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      • KCI등재

        Evaluation of an Immune-privileged Scaffold for In vivo Implantation of Tissue-engineered Trachea

        Shu Pan,Fei Sun,Hongcan Shi,Fangbiao Zhang,Xingchen Liu,Weidong Zhang 한국생물공학회 2014 Biotechnology and Bioprocess Engineering Vol.19 No.5

        Forty tracheas were harvested from donor NewZealand rabbits. Thirty of the tracheas were randomlydivided into four treatment groups corresponding to 4, 5, 6,or 7% NaClO4 and one untreated group (n = 6 each group). Scanning electron microscopy distinctly revealed thecilium of epithelial cells in the fresh trachea. The internalsurface of the trachea was rough in the 4% treatment groupand smooth in the 5% treatment group, whereas the matrixwas fractured in the 6% treatment group and highlyfractured in the 7% treatment group. We observed that thenumber of nuclei in the cells of the 4, 5, 6, and 7%treatment groups decreased compared to the cells of theuntreated group (p < 0.05). Although there was a significantdecrease in maximum tensile strength, tensile strain atfracture and the elastic modulus (p < 0.05) with increasingconcentrations of NaClO4, the content of glycosaminoglycans(GAGs) did not significantly decline (p > 0.05) in the 5%treatment group. In addition, histopathological analysisshowed that the fiber component and basement membraneof the matrix in the 5% treatment group were retained afteroptimal decellularization. Despite the preserved cartilage,in vitro immunohistochemical analysis revealed that thematrix did not show the presence of major histocompatibilitycomplex (MHC) antigens. The remaining ten donor tracheas,which were divided into a positive control group and anoptimal decellularized group, were used for allogeneictransplantation. Blood samples were taken regularly, andhistologic examinations were performed at 30 days postimplantation,which showed no significant immune rejection. In conclusion, we surveyed the structural integrity throughmorphological observation and compared the biomechanicaland immunogenic changes in the tracheal matrix under thedifferent treatments. The optimal decellularized trachealmatrix with preserved cartilage, which was acquired via 5%NaClO4 treatment, exhibited structural integrity, antigen cellremoval and immune privilege and would be suitable for useas a tissue-engineered trachea for in vivo transplantation inrabbit models.

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        A Comparative Study of the Effects of Different Decellularization Methods and Genipin-Cross-Linking on the Properties of Tracheal Matrices

        Yi Zhong,Ai Jiang,Fei Sun,Yuanfan Xiao,Ying Gu,Lei Wu,Yujie Zhang,Hongcan Shi 한국조직공학과 재생의학회 2019 조직공학과 재생의학 Vol.16 No.1

        BACKGROUND: Different decellularization methods can affect the integrity and the biomechanical and biocompatible properties of the tracheal matrix. Natural cross-linking with genipin can be applied to improve those properties. The goals of this study were to evaluate the effects of different decellularization methods on the properties of genipin-cross-linked decellularized tracheal matrices in rabbits. METHODS: The tracheas of New Zealand rabbits were decellularized by the Triton-X 100-processed method (TPM) and the detergent-enzymatic method (DEM) and were then cross-linked with genipin. Mechanical tests, haematoxylin–eosin staining, Masson trichrome staining, Safranin O staining, DAPI staining, scanning electronic microscopy (SEM), and biocompatibility tests were used to evaluate the treatment. The bioengineered trachea and control trachea were then implanted into allogeneic rabbits for 30 days. The structural and functional analyses were performed after transplantation. RESULTS: The biomechanical tests demonstrated that the biomechanical properties of the decellularized tracheas decreased and that genipin improved them (p\0.05). The histological staining results revealed that most of the mucosal epithelial cells were removed and that the decellularized trachea had lower immunogenicity than the control group. The analysis of SEM revealed that the decellularized trachea retained the micro- and ultra-structural architectures of the trachea and that the matrices cross-linked with genipin were denser. The biocompatibility evaluation and in vivo implantation experiments showed that the decellularized trachea treated with the DEM had better biocompatibility than that treated with the TPM and that immunogenicity in the cross-linked tissues was lower than that in the uncross-linked tissues (p\0.05). CONCLUSIONS: Compared with the trachea treated with the TPM, the rabbit trachea processed by the DEM had better biocompatibility and lower immunogenicity, and its structural and mechanical characteristics were effectively improved after the genipin treatment, which is suitable for engineering replacement tracheal tissue.

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