Compatibility Of Human Nasoseptal Chondrocytes And Porcine Acellular Dermal Matrix

Objectives:For human cartilage tissue,the application of tissue engineering provides a new surgical plan for the treatment of cartilage trauma.The principle is mainly to cultivate chondrocytes in vitro and obtain large cartilage tissue through in vitro propagation of chondrocytes to repair the lesions of cartilage tissue.The septal cartilage,which is the easiest cartilage tissue in the human body,can be used for tissue engineering research.In this experiment,human nasal septal chondrocytes were perfectly combined with porcine acellular dermal matrix(PADM)to study their compatible properties and provide a theoretical basis for cartilage transplantation.Methods:1.Human chondrocytes were isolated from the nasal septum and the cultured human nasal septal chondrocytes were cultured and subcultured.The growth state of the chondrocytes was observed periodically and the growth curve was depicted.Immunofluorescence staining with DAPI,Toluidine Blue,and Collagen II was performed to observe the cell morphology.And growth conditions.2.The human nasal septum cells with good growth state after passaging were planted on PADM and their compatibility was studied.After cells were grown on PADM for a certain period of time,histochemical staining and field emission scanning electron microscopy were performed to observe the morphology of human nasal septal chondrocytes on PADM.At the same time,RT-PCR was performed to determine the function of cells on PADM.Results :The human nasal septal chondrocytes were successfully isolated and cultured.The mature chondrocytes were observed to be polygonal and slightly spindle-shaped.After the third-generation human nasal septal chondrocytes cultured for one week in vitro,immunofluorescence staining of type II collagen showed blue and red positive staining of nuclei and cytoplasm.The DAPI staining showed that the cells on the surface of the culture dish had a round shape,a more regular shape,and a uniform density.Toluidine blue staining revealed a large number of blue-stained irregular polygonal cells on the surface of the slide.The MTT method showed that the third generation human nasal septal chondrocytes had normal proliferation patterns.After the third generation of human nasal septal chondrocytes were cultured on PADM for one week and stained with DAPI,a large number of cells were distributed on the PADM structure under a microscope,and the density was uniform.Scanning electron microscopy showed that the nasal septal chondrocytes adhered,stretched and grew well on the surface and in the pores of the PADM,and adjacent cell protrusionscould connect to each other.RT-PCR results showed that there was no significant difference in the expression of Collagen X,Collagen X,SOX9,Aggrecan,and osteoblast-associated Collagen I in human nasal septal chondrocytes from PADM compared with human nasal septal chondrocytes alone(P>0.05).Conclusions:Through this experiment,we successfully isolated and subcultured human nasal septal chondrocytes,and preliminary understanding of cells was performed by morphological observation and histochemical staining.At the same time,the proliferation of human nasal septal chondrocytes was detected by MTT assay,which proved that human nasal septal chondrocytes grew well in vitro.The most stable third generation human nasal septal chondrocytes were then seeded on PADM.After culture,histochemical staining,scanning electron microscopy and RT-PCR showed that the third generation of human nasal septal chondrocytes cultured on PADM had good morphology and growth status.The above studies confirm that human nasal septal chondrocytes can grow normally and divide in vitro.When cultured on PADM,they can still maintain normal reproductive ability and have good biocompatibility with PADM,providing basic research for cartilage tissue engineering.