肝细胞生长因子对关节软骨缺损修复作用的实验研究

目的 探讨肝细胞生长因子＊ｈｅｐａｔｏｃｙｌｅ ｇｒｏｗｔｈ ｆａｃｔｏｒ,ＨＧＦ）对创伤性关节软骨损伤的修复作用。方法 选 用大耳白 兔２５只,随机分成５组,每组５只。在两侧髌股关节股骨侧的关节面中心分别造成直径３．５ｍｍ的全层关节软骨缺损。造反右侧为实验组,自术后起,膝关节内注射１０μｇ／ｍｌ的ＨＧＦ０．１ｍｌ,每周３次,共４周;左侧为对照组,与实验组同一时间注射等量生理盐水。于术后４、６、８     

bFGF对关节软骨缺损的修复作用

目的观察碱性成纤维细胞生长因了对关节软骨缺损修复的影响。方法在兔双侧股骨髁间窝造成骨软骨缺损．一侧应用碱性成纤维细胞生长因子．另一侧作对照。术后3个月．利用组织切片及扫描电镜等方法，观察两侧骨软骨缺损修复情况。结果修复3个月后．对照侧软骨缺损难以完全修复．修复细胞形态多样，似成纤维细胞。蛋白多糖颗粒较少。应用碱性成纤维细胞生长因子删软骨缺损基本修复．修复细胞似软骨细胞．有较多的蛋白多糖颗粒与胶原纤维结合。缺损修复组织评分。碱性成纤维细胞生长因子组优于对照组。结论碱性成纤维细胞生长因子可促进骨软骨缺损的修复。     

FGF对兔关节软骨细胞基质胶原和蛋白多糖的影响

关节软骨的损伤与修复是目前骨科临床基础研究的一个重要课题。关节软骨损伤后的修复是极其有限的〔１〕。许多多肽生长因子在关节软骨损伤与修复中起着重要的调节作用，如成纤维细胞生长因子（ｆｉｂｒｏｂｌａｓｔｇｒｏｗｔｈｆａｃｔｏｒ，ＦＧＦ）对中胚层来源的细胞...     

兔膝关节内骨折后关节面塑形能力的实验研究

目的观察兔膝关节内骨折后不同形式关节面不平整的塑形能力。方法在兔胫骨平台造成关节内骨折,骨折移位0.5、1.0、2.0 mm,形成关节面凹陷或台阶样结构。术后3个月取胫骨平台标本进行组织切片、HE染色,观察不同形式关节面不平整的塑形能力。结果兔胫骨平台凹陷性或台阶性不平整关节面移位0.5 mm时,通过凹侧软骨增生肥厚等塑形,可恢复关节面的平整。关节面移位1.0 mm及2.0 mm时,通过凹侧软骨增生肥厚及来自髓腔的增生修复组织,凹陷性不平整仍可恢复关节面的平整,但台阶性不平整难以通过关节面的重新塑形恢复关节面的平整。结论关节内骨折后形成关节面不平整,凹陷性骨折的再塑形能力可能优于台阶性骨折。     

新鲜同种异体骨软骨移植修复软骨缺损

目的联合应用新鲜同种异体骨软骨移植,和局部注射碱性成纤维细胞生长因子(basic fibroblast growthfactor,bFGF),探讨能否促进关节软骨缺损区新生软骨的形成,提高软骨缺损修复的成功率。方法48只青紫兰兔,96个实验关节,随机分为A、B、C、D组。无菌条件下制作骨软骨缺损模型。在A组缺损区单纯植入新鲜的同种异体骨软骨,B组单纯局部注射重组人bFGF,C组局部注射bFGF后同时植入新鲜的同种异体骨软骨,D组用作空白对照。术后第4、8、12周作大体观察、X线摄片、组织学检查及免疫组化检查。结果移植加注射bFGF组促进软骨缺损修复的效果均好于其他组,图像分析仪进行软骨细胞记数有显著差异(P<0.05),有统计学意义。修复软骨型胶原免疫组化染色强阳性。结论采用新鲜的同种异体骨软骨移植及联合应用碱性成纤维细胞生长因子,二者能起交互作用,促进了新生软骨的形成。     

