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?     

异种脐血干细胞移植修复兔全层关节软骨缺损的初步研究

目的 :探讨人脐血干细胞对兔全层关节软骨缺损的修复作用及免疫反应。材料和方法 :取人脐带血中脐血干细胞及幼兔的骨髓基质细胞 ,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 0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 ,将两种细胞 -PLA复合物分别植入关节软骨缺损处。植入异种脐血干细胞 -PLA复合物为实验组 ,植入同种异体骨髓基质细胞 -PLA复合物为阳性对照组 ,缺损不处理为阴性对照组。术后 6周、1 2周观察缺损修复情况、新生组织类型及有无免疫反应。结果 :脐血干细胞组 6周时标本为纤维组织修复 ,内有少量软骨细胞 ;1 2周时 40 %标本为软骨样组织修复 ,较薄 ;60 %标本为纤维组织修复。移植物周围无明显淋巴细胞聚集 ,部分滑膜有炎症反应。骨髓基质细胞组(阳性对照)为软骨样组织修复 ;滑膜无明显炎症反应。阴性对照组为纤维组织修复 ,无软骨形成。结论 :异种脐血干细胞移植修复软骨缺损优于缺损不处理组 (阴性对照) ,但明显差于同种骨髓基质细胞组 (阳性对照 )。脐血干细胞有可能成为软骨修复的新的种子细胞。由于种属差异的影响 ,脐血干细胞组可能存在免疫反应 ,结果需进一步研究     

人半月板纤维软骨细胞培养及生物学特性研究

为探索分离培养人半月板纤维软骨细胞的简便实用方法，采用胰蛋白酶和胶原酶联合消化的方法，简便快速地获得大量成活率高的人半月板纤维软骨细胞，在F－12培养液中进行原代和传代培养，并对传代培养纤维软骨细胞进行了免疫组化鉴定，光镜及超微结构的观察，结果显示，光镜下，原代培养细胞呈线形，单层排列，电镜可见细胞内有丰富的粗面内质网及线粒体，细胞呈多极性，表面有突起，纤维软骨细胞免疫组化Ⅱ型胶原染色阳性，提示本实验建立的人纤维软骨细胞分离培养方法是一种可行的方法。，     

骨髓基质细胞移植治疗骨缺损的实验研究

目的 探讨以Ⅰ型胶原海绵为载体的自体骨髓基质细胞移植治疗骨缺损的效果。方法 32只成年新西兰白兔随机分成Ⅰ、Ⅱ两组，分别自股骨大转子及胫骨结节抽取骨髓基质细胞进行培养，扩增后种植于Ⅰ型胶原海绵上，继续培养2周后植入1．5cm长桡骨缺损。Ⅰ组一侧桡骨缺损植入Ⅰ型胶原 自体骨髓基质细胞复合物(A处理)，对侧桡骨不做任何植入(B处理)。Ⅱ组一侧桡骨缺损做A处理，对侧桡骨缺损植入Ⅰ型胶原海绵(C处理)，分别于术后8周和12周取材，观察骨缺损的修复情况。结果 术后12周，A处理组全部骨性愈合，B处理组由纤维组织填充，C处理组有少量骨痂形成。结论 以Ⅰ型胶原海绵为载体的自体骨髓基质细胞移植能有效修复骨缺损。     

同种异体关节软骨细胞分离体外培养再移植修复关节软骨缺损实验研究

目的关节软骨一旦缺损，其修复能力有限。本实验通过移植同种异体兔软骨细胞，修复关节软骨缺损。方法软骨细胞取自出生后３周雄性新西兰幼兔，体外培养，移植于成年雌性新西兰兔膝关节。股骨内髁缺损区为移植细胞组，左股骨外髁缺损为移植胶原凝胶组，右股骨外缺损为空白对照组。术后所有动物不加外固定任意自由活动，于术后４，８，１２周处死，进行肉眼、组织学、透射电镜观察，标本用带有Ｙ染色体的Ｓｅｘ－ｄｅｔｅｒｍｉｎｇ－ｒｅｇｉｏｎ－Ｙ（ＳＲＹ）基因进行杂交。结果移植软骨细胞组术后股骨内髁缺损区得到修复，而移植胶原凝胶组和空白对照组术后股骨外髁缺损未得到修复。ＳＲＹ基因性别鉴定也证明了以上结果。结论关节软骨的缺损及骨性关节炎通过软骨细胞移植方法进行治疗，可再生出新的透明软骨。     

