Effect of chemical treatments on tendon cellularity and mechanical properties

Abstract

Removal of cells may decrease the antigenicity and risk of disease transmission associated with tendon al-lografts and xenografts. An ideal cell removal method would not compromise graft structure and mechanical properties. This study compared the effects of three extraction chemicals [t-octyl-phenoxypolyethoxyethanol (Triton X-100), tri(n-butyl)phosphate (TnBP), and sodium dodecyl sulfate (SDS)] on tendon cellularity, structure, nativity, and mechanical properties. Rat tail tendons were soaked in extraction solutions for various time periods (12-48 h) and concentrations (0.5-2%), then they were rinsed with distilled water and ethyl alcohol. Histological analysis and tensile tests were performed on control and chemically treated tendons. Changes in collagen nativity were estimated by mechanical testing following incubation in a trypsin solution. Treatment of tendons with 1% Triton X-100 for 24 h disrupted the collagen fiber structure and did not remove cells. Treatment with 1% SDS for 24 h or 1% TnBP for 48 h resulted in an acellular tendon matrix with retention of near normal structure and mechanical properties. Consistent with previous studies demonstrating cell removal from other tissue types using SDS and TnBP, our preliminary results suggest these treatments are potentially useful for removing cells from tendon allografts or xenografts without compromising the graft structure or mechanical properties.

Chemical decellularization: a promising approach for preparation of extracellular matrix

“…Some detergents can induce alterations in the collagen fibers resulting in decreased mechanical stability of the tissue 52,53 .…”

Antibacterial activity and composition of decellularized goat lung extracellular matrix for its tissue engineering applications

“…Many organs, e.g. urinary bladder [6], small intestinal [7][8][9][10][11], heart valves [12][13][14][15], skin [16,17], liver [18], anterior crucial ligament bone [19], kidney [20], pericardium [21], blood vessels [22], nerves [23], skeletal muscle [24], tendon [25] and vocal cord [26,27] from bovine and porcine tissues, have been used for fabricating the ECMbased scaffold. But, the use of goat-lung has not been focused for tissue engineering applications.…”

Freeze-Thaw Cycles Enhance Decellularization of Large Tendons

“…10 The ideal scaffold should not only be biocompatible and match the natural biomechanical properties, but should also comprise naturally structured, extracellular matrix proteins that can interact with the repopulating cells and direct them toward tenogenic differentiation and matrix remodeling. 10,11 Decellularization of tendon tissue offers the unique opportunity of obtaining a scaffold with a natural extracellular matrix structure that is hypoimmunogenic 13,14 and displays biomechanical properties very similar to the original tissue, [15][16][17][18] providing great advantages for potential clinical application and research use.…”

“…[15][16][17][18][19][20][21] Results obtained after applying different detergent-based protocols remain conflicting. While t-octyl-phenoxypolyethoxyethanol (Triton X-100) did not lead to satisfactory results in an early study, 15 it was found to be most effective by others. 19 Sodium dodecyl sulfate (SDS) was reported to be effective but to reduce cytocompatibility due to matrix alterations; however, the protocol used in these studies did not only include incubation in SDS, but also in Triton X-100.…”