PATHOLOGICAL AND MORPHOLOGICAL RESEARCH OF EXPERIMENTAL ACETABULAR DYSPLASIA

Objective To investigate the pathological mechanism of hip dysplasia. Methods The left knee joints of eighteen rabbits were fixed in extending position with plaster cylinder for four weeks, but their hip joints were flexed. The right side served as control. Roentgenogram was made in all animals. The changes of the xray films and the pathological findings between left and right hips were compared. Results Appearance of hip dysplasia was obvious at four weeks after plaster fixation. There were pathological changes, including shallow acetabulum and fiat femoral head, increased acetabular index and decreased acetabular head index on the x-ray films. Conclusion The hip dysplasia is the result of prolonged extending position of the knee joint. Abnormal knee posture seems to be one of the important factors of hip dysplasia. This kind of deformation may be worsened with time.     

ULTRASTRUCTURAL INVESTIGATION ON THE ARTICULAR CARTILAGE FROM RABBIT KNEE JOINT AFTER IMMOBILIZATION IN CAST

The articular cartilages from the left knee of 30 rabbits immobilized in casts in functional position were studied under transmission electron microscope. The pathological changes which begin at early stage of immobilization included both degeneration and repair of cartilage. The striking morphological features observed were: (1) most chondrocytes in various layers of cartilage underwent progresive degeneration; (2) the concomitant abnormal proliferation of some chondrocytes happened in a brief period. The changes in mechanical stress caused by immobilization affected the nutrition of cartilage and finally led to its reactive structural reformation.     

Gene expression of fibrinolytic factors urokinase plasminogen activator and plasminogen activator inhibitor-1 in rabbit temporo-mandibular joint cartilage with disc displacement

Background The urokinase plasminogen activator system is believed to play an important role in degradation of the extracellular matrix associated with cartilage and bone destruction; however its precise roles in temporomandibular disorders have not yet been clarified. The aims of this study were to investigate the gene expression of fibrinolytic factors urokinase plasminogen activator (uPA) and plasminogen activator inhibitor-1(PAI-1) in the articular cartilage of rabbit temporomandibular joint (TMJ) with disc displacement (DD) and to probe the relationship between fibrinolytic activity and cartilage remodeling.Methods Disc displacement of right joints was performed in 36 of 78 rabbits under investigation. The animals were sacrificed at 4 days and 1, 2, 4, 8 and 12 weeks after surgery, respectively. The right joints of these animals were harvested and processed for the examination of mRNA expression of uPA and PAI-1 in articular cartilage using in situ hybridization techniques.Results The expression of uPA and PAI-1 was co-expressed weakly in the chondrocytes from transitive zone to hypertrophic zone and mineralized zone, while no hybridizing signals were shown in proliferative zone and superficial zone in control rabbits. The most striking was the up-regulation of uPA and PAI-1 mRNA in 4-day rabbits postoperatively at the onset of cartilage degeneration. The strongest hybridizing signals for uPA and PAI-1 were seen in 2-week rabbits postoperatively. After 2 weeks, the expression of uPA and PAI-1 began to decrease and reached nearly normal level at 12 weeks.Conclusions The expression of the uPA/PAI-1 system coincides with the pathological changes in condylar cartilage after DD. The uPA/PAI-1 system may be one of the essential mediators in articular cartilage remodeling.     

Degenerative mechanism of articular cartilage induced by low stress. A morphological study.

Immobilization leads not only to diminished joint movement but also low stress of articular cartilage. The present investigation was undertaken to observe the morphological changes which arose in articular cartilage with low stress. The joint motion remained intact. Articular cartilage from the left knees of 66 rats whose left calcaneal tendons had been transected was examined under transmission electron microscope and light microscope. The degenerative changes were observed: decreased functional activity of chondrocytes progressively degenerated cartilage and lack of compensatory proliferation of chondrocytes at the early stage. We propose that the degeneration of articular cartilage induced by immobilization is the result of combination of low stress and lack of joint motion. The following degenerative mechanism begins with chondrocytes. Chondrocyte and matrix influence each other in a vicious cycle. Low stress may restrain the repair activities.     