人胰岛素样生长因子Ⅰ基因增强组织工程修复全层半月板缺损

目的 建立全层无血运区半月板缺损模型,观察人胰岛素样生长因子Ⅰ (human insulin-like growth factor Ⅰ,hIGF-Ⅰ)基因修饰骨髓间充质干细胞(bone-marrow mesenchymal stem cells,BMSCs)复合可注射藻酸钙凝胶修复半月板缺损的效果. 方法 制造成年山羊半月板前角无血运区全层缺损模型.实验分为四组,即基因增强组织工程(gene-enhanced tissue engineering,GETE)组(GETE组):用hIGF-Ⅰ基因转染的BMSCs复合可注射藻酸钙凝胶修复半月板缺损；BMSCs组:用BMSCs复合藻酸钙凝胶修复缺损；空载体组:用藻酸钙凝胶空载体修复缺损；对照组:缺损旷置,不做任何修复.术后4,8,16周时相点行大体观察；光镜、电镜观察修复组织变化；测定修复组织中蛋白聚糖含量. 结果 GETE组修复半月板缺损在4～ 16周效果逐渐改善,缺损完全被修复组织填充,色白质韧,结合紧密,与正常半月板组织相似,大体观察优于其他各组；光镜下可见细胞随凝胶纤维排列分布,载体纤维间隙大多为细胞分泌基质所充填,细胞排列密集,基质分布均匀；扫描电镜观察到纤维排列规整紧密,纤维间缝隙为致密的细胞外基质填充；修复组织中蛋白聚糖含量较高,但与正常半月板仍有差异. 结论 转染hIGF-Ⅰ基因的BMSCs复合可注射藻酸钙凝胶可改善全层半月板缺损的修复效果.     

胰岛素样生长因子-Ⅰ对软骨细胞移植修复关节软骨缺损的作用

目的：观察胰岛素样生长因子－Ⅰ（insulin-like growth factor-Ⅰ,IGF-Ⅰ)对关节软骨缺损修复的作用。方法：采用组织工程方法制备骨基质明胶（BMG）－软骨细胞移植物。40只4－5个月龄的新西兰兔均分为软骨细胞实验组及软骨细胞对照组,BMG对照组,空白对照组。实验组兔膝关节注射IGF－Ⅰ,各对照组均注射等渗盐水。术后行大体,组织学观察及免疫组化检测。结果：实验组软骨细胞生长较快,术后24周修复的软骨组织Safranin-O染色,Ⅱ型胶原染色与正常关节软骨一致,软骨细胞呈柱状排列,软骨细胞对照组术后24周修复软骨组织Safranin-O染色较淡,部分软骨细胞呈柱状排列,BMG,空白对照组未修复。结论：IGF－Ⅰ能促进关节软骨缺损修复。     

体外器官培养系统中纤维软骨组织修复的实验研究

半月板是一种纤维软骨组织，由纤维软骨细胞与以Ｉ型胶原为主的基质构成。以往认为纤维软骨细胞缺乏再生能力，使半月板损伤不能愈合。近来有研究表明脱离了基质的纤维软骨细胞在体外单层培养条件下有很强的增生分化能力。因此我们在体外器官培养并提供纤维蛋白支架的条件...     

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。     

异体滑膜及青少年软骨颗粒移植修复兔半月板“白区”缺损及移植细胞的转归研究

目的：探讨异体滑膜和青少年软骨颗粒移植在半月板“白区”缺损修复上的协同作用;研究移植细胞的转归（存活和分化）;研究滑膜间充质干细胞和软骨细胞间的相互作用。方法：选取2只未成年雄性兔作为滑膜及青少年软骨颗粒供体。选取24只成年雌性兔（共48个膝关节）制备半月板缺损模型,分为以下几组：空白组,半月板缺损无任何处理;单纯纤维蛋白胶组,在半月板缺损处填充单纯纤维蛋白胶;纤维蛋白胶+滑膜移植组,纤维蛋白胶包裹异体滑膜碎片填充到半月板缺损处;纤维蛋白胶+滑膜+异体青少年软骨颗粒移植组,纤维蛋白胶包裹异体滑膜和异体青少年软骨颗粒填充到半月板缺损处,每组样本量为12个膝关节。分别在术后8周和16周取材,通过组织学和免疫组织化学染色评估半月板缺损修复程度,针对雄性个体特有的Y染色体性别决定区基因（SRY）进行核酸原位杂交,并结合免疫荧光共定位评估移植细胞的存活和分化。其次,建立体外兔滑膜间充质干细胞和软骨细胞共培养系统,检测半月板纤维软骨细胞相关基因转录水平,并探索细胞间相互作用的可能机制。结果：在术后8、16周,相较于空白组、单纯纤维蛋白胶组及纤维蛋白胶+滑膜移植组,纤维蛋白胶+滑膜+异体青少年软骨颗...     

