中华创伤骨科杂志
中華創傷骨科雜誌
중화창상골과잡지
CHINESE JOURNAL OF ORTHOPAEDIC TRAUMA
2012年
3期
232-235
,共4页
郑帅%王吉兴%任海龙%张继业%姜欢畅
鄭帥%王吉興%任海龍%張繼業%薑歡暢
정수%왕길흥%임해룡%장계업%강환창
腰椎%关节%软骨细胞%压力%胶原Ⅱ型
腰椎%關節%軟骨細胞%壓力%膠原Ⅱ型
요추%관절%연골세포%압력%효원Ⅱ형
Lumbar vertebrae%Joints%Chondrocytes%Pressure%Collagen type Ⅱ
目的 探讨不同强度持续静态压力对体外培养的兔腰椎小关节软骨细胞活性的影响. 方法 分离新西兰白兔腰椎小关节软骨细胞,体外培养.将培养的第3代软骨细胞分为4组:对照组、30 kPa组、60 kPa组和90 kPa持续静态加压组.显微镜下观察其形态,分别用免疫组化、噻唑蓝(MTT)、酶联免疫吸附法(ELISA)法鉴定软骨细胞,检测细胞增殖及Ⅱ型胶原的合成情况. 结果 第3代软骨细胞Ⅱ型胶原免疫组织化学染色,显示为阳性.软骨细胞增殖OD值,1d除了对照组与30 kPa组相比差异无统计学意义外,其他各组两两比较差异均有统计学意义(P ＜0.05);2～4 d:除了 30 kPa组与60 kPa组相比差异无统计学意义外,其他各组两两比较差异均有统计学意义(P＜0.05);5～10 d:各组间两两比较差异均有统计学意义(P＜0.05).空白对照组的Ⅱ型胶原含量最高[(7.517±0.328)μg/L],其次是60 kPa组[(6.035±0.075) μg/L],90 kPa组含量最低[(2.873±0.127) μg/L],30 kPa组的Ⅱ型胶原含量为(4.846±0.093)μg/L,各组间两两比较差异均有统计学意义(P＜0.05).结论 兔腰椎小关节软骨细胞在持续静态压力下容易发生退变,细胞增殖率和Ⅱ型胶原含量均下降.
目的 探討不同彊度持續靜態壓力對體外培養的兔腰椎小關節軟骨細胞活性的影響. 方法 分離新西蘭白兔腰椎小關節軟骨細胞,體外培養.將培養的第3代軟骨細胞分為4組:對照組、30 kPa組、60 kPa組和90 kPa持續靜態加壓組.顯微鏡下觀察其形態,分彆用免疫組化、噻唑藍(MTT)、酶聯免疫吸附法(ELISA)法鑒定軟骨細胞,檢測細胞增殖及Ⅱ型膠原的閤成情況. 結果 第3代軟骨細胞Ⅱ型膠原免疫組織化學染色,顯示為暘性.軟骨細胞增殖OD值,1d除瞭對照組與30 kPa組相比差異無統計學意義外,其他各組兩兩比較差異均有統計學意義(P ＜0.05);2～4 d:除瞭 30 kPa組與60 kPa組相比差異無統計學意義外,其他各組兩兩比較差異均有統計學意義(P＜0.05);5～10 d:各組間兩兩比較差異均有統計學意義(P＜0.05).空白對照組的Ⅱ型膠原含量最高[(7.517±0.328)μg/L],其次是60 kPa組[(6.035±0.075) μg/L],90 kPa組含量最低[(2.873±0.127) μg/L],30 kPa組的Ⅱ型膠原含量為(4.846±0.093)μg/L,各組間兩兩比較差異均有統計學意義(P＜0.05).結論 兔腰椎小關節軟骨細胞在持續靜態壓力下容易髮生退變,細胞增殖率和Ⅱ型膠原含量均下降.
목적 탐토불동강도지속정태압력대체외배양적토요추소관절연골세포활성적영향. 방법 분리신서란백토요추소관절연골세포,체외배양.장배양적제3대연골세포분위4조:대조조、30 kPa조、60 kPa조화90 kPa지속정태가압조.현미경하관찰기형태,분별용면역조화、새서람(MTT)、매련면역흡부법(ELISA)법감정연골세포,검측세포증식급Ⅱ형효원적합성정황. 결과 제3대연골세포Ⅱ형효원면역조직화학염색,현시위양성.연골세포증식OD치,1d제료대조조여30 kPa조상비차이무통계학의의외,기타각조량량비교차이균유통계학의의(P ＜0.05);2～4 d:제료 30 kPa조여60 kPa조상비차이무통계학의의외,기타각조량량비교차이균유통계학의의(P＜0.05);5～10 d:각조간량량비교차이균유통계학의의(P＜0.05).공백대조조적Ⅱ형효원함량최고[(7.517±0.328)μg/L],기차시60 kPa조[(6.035±0.075) μg/L],90 kPa조함량최저[(2.873±0.127) μg/L],30 kPa조적Ⅱ형효원함량위(4.846±0.093)μg/L,각조간량량비교차이균유통계학의의(P＜0.05).결론 토요추소관절연골세포재지속정태압력하용역발생퇴변,세포증식솔화Ⅱ형효원함량균하강.
Objective To investigate the effects of different continuous hydrostatic pressures on the chondrocytes of the lumbar facet joint cultured in vitro in rabbits. Methods The cartilage cells of the lunbar facet joint in New Zealand white rabbits were isolated and cultured in vitro.Immunohistochemistry was used to identify the cell type.The third generation of the cartilage cells were put into a self-designed pressure chamber and subjected to different continuous hydrostatic pressures (0,30,60,and 90 kPa).The morphological changes of the cells were observed in all groups by the Laser Scanning Confocal Microscopy.3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was adopted to test the increment rate of the cartilage cells.The enzyme-linked immunosorbent assay (ELISA) was performed to measure contents of collagen Ⅱ in the supernatant fluid of chondrocytes in different groups. Results Immunohistochemistry showed a positive staining of collagen Ⅱ of the third generation of the cartilage cells.MTT showed significant differences(P ＜ 0.05) between either 2 groups except between 0 and 30 kPa groups on day one,except between 30 and 60 kPa groups on days 2 to 4,and with no exception on days 5 to 10.ELISA showed that the 0 kPa group had the highest content of collagen Ⅱ (7.517 ±0.328 μg/L),followed by the 60 kPa group (6.035 ±0.075 μg/L),the 30 kPa group (4.846 ±0.093 μg/L) and the 90 kPa group (2.873 ±0.127μg/L),with significant between-group differences ( P ＜ 0.05). Conclusion A continuous hydrostatic pressure,especially an excessively low or high one,can result in a significant decrease in proliferation of the chondrocytes of the lumbar facet joint cultured in vitro in rabbits and in collagen Ⅱ content as well which may induce degeneration of the chondrocytes.