骨髓间充质干细胞复合纤维蛋白凝胶修复兔关节软骨缺损

目的观察骨髓间充质干细胞(MSCs)复合纤维蛋白凝胶(FG)对兔关节软骨缺损的修复效果。方法12只兔24个膝关节按左右分为实验组与对照组,抽取兔骨髓,密度梯度离心法分离间充质干细胞并行体外培养扩增,实验组以MSCs与FG及转化生长因子β1(TGFβ1)复合后填充到兔膝关节全厚软骨缺损模型中,而对照组以FG/TGFβ1填充,于术后12周取材,观察检测软骨缺损的修复情况。结果术后12周,实验组缺损由类透明软骨或透明软骨修复,Ⅱ型胶原免疫组化染色阳性,甲苯胺蓝异染性明显,修复面较平整光滑;而对照组则以纤维软骨及纤维组织修复,Ⅱ型胶原免疫组化染色阴性。结论MSCs复合FG及TGFβ1能够修复兔关节软骨缺损。     

慢病毒介导的Sox9基因在兔骨髓间充质干细胞的过表达促进软骨损伤修复

目的:明确在体内Sox9基因过表达的兔骨髓间充质干细胞对于关节软骨损伤修复的作用。方法:以慢病毒介导的Sox9基因转染兔骨髓间充质干细胞(BMSCs),体外检测软骨特异性分子,将新西兰大白兔24只48个膝关节随机分为3组,动物麻醉后,双侧股骨滑车处的关节面上用直径4 mm的钻头钻孔,深度3 mm,穿透软骨下骨,造成全层关节软骨损伤,将转染后的细胞植入体内用以修复全层关节软骨损伤,实验组植入BMSCs-(Lenti-Sox9-EGFP)-藻酸钙复合物,实验对照组植入BMSCs-藻酸钙复合物,空白对照组只钻孔。术后6、12周分别进行光镜、电镜观察,以及HE、免疫组织化学染色检测软骨的修复程度。结果:经Sox9基因转染后的细胞在3 d时,Sox9基因表达最高,随后下降。转染后3 d,Ⅱ型胶原开始表达,到14 d时达到最高。表明Sox9过表达启动了兔骨髓间充质干细胞的软骨分化。组织学观察显示,实验组术后6周缺损处有透明软骨样组织填充,术后12周缺损处软骨和软骨下骨修复良好。两对照组,缺损处由纤维组织填充。免疫组织化学显示,修复组织内Ⅱ型胶原,免疫组化染色结果阳性强于两对照组。组织学评分结果显示实验组软骨损伤修复各时间点效果明显优于两对照组,差异有统计学意义。结论:Sox9基因过表达的兔骨髓间充质干细胞(BMSCs)促进软骨损伤的修复。     

血管内皮生长因子对兔软骨修复的影响

已证实血管内皮生长因子(VEGF)可以促进血管的增殖,本实验旨在探讨通过VEGF增加损伤区域的血运,观察其是否可以促进软骨的修复. 一、材料与方法 24只新西兰大白兔,随机分为2组.静脉麻醉、消毒后,取膝关节外侧入路,暴露股骨滑车负重区的软骨面,制作直径4 mm、深2 mm的全层软骨损伤的模型.实验组关节内注射30 μg VEGF-A混合0.2 ml生理盐水;对照组注射0.5 ml生理盐水.术后3、6和12周随机选取实验组和对照组的4只兔处死,切取标本,进行组织学观察.     

