不同细胞移植修复兔全层关节软骨缺损的比较研究

目的 :比较软骨细胞、骨髓基质细胞及成纤维细胞对全层关节软骨缺损的修复作用。材料和方法 :取幼兔的软骨细胞、骨髓基质细胞及成纤维细胞 ,共 3种有生成软骨潜力的细胞进行体外分离培养 ;以聚乳酸 (PLA)为载体 ,将培养的原代细胞植入PLA支架上 ,形成细胞 -PLA复合物。于 2 8只成年新西兰大白兔的股骨滑车关节面上造成直径 4 5mm、深 3 0mm的全层关节软骨缺损 ,将 3种细胞 -PLA复合物分别植入关节软骨缺损处。植入细胞 -PLA复合物为实验组 ,单纯植入PLA支架为对照组。术后 6周、12周观察缺损修复情况及新生组织类型。结果 :软骨细胞移植组为软骨样组织修复 ,分界明显 ,甲苯胺兰及Ⅱ型胶原染色阳性 ;软骨下骨部分重建 ;细胞排列紊乱。骨髓基质细胞移植组为软骨样组织修复 ,分界不明显 ,甲苯胺兰及Ⅱ型胶原染色阳性 ;软骨下骨重建良好 ,软骨下潮线恢复 ;细胞排列趋于正常。成纤维细胞移植组为纤维组织修复 ,甲苯胺兰及Ⅱ型胶原染色阴性 ;软骨下潮线消失。对照组为纤维组织修复。结论 :软骨细胞、骨髓基质细胞移植修复软骨缺损明显优于成纤维细胞及对照组。骨髓基质细胞与软骨细胞移植组的修复结果无统计学差异 ,但骨髓基质细胞修复组织的细胞排列有序 ,软骨下骨重建良好 ,与周围组织融合密切 ,更接近正?     

载体复合软骨细胞在同种动物体内异位成软骨的实验观察

通过观察软骨细胞复合于载体后在同种异体动物 体内异位成软骨情况,为组织工程方法修复关节软骨缺损的研究创造条件。选用一定浓度的海藻酯钠溶液与葡萄糖酸钙溶液按比例混合均匀,得到具有一定形状及韧性的藻酸钙凝胶,将其与经培养的软骨细胞 复合后植入动物体内,分别于术后2、4周时取材进行组织学观察,发现软骨细胞在体内生长增殖良好,4周时可见新生毛细血管软骨陷窝形成,说明软骨细胞复合于藻酸钙可在异体动物体内生长增殖,并形成有营养供应的软骨细胞块。     

转胰岛素样生长因子-Ⅰ基因软骨细胞体内构建组织工程软骨

目的探讨转胰岛素样生长因子-Ⅰ(insu lin like growth factorⅠ,IGF-Ⅰ)基因软骨细胞与交联透明质酸复合后在裸鼠体内形成组织工程软骨的能力。方法分离培养人关节软骨细胞,将IGF-Ⅰ基因转染人关节软骨细胞,转基因细胞(转染组)、未转基因的软骨细胞(对照组)分别与交联透明质酸材料在体外孵育2 d后,移植裸鼠皮下。移植后6周,取出裸鼠皮下新生组织进行大体形态学观察(HE染色、阿尔新蓝染色),逆转录-聚合酶链反应(RT-PCR)法检测新生组织中Ⅰ型、Ⅱ型前胶原和IGF-Ⅰ基因mRNA的表达,判断裸鼠体内新生组织的结构和功能。结果转染组裸鼠体内新生组织6周仍能表达IGF-ⅠmRNA,而对照组不能;转染组和对照组在6周后均能形成软骨组织陷窝,但对照组的陷窝数量明显低于转染组;转染组的Ⅱ型前胶原mRNA的相对表达量为1.204±0.139,高于对照组(P<0.05);Ⅰ型前胶原mRNA的相对表达量为0.069±0.019,低于对照组(P<0.01)。结论转IGF-Ⅰ基因软骨细胞与交联透明质酸在体内能构建出软骨样组织,其结构更接近于自然软骨。     

