戊二醛/1-(3-二甲基胺丙基)-3-乙基碳二亞胺交聯的明膠支架於腸平滑肌細胞組織工程之研究

Abstract

Short bowel syndrome (SBS) patients is massive resection of the small intestine. The patients require total parenteral nutrition (TPN), medication, and surgical operation to different degrees, but these therapies have side effects. This study aims to crosslink gelatin with glutaraldehyde (GA) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) to fabricate a tubular scaffold for small intestine tissue engineering. This study aims to crosslink 10, 15, 20 wt% difference concentration gelatin with glutaraldehyde (GA) and 1-ethyl-3-3-dimethylaminopropyl carbodiimide hydrochloride (EDC) than use freeze dryer to fabricate a outside 0.3 cm, inside 0.8 cm higher 3 cm tubular scaffold for small intestine tissue engineering. The scaffold’s physical and chemical properties were characterized. Its biocompatibility was evaluated by observing smooth muscle cell (SMC) seeding on the scaffold and in vivo, in vitro study. Use Scanning Electron Microscope observe the glutaraldehyde crosslinked scaffold had a pore size range of 100-300 μm, Ninhydrin test get 30 % crosslinking index, Use ddH2O make Swelling tset get 600 % swelling ratio, Archimedes test get porosity of 24-40 % and the machine test get Young’s modulus of 64-102 MPa. The results obtained from in vitro degradation study showed that the GA groups degrade before 24 hours. The biocompatibility results also show GA groups were cytotoxic to L929 cells, under the scaffold’s cell cytotoxic ratio is 70%. In vivo study observed many immunization cell around scaffold after implantation 1 week. Meanwhile, the EDC crosslinked scaffolds resulted to a pore size range of 100-300 μm, 80 % crosslinking index, 320-500 % swelling ratio, porosity of 40-64 % and a Young’s modulus of 55 MPa, scaffold can maintain their structure in PBS more than 14 days, scaffold can maintain there structure in collagenase PBS 7 days. In vivo study observed few immunization cell around scaffold after implantation 1 week. Cells were seeded on the EDC crosslinked scaffolds using centrifugal cell immobilization (CCI) method. The seeding efficiency was 71 %. The cell’s morphology and growth on the scaffold were observed with the use of SEM, immunofluorescence (IF) and Immunohistochemistry (IHC) after static growth. At future will implantation in biological’s body to evalulate for intenstine’s regeneration.