Due to the poor repair ability of cartilage tissue,regenerative medicine still faces great challenges in the repair of large articular cartilage defects.Quercetin is widely applied as a traditional Chinese medicine in...Due to the poor repair ability of cartilage tissue,regenerative medicine still faces great challenges in the repair of large articular cartilage defects.Quercetin is widely applied as a traditional Chinese medicine in tissue regeneration including liver,bone and skin tissues.However,the evidence for its effects and internal mechanisms for cartilage regeneration are limited.In the present study,the effects of quercetin on chondrocyte function were systematically evaluated by CCK8 assay,PCR assay,cartilaginous matrix staining assays,immunofluorescence assay,and western blotting.The results showed that quercetin significantly up-regulated the expression of chondrogenesis genes and stimulated the secretion of GAG(glycosaminoglycan)through activating the ERK,P38 and AKT signalling pathways in a dose-dependent manner.Furthermore,in vivo experiments revealed that quercetin-loaded silk protein scaffolds dramatically stimulated the formation of new cartilage-like tissue with higher histological scores in rat femoral cartilage defects.These data suggest that quercetin can effectively stimulate chondrogenesis in vitro and in vivo,demonstrating the potential application of quercetin in the regeneration of cartilage defects.展开更多
Objective To immortalize human articular chondrocytes ( HACs) using gene transfection and to maintain stable phenotype of transformed HACs after induction.Methods HACs were transfected with the retroviral vector pLXSN...Objective To immortalize human articular chondrocytes ( HACs) using gene transfection and to maintain stable phenotype of transformed HACs after induction.Methods HACs were transfected with the retroviral vector pLXSN encoding human papillomavirus 16E7 (HPV16E7), and the transformed clones were sorted and proliferated. Karyotype analysis, clone forming tests and nude mice tumor forming tests were applied to check the characteristics of the transformation. Type II collagen of transformed chondrocytes was inducted with free serum medium (FSM) supplemented with nutridoma-sp and ascorbate.Results Immortalized HACs were isolated with fifty passages achieved. The HPV16E7 transformed cells were confirmed to be benign. Induction of FSM with nutridoma-sp and ascorbate promoted type II collagen of transformed chondrocytes to the high levels of normal chondrocytes.Conclusion HACs transformed with HPV16E7 survive for long periods in vitro, and type Ⅱ collagen can maintain stability after induction.展开更多
基金supported by the National Natural Science Foundation of China(81771038,82071082)Sciece and Technology Commission of Shanghai Municipality(18441902900,19142202200)。
文摘Due to the poor repair ability of cartilage tissue,regenerative medicine still faces great challenges in the repair of large articular cartilage defects.Quercetin is widely applied as a traditional Chinese medicine in tissue regeneration including liver,bone and skin tissues.However,the evidence for its effects and internal mechanisms for cartilage regeneration are limited.In the present study,the effects of quercetin on chondrocyte function were systematically evaluated by CCK8 assay,PCR assay,cartilaginous matrix staining assays,immunofluorescence assay,and western blotting.The results showed that quercetin significantly up-regulated the expression of chondrogenesis genes and stimulated the secretion of GAG(glycosaminoglycan)through activating the ERK,P38 and AKT signalling pathways in a dose-dependent manner.Furthermore,in vivo experiments revealed that quercetin-loaded silk protein scaffolds dramatically stimulated the formation of new cartilage-like tissue with higher histological scores in rat femoral cartilage defects.These data suggest that quercetin can effectively stimulate chondrogenesis in vitro and in vivo,demonstrating the potential application of quercetin in the regeneration of cartilage defects.
基金This study was supported by a grant from the National Natural Science Foundation of China (No. 39830100).
文摘Objective To immortalize human articular chondrocytes ( HACs) using gene transfection and to maintain stable phenotype of transformed HACs after induction.Methods HACs were transfected with the retroviral vector pLXSN encoding human papillomavirus 16E7 (HPV16E7), and the transformed clones were sorted and proliferated. Karyotype analysis, clone forming tests and nude mice tumor forming tests were applied to check the characteristics of the transformation. Type II collagen of transformed chondrocytes was inducted with free serum medium (FSM) supplemented with nutridoma-sp and ascorbate.Results Immortalized HACs were isolated with fifty passages achieved. The HPV16E7 transformed cells were confirmed to be benign. Induction of FSM with nutridoma-sp and ascorbate promoted type II collagen of transformed chondrocytes to the high levels of normal chondrocytes.Conclusion HACs transformed with HPV16E7 survive for long periods in vitro, and type Ⅱ collagen can maintain stability after induction.
