Feasibility Study About The Three Dimensional Scaff-old Made Of Hydroxyapatite And N-carboxyethylchi-Tosan To Construct Tissue Engineering Trachea

Objective:To explore the feasibility about the three dimensional scaffold which is made of hydroxyapatite and N-carboxyethylchitosan constructed tissue engineering trachea.1.Through primary culture and appraisement of chondrocyte to explore the feasibility about the chondrocyte as seeds cell.2.To prepare the three dimensional scaffold of hydroxyapatite based on N-carboxyethylchitosan,and seed the chondrocyte on the surface of the scaffold,then through observeing the growth of the chondrocyte to explore the feasibility about the three dimensional scaffold which is made of hydroxyapatite and N-carboxyethylchitosan constructed tissue engineering trachea.Methods:(1)Obtains the rabbit auricle original generation of cartilage cell through two step enzyme digestion and transfer of generation raise.(2) Through the immunity histochemistry, the immunity fluorescence as well as the toluidine blue dye to examinate the collagen-Ⅱ and the osamine polyose which are Secreted from the cartilage cell for reaching the appraisal purpose.(3) The Three dimensional NCECS/n-HA porous scaffold were prepared by means of vacuum freeze-drying method,and examinated the porosity and the aperture size of the scaffold’s surface with scanning electron microscope (SEM).(4) In vitro plants the second generation of cartilage cell to the NCECS/n-HA scaffold surface raises for one week,and examination cartilage cell in support surface growth situation with SEM.Results：(1) To obtain the relatively pure chondrocyte through the collagenase-Ⅱ and trypsogen,The trypsogen difference digestion law and the machinery blow the division to be helpful to purificated the original generation of chondrocyte.(2) The collagen-Ⅱ immunity histochemistry,the immunity fluorescence as well as the toluidine blue dyeing demonstration obtains the cell could specificity secrete collagen and the osamine polyose,so the obtains cell is cartilage cell.(3) The scaffold surface which make through the vacuum freezing drying method becomes the porosity to reach as high as90%,the aperture is70-200nm,so it is Suit to grow for the cartilage cell.(4) The cartilage cell and the NCECS/n-HA scaffold altogether raise one week later. SEM demonstrates that the cartilage cell can good grow on the scaffold surface.Conclusion:1.To gain the cell which is chondrocyte through the collagenase-Ⅱ and trypsogen to digest the rabbit’s cartilago auricularis,so it can made as seeds cell to constructed tissue engineering trachea.2.The three dimensional scaffold which is made of hydroxyapatite and N-carboxyethylchitosan has high porosity,and the aperture is suit to the growth of the chondrocyte,so the scaffold can constructe tissue engineering trachea as supporter of the chondrocyte.