Genipin cross-linked decellularized scaffold for allogenic transplantation in situ

Abstract

Objective To explored the bio-compatibility and cartilage regeneration of the rabbits genipin cross-linked decellularized scaffold, to provide experimental and theoretical support for the clinical application of genipin cross-linked decellularized scaffold. Methods Detergent-enzyme method was used to prepare decellularized tracheal scaffolds. Cellular content of native trachea and decellularized trachea were compared by 4′, 6-diamidino-2-phenylindole(DAPI) staining. Masson trichrome staining was used to compare the histological structure of the progenitor tube, decellularized trachea, and genipin cross-linked decellularized trachea. Nine adult New Zealand white rabbits were randomly divided into autologous tracheal transplantation group (negative control group), allogeneic tracheal transplantation group (positive control group), and genipin cross-linked decellularized tracheal transplantation group (experimental group). Autologous bone marrow mesenchymal stem cells were implanted on the surface of trachea in each group. The blood cells and type II collagen were detected to compare the inflammatory response and chondrocyte regeneration after tracheal orthotopic transplantation in the three groups. Results After DAPI staining and light microscope observation (×200), the cell content of the acellular 7-cycle trachea [(143.0 ± 71.1) cells/field] was significantly lower than that of the native trachea [(853.5 ± 149.6) cells/field], and the difference was statistically significant (P<0.001). Masson′s trichrome staining showed that the tissue structure of genipin cross-linked decellularized trachea was more complete. Blood cell analysis and type II collagen test results showed that genipin cross-linked decellularized trachea transplanted with bone marrow mesenchymal stem cells after transplantation in situ has little rejection and can be converted into chondrocytes by the action of related growth factors in vivo. Conclusions Genipin cross-linked decellularized tracheal scaffold combined with stem cell transplantation can successfully construct a tracheal in situ replacement model. This study provides a strong support for the research of tissue engineering trachea. Key words: Tissue engineering; Trachea; Transplantation; Genipin cross-linked; Mesenchymal stem cells