组织工程重建兔颞下颌关节盘软骨

目的 应用组织工程学方法重建颞下颌关节盘软骨。方法 分离6只日本大耳白兔髁状突软骨细胞。进行细胞的微载体大规模扩增,将扩增后的软骨细胞接种于组织引导再生胶原膜,体外适当培养后植入4只同种成年兔皮下,植入后12周,对所获组织进行组织形态学观察。结果 髁状[突软骨细胞在胶原膜内生长良好,植入动物体内12周后可形成乳白色类软骨样组织,其表面光滑,有弹性。甲苯胺蓝染色,细胞周围基质呈异染性。结论 应用胶原膜结合软骨细胞共同培养,可形成软骨样组织,该方法将有可能成为软骨缺损及关节盘破损修复的有途径。     

采用温敏性壳聚糖水凝胶体外构建组织工程化软骨的实验研究

目的 验证自行制备的温敏性壳聚糖水凝胶作为软骨细胞支架材料构建组织工程化软骨的可行性.方法 以壳聚糖、β-甘油磷酸钠和羟乙基纤维素为原料,制备温敏性壳聚糖水凝胶,通过SD大鼠肌内注射植入,进行组织相容性检测.在此基础上,将其与软骨细胞复合构建组织工程化软骨,观察软骨细胞在壳聚糖水凝胶中的存活情况,并于体外培养3周后,作相关组织形态学检测.结果 制备的壳聚糖水凝胶具有温度敏感性,即室温时为液态,37℃时10～15min可发生交联反应成为固态凝胶.SD大鼠肌内注射不同时间点组织相容性检测表明,该材料具有良好的组织相容性,植入体内2、4周时有少量炎性细胞浸润,6周时材料降解明显,8周时已经基本降解.采用该材料构建的组织工程化软骨体外培养3周后取材,通过组织学检查,HE染色可观察到软骨陷窝样结构,甲苯胺蓝染色、番红O染色及免疫组化染色结果显示软骨细胞具有分泌细胞外基质的功能.结论 本研究自行制备的温敏性壳聚糖水凝胶材料具有良好的组织相容性,将其与软骨细胞复合后,可以在体外再造组织工程化软骨,是一种有广泛应用前景的软骨组织工程支架材料.     

髓髓基质成骨细胞—松质骨基质复合人工骨肌袋移植的成骨作用

成年新西兰兔骨髓细胞体外培养、诱导后，经混匀、 接种、固化形成 骨髓基质成骨细胞－藻酸盐－松质骨基质复合人工骨，并植回取材兔肌袋内，对照组分别植入藻酸盐－松质骨基质复合物和松质骨基质。植入4、8周取材，行X线摄 片、组织学检查，观察骨形成情况。结果显示，骨髓基质成骨细胞－松质骨基质复合人工骨肌袋内成骨效果明显优于藻酸盐－松质骨基质复合物组和松质骨基质组。复合人工骨标本兼有膜内成骨和软骨成骨，以膜内成骨为主；对照组仅见少量软骨成骨。提示，采用组织工程方法形成的复合人工肌袋内成骨作用明显，用该方法所形成的组织工程骨展现出良好的应用前景。     

以PGA为三维支架同种异体工程化软骨的构建

为探讨以聚羟基乙酸（PGA）为三维支架材料的同种异体软骨细胞构建组织工程化软骨的能力,采用胶原酶消化方法分离乳兔肋软骨获取种子细胞,收集体外培养传2-3代的软骨细胞,接种于经多聚赖氨酸处理的PGA支架材料上,将细胞-材料复合物种植在成兔皮下,一定时间取材,对获得的同种异体工程化软骨进行组织学评价。结果显示,软骨细胞体外培养1天后,在PGA支架上分布均匀;培养1周左右,PGA纤维间有呈蜘蛛网状基质产生。种植复合物4周取材可见PGA纤维未完全降解,软骨细胞不成熟,周边存在一定程度炎细胞浸润;8周取材,PGA纤维消失,软骨基本成熟,炎细胞浸润不明显。软骨基质含量丰富,分布与正常肋软骨组织学特征相似,提示以PGA为支架材料同种异体软骨细胞在有免疫力的动物体内可形成工程化软骨,无明显免疫排斥反应。     

纤维软骨细胞-胶原复合物修复半月板损伤的实验研究

为了探讨纤维软骨细胞－胶原复合物对半月板损伤的修复作用，分离提取狗半月板纤维软骨细胞，体外扩增培养后种植于自制胶原模板上形成细胞－胶原复合物，将18只狗随机分为3组，在狗的外侧半月板作一楔形缺损，缺损内分别植入细胞－胶原复合物、胶原或不作处理，在不同时间取材进行大体和组织学观察。结果显示，植入的复合物形成与正常半月板相似的组织，胶原支架逐渐降解，而胶原对照组及空白对照组缺损区仅少部分修复或无修复。提示，这一新的组织工程学方法具有一定的指导意义及实用性。     

