Chemical extraction method versus detergent-enzymatic method for the preparation of tissue-engineered trachea matrix / 中国组织工程研究

ABSTRACT

BACKGROUND:Acel ular tracheal matrix is similar to the native trachea with structure and biological performance preserved after decel ularization, and it is an important aim in tissue engineering to find an effective method of decel ularization. OBJECTIVE:To select the optimal decel ularization method through comparing chemical extraction method with detergent-enzymatic method for preparing rabbit tissue engineering trachea matrix. METHODS:Thirty tracheas from New Zealand white rabbits were randomly divide into three groups. Twenty of rabbit tracheas were subjected to decel ularization using 2%TritonX-100 combined with deoxyribonuclease I and ribonuclease (chemical extraction method group), and sodium deoxycholate combined with deoxyribonuclease I (detergent-enzymatic group), respectively. The other ten were given no intervention as controls. Samples were col ected and observed by hematoxylin-eosin staining, Masson-trichrome staining, safranin O staining, DAPI staining and scanning electronic microscope. RESULTS AND CONCLUSION:Hematoxylin-eosin staining demonstrated that compared with the control group, almost al cel ular components of the mucosal epithelium were removed in the detergent-enzymatic and chemical extraction groups, and there were few remnant chondrocytes. Masson-trichrome staining indicated that compared with the control group, components of the mucosal layer chondrocytes in the chemical extraction and detergent-enzymatic groups were completely removed, with only part of remained chondrocytes in the cartilage zone. Glycosaminoglycan was slightly decreased both in the chemical extraction and detergent-enzymatic groups shown by Safranin O staining, but more reduction was found in the chemical extraction group. DAPI staining reveled that only a smal amount of cartilage cel s remained in the dense layer of cartilage and lacuna both in this two methods. Scanning electronic microscope showed that using the detergent-enzymatic method there were the hierarchical structures of native trachea, but slight disruption using the chemical extraction method. In conclusion, decel ularized rabbit trachea matrix obtained by detergent-enzymatic method is better, with little disruption to the matrix.