Ultrastructural study of hip joint osteoarthritis. Scanning electron microscopic study of articular cartilage of femoral head.

The surface structure of four different articular cartilages of the femoral head was studied with scanning electron microscope in 12 cases of hip joint osteoarthritis. These articular cartilages were mink white, yellow, dusky red and hyperplastic and thickened. The osteoarthritic articular cartilage surface was uneven with puckerings of various height, width and orientation. The puckerings were covered with fibril network. The fibrils were exposed collagen fibrils and of different diameters. Wider fiber bundles without orientation were found on the fibril network. On the surface of the hyperplastic articular cartilage observed were many deep and oval spaces left behind after breakdown of the lacunae of cells in the clusters of articular chondrocytes. In the spaces, remnants of articular chondrocytes were seen.

     

Glucan HBP-A Increase Type Ⅱ Collagen Expression of Chondrocytes in Vitro and Tissue Engineered Cartilage in Vivo

Objective:Although chondroprotective activities have been documented for polysaccharides,the potential target of different polysaccharide may differ.The study was aimed to explore the effect of glucan HBP-A in chondrocyte monolayer culture and chondrocytes-alginate hydrogel constructs in vivo,especially on the expression of type Ⅱ collagen.Methods:Chondrocytes isolated from rabbit articular cartilage were cultured and verified by immunocytochemical staining of type Ⅱ collagen.Chondrocyte viability was assessed after being treated with HBP-A in different concentrations.Morphological status of chondrocytes-alginate hydrogel constructs in vitro was observed by scanning electron microscope(SEM).The constructs were treated with HBP-A and then injected to nude mice subcutaneously.Six weeks after transplantation,the specimens were observed through transmission electron microscopy(TEM).The mRNA expressions of disintegrin and metalloproteinase with thrombospondin motifs 5(ADAMTs-5),aggrecan and type Ⅱ collagen in both monolayer culture and constructs were determined by real time polymerase chain reaction(PCR).The expression of typeⅡ collagen and matrix metalloproteinases-3(MMP-3) in chondrocyte monolayer culture was also tested through Western blot and enzyme linked immunosorbent assay(ELISA),respectively.Results:MMP-3 secretion and ADAMTs-5 mRNA expression in vitro were inhibited by HBP-A at 0.3 mg/mL concentration.In morphological study,there were significant appearance of collagen in those constructs treated by HBP-A.Accordingly,in both chondrocyte monolayer culture and chondrocytes-alginate hydrogel constructs,the expression of type Ⅱcollagen was increased significantly in HBP-A group when compared with control group(P0.001).Conclusions:The study documented that the potential pharmacological target of glucan HBP-A in chondrocytes monolayer culture and tissue engineered cartilage in vivo may be concerned with the inhibition of catabolic enzymes MMP-3,ADAMTs-5,and increasing of type Ⅱ collagen expression.     

Expression of miRNA-140 in Chondrocytes and Synovial Fluid of Knee Joints in Patients with Osteoarthritis

Objective To investigate the expression of miRNA-140 in chondrocytes and synovial fluid of osteoarthritis (OA) patients, and explore the relationship between the miRNA-140 expression and OA severity.
Methods This study enrolled 30 OA patients who underwent total knee arthroplasty for chondrocytes sampling and 30 OA patients who underwent intra-articular injection for synovial fluid sampling. All OA patients were grouped intomild [Kellgren and Lawrence (KL) grade 1-2], moderate (KL grade 3) and severe (KL grade 4), with 10 in each subgroups for each sampling purposes. 7 non-OA patients and 10 patients with knee injury were collected for cartilage and synovial fluid sampling respectively as control groups. Chondrocytes were isolated from the cartilage tissue and culturedin vitro. Quantitative real time PCR for miRNA-140 in chondrocytes and synovial fluid were performed, and the U6 snRNA was used as internal control. The expression difference of miRNA-140 among groups and correlation between the expression and the KL grade of OA were analysed using one-way ANOVA and Spearman test respectively.
Results The expression of miRNA-140 in chondrocytes of knees in OA patients was reduced than that in normal knees, and the between-group difference was statistically significant (F=305.464,P<0.001). miRNA-140 could be detected in synovial fluid of both normal knees and OA knees, its relative expression level was reduced in synovial fluid of OA group compared with normal group, and the between-group difference was statistically significant as well (F=314.245,P<0.001). The relative expression level of miRNA-140 in both chondrocytes and synovial fluid were negatively correlated with the KL grade of OA(r=-0.969,P<0.001;r=-0.970,P<0.001).
Conclusion miRNA-140 could be detected in chondrocytes and synovial fluid of OA patients, and its expression was negatively correlated with the severity of OA.     