Replacement of the knee meniscus by a porous polymer implant: a study in dogs

BACKGROUND: Meniscectomy will lead to articular cartilage degeneration in the long term. Therefore, the authors developed an implant to replace the native meniscus. HYPOTHESIS: The porous polymer meniscus implant develops into a neomeniscus and protects the cartilage from degeneration. STUDY DESIGN: Controlled laboratory study. METHODS: In a dog model, a porous polymer scaffold with optimal properties for tissue infiltration and regeneration of a neomeniscus was implanted and compared with total meniscectomy. The tissue infiltration and redifferentiation in the scaffold, the stiffness of the scaffold, and the articular cartilage degeneration were evaluated. RESULTS: Three months after implantation, the implant was completely filled with fibrovascular tissue. After 6 months, the central areas of the implant contained cartilage-like tissue with abundant collagen type II and proteoglycans in their matrix. The foreign-body reaction remained limited to a few giant cells in the implant. The compression modulus of the implant-tissue construct still differed significantly from that of the native meniscus, even at 6 months. Cartilage degeneration was observed both in the meniscectomy group and in the implant group. CONCLUSION: The improved properties of these polymer implants resulted in a faster tissue infiltration and in phenotypical differentiation into tissue resembling that of the native meniscus. However, the material characteristics of the implant need to be improved to prevent degeneration of the articular cartilage. CLINICAL RELEVANCE: The porous polymer implant developed into a polymer-tissue construct that resembled the native meniscus, and with improved gliding characteristics, this prosthesis might be a promising implant for the replacement of the meniscus.     

Arthroscopic treatment of lateral meniscal cysts using an outside-in technique

This paper describes 19 cases of lateral meniscal cysts treated arthroscopically using an outside-in technique. In all patients, a horizontal or radial lesion (or both) of the meniscus was present. After the meniscal lesion was arthroscopically removed, the cyst was decompressed both from inside and percutaneously from outside with a motorized instrument introduced through a transmeniscal approach. The follow-up ranged from 2 to 5 years with an average of 3.3 years. On the basis of the evaluation scale developed by Cerullo et al. (1991), the results were rated excellent or good in 17 patients and fair in 2. No patient had cyst recurrence. The computed tomography follow-up examination, done in 10 of the 19 patients after an average of 3 years, showed a good remodeling of the meniscus. One of the two patients with a fair result had hypermobility of the posterior horn of the lateral meniscus due to the resection of the meniscal tissue overlying the popliteus tendon, whereas the other had pain and patellar crepitation owing to a preexisting patellofemoral abnormality.     

Epidemiologic, clinical and arthroscopic study of the discoid meniscus variant in Greek population

This retrospective study was aimed to investigate the epidemiologic, clinical and arthroscopic features of discoid meniscus variant in Greek population. We reviewed the cases of 2,132 patients who underwent knee arthroscopy between 1986 and 2004 and diagnosis of discoid lateral meniscus was established in 39 patients with mean age of 31.7 ± 9.4 years old. Incidence of the discoid lateral meniscus variant was recorded at rate of 1.8% presenting no significant differences according to patient gender or lesion body side. Regarding the type of discoid dysmorphy, 23 cases attributed to complete type, 15 were incomplete and in one case, Wrisberg type was observed. Predictive values of the most commonly recorded physical signs in the clinical diagnosis of the discoid meniscus were analysed. Comparative evaluation of the long-term results of arthroscopic partial meniscectomy performed in patients with intact or torn discoid lateral meniscus and torn normally shaped lateral meniscus was carried out using Lysholm and IKDC scoring systems. Also, we investigated any correlation between dysmorphy type and tear pattern analysing the arthroscopic findings. Results demonstrated that the discoid meniscus lesion represents an atypical clinical entity in adults and no significant predictive value of the signs encountered in the clinical examination of the patients with discoid meniscus was observed. Clinical outcome after arthroscopic partial meniscectomy regarding the intact discoid meniscus group was superior in comparison with that of torn discoid meniscus cases. On other hand, no difference in the result of partial meniscectomy between discoid and normal lateral meniscus tear groups was found. No statistically significant relationship between the type of discoid menisci and tear pattern or incidence rate of concomitant intraarticular lesions was confirmed.     

Meniscal regeneration with copolymeric collagen scaffolds. In vitro and in vivo studies evaluated clinically, histologically, and biochemically.

