DEVELOPMENT OF COATING MATERIAL FOR ARTIFICIAL TRACHER

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The outgrowth culture method was used to grow NHBEC.A normal human bronchus was dissected into 3mm squares and these bronchial segments were placed with the mucosal surface up on a polystirene dish and a Marlex mesh used as an artificial tracheal that was coated with one or more extracellular matrices that are described below.Then explant culture was performed using a serum free culture, LHC-9. A phase contrase microscope was used to examine the outgrowth of NHBEC in the periphery of the bronchial segments, and the surface area on completion of the experiment was masured and used as a growth indicator.Experiment 1 : It was examined as to which type of collagen (type I,II,III or IV) would promote the growth of NHBEC.Experiment 2 : Based on the result obtained in experiment 1, type IV collagen and two cellular adhesive proteins, laminin and fibronectin, were used to coat a culture dish for comparison. Experiment 3 : In group A,a Marlex mesh was coated with type I collagen only, and in group B,a Marlex mesh was coated with type I collagen, type IV collagen, laminin and fibronectin to examine the growth of NHBEC.The results obtained in experiment I showed that the growth of NHBEC in the polystirene dish coated with type IV collagen was most stable when compared to the other types of collagen, in experiment 2 showed that the combination of type IV collagen, laminin and fibronectin promoted the growth of NHBEC,in experiment 3 showed that the mesh coated with type IV collagen, laminin and fibronectin promoted the growth of NHBEC.From these findings, it was suggested that the combination of type IV collagen, laminin and fibronectin could beused as an effective base to cover the interior surfase of artificial trachea with bronchial epithelial cells.