The aim of this study was to investigate the tissue damage created by the two commonly used percutaneous tracheostomy methods of multiple dilatational tracheostomy and the controlled rotating dilation technique. A total of 21 sheep trachea samples were obtained from the slaughterhouse. The tracheas were separated into 3 groups of 7 samples in each. In Group 1, only an incision was made. In Group 2, the multiple dilatational tracheostomy method was applied, and in Group 3, the controlled rotating dilation technique. The tissue samples taken were separated into 2 equal pieces. One piece was used for histopathological examination and the other was examined under electron microscope. When the hematoxylin-eosin staining results and the electron microscope images of the sheep trachea slices were evaluated, cartilage integrity was observed to have been preserved in Groups 1 and 2. In Group 3, there was seen to be epithelial injury and disruption of the cartilage integrity. To reduce the complications of percutaneous tracheostomy techniques to a minimum, the protection of tissue integrity is important. The multiple dilatational tracheostomy method was seen to be superior to the controlled rotating dilation technique in respect of protecting tissue integrity.