ABSTRACT
BACKGROUND:Recently,the development of tissue-engineered technique lms broadened the study of artificial esophagus.Some investigators have inoculated esophageal epithelial cells cultured in vitro onto compound polymer material and successfully constructed tissue-engineered esophagus.OBJECTIVE:To investigate the feasibility of tissue-engineered artificial esophagus by combining dog esophageal epithelail cells and an acelhilarized porcine thoracic aorta allogenic matrix.DESIGN:An experimental observation.SETTING:Central Laboratory,Taishan Medical College.MATERIALS:This study was carried out in the Central Laboratory.Talshan Medical College from June to December in 2004.Three hybrid dogs,24-hour-old,were provided by the Laboratory Animal Center,Taishan Medical College.The protocol was performed in accordance with ethical guidelines for the use and care of animals.The experimenhal instruments and reagents were as follows:CO2 incubator(MCO-15AC,SANYO),hypothermal high-speed centrifuge(RC-26,Dupont),trypsin,transferrin,type Ⅱ collagenase(Gibco),dulbecco's modified eagle's medium(DMEM),DispaseⅡ isolated enzyme,and rat monoclonal anti-keratin antibody(Sigma).METHODS:Acellularization of porcine aortas was performed by a method of enzyme-detergent.Esophageal epithelial cells of hybrid dogs were in vitro isolated,cultured and proliferated.Next,they were inoculated onto an acellularized porcine thoracic aortas allogenic matrix seaffold.Three and seven days later,the growth of esophageal epithelial cells on the acellularized matrix was observed under an electron microscope.MAIN OUTCOME MEASURES:Morphology of esophageal epithelial cells cultured in vitro;Biocompatibility of acellular matrix and dog esophageal epithelial cells.RESULTS:The acellularized procedure resulted in an almost complere removal of the cells and the loose three-dimensional matrix.The acellular matrix could be reseeded with expended esophageal epithelial cells in vitro.and esophageal epithelial cells had the potential of spread and proliferation.CONCLUSION:Acellular matrix possesses satisfactory biocompatibility for allogenic esophageal epithelial cells.Tissue-engineered artificial esophagus can be generated in vitro by a combination of esophageal epithelial cells and allogenic aceilularized matrix.