Dedifferentiation of chondrocyte greatly restricts its function and application,however,it is poorly understood except a small number of canonical markers.The non-cell-adhesive property endows polysaccharide hydrogel ...Dedifferentiation of chondrocyte greatly restricts its function and application,however,it is poorly understood except a small number of canonical markers.The non-cell-adhesive property endows polysaccharide hydrogel with the ability to maintain chondrocyte phenotype,which can serve as a platform to identify new molecular markers and therapeutic targets of chondrocyte dedifferentiation.In this study,the high-throughput RNA sequencing(RNA-seq)was first performed on articular chondrocytes at primary(P0)and passage 1(P1)stages to explore the global alteration of gene expression along with chondrocyte dedifferentiation.Significantly,several potential marker genes,such as PFKFB3,KDM6B,had been identified via comparatively analyzing their expression in P0 and P1 chondrocytes as well as in 3D constructs(i.e.chondrocyte-laden alginate hydrogel and HA-MA hydrogel)at both mRNA and protein level.Besides,the changes in cellular morphology and enriched pathway of differentially expressed genes during chondrocyte dedifferentiation was studied in detail.This study developed the use of hydrogel as a platform to investigate chondrocyte dedifferentiation;the results provided new molecular markers and potential therapeutic targets of chondrocyte dedifferentiation.展开更多
基金The work is financially supported by the China Postdoctoral Science Foundation(Grant No.2020M671993)Start-Up Grant for Professor(SGP 9380099 to D.-A.W.)City University of Hong Kong,and the National Natural Science Foundation of China,China(Grant No.51973180 and 21604045).
文摘Dedifferentiation of chondrocyte greatly restricts its function and application,however,it is poorly understood except a small number of canonical markers.The non-cell-adhesive property endows polysaccharide hydrogel with the ability to maintain chondrocyte phenotype,which can serve as a platform to identify new molecular markers and therapeutic targets of chondrocyte dedifferentiation.In this study,the high-throughput RNA sequencing(RNA-seq)was first performed on articular chondrocytes at primary(P0)and passage 1(P1)stages to explore the global alteration of gene expression along with chondrocyte dedifferentiation.Significantly,several potential marker genes,such as PFKFB3,KDM6B,had been identified via comparatively analyzing their expression in P0 and P1 chondrocytes as well as in 3D constructs(i.e.chondrocyte-laden alginate hydrogel and HA-MA hydrogel)at both mRNA and protein level.Besides,the changes in cellular morphology and enriched pathway of differentially expressed genes during chondrocyte dedifferentiation was studied in detail.This study developed the use of hydrogel as a platform to investigate chondrocyte dedifferentiation;the results provided new molecular markers and potential therapeutic targets of chondrocyte dedifferentiation.