不同年龄阶段家兔髂软骨组织学特点

目的:观察比较不同年龄阶段新西兰大白兔髂软骨组织学形态异同。方法:选取1月龄、3月龄、4月龄、6月龄、12月龄的新西兰大白兔各3只。取其髂软骨及少量软骨下骨做HE染色、甲苯胺蓝染色及I型、II型胶原免疫组织化学染色;同时取1月龄、3月龄和12月龄组膝关节股骨滑车软骨做相同染色用于对比。结果:①幼年期(1月龄),髂软骨全部由透明软骨组成,随着年龄的增长,4月龄部分表层软骨细胞逐渐肥大、骨化,至成年期全层软骨细胞几乎完全骨化。甲苯胺蓝染色、II型胶原免疫组化染色均显示由幼年时的阳性逐渐减弱为成年时的弱阳性。I型胶原只在骨化区、软骨下骨及肌腱部位着色阳性。②随着年龄增长,关节软骨细胞密度逐渐下降,软骨层厚度逐渐变薄。细胞肥大及骨化发生在钙化层。结论:家兔幼年期髂软骨类似关节透明软骨,随着年龄增长髂软骨细胞逐渐肥大、骨化,至成年期仍有少量透明软骨细胞特有的II型胶原蛋白及蛋白多糖的合成。     

端粒酶反转录酶基因介导的人骨髓基质干细胞永生化研究

目的 建立永生化人骨髓基质干细胞系,为组织工程提供种子细胞库及标准化细胞系.方法 取单克隆培养的人骨髓基质干细胞,采用脂质体转染法将人端粒酶反转录酶(hTERT)基因导入细胞,G418筛选后获得阳性克隆,体外连续扩增培养,RT-PCR检测转染细胞内hTERT基因的整合与表达.取转染后第100代细胞,用成骨诱导条件培养基(普通培养基加地塞米松、β-甘油磷酸、维生素C)、软骨诱导条件培养基(普通培养基加地塞米松、1bF-β、维生素C)、心肌细胞诱导条件培养基(普通培养基加5-氮胞苷)分别向成骨、软骨、心肌细胞诱导培养.于培养后第7、14、21、28天,采用免疫细胞化学染色观察成骨细胞诱导Ⅰ型胶原、骨钙素的表达;采用茜素红染色观察钙结节形成;采用免疫细胞化学染色观察软骨细胞诱导Ⅱ型胶原的表达,同时进行甲苯胺蓝染色;采用免疫细胞化学染色观察心肌细胞诱导横纹肌肌动蛋白的表达,分析细胞系向成骨、软骨及心肌细胞分化的能力.结果 hTERT稳定转染入人骨髓基质干细胞,转化细胞端粒酶表达阳性.转化细胞在体外长期连续培养下生长良好,已传至136代.取第100代细胞进行诱导分化,结果显示其能向成骨、软骨及心肌细胞分化.结论 外源性hTERT的导入可致人骨髓基质干细胞永生化,并维持成体干细胞的多向分化特性,可为组织工程提供种子细胞库并为组织工程种子细胞研究提供标准化细胞.     

Transplanted articular chondrocytes co-overexpressing IGF-I and FGF-2 stimulate cartilage repair in vivo

Purpose

The combination of chondrogenic factors might be necessary to adequately stimulate articular cartilage repair. In previous studies, enhanced repair was observed following transplantation of chondrocytes overexpressing human insulin-like growth factor I (IGF-I) or fibroblast growth factor 2 (FGF-2). Here, the hypothesis that co-overexpression of IGF-I and FGF-2 by transplanted articular chondrocytes enhances the early repair of cartilage defects in vivo and protects the neighbouring cartilage from degeneration was tested.

Methods

Lapine articular chondrocytes were transfected with expression plasmid vectors containing the cDNA for the Escherichia coli lacZ gene or co-transfected with the IGF-I and FGF-2 gene, encapsulated in alginate and transplanted into osteochondral defects in the knee joints of rabbits in vivo.

