Tuesday, 05 September 2017 10:21

3D micro CT imaging of the human peripheral nerve fascicle

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Abstract: Autologous nerve grafting is the gold standard for the treatment of peripheral nerve injury; however, it involves certain complications. With the maturity of three-dimensional (3D) bioprinting techniques, the 3D bioprinting of nerve grafts has become theoretically feasible. The primary 3D bioprinting of nerve grafts results in a 3D nerve structure, in particular, a fascicular 3D structure. The aim of this study was to identify a method for enhancing Xray micro computed tomography (CT) to reconstruct 3D nerve structures. Here, we used three techniques, Lugol’s iodine solution (I2KI) enhancement, calcium chloride enhancement and freeze-drying (n=6), to identify a better method for obtaining high-resolution 3D images of a normal human peripheral nerve fascicle. Contrast differences were employed to compare the homologous nerve structure with the fascicular structure in each group. Unlike hematoxylin and eosin (H&E) staining and scanning electron microscopy (SEM), the micro CT approach produced continuous serial images, which are necessary for 3D reconstruction. The fascicular increases in the contrast levels were 211.74±31.44%, -6.51±1.46% and 125.41±27.14% for the H&E, SEM and micro CT methods, respectively.

The calcium chloride enhancement resulted in an excellent contrast between the human nerve fascicle and the following tissues: the perineurium, connective tissue, epineurium, and endoneurium (p<0.001). These findings indicate that the anatomical structures were clearly identified. This is the first use of micro CT to reconstruct a 3D image of a human peripheral nerve fascicle using three different pretreatment methods. Compared to H&E and SEM, our micro CT approach has the advantage of continuous serial imaging. In addition, compared to the iodine enhancement and freeze-drying methods, the calcium chloride-enhanced contrast method for micro CT scanning yielded the highest-quality 3D images of a human peripheral nerve fascicle.

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