The present study aims to systematically investigate the freezing-induced changes that occur at multiple levels of organization of collagen nanostructure in the engineered tissues (ET). Collagen is a major constituent of the extracellular matrix (ECM) of biological tissues, and is also used for scaffold of engineered tissues and biomaterials [1, 2]. Given its abundance and widespread physiological function in vivo, a proper understanding of the relationships between the collagen’s structure, properties, and function is essential for the improvement of current tissue cryopreservation protocols that suffer from highly variable and tissue specific outcomes [3, 4].

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