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Developing staining protocols for visualization of tissue-engineering scaffolds using micro computed tomography in native wet state

BNC-alginate and silk fibroin tissue-engineering scaffolds were stained with X-ray contrast agents in order to visualize internal microstructure in the native wet state with microcomputed tomography. A successful protocol employing amphiphilic contrast agents (CAs) dissolved in a water-based staining solution was used. The CAs were then fixed to the scaffold by neutralizing their charged functional groups, increasing their hydrophobicity and retention on the scaffold surface in water. While some unresolved issues concerning homogeneous staining and strength of contrast remain, these first successes constitute an important milestone by identifying good contrast agent candidates and staining protocols for longitudinal monitoring of tissue-engineering studies.