A Comparative Examination of Two Methods of Sciatic Nerve Decellularization and Their Histological Analysis

Objective: This study aimed to decellularize bovine sciatic nerves using a combined Triton X-100 and SDS protocol and to compare the histological characteristics of the resulting scaffolds after processing by frozen sectioning versus paraffin sectioning, using hematoxylin and eosin (H&E) staining as the sole evaluation method. Methods: Bovine sciatic nerves were subjected to sequential exposure to 1 Triton X-100 for 24 h and 3% SDS for 48 h, followed by extensive washing to remove residual detergents. Native and decellularized nerve segments were processed in parallel using frozen sectioning (8 µm) or paraffin sectioning (5–6 µm). All sections were stained with H&E and examined by light microscopy. Results and Discussion: Macroscopically, decellularized nerves appeared white and semi-translucent, in contrast to the dense, opaque appearance of native tissue. H&E staining demonstrated complete removal of nuclear material in decellularized scaffolds with both processing approaches. Frozen sections yielded more intense staining and sharper visualization of microanatomical features, whereas paraffin sections showed lighter staining and slight compression or distortion of extracellular matrix elements. These discrepancies reflect processing-related artifacts—such as dehydration, heat exposure, and reduced section thickness in paraffin processing—rather than actual differences in extracellular matrix preservation. Conclusions: When routine histological evaluation of decellularized peripheral nerve scaffolds relies solely on H&E staining, frozen sectioning provides superior morphological clarity and minimizes processing artifacts compared with conventional paraffin sectioning.