注射型软骨组织工程活性材料修复关节软骨缺损的实验研究

目的 利用注射型活性材料修复兔关节软骨,比较碱性成纤维细胞生长因子(Basic fibroblastic growth factor,bFGF)和骨形态发生蛋白(Bone morphogenic protein,BMP)的治疗效果。方法 取18只14周龄成年大白兔随机分为3组,每只动物于双侧膝关节股骨髌股关节面制备直径4mm全层软骨缺损模型,钻通骨髓腔。准备注射型bFGF和BMP生物蛋白胶材料,三组分别接受如下治疗。A组注射型碱性成纤维细胞生长因子,B组注射型骨形态发生蛋白,C组单纯生物蛋白胶治疗。于8周,12周,24周观察大体和形态学特征,组织学评分。结果 A组于8,12周时,软骨缺损被白色半透明坚硬组织平滑修复,富有光泽,与正常软骨边界清楚,24周时修复组织与正常软骨边界模糊,质地坚硬,表面平滑光润。B组于8,12,24周时候均未将缺损表面修复平滑,为疏松组织缺损底部,凹陷明显残留。C组全程均未见软骨修复。组织评分A组明显优于B组C组(P<0.05),而B组和C组间无差异(P>0.05)。结论 注射型碱性成纤维细胞生长因子可以有效修复兔关节软骨的缺损,效果明显优于BMP治疗组,为关节镜等微创治疗方法提供了实验参考。     

透明质酸钠与强的松龙治疗兔实验性颞颌关节骨关节炎的研究

目的　探讨透明质酸钠关节内注射治疗兔实验性颞颌关节骨关节炎的效果 ,并与强的松龙进行比较。另对透明质酸钠的治疗机制进行初步探讨。方法　选择 12只日本大耳白兔 ,在双侧颞颌关节上腔各注射1.6 %木瓜蛋白酶溶液 0 .2 ml,3天后右侧再注射 0 .2 ml以诱发程度不同的骨关节炎。除 1只兔意外死亡外 ,11只兔均存活至预期时间。于末次注射后 1周 ,用 6只兔每周在双侧关节上腔各注射透明质酸钠 1.3mg作为实验组。另 5只兔双侧关节上腔各注射强的松龙 1.6 m g作为对照组。连续 4次。疗程结束后第 3、5及 7周分批处死动物 ,对关节进行病理学观察。结果　疗程结束后第 3、5及 7周 ,对照组双侧关节均可见明显结构破坏 ,以右侧较重 ;主要表现为纤维软骨变性 ,软骨变薄或脱落 ,关节面可见以不成熟的纤维细胞和成纤维细胞为主的不完全修复。实验组纤维软骨病变多表现为细胞轻度减少 ,纤维变性 ,甚至局部表层软骨缺损 ,周围软骨细胞簇状增生 ,数量增多 ,骨关节炎病理记分显著低于对照组。结论　透明质酸钠有修复和保护软骨的作用 ,强的松龙无助于软骨的修复 ,反而加重骨关节炎病变     

肝细胞生长因子对膝关节软骨影响的临床意义

目的 研究前列腺素Ｅ２（ＰＧＥ２）参与关节软骨退变的进程机理,以及肝细胞生长因子（ＨＧＦ）对膝关节软骨损伤修复的作用机理。方法 对符合临床诊断标准的３０个膝关节,分别给予膝关节腔注射肝细胞生长因子,每天１次,每次２ｍｌ,观察ＰＧＥ２、尿中１型胶原（ｃｒｏｓｓｌａｐｓ）、碱性磷酸酶（ＡＫＰ）以及临床症状的改善程度,并做了给药６周后与给药有的对比研究。结果 给药６周后ｃｒｏｓｓｌａｐｓ和ＰＧＥ２与给药     

The Healing of Cartilage Injuries Under the Influence of Joint Immobilization and Repeated Hyaluronic Acid Injections: An Experimental Study

In an experimental animal model the influence of intraarticularly injected high-molecular hyaluronic acid on the healing of superficial and deep lesions of the articular cartilage in freely mobile and immobilized joints was investigated. In the right knee joint in 42 adult rabbits two intracartilaginous lesions were produced in one of the femoral condyles and an osteochondral lesion in the other. In half of the animals the injured knee joint was immobilized in plaster of Paris, while the other half were allowed to move the joint freely. Half of the injured joints in each group were then injected with high-molecular hyaluronic acid once a week for 8 weeks. After 2 months the knee joints were examined histologically, histochemically and microangio-graphically and the water content of the articular cartilage was determined. The intracartilaginous lesions remained unchanged in all knee joints. The wound margins of the osteochondral lesions showed better closure in the immobilized animals. As a rule the osteochondral lesion healed with fibrous connective tissue. In a few joints, however, hyaline cartilage, with production of chondroitin sulphate, developed during healing of the osteochondral lesion. In the vicinity of the joint surface this tissue was transformed into fibrocartilage. Immobilization of the joint invariably led to pannus formation. The hyaluronic acid injections appeared to have no effect, either positive or negative, on the healing of intracartilaginous and osteochondral joint lesions. In mobile joints, however, these injections prevented a reduction of the water content of the articular cartilage, which was regarded as favourable.     