Results

After 3 weeks, co-overexpression of IGF-I/FGF-2 improved the macroscopic aspect of defects without affecting the synovial membrane. Immunoreactivity to type-I collagen, an indicator of fibrocartilage, was significantly lower in defects receiving IGF-I/FGF-2 implants. Importantly, combined IGF-I/FGF-2 overexpression significantly improved the histological repair score. Most remarkably, such enhanced cartilage repair was correlated with a 2.1-fold higher proteoglycan content of the repair tissue. Finally, there were less degenerative changes in the cartilage adjacent to the defects treated with IGF-I/FGF-2 implants.

Conclusion

The data demonstrate that combined gene delivery of therapeutic growth factors to cartilage defects may have value to promote cartilage repair. The results also suggest a protective effect of IGF-I/FGF-2 co-overexpression on the neighbouring articular cartilage. These findings support the concept of implementing gene transfer strategies for articular cartilage repair in a clinical setting.     

In vitro uptake 153gadolinium and gadolinium complexes by hyaline articular cartilage.

This in vitro study evaluated whether Gadolinium (Gd) penetrates into hyaline cartilage and would be incorporated into vital chondrocytes. Hyaline joint cartilage of rabbits was exposed to radioactive 153GdCl3 and to a radioactive 153Gd-DTPA-BSA-complex (DTPA, diethylene-triaminepentaacetic acid; BSA, bovine serum albumine). In addition an exchange experiment with radioactive 153GdCl3 versus Gd-DTPA-di-N-methylglucamine (Magnevist) was performed. Incorporation of 153GdCl3 into neuroblastoma cells, connective tissue cells and chondrocytes was tested. The results showed that the depth and extent of incorporation of Gd depends on the molecular mass and time of exposure. 153Gd-DTPA-BSA complexes exhibited an incorporation rate of maximal 11% +/- 2.8% up to the middle third of the cartilage within 24 h with almost no incorporation (2 +/- 1.9%) for the deep layer. The exchange experiment revealed no uptake of Gd for the deep layer. The maximal incorporation rate of 153GdCl3 into vital chondrocytes was 6.3%. These data indicate that under the condition of MR-arthrography, Gd-DTPA-di-N-methylglucamine will not be absorbed into the deep layers of hyaline cartilage and will not be incorporated into vital chondrocytes.     

Effect of in vitro culture on a chondrocyte-fibrin glue hydrogel for cartilage repair

Research in tissue engineering has been focused on articular cartilage repair for more than a decade. Some pioneristic studies involved the use of hydrogels such as alginate and fibrin glue which still possess valuable potential for cartilage regeneration. One of the main issues in cartilage tissue engineering is represented by the ideal maturation of the construct, before in vivo implantation, in order to optimize matrix quality and integration. The present study was focused on the effect of in vitro culture on a fibrin glue hydrogel embedding swine chondrocytes. We performed an evaluation of the immunohistochemical and biochemical composition and of the biomechanical properties of the construct after 1 and 5 weeks of culture. We noticed that chondrocytes survived in the fibrin glue gel and enhanced their synthetic activity. In fact, DNA content remained stable, while all indices of cartilage matrix production increased (GAGs content, immunohistochemistry for collagen II and safranin-o staining). On the other hand, the biomechanical properties remained steady, indicating a gradual substitution of the hydrogel scaffold by cartilaginous matrix. This demonstrates that an optimal preculture could provide the surgeon with a better engineered cartilage for implantation. However, whether this more mature tissue will result in a more efficient regeneration of the articular surface still has to be evaluated in future investigations.     

MRI evaluation of a new scaffold-based allogenic chondrocyte implantation for cartilage repair

Aim

The present study was designed to evaluate the implantation of alginate beads containing human mature allogenic chondrocytes for the treatment of symptomatic cartilage defects of the knee. MRI was used for the morphological analysis of cartilage repair. The correlation between MRI findings and clinical outcome was also studied.

Methods

A biodegradable, alginate-based biocompatible scaffold containing human mature allogenic chondrocytes was used for the treatment of symptomatic chondral and osteochondral lesions in the knee. Twenty-one patients were prospectively evaluated with use of the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and the Visual Analogue Scale (VAS) for pain preoperatively and at 3, 6, 9 and 12 months of follow-up. Of the 21 patients, 12 had consented to follow the postoperative MRI evaluation protocol. MRI data were analyzed based on the original MOCART (Magnetic Resonance Observation of Cartilage Repair Tissue) and modified MOCART scoring system. The correlation between the clinical outcome and MRI findings was evaluated.

