The Characteristics Of The Human Umbilical Cord Wharton’s Jelly Extracellular Matrix And The Human Articular Cartilage Extracellular Matrix Prepared By Differential Centrifugation And Their Effect On The Rabbit Chondrocyte Cell Proliferation And Phenoty

Objective: make a comparison between the physical and physical chemistry of preparedhACECM. To observe the proliferation and phenotype of rabbit chondrocytes on thehuman Wharton’s jelly extracellular matrix and human articular cartilage extracellularmatrix during in vitro expansion.Methods: We prepared the human cartilage extracellular matrix by the simpledifferential centrifugation and the physics and physical chemistry separately. We made aseries of comparison by DNA&GAG quantification and hydroxyproline quantificationand the MTT assay. The hWJECM and hACECM was prepared with the differentialcentrifugation. Chondrocytes were isolated from articular cartilage of New Zealandwhite rabbits and cultured on hWJECM-coated plates and hACECM-coated plates.These chondrocytes were cultured on the hWJECM-plated wells and hACECM-platedwells as the experiment groups. The group of Simple medium was set as the controlgroups. Cells were analyzed regarding cell growth, morphology using an invertedmicroscope, toluidine blue staining, Cell Counting Kit-8(CCK-8) assay, DNA/GAGquantification, hydroxyproline quantification, and atomic force microscope. Weevaluated the cytotoxicity of hWJECM and hACECM by MTT assay.Results: hWJECM and hACECM were prepared by the differential gradientcentrifugation. And they were decellularized completely by hochest33258staining. The hWJECM and hACECM contain large amounts of glycosaminoglycans by toluidineblue staining and safranin O staining. We can find the collagen of two kinds of ECMmay arranged orderly by the SEM images. The amount of DNA of the cells of thehWJECM and hACECM after the cell removal was significantly less than before(p<0.05). hWJECM and hACECM remain a large number of GAG and collagen. Thesurface of the plates coated by the hWJECM and hACECM go ups and downsobviously by the atomic force microscopy images for the chondrocytes adhesion.By inverted microscope, the cell proliferation in experimental groups in5,10,15days are better than the control groups. The results of toluidine blue staining also provedthis point. The cell proliferation of the hWJECM groups and hACECM groups in the5,10,15days was better than the control groups by the inverted microscope, the results oftoluidine blue staining5,10,15days also proved this point. The proliferation of thehWJECM groups in the5,7days was significantly better than control group (p=0.0001;p=0.0006) by the CCK8cell proliferation assay. CCK-8cell proliferation assaysshowed that the cell proliferation of hACECM groups in5days were significantly betterthan the control groups (P=0.0298<0.05). The chondrocytes seeded on thehWJECM-coated plates and hACECM-coated plates were significantly more than thechondrocytes of the control groups in the amount of DNA (p <0.05)&GAG (p <0.05)quantification and in the amount of hydroxyproline quantification (p <0.05).Conclusion: The ECM by the physical method can remove the cells and debris. TheECM retained a large degree of GAG and collagen. Therefore, the hWJECM can be agood substitute for hACECM as a scaffold for cartilage tissue engineering. The ECMscaffolds made from human umbilical cord, which is better than the biomechanicalproperties of human cartilage ECM, as well as materials GAG content, biomechanicalstrength of the relationship with the type and content of collagen produced. It needssome further study.