Effect of Anluohuaxian Tablet Combined with y-IFN on Schistosomal Liver Fibrosis

The therapeutic effects of anluohuaxian tablet combined with γ-IFN on schistosomal liver fibrosis and its mechanism were studied in a murine model and clinical cases of schistosomal liver fibrosis.Fifty Kunming mice were randomly divided into 5 groups:normal control group,infection control group,anluohuaxian tablet-treated group,γ-IFN-treated group and combined treatment (anluohuaian tablet+γ-IFN) group.Pathologic changes in liver,including hepatic pigmentation and the size of schistosomal egg granuloma,were observed by HE staining after treatment for 8 weeks.The expression of the type Ⅰ and Ⅲ collagen,and TIMP-1 was detected by immunohistochemistry.TGF-β1 mRNA expression was examined by real-time fluorescent quantitative PCR.Sixty patients with schistosomal liver fibrosis were divided into treatment group and control group.The patients in treatment group were treated with anluohuaxian tablet in combination with γ-IFN for 6 months.Be-fore and after treatment,the changes of symptoms and signs,liver function,serum liver fibrosis in-dexes and imaging indexes were observed.The results showed that as compared with infection con-trol group,all forms of treatments relieved the hepatic pathological injury with apparently diminished size of schistosomal egg nodules and decreased percentage of pigmentation (P<0.05).Furthermore,the expression of collagen Ⅰ and Ⅲ,TIMP-Ⅰ,and TGF-β1 mRNA in combined treatment group was significantly decreased as compared with anluohuaxian tablet-treated and γ-IFN-treated groups (P<0.05).In the clinical observation,the serum liver fibrosis indexes,the portal vein width as well as the spleen thickness was significantly reduced in treatment group as compared with control group (P<0.05).It was concluded that the combined use of anluohuaxian tablet with γ-IFN in schistosomal liver fibrosis could protect liver function,alleviate liver fibrosis,and could be used as a choice in treating patients with schiatosomal liver fibrosis.     

Ventricular remodeling and intervention with losartan following rabbit chronic hibernating myocardium