We sought to create a regeneration template for the meniscal cartilage of the knee to induce complete meniscal regeneration, and to develop the technique for implanting the prosthetic appliance in vivo. We designed a resorbable collagen-based scaffold and conducted in vitro and in vivo studies. In vivo, the scaffold was implanted in the knees of immature swine and mature canines and evaluated clinically, histologically, and biochemically. Because the canine stifle joint meniscus is more clinically relevant to the human meniscus, this paper emphasizes those results. We studied 24 mixed breed dogs (14 males and 10 females) with an average weight of 25.5 kg (range, 20 to 35) that were obtained from a USDA-licensed supplier. The dogs were deemed clinically and radiographically skeletally mature. None of the dogs had a preexisting knee joint abnormality. All dogs underwent an 80% subtotal resection of the medial meniscus bilaterally. A collagen template was implanted in one stifle (N = 24). The contralateral side served as a control: 12 dogs had a total resection alone and the other 12 dogs had an immediate replantation of the autologous meniscus. Results were tabulated at 3, 6, 9, and 12 months. At final evaluation, before the animals were euthanized, the results were submitted for statistical analysis as well as histologic and biochemical analyses. The results demonstrated that a copolymeric collagen-based scaffold can be constructed that is compatible with meniscal fibrochondrocyte growth in vitro and in vivo, that does not inhibit meniscal regeneration in an immature pig, and that may induce regeneration of the meniscus in the mature dog.(ABSTRACT TRUNCATED AT 250 WORDS)     

Meniscal repair with fibrocartilage engineering

Injuries to the knee meniscus, particularly those in the avascular region, pose a complex problem and a possible solution is tissue engineering of a replacement tissue. Tissue engineering of the meniscus involves scaffold selection, addition of cells, and stimulation of the construct to synthesize, maintain, or enhance matrix production. An acellular collagen implant is currently in clinical trials and there are promising results with other scaffolds, composed of both polymeric and natural materials. The addition of cells to these constructs may promote good matrix production in vitro, but has been studied in a limited manner in animal studies. Cell sources ranging from fibroblasts to stem cells could be used to overcome challenges in cell procurement, expansion, and synthetic capacity currently encountered in studies with fibrochondrocytes. Manipulation of construct culture with exogenous growth factors and mechanical stimulation will also likely play a role in these strategies.     

碱性成纤维细胞生长因子与纤维蛋白黏合剂对半月板无血运区损伤的修复作用

目的 研究碱性成纤维细胞生长因子（ｂＦＧＦ）与纤维蛋白黏合剂（ＦＴＡ）对半月板无血运区损伤的修复作用。方法 选用健康成年青紫兰兔６只,在兔半月板上造成统一的无血运区损伤,随机将兔分成３组,分别为空白且、ＦＴＡ治疗组和ＦＧＦ／ＦＴＡ治疗组。术后２,６,１２周分批处死动物,进行大体形态观察和组织学检查。结果 ＦＴＡ治疗组能形成癜痕组织愈合;ＦＧＦ／ＦＴＡ治疗组为纤维软骨样组织愈合,但其愈合组织与正常半     

膝关节盘状半月板分型与撕裂类型相关性的MRI分析

目的:进行盘状半月板的流行病学研究,探讨膝关节盘状半月板的诊断标准、分型及其与年龄、撕裂类型的关系,以提高对盘状半月板及撕裂MRI表现的认识。方法:对842例诊断为盘状半月板的患者按年龄分为≤19岁、20³9岁、4039岁、40⁵9岁、≥60岁组。对全部患者冠状面髁间棘层面半月板宽度与胫骨平台宽度之比(板面比)、矢状面"领结样"改变层面中半月板后角最厚层面的厚度及矢状面"领结样"改变层数进行测量、分析。根据盘状半月板MRI表现,分为板型、楔型、肥角型。分析盘状半月板分型、年龄与撕裂类型的关系。结果:盘状半月板以外侧多见,女性发病率是男性的1.64倍。842例中,板型535例、楔型274例、肥角型33例。半月板撕裂354例,撕裂率为42.0%。842例板/面比均≥0.20,矢状面"领结样"改变层面中半月板后角最厚层面的厚度≥4.40mm。盘状半月板分型与撕裂类型及年龄分布有关系,而年龄分布与撕裂类型无明显相关。结论:盘状半月板多见于外侧,常伴半月板撕裂。板面比≥0.20、矢状面"领结样"改变层面中半月板后角最厚层面的厚度≥4.40mm、矢状面连续≥3层"领结样"改变,为盘状半月板的MRI诊断标准;盘状半月板分型与撕裂类型有关系;年龄可影响盘状半月板分型。     

