Development of Methods to Obtain Grafts for the Anterior Abdominal Wall in Surgical Treatment of External Abdominal Hernias

Abstract

INTRODUCTION: Currently, tissue engineering is one of the most promising fields of medicine dealing with search for solutions to the problems of creation of biocompatible structures capable of partial or complete replacement of organs or tissues of an organism. An advanced method of tissue engineering implies using an extracellular matrix (ECM) cleared from the cellular material. The main problem of obtaining an ECM consists in the difficulty of selecting a suitable decellularization method and exposure time.

AIM: Development of a method for obtaining grafts for the anterior abdominal wall (AAW) and evaluation of the histological properties of the ECM to determine the optimal protocol for donor tissue decellularization providing a complete removal of cellular material without any significant damage to the scaffold.

MATERIALS AND METHODS: The experiments were conducted on 10 White Flemish Giant male rabbits. All the animals were withdrawn from the experiment, after which, on autopsy, AAW grafts were obtained for evaluation of the decellularization properties of three detergents (alkaline solutions): 2% SDS (sodium dodecyl sulfate) solution, 2% CHAPS (zwitterionic surfactant, 3-[(3-cholamidopropyl)-dimethylammonio]-2-hydroxy-1-propansulfonate) solution, and 1% Triton-X 100 solution. The decellularization quality was controlled in histological examination.

RESULTS: Morphological evaluation of tissue samples revealed that after decellularization of the material with 2% SDS solution and 2% CHAPS solution, the number of cells remaining in the preparations was comparable and significantly less (Me = 10 in a microscope field) than after treatment with 1% Triton-X 100 solution (Me = 45 in a microscope field). However, decellularization with 2% CHAPS solution led to fiber ruptures and tissue edema manifested by an increase in the distance between the fibers (Me = 163 µm).

CONCLUSIONS: Decellularization of the anterior abdominal wall tissues with 2% SDS solution provides a sufficient cell elimination while preserving the fiber integrity and ECM structure. 2% CHAPS solution was comparable to 2% SDS solution in terms of the quality of cell elements removal, but its more aggressive effect on collagen fibers was noted (their multiple ruptures and a more pronounced edema). 1% Triton-X 100 solution appeared to be of low efficiency in elimination of cellular material, although it had a minimal effect on the integrity of ECM network.

Keywords

tissue engineering / decellularization / extracellular matrix / sodium dodecyl sulfate / Triton-Х 100 / CHAPS

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