Effects of various decellularization methods on histological and biomechanical properties of rabbit tendons

Exp Ther Med. 2014 Aug;8(2):628-634. doi: 10.3892/etm.2014.1742. Epub 2014 May 28.

Abstract

The aim of the present study was to investigate the effects of various decellularization methods on the histological and biomechanical properties of rabbit tendons. In total, six chemical reagents, including 1% t-octyl-phenoxypolyethoxyethanol (Triton-X 100), 0.5% sodium dodecyl sulfate (SDS), 1% tri-n-butyl phosphate (TnBP), 1% Triton-X 100 + 0.5% SDS, 1% TnBP + 0.5% SDS and 1% TnBP + 1% Triton-X 100, were used on rabbit semitendinosus muscles and flexor digitorum tendons for 24 h to remove cells. Hematoxylin and eosin staining was applied for histological observation, while tension testing was used for biomechanical studies. The effects of the various decellularization methods on the histological structure and biomechanical properties of rabbit tendons were evaluated. A group of fresh tendons treated with phosphate-buffered saline served as controls. The various decellularization methods resulted in different effects on the tendons. All the treatment groups exhibited a decrease in tendon biomechanical properties, but no statistically significant differences were observed among the experimental groups. The extensibility of the 1% TnBP-treated group was found to be greater than that of the other groups; however, the difference was not statistically significant. Histologically, the 1% TnBP + 0.5% SDS treatment was shown to have the least impact on the rabbit tendon structure, with good decellularization and no clear cellular remnants observed. The 1% Triton-X 100 + 0.5% SDS treatment had a pronounced effect on the tendon collagen structure and a number of collagen ruptures were observed. Overall, 1% TnBP + 0.5% SDS was found to be the most effective compared with the other treatments, as this treatment preserved the tendon collagen structure while completely removing the cells. Tendons treated with 1% TnBP + 0.5% SDS were histologically similar to normal tendon tissue and biomechanically similar to the tendons in the control group.

Keywords: acellular; biomechanics; collagen; extracellular matrix; tendon.