The Study Of Synovial Mesenchymal Stem Cells/Chondrocytes And ECM-PCL-HA Implanted In Articular Cartilage In Vitro

Objective:In this study,we observed the ability of synovial mesenchymal stem cells/chondrocyte-rich decellularized scaffolds containing extracellular matrix to form cartilage under three-dimensional conditions.Methods:(1)The synovial tissue,cartilage tissue,and inguinal adipose tissue of rabbits were isolated and cultured and expanded in vitro.(2)The mesenchymal stem cells of the well-growing first-generation adipose-derived transfected with lentivirus-mediated TGF-?3 and inoculate them into a 3D printed polycaprolactone/hydroxyapatite(PCL-HA)composite scaffold material.(3)The synovial mesenchymal stem cells/chondrocytes were seeded in a constructed scaffold material rich in extracellular matrix(ECM)to construct articular cartilage in vitro.(4)The effects of COL-I,COL-II,SOX-9,and Aggrecan gene expression on RNA levels were detected by RT-PCR.(5)The expression of cartilage-specific protein was detected by immunoblotting.Results:(1)After culturing the scaffold complex of transfected adipose-derived mesenchymal stem cells for 14 days,laser confocal scanning showed that the cells were uniformly distributed on the surface of the scaffold,and the cells gradually decreased after scanning layer by layer.(2)After synovial mesenchymal stem cells/chondrocytes were mixed and cultured in the ECM-PCL-HA scaffold material,and then cultured for 1 week,2 weeks and 4 weeks in vitro,synovial mesenchymal stem cells/Chondrocytes were inoculated in the ECM-PCL-HA group,and only synovial mesenchymal stem cells or chondrocytes were inoculated in the ECM-PCL-HA group.Aggrecan and COL-II can be seen from the time(1 week,2 weeks and 4 weeks),SOX-9 and COL-1 all showed an upward trend,and there had significant difference(P<0.05).Comparing the experimental group with the two control groups,it can be seen that the mRNA expression of Aggrecan,COL-II and SOX-9 increased significantly(P<0.05);but there was no significant difference in the expression of COL-I mRNA gene between the two groups(P>0.05);(3)Synovial mesenchymal stem cells/chondrocytes were cultured in ECM-PCL After 4 weeks of culturing the HA scaffold material,the protein level of COL-II was higher than that of synovial mesenchymal stem cells cultured in the ECM-PCL-HA scaffold material group(P<0.05)and chondrocytes cultured in the ECM-PCL In the HA scaffold material group(P<0.01),the difference was statistically significant.Conclusion: The two groups of cells were mixed and cultured in the extracellular matrix-rich scaffold material,and there was expression of cartilage-related genes or proteins,and significantly higher than that of simple synovial mesenchymal stem cells or pure chondrocytes inoculated into ECM-PCL-HA Scaffolds.