicted and measured meniscus composition, thus suggesting that spectral data in the NIR range can be utilized for estimating the matrix composition of human meniscus. In conclusion, optical spectroscopy, particularly in the NIR spectral range, is a potential method for evaluating the composition of human meniscus. This presents a promising technique for rapid and nondestructive evaluation of meniscus integrity in real\u2010time during arthroscopic surgery. \u00a9 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. 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