CAJ | 학술논문

目的探讨新生大鼠髋脱位在自然演变过程中关节软骨细胞的增殖能力和凋亡的变化及意义.方法将新生大鼠按数字随机法随机分成实验组和对照组,实验组模拟襁褓位固定双下肢伸髋伸膝位持续10天.解除外固定后每鼠龄2、4、8周处死并离断髋关节,一侧用于大体形态测量与番红O-固绿染色评估;另一侧用于体外原代细胞培养,流式分析增殖指数与凋亡率;CCK-8计算增殖速率.采用t检验进行统计分析,P＜0.05为差异有统计学意义.结果 ①大体形态:实验组大鼠去除外固定后未获得复位,股骨头一直脱出于髋臼外上方,对照组髋关节臼深指数随着生长发育逐渐增大,而实验组在4周时明显增大后至8周时却又明显变小,并且与不同时段相比明显小于对照组,差异有统计学意义(P＜0.01).对照组股骨头形态指数随生长发育无明显变化,而实验组逐渐减小,并明显小于对照组.②组织形态学:番红-固绿染色可见对照组随着生长发育,关节软骨逐渐变薄,且层次清晰;实验组在4周软骨形态失常,且浅层基质淡染,细胞表现出排列紊乱,局部出现细胞聚集成簇;8周浅层软骨出现大面积的失染,浅层出现软骨细胞丢失,伴随裂隙和溃疡,移行层细胞却增多;Mankin评分显示在4、8周阶段实验组的评分显著高于对照组(P＜0.01).③细胞增殖能力分析:流式分析结果显示不同时段实验组的增殖指数均明显高于对照组;CCK-8也显示在不同时段实验组的增殖速率高于对照组,差异有统计学意义(P＜0.01).④凋亡检测:流式分析显示在鼠龄2、8周阶段实验组凋亡率明显高于对照组(P＜0.01);而在4周阶段却无显著差异.结论新生大鼠髋脱位关节软骨细胞表现出较强的增殖活性与异常增高的凋亡对早期发生退行性改变有着重要意义.
목적 탐토신생대서관탈위재자연연변과정중관절연골세포적증식능력화조망적변화급의의.방법 장신생대서안수자수궤법수궤분성실험조화대조조,실험조모의강보위고정쌍하지신관신슬위지속10천.해제외고정후매서령2、4、8주처사병리단관관절,일측용우대체형태측량여번홍O-고록염색평고;령일측용우체외원대세포배양,류식분석증식지수여조망솔;CCK-8계산증식속솔.채용t검험진행통계분석,P＜0.05위차이유통계학의의.결과 ①대체형태:실험조대서거제외고정후미획득복위,고골두일직탈출우관구외상방,대조조관관절구심지수수착생장발육축점증대,이실험조재4주시명현증대후지8주시각우명현변소,병차여불동시단상비명현소우대조조,차이유통계학의의(P＜0.01).대조조고골두형태지수수생장발육무명현변화,이실험조축점감소,병명현소우대조조.②조직형태학:번홍-고록염색가견대조조수착생장발육,관절연골축점변박,차층차청석;실험조재4주연골형태실상,차천층기질담염,세포표현출배렬문란,국부출현세포취집성족;8주천층연골출현대면적적실염,천층출현연골세포주실,반수렬극화궤양,이행층세포각증다;Mankin평분현시재4、8주계단실험조적평분현저고우대조조(P＜0.01).③세포증식능력분석:류식분석결과현시불동시단실험조적증식지수균명현고우대조조;CCK-8야현시재불동시단실험조적증식속솔고우대조조,차이유통계학의의(P＜0.01).④조망검측:류식분석현시재서령2、8주계단실험조조망솔명현고우대조조(P＜0.01);이재4주계단각무현저차이.결론 신생대서관탈위관절연골세포표현출교강적증식활성여이상증고적조망대조기발생퇴행성개변유착중요의의.
Objective To explore the proliferation and apoptosis of chondrocytes during the de genration process resulted from dislocation of the hip in neonatal rat model.Methods Neonatal Wistar rats were randomly divided into two groups (experimental and control group).Developmental dislocation of the hip (DDH) model was mimicked by swaddling position,which was to immobilize the bilat eral hips and legs at extension with medical tape for ten days.The rats were sacrificeal at 2,4 and 8 weeks of age and hips were isolated for gross measurement and sanfranin O-fast green staining.Chondrocytes were isolated for proliferation index (PI) and apoptosis ratio (AR) by flow cytometry (FCM) and CCK 8 test.Student's-t test was applied for statistical analysis.Results The gross sample showed the abnormal relationship of the hips,proving the success of model.The anatomy didn't get reduction with development.Acetabula depth ratio (ADR) of control group increased along with development,however,ADR in DDH group showed an obvious increase at 4-weeks with a deep decrease at 8-weeks and were smaller than those in the control group at each stage.Femoral head ratio (FHR) in control group didn't show significant changes along with growing up; however,FHR in DDH group decreased along with development and was smaller than those in the control group.The sanfranin O-fast green staining revealed that the cartilage in control group became thinner and showed distinct layers along with development,whereas the shape of cartilage in DDH group deformed from 4 weeks after birth and extracellular matrix (ECM) became less stained,chondrocytes disordered and clusters presented.Mankin score showed significant difference at 4 and 8 weeks after birth (P＜0.01).PI of DDH group showed higher than those in the control group at each stage,as well as proliferative ratio by CCK-8 test.AR in DDH group was higher than that in control group at 2 and 8 weeks,while no sig nificance was presented at 4 weeks.Conclusions Proliferation and apoptosis of chondrocytes are evident during the process from dislocational hip to early degeneration in neonatal rat.