Objectives To explore the changes of myocyte and the interstitial collagen in a model of chronic hibernating myocardium (CHM) in rabbits, and to determine whether these alterations affect the cardiac function, and to further observe the effects of losartan on ventricular remodeling. Methods A left anterior descending (LAD) coronary artery stenosis was created and maintained for 4 weeks to create a CHM model in rabbits. Thirty-six rabbits were assigned to the following three groups(12 rabbits per group): CHM for 4 weeks(CHM group), low-dose of losartan intervention group for 4 weeks(LTl group, 10 mg·kg-1·d-1), high-dose of losartan intervention group for 4 weeks (LT2 group, 30 mg·kg-1·d-1); and 12 sham-operated rabbits served as controls (SO group). A microscopic imaging system (Image-Pro Plus, Olympus) was used to assess the Interstitial collagen volume fraction (ICVF) in myocardial sections with picrosirius-red staining, and a polarized light microscopy to qualitatively analysis the changes in the type and the proportion of collagen fibers, to semi-quantitatively score the proportion of collagen I to collagen III(PI/III). The expression of MMP-2, 9 and TIMP-2 was assessed by immunohistochemistry and western bloting. The myocyte apoptosis rate (Rapo) was calculated with TUNEL-staining. And echocardiography was performed to measure left ventricular end-systolic and end-diastolic diameter (LVESD, LVEED), and left ventricular short-axis fraction shortening (LVFS) and ejection fraction (LVEF). Results The animal model of CHM was induced successfully in 36 out of 39-rabbits and maintained for 28 days. Compared with the sham group, ICVF) was significantly increased (P<0.01) in CHM group; compared with CHM group, ICVF was significantly decreased (P<0.01,each) in LTl group and LT2 group, and the change were more remarkable in LT2 group, compared to LTl group(P<0.05). Compared with sham group, PI/IH was significantly increased (P < 0.01) in CHM group; compared with CHM group, PI/III was significantly decreasedin the losartan intervention groups (P<0.01,each), and the change was more remarkable in LT2 group compared to LTl group (P<0.05). Compared with sham group, the expression of MMP-2 and MMP-9 were greatly increased (P<0.01) in CHM group, while TIMP-2 were greatly decreased(P< 0.01); compared with CHM group, the expression of MMP-2 and MMP-9 were significantly decreased (P<0.01,each) .while TIMP-2 were significantly increased (P<0.01,each) in the losartan intervention groups, and the change was more remarkable in LT2 group than in LT1 group (.P<0.05). Compared with sham group, myocyte Rapo was markedly increased (P<0.01) in CHM group; compared with CHM group, myocyte Rapo was significantly decreased (P<0.01,each) in the losartan intervention groups, and the change was more remarkable in LT2 group than in LT1 group (P<0.05). Compared with sham group, LVEF and LVFS were significantly reduced in CHM group (P <0.01), compared with CHM group, LVEF and LVFS were higher (P<0.01,each) in the losartan intervention groups ,and the changes were more remarkable in LT2 group than in LTl group (P<0.05). Conclusions CHM underwent interstitial collagen proliferation and myocyte apoptosis, leading to ventricular remodeling and ventricular functional impairment, Losartan intervention reduces myocyte apoptosis and interstitial collagen proliferation, and improve ventricular function.     

Effect of Anluohuaxian Tablet Combined with γ-IFN on Schistosomal Liver Fibrosis

The therapeutic effects of anluohuaxian tablet combined with γ-IFN on schistosomal liver fibrosis and its mechanism were studied in a murine model and clinical cases of schistosomal liver fibrosis, Fifty Kunming mice were randomly divided into 5 groups： normal control group, infection control group, anluohuaxian tablet-treated group, γ-IFN-treated group and combined treatment （anluohuaian tablet＋γ-IFN） group. Pathologic changes in liver, including hepatic pigmentation and the size of schistosomal egg granuloma, were observed by HE staining after treatment for 8 weeks. The expression of the type Ⅰ and Ⅲ collagen, and TIMP-1 was detected by immunohistochemistry. TGF-β1 mRNA expression was examined by real-time fluorescent quantitative PCR. Sixty patients with schistosomal liver fibrosis were divided into treatment group and control group. The patients in treatment group were treated with anluohuaxian tablet in combination with γ-IFN for 6 months. Before and after treatment, the changes of symptoms and signs, liver function, serum liver fibrosis indexes and imaging indexes were observed. The results showed that as compared with infection control group, all forms of treatments relieved the hepatic pathological injury with apparently diminished size of schistosomal egg nodules and decreased percentage of pigmentation （P〈0.05）. Furthermore, the expression of collagen Ⅰ and Ⅲ, TIMP-1, and TGF-β1 mRNA in combined treatment group was significantly decreased as compared with anluohuaxian tablet-treated and γ-IFN-treated groups （P〈0.05）. In the clinical observation, the serum liver fibrosis indexes, the portal vein width as well as the spleen thickness was significantly reduced in treatment group as compared with control group （P〈0.05）. It was concluded that the combined use of anluohuaxian tablet with γ-IFN in schistosomal liver fibrosis could protect liver function, alleviate liver fibrosis, and could be used as a choice in treating patients with schiatosomal liver fibrosis.     