软骨脱细胞基质-Ⅱ型胶原纳米支架复合BMSC修复兔关节软骨缺损的实验研究

目的 观察软骨脱细胞基质(Cartilage acellular extracellular matrix,CAEM)-Ⅱ型胶原(CollagenⅡ,COLⅡ)纳米支架,复合骨髓基质干细胞(Bone marrow stem cells,BMSCs)修复兔关节软骨缺损的效果。方法 CAEM和COLⅡ按质量比1∶1混合,通过静电纺丝技术制备组织工程纳米支架。将第二代BMSCs种植到该支架上,培养箱内静置2 h。12只日本大耳白兔随机分为实验组和对照组,将细胞支架复合物植入实验组兔膝关节软骨缺损处,对照组仅行膝关节软骨缺损建模。12周后实验动物取材,大体观察修复效果,并行HE染色、Ⅱ型胶原染色观察。结果 大体观察见实验组软骨缺损修复良好,对照组软骨缺损处由肉芽样组织充填。HE染色显示,实验组关节软骨缺损处可见软骨陷窝形成,对照组关节软骨缺损处仅有纤维组织充填。实验组修复区Ⅱ型胶原染色为阳性,对照组为阴性。结论 CAEM-COLⅡ纳米支架复合BMSC,对兔关节软骨缺损具有较好的修复能力,具有潜在的临床应用价值。     

One intra‐articular injection of hyaluronan prevents cell death and improves cell metabolism in a model of injured articular cartilage in the rabbit

The purpose of this study was to determine the effect of one intra‐articular injection of hyaluronan on chondrocyte death and metabolism in injured cartilage. Twenty‐three 6‐month‐old rabbits received partial‐thickness articular cartilage defects created on each medial femoral condyle. In order to examine the effect on articular cartilage surrounding iatrogenic cartilage lesions, which can occur during arthroscopic procedures, Study 1 was performed: in 14 rabbits both knees were immediately rinsed with 0.9% NaCl. Experimental knees were treated with hyaluronan. Six rabbits were sacrificed at 2 days; eight rabbits 3 months postoperatively. Histomorphometric analysis was used for studying cell death in cartilage next to the defect. In order to examine the effect on longer lasting lesions, more reflecting the clinical situation, Study 2 was performed: after 6 months knee joints of nine rabbits were (i) irrigated with 0.9% NaCl, (ii) treated with hyaluronan after irrigation with 0.9% NaCl, or (iii) sham‐treated. After 7 days patellas were used to study the chondrocyte metabolism by measuring the [³⁵S]sulfate incorporation. Study 1: Two days postoperatively, in hyaluronan‐treated cartilage the percentage of dead cells was 6.7%, which was significantly lower compared to 16.2% in saline‐treated cartilage. After 3 months the percentages of dead cells in both groups were statistically similar. Study 2: Hyaluronan treatment resulted in significantly higher [³⁵S]sulfate incorporation compared to knees irrigated with 0.9% NaCl. These results suggest a potential role for hyaluronan in preventing cell death following articular cartilage injury. One injection of hyaluronan improved cartilage metabolism in knees with 6‐month‐old cartilage defects. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 26:624–630, 2008     

Elevation in urinary levels of pyridinium cross-links of collagen following chymopapain-induced degradation of articular cartilage in the rabbit knee provides evidence of metabolic changes in bone