Magnetic resonance imaging as a tool to predict reparability of longitudinal full-thickness meniscus lesions

Prediction of meniscus reparability is useful for surgeons to optimise surgical scheduling and to inform patients about postoperative management. This study was designed to determine the accuracy of MRI in predicting the reparability of longitudinal full-thickness meniscus lesions. We studied 100 MRIs of longitudinal full-thickness medial or lateral meniscus lesions located from the meniscosynovial junction to the axial part of the meniscus. The MRI criteria of reparability were a peripheral rim smaller than 4 mm and a lesion longer than 10 mm. At arthroscopy the menisci were considered as reparable when the lesion was located in red–red or red–white zones and when it was more than 1 cm in length. A heterogeneous or homogeneous aspect of the meniscus body was also considered. The length of the meniscus lesion averaged 17.6 mm and the thickness of the rim 3.4 mm. A heterogeneous signal of the axial fragment and of the rim was found in 11 cases and in 48 cases, respectively. In 90 cases, there was no difference between the MRI-predicted reparability and the arthroscopic findings. Global sensitivity of MRI to determine reparability of full-thickness meniscus lesions was 94%, increasing to 96% for the medial meniscus and dropping to 83% for the lateral meniscus. Global specificity was 81%, and was higher for the lateral meniscus (90%) than for the medial one (82%). Global positive predictive value was 0.94 and global negative predictive value was 0.82. A heterogeneous aspect of the rim did not compromise arthroscopic reparability whereas a heterogeneous aspect of the axial fragment indicated an irreparable meniscus. The described MRI criteria can predict meniscus reparability. Their accuracy is limited for lateral meniscus lesions close to the popliteal hiatus and for very young active patients in whom repair of white–white lesions can be attempted. Longitudinal full-thickness meniscus lesions are a good indication for repair in young active patients.     

MR of the knee: the significance of high signal in the meniscus that does not clearly extend to the surface

On MR images of the knee it is sometimes impossible to determine with confidence if a focus of high signal in the meniscus is confined to the substance of the meniscus or if it extends to involve the surface. This is a critical differentiation because the latter represents meniscal tears that can be found and treated at arthroscopy, whereas the former represents degeneration, tears, or perhaps normal variants that cannot be detected or treated arthroscopically. We make an equivocal diagnosis of a tear when it is difficult to decide if signal in a meniscus involves the meniscal surface. We studied MR scans of the knee in 142 consecutive patients for the presence of such equivocal tears. Their prevalence was 14% (20/142); 17 were in the posterior horn of the lateral meniscus and three were in the posterior horn of the medial meniscus. In 13 cases with arthroscopy/arthrotomy correlation, no tears were found. In one of the 20 patients in whom the meniscus was removed during arthroplasty, histologic examination of the meniscus showed separation of collagen bundles, which was caused by meniscal degeneration confined to the substance of the meniscus. These results suggest that a meniscal tear is unlikely when MR scans show a focus of high signal in a meniscus that does not unequivocally extend to involve the surface of the meniscus.     

Healing results of meniscal tears left in situ during anterior cruciate ligament reconstruction: a review of clinical studies

The presence of peripheral meniscal tears is common at the time of anterior cruciate ligament (ACL) reconstruction. Techniques to preserve the maximum amount of meniscal tissue include “non operative” management (tear left alone, without repair or removal), meniscus repair, partial meniscectomy. There is a lack of consensus guidelines about the management of peripheral stable meniscal tears. When to remove, let alone or repair? We performed an evidence-based review of the outcomes of stable meniscal tears left in situ during ACL surgery, in order to assess the effectiveness of this popular procedure. Clinical and anatomical results (arthrography or second look arthroscopy) were analyzed. Our literature search yielded ten relevant studies (9 level IV, 1 level III). The mean time of follow-up was 16 months. Pain or mechanical symptoms related to the medial tibiofemoral joint were reported in 0–66% of cases. Subsequent medial meniscectomy or repair were performed in 0–33% of cases. Pain or mechanical symptoms related to the lateral tibiofemoral joint were reported in 0–18% cases. Subsequent lateral meniscectomy or repair were performed in 0–22% cases. A complete healing occurred in 50–61% cases for the medial meniscus and in 55–74% cases for the lateral meniscus. No definite conclusion can be made with regard to these results. The conservative approach is more effective for lateral menisci. The rate of bad results for the medial meniscus remains high when a conservative treatment is used. For the medial meniscus, repair of stable peripheral tears may be always indicated to decrease the risk of postoperative pain or subsequent meniscectomy.