髋关节发育不良髋臼软骨中Ⅱ型胶原、基质金属蛋白酶7表达的改变

目的 检测Ⅱ型胶原、基质金属蛋白酶7(MMP-7)在发育性髋关节发育不良(DDH)早期髋臼软骨中表达的改变情况,探讨Ⅱ型胶原、MMP-7与软骨退变的相关性.方法 兔左下肢作为实验侧,膝关节伸直位、长腿管形石膏固定;右下肢作为对照侧,5周后经X线证实构建DDH动物模型8只.观察实验组和对照组髋臼软骨形态学改变;免疫组化两步法、免疫印迹方法检测Ⅱ型胶原、MMP-7在髋臼软骨中表达的改变.结果 实验组可见有局部软骨细胞簇集,细胞外基质甲苯胺蓝染色有失染现象.免疫组化染色结果显示实验组Ⅱ型胶原和MMP-7阳性染色细胞明显高于对照组.免疫印迹结果显示实验组Ⅱ型胶原和MMP-7蛋白表达量均升高,组间比较差异均有统计学意义(t=2.18,t=2.334,P<0.05).结论 DDH早期髋臼软骨发生退行性变,Ⅱ型胶原、MMP-7高表达,提示Ⅱ型胶原、MMP-7的数量和强度可能与软骨退行性变的程度差异有关.     

Characterization of Human Primary Chondrocytes of Osteoarthritic Cartilage at Varying Severity

Background  There is a difficulty in evaluating the in vivo functionality of individual chondrocytes, and there is much heterogeneity among cartilage affected by osteoarthritis (OA). In this study, in vitro cultured chondrocytes harvested from varying stages of degeneration were studied as a projective model to further understand the pathogenesis of osteoarthritis.

Methods  Cartilage of varying degeneration of end-stage OA was harvested, while cell yield and matrix glycosaminoglycan (GAG) content were measured. Cell morphology, proliferation, and gene expression of collagen type I, II, and X, aggrecan, matrix metalloproteinase 13 (MMP-13), and ADAMTS5 of the acquired chondrocytes were measured during subsequent in vitro culture.

Results  Both the number of cells and the GAG content increased with increasing severity of OA. Cell spreading area increased and gradually showed spindle-like morphology during in vitro culture. Gene expression of collagen type II, collagen type X as well as GAG decreased with severity of cartilage degeneration, while expression of collagen type I increased. Expression of MMP-13 increased with severity of cartilage degeneration, while expression of ADAMTS-5 remained stable. Expression of collagen type II, X, GAG, and MMP-13 substantially decreased with in vitro culture. Expression of collagen type I increased with in vitro cultures, while expression of ADAMTS 5 remained stable.

Conclusions  Expression of functional genes such as collagen type II and GAG decreased during severe degeneration of OA cartilage and in vitro dedifferentiation. Gene expression of collagen I and MMP-13 increased with severity of cartilage degeneration.

     

点种法同种异体兔软骨移植修复关节软骨缺损的研究

将 30只新西兰纯种兔共 60个膝关节随机分为 3组 :软骨块点种法移植组、软骨细胞点种法移植组和对照组。分别于术后 2、4、1 2、2 4周取标本行大体、光镜、电镜的组织学动态观察 ,并同时对修复组织进行组织学和组织化学质量评估 ,观察点种法软骨移植术修复透明软骨的效果。结果显示 :2种点种法移植组均能获得透明软骨修复 ,而对照组缺损区仅为纤维组织填充 ,并且点种法移植组兔各期平均组织学和组织化学得分均高于对照组 (P <0 .0 1 )。提示 :点种法软骨移植能获得透明软骨修复 ,尤其适用于大面积软骨缺损。     