Results

A statistically significant clinical improvement became apparent after 6 months and patients continued to improve during the 12 months of follow-up. One of the two MRI scoring systems that were used, showed a statistically significant deterioration of the repair tissue at 1 year of follow-up. Twelve months after the operation complete filling or hypertrophy was found in 41.6%. Bone-marrow edema and effusion were seen in 41.7% and 25% of the study patients, respectively. We did not find a consistent correlation between the MRI criteria and the clinical results.

Discussion

The present study confirmed the primary role of MRI in the evaluation of cartilage repair. Two MOCART-based scoring systems were used in a longitudinal fashion and allowed a practical and morphological evaluation of the repair tissue. However, the correlation between clinical outcome and MRI findings was poor. Further validation of these scoring systems is mandatory. The promising short-term clinical outcome of the allogenic chondrocytes/alginate beads implantation was not confirmed by the short-term MRI findings.     

Biochemical and biomechanical properties of lesion and adjacent articular cartilage after chondral defect repair in an equine model

BACKGROUND: Chondral defects may lead to degradative changes in the surrounding cartilage, predisposing patients to developing osteoarthritis. PURPOSE: To quantify changes in the biomechanical and biochemical properties of the articular cartilage adjacent to chondral defects after experimental defect repair. STUDY DESIGN: Controlled laboratory study. METHODS: Specimens were harvested from tissue within (lesion), immediately adjacent to, and at a distance from (remote area) a full-thickness cartilage defect 8 months after cartilage repair with genetically modified chondrocytes expressing insulin-like growth factor-I or unmodified, control chondrocytes. Biomechanical properties, including instantaneous Young's and equilibrium aggregate moduli, were determined by confined compression testing. Biochemical properties, such as water and proteoglycan content, were also measured. RESULTS: The instantaneous Young's modulus, equilibrium modulus, and proteoglycan content increased, whereas water content decreased with increasing distance from the repaired lesion. The instantaneous Young's and equilibrium moduli of the adjacent articular cartilage were 80% and 50% that of remote area samples, respectively, whereas water content increased 0.9% and proteoglycan content was decreased by 35%. No significant changes in biomechanical and biochemical properties were found either in the lesion tissue or in adjacent cartilage with genetic modification of the chondrocytes. CONCLUSION: Articular cartilage adjacent to repaired chondral defects showed significant remodeling 8 months after chondral defect repair, regardless of whether genetically modified or unmodified cells were implanted. CLINICAL RELEVANCE: Changes in the biochemical and biomechanical properties of articular cartilage adjacent to repaired chondral defects may represent remodeling as part of an adaptive process or degeneration secondary to an altered distribution of joint forces. Quantification of these changes could provide important parameters for assessing progress after operative chondral defect repair.     

In vitro bonding of pre-seeded chondrocytes

Abstract This study investigated the capacity of seeded chondrocytes to join separate cartilage disc matrices in an in vitro model. Articular cartilage discs were harvested from pigs and devitalized by multiple freeze/thaw cycles. The devitalized cartilage discs were incubated in the presence (experimental group) or absence (control group) of chondrocytes for 10 days in order to allow chondrocytes to adhere to the matrix. After culturing, pairs of cartilage discs were held in apposition in a 48-multiwell plate and cultured for two and eight weeks. Twelve experimental composites (with cells) and twelve controls (without cells) were prepared per each time point. Samples were retrieved from culture and grossly inspected for adherence and processed for histological evaluation. Histological sections demonstrated the presence of new cartilage matrix formed by seeded chondrocytes bonding the two matrix discs together and producing glycosaminoglycans (GAG) able to diffuse within the devitalized tissue. Generally, gross adherence between the discs was demonstrated in the experimental samples, while the controls did not show any bonding. We conclude that isolated and seeded chondrocytes produce a new cartilaginous matrix, capable to join devitalized cartilage discs in vitro.     

