Preparation and performance evaluation of a novel detrgent-enzymatic tissue-engineered tracheal matrix in rabbits / 中华胸心血管外科杂志

ABSTRACT

Objective To determine the detergent-enzymatic cycles and evaluate the biomechanical characteristics as well as extracellular matrix integrity of the decellularized tracheal scaffold in rabbit.Methods Forty tracheal segments were harvested from New Zealand white rabbits.Thirty-five of tracheas were subjected to a detergent-enzymatic method of decellularization for 1/3/5/6/7/8/9 cycles,respectively,and other five were stored in phosphate-buffered saline at 4℃ as a control.Comparative examinations were performed by the macroscopic view,histological view(hematoxylin and eosin stain,Movat Pentachrome stain,4-6-diamidino-2-phenylindole),scanning electron microscope (SEM) and biomechanical properties between decellularized groups and control group.Results After 7 detergent-enzymatic cycles,almost complete decellularized tracheae,retaining the hierarchical and mechanical properties of the native tissues,could be obtained.Histological and molecular biology analysis demonstrated that all cellular components and nuclear material were removed.SEM analysis revealed that the decellularized matrices retained the hierarchical structures of native trachea,and biomechanical tests showed that decellularization approach did not led to any influence on tracheal morphological and mechanical properties.Immunofluorescence analysis show a significant reduction of nuclear material in decellularized tracheas (P ＜ 0.05).Conclusion In conclusion,this work suggests that 7 cycles of the modified DEM generates a bioengineered rabbit tracheal matrix that is structurally and mechanically similar to native trachea which could be a better selection for tracheal reconstruction with tissue engineering method.