大鼠DDH关节软骨早期退变和X型胶原、基质金属蛋白酶13表达相关性的研究

目的：检测 X 型胶原、基质金属蛋白酶13（MMP-13）在发育性髋关节发育不良（DDH）大鼠模型不同时期关节软骨中表达的改变情况,探讨 X 型胶原、MMP-13与软骨早期退变的相关性。方法选取新生 Wistar 大鼠80只,随机均等分成 DDH 组和对照组。DDH 组新生大鼠后肢襁褓位固定10 d 后解除固定继续饲养,分别于鼠的2、4、6、8周龄处死,离断髋关节,观察髋关节软骨的大体形态。用免疫组化和 qRT-PCR 法检测 X 型胶原、MMP-13的表达。结果 DDH模型鼠的尸体解剖样本显示关节囊增厚,髋臼表面呈不规则、不平整,股骨头扁平且发育较小,头臼包容差,尤其4周后更明显;与对照组相比,在髋关节软骨浅表区域有较高的X 型胶原和 MMP-13表达（P <0.05）,并且这种增长是随着年龄增长而增长的,X 型胶原和 MMP-13的 mRNA 表达显示出相似的结果（P <0.05）。结论 DDH 模型鼠在较早阶段出现软骨退化,随着年龄逐渐加重;X 型胶原和 MMP-13表达与 DDH 软骨早期退变有密切的关系。     

垛状髋臼扩大术治疗髋臼发育不良

目的丰富髋关节发育不良及大龄发育性髋关节半脱位的治疗手段。方法采用关节囊外的沟槽植骨,外板夹卡,垛状髋臼扩大术对发育性髋关节发育不良,大龄髋关节半脱位的患儿15例（20个髋,5例为双髋）进行治疗。采用先天性髋脱位疗效评定标准,根据患儿主观感觉,临床检查及X线摄片检查评定为依据进行评分。用Charnley评分标准及Harris评分标准对照。结果垛状髋臼扩大术采用交叉网状外板夹卡植骨方式,可大大改善髋臼包容,20髋中最大改善度由65％增大到85％,最小改善度由75％到90％。髋臼覆盖率超过85％,髋臼指数小于25。。结论垛状髋臼扩大术是一种安全、复合解剖结构而设计的一种外板夹卡交叉网状植骨造盖手术,有效扩大髋臼包容面积更加稳定。     

发育性髋关节脱位髋臼形态的MRI研究

目的 通过对发育性髋关节脱位患儿患侧髋臼与正常侧髋臼磁共振成像(MRI)表现的观察,探究发育性髋关节脱位患儿髋臼的病理改变。方法 对12例发育性髋关节脱位的患儿进行髋关节MRI检查,其中单侧脱位8例,双侧脱位4例,共8个正常髋关节(对照组)、16个脱位髋关节(病变组)。测量双侧髋关节骨性髋臼指数(BAI)、软骨性髋臼指数(CAI),骨性髋臼前倾角(BAAV)及软骨性前倾角(CAAV),对所得结果进行统计学分析。结果 BAI与CAI之间存在相关性,对照组两指数间相关系数(0.945)高于病变组的相关系数(0.814)。病变组BAAV与CAAV均明显大于对照组(P<0.01),病变组髋关节的CAAV明显大于BAAV(P<0.01)。所有髋关节BAI与BAAV,CAI与CAAV之间均存在相关性(相关系数分别为0.884和0.822)。结论 发育性髋关节脱位的髋臼病理改变不仅体现在髋臼指数的改变,还体现在髋臼朝向的改变,这两种改变的程度存在着相关性。发育性髋关节脱位髋臼形态的畸形不仅体现在骨性结构,还体现在软骨结构上。两种结构的病理变化并不是完全同步的。采用MRI成像对DDH患儿的软骨结构进行评估可用于指导DDH的术式选择和手术操作。     

