自体骨膜移植治疗关节软骨损伤的研究现状

关节软骨损伤修复大致分两类，即内源性修复和外源性修复。内源性修复也称骨髓刺激术，外源性修复包括生物移植和组织工程化关节软骨，前者包括骨膜、软骨膜移植，软骨细胞移植和骨-软骨移植；后者是体外构建种子细胞-载体复合物注入缺损区或者利用种子细胞悬液注入缺损区再用骨膜或软骨膜覆盖封闭软骨缺损的开口。两者均涉及到骨膜的成软骨作用。骨膜移植应用于临床治疗软骨损伤已有近20a历史，它有取材方便、对机体损害小等特点，但存在许多影响因素，限制了它的应用。本文就骨膜移植治疗关节软骨损伤的现状作一综述。     

组织细胞移植修复关节软骨缺损的研究现状

关节软骨组织的自身修复能力很差,临床治疗中要使修复或再生的软骨能恢复到滑膜关节正常的无痛运动,修复的新组织须在结构、组成、机械性能和持久耐用方面与正常关节软骨相似。目前关节软骨修复损伤的方法主要有两大类：一类为刺激关节软骨自身修复的方法,另一类为组织细胞移植方法。前者包括,清创术和灌洗法、软骨下骨钻孔术、微骨折、截骨术等,后者包括软骨移植、骨膜／软骨膜移植、软骨细胞及间充质细胞移植等［１］。本文就组织细胞移植修复关节软骨缺损的研究进展进行了综述。一、关节软骨移植关节软骨移植是用完整的正常关节软骨…     

自体骨膜游离移植的实验研究--不同龄动物大块关节软骨