[Establishment of chondrocyte degeneration model in vitro by rat serum].
To establish a model of chondrocyte degeneration in vitro.



Chondrocytes were isolated from articular cartilages of newly born SD rats by digestion with typeⅡ collagenase. The chondrocytes were cultured with H-DMEM medium containing 10%FBS, 50 ng/mL IL-1β+10%FBS, 2.5% rat serum and 5% rat serum, respectively; and the chondrocytes at passage one were used in the experiments. The morphology changes were investigated under phase contrast microscope after chondrocytes were treated with rat serum and IL-1β. Proliferation of chondrocytes was detected by MTT method. The protein expression levels of PCNA, typeⅡ collagen and MMP-13 were examined by Western blotting. The levels of ADAMTS5, MMP-9, Aggrecan and SOX-9 mRNA were detected by real-time PCR.



The cell morphology was changed from polygon to spindle in both rat serum groups and IL-1β group, and the proliferation of chondrocytes in these groups was much higher than that in control group. The results showed that the expression levels of typeⅡ collagen, Aggrecan and SOX-9 decreased while the expression levels of MMP-13, MMP-9 and ADMATS5 were up-regulated in rat serum and IL-1β-treated groups compared with control group.



The results indicate that rat serum can induce chondrocyte degeneration and may be used for osteoarthritis model in vitro.
Huzhou Hospital Affiliated to Zhejiang Chinese Medical University, Huzhou 313000, China;2. Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai TCM University, Shanghai 201203, China.
Xiao-jun
XJ
Wang
Huzhou Hospital Affiliated to Zhejiang Chinese Medical University, Huzhou 313000, China;2. Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai TCM University, Shanghai 201203, China.
Hao
H
Zhang
Huzhou Hospital Affiliated to Zhejiang Chinese Medical University, Huzhou 313000, China;2. Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai TCM University, Shanghai 201203, China.
Hong-sheng
HS
Zhan
Huzhou Hospital Affiliated to Zhejiang Chinese Medical University, Huzhou 313000, China;2. Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai TCM University, Shanghai 201203, China.
Dao-fang
DF
Ding
D051722:ADAM Proteins / Q000378:metabolism; D000071116:ADAMTS5 Protein; D053674:Aggrecans / Q000378:metabolism; D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000473:pathology*; D024043:Collagen Type II / Q000378:metabolism; D004195:Disease Models, Animal; D053583:Interleukin-1beta / Q000494:pharmacology; D053509:Matrix Metalloproteinase 13 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D010003:Osteoarthritis / Q000473:pathology*; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D012333:RNA, Messenger; D051381:Rats; D017207:Rats, Sprague-Dawley; D055758:SOX9 Transcription Factor / Q000378:metabolism; D044967:Serum; D015854:Up-Regulation
D053674:Aggrecans; D024043:Collagen Type II; D053583:Interleukin-1beta; D018809:Proliferating Cell Nuclear Antigen; D012333:RNA, Messenger; D055758:SOX9 Transcription Factor; D051722:ADAM Proteins; D000071116:ADAMTS5 Protein; C518696:Adamts5 protein, rat; D053509:Matrix Metalloproteinase 13; C507083:Mmp13 protein, rat; D020780:Matrix Metalloproteinase 9; C572337:Mmp9 protein, rat