人颈椎椎体终板软骨细胞退变模型的建立及其意义

目的 建立人颈椎椎体终板软骨细胞退变模型,观察人正常颈椎椎体和退变颈椎椎体终板软骨细胞的形态及表征.方法 选择2010年7月至2011年7月49例颈椎骨折、脱位(19例)及颈椎病(30例)患者术中取出的颈椎终板软骨,用酶消化法分别分离培养人正常颈椎椎体终板软骨细胞(对照组)和退变颈椎椎体终板软骨细胞(颈椎病组);用倒置显微镜和HE染色法观察细胞形态学变化;四甲基偶氮唑蓝(MTT)法绘制细胞生长曲线;甲苯蓝染色及反转录-PCR(RT-PCR)法对终板软骨细胞进行鉴定;RT-PCR法检测终板软骨细胞特征性基因蛋白多糖、Ⅱ型胶原及Ⅰ型胶原的表达.结果 人颈椎椎体终板软骨细胞表达特征性蛋白多糖、Ⅱ型胶原及Ⅰ型胶原,其生长情况及细胞表型类似于关节软骨细胞.对照组原代终板软骨细胞以多角形为主,增殖速度较快;而颈椎病组原代终板软骨细胞以梭形为主,细胞增殖速度较慢.颈椎病组原代终板软骨细胞表达的蛋白多糖基因(0.695 ±0.052)和Ⅱ型胶原基因(0.726 ±0.035)均低于对照组(0.950±0.032、0.907±0.078,t=7.263、3.681,P=0.002、0.021),Ⅰ型胶原基因则高于对照组(0.795±0.028比0.552±0.070,t=-5.560,P=0.005).结论 成功建立了人颈椎椎体终板软骨细胞退变模型,为椎间盘退变机制研究提供了较好的细胞学基础,解决了以前一直以动物细胞模型为研究对象的局限性.     

髋关节发育不良软骨退行性变患者软骨组织胶原蛋白、基质金属蛋白酶及转化生长因子的变化研究

目的分析研究髋关节发育不良软骨退行性变患者软骨组织胶原蛋白、基质金属蛋白酶及转化生长因子的变化情况。方法选取2010年2月~2012年11月于山东省聊城市第二人民医院及复旦大学附属儿科医院进行诊治的26例髋关节发育不良软骨退行性变患者为观察组,然后选取同期的26例车祸致伤患者为对照组,将两组患者的软骨组织胶原蛋白、基质金属蛋白酶及转化生长因子阳性率进行统计及比较,并将观察组中不同Crowe分期者的阳性率进行比较。结果观察组的各型胶原蛋白、基质金属蛋白酶及转化生长因子阳性表达率均高于对照组,而观察组中Crowe分期Ⅳ期阳性率高于Ⅰ～Ⅲ期患者,Ⅲ期高于Ⅰ、Ⅱ期患者（均P〈0.05）。结论髋关节发育不良软骨退行性变患者胶原蛋白、基质金属蛋白酶及转化生长因子均呈现高表达状态,因此认为对其检测有利于了解患者的疾病状态。     

胶原-纤维蛋白混合凝胶对关节软骨细胞合成Ⅱ型胶原的影响

目的：以胶原蛋白和人血纤维蛋白混合物为载体在体外进行关节软骨细胞三维立体培养,构建人工软骨组织,研究载体对软骨细胞在其中的表达及合成Ⅱ型胶原的影响。方法：取2周龄的新生兔关节软骨,经消化,将获得的软骨细胞与牛Ⅰ型胶原、人血冻干纤维蛋白原、凝血酶按一定比例混合,制成软骨培养物并在体外培养。培养第3周时,取材进行Masson染色、Ⅱ型胶原寡核苷酸探针原位杂交和Ⅱ型胶原的免疫组化观察。 结果：体外培养3周,培养物内细胞大部分存活, Masson染色形成软骨陷窝,同源性细胞簇出现,细胞周围有蓝色物质环绕存在,原位杂交证实其软骨细胞表达Ⅱ型胶原mRNA,免疫组化证实软骨细胞分泌Ⅱ型胶原。 结论：用胶原蛋白和人血纤维蛋白为载体支架体外培养软骨细胞,可促进关节软骨细胞合成分泌Ⅱ型胶原,Ⅱ型胶原是软骨组织最主要的细胞外基质成分。