关节软骨修复细胞基因治疗中软骨细胞支架载体特点

目的 探讨四种不同类型三维支架中培养的关节软骨细胞活性状况及其转基因特性.方法 原代培养兔膝关节软骨细胞,并且以表达绿色荧光蛋白(GFP)和萤火虫荧光素酶(GL3)的腺病毒载体AdGFP和AdGL3进行感染.在I型胶原海绵、纤维蛋白胶、透明质酸和聚乳酸四种类型的支架材料中,以荧光显微镜和荧光素酶分析法检测细胞的活性状况及其转基因特性;以阿辛蓝染色评价软骨基质的产生.结果 表达绿色荧光蛋白的腺病毒(AdGFP)成功感染兔关节软骨细胞,并且持续表达绿色荧光蛋白.被感染的软骨细胞在所有被测试的细胞支架上均能存活,并能表达GFP和荧光素酶报告基因.与其余三种支架比较,聚乳酸支架中转基凶表达率最高(P<0.01).而且聚乳酸培养体系中,4周时可以检测到阿辛蓝染色阳性的基质材料.结论 在关节软骨修复的细胞基因治疗领域,聚乳酸可能是一种合适的支架材料.     

兔膝内侧副韧带修复及内侧半月板切除对膝骨关节病发病过程的影响

通过经典的建立膝骨关节病运动模型的方式，设计了在切除内侧半月板的情况下，将内侧副韧带（ＭＣＬ）切断和切断后再缝合恢复张力的两组动物模型。观察术后５、１０、１５、２０天，关节软骨的组织学、组织化学及扫描电镜下形态学方面的变化，旨在探讨切除内恻半月板时，恢复ＭＣＬ的张力对膝骨关节病（ＯＡ）发病过程的影响。实验结果显示，ＭＣＬ切断无张力组和ＭＣＬ缝合有张力组于术后５天，在光镜下，关节软骨即出现某此变化。包括敕骨细胞排列紊乱，增生层有软骨细胞簇集现象、族集细胞的周围甲苯胺兰深染。增生层和肥大层一些软骨陷窝有细胞消失现象，其周围淡染。术后５、１０天扫描电镜观察，两实验组关节软骨表面未见异常改变。术后１５、２０天，光镜下，两实验组关节软骨退行性改变进一步加重，扫描电镜观察，两实验组软骨表面正常结构消失、胶原纤维断裂，软骨显微骨折、出现部分软骨细胞裸露及细胞消失现象。韧带无张力组及有张力组，在软骨退行性变化方面未见明显程度上的差别，结果表明，只要兔半月板切除，无论ＭＣＬ张力存在与否，都不能阻止ＯＡ的产生，亦不能减轻其发生的程度。     

Wnt3a在人骨性关节炎软骨中的表达及相关性研究

目的 Wnt3a参与Wnt经典通路信号的传递,而Wnt通路又参与关节软骨及软骨细胞的发生。本实验目的是检测Wnt3a在骨性关节炎（OA）中的表达及发病风险的相关性。方法 40例人类关节软骨标本,10例正常关节软骨标本（取自外伤、肿瘤截肢术患者）,30例膝OA关节软骨标本（取自膝OA患者）.将取下的软骨标本分为四组：正常关节软骨组、轻度软骨退变组、中度软骨退变组、重度软骨退变组。根据Mankin关节软骨病理评分标准分别打分,按评分结果分组：正常关节软骨标本5份（A组）,OA软骨轻度退变标本6份（B组）,OA软骨中度退变标本13份（C组）,OA软骨重度退变标本16份（D组）。对四组切片行Wnt3a相关染色,根据免疫反应积分IRS打分法进行评分,采用SPSS17.0软件对数据进行统计学分析。统计分析Wnt3a在正常组与非正常组的差异性、表达程度及软骨退变程度的相关性。结果 Wnt3a在三组人膝骨性关节炎软骨呈现一致的高表达,其表达的平均值分别为：3.33±0.516（B组,轻度）,4.85±1.68（C组,中度）,9.44±2.31（D组,重度）。正常组（A组,表达均值和标准差0.0±0.0）与中度、重度退组两两比较,轻度组与中度、重度组,中度与重度组比较显示一致的显著性差异。Wnt3a表达值与其相应的Mankin病理评分呈显著正相关（B组皮尔森相关系数=0.74,P=0.047;C组皮尔森相关系数=0.81,P=0.000 36;D组皮尔森相关系数=0.94,P=2.75e-08）,即关节软骨中Wnt3a的表达与原发性膝骨关节炎（OA）发生与发展程度呈显著的正相关。结论 1在膝骨性关节炎软骨中Wnt3a表达升高;2Wnt3a表达与骨性关节炎关节软骨退变程度呈正相关,可作为OA病程进展的监测指标。