To measure urinary levels of two pyridinium cross-links of collagens at various times, rabbits were injected in the left knee with 0.2 mg (N = 11) or 2.0 mg chymopapain (N = 11). Urinary levels of the bone-specific deoxy-pyridinoline cross-link and of the hydroxylysyl pyridinoline cross-link, found in large amounts in articular cartilage and bone, were measured by high performance liquid chromatography (HPLC). The urinary levels were correlated with histological evidence of progressive articular cartilage destruction in animals injected with 2.0 mg chymopapain and of successful repair in rabbits injected with the lower dose. Rabbits injected with 2.0 mg chymopapain showed a very large increase in urinary levels of deoxy-pyridinoline during the first 30 days after the injection. This rise was not seen in animals injected with the lower dose. A rise in urinary level of the hydroxylysyl pyridonline cross-link was observed in all rabbits but was more pronounced and lasted longer in animals receiving the higher dose. The failure of rabbits injected with 2.0 mg chymopapain to repair the damaged articular cartilage was strongly associated with a marked increase in the catabolism of the bone-specific deoxy-pyridinoline cross-link and to a lesser extent with the catabolism of the hydroxylysyl pyridinoline found in bone and in articular cartilage.     

Articular cartilage changes following meniscal lesions. Repair and meniscectomy studied in the rabbit knee

A well-defined, longitudinal lesion in the avascular part of the medial meniscus of the right knee was made in 30 rabbits. After 3 months, the lesion was repaired surgically in 12 rabbits, was untreated in 12 rabbits, and a meniscectomy was performed in 6 rabbits. The articular cartilage was studied macroscopically and microscopically at 3-month intervals. Cartilage changes 3 months after meniscectomy were more pronounced than after meniscal repair or than in untreated lesions. However, meniscal repair did not reverse the cartilage changes.     

自体骨膜延迟游离移植修复成年后关节软骨缺损的实验研究

目的：研究年龄对自体骨膜游离移植修复关节软骨缺损的影响,探讨延迟游离移植能否提高成年后骨膜修复软骨能力。方法：选中国白兔,成年兔20只,幼兔10只,分3组。A组：成年兔左膝骨膜直接游离移植组;B组：成年兔右膝骨膜延迟游离移植组;C组：幼兔骨膜直接游离移植组,取骨膜或骨膜新生组织、行光镜、电镜组织学观察比较。结果：移植前B、C组骨膜厚度、细胞计数及细胞活跃程度均优于A组（均为P＜0．01）,移植后12周3组关节软骨缺损获得不同程度修复,C组优于A组（P＜0．01）及B组（P＜0．05）,B组优于A组（P＜0．01）。结论：自体骨膜局部剥离、原位激活,体内培养、延迟游离移植可提高成年骨膜成软骨能力,更好地修复成年后关节软骨缺损。     

孔数不同的骨钻孔术对兔软骨缺损远期修复效果的实验比较

目的: 评价孔数不同的钻孔术对软骨缺损的远期修复效果。方法: 用中国白兔40只, 在股骨髁关节面制造6mm×8mm全层软骨缺损, 分别施行10孔及5孔钻孔术, 孔径1mm, 于术后13个月取材做组织学及电镜观察,并进行评估。结果: (1) 10孔、5孔和对照组中, 优势修复组织为透明软骨者分别占75%、70%、0%。(2) 修复组织厚度: 10孔与5孔无显著性差异, 已接近毗邻软骨厚度。(3) 修复组织覆盖缺损的面积: 10孔>5孔>对照组。结论: 软骨下骨钻孔对关节软骨缺损的远期修复效果良好, 能长期适应关节的运动和负重, 10孔比5孔的修复效果好。     

The effect of hyaluronan combined with microfracture on the treatment of chondral defects: an experimental study in a rabbit model

Because articular hyaline cartilage has low potential for regeneration, numerous methods and techniques have been proposed to induce the reparation process. Microfracture is a convenient procedure for this purpose. However, the quality of the new cartilage after microfracture is still not as proper as original. In this experimental study, we used microfracture in combination with intraarticular application of hyaluronan in rabbit knee articular defect. Bilateral knee arthrotomies, chondral defects, and microfracture were created on each intercondylar notch in thirty rabbits. Rabbits received intraarticular injections of hyaluronan once a week for 3 weeks in the right knee, started from 1 week after injury. The left knees, which served as controls, were injected with normal saline. Biopsy was taken from both knees at the 4th and 6th weeks. In comparison with the control group, after 6 weeks we observed a higher potential for healing in the experimental group, with thicker and more organized repair tissue filling the defect. The current study reveals that application of hyaluronan after the microfracture might be beneficial in inducing articular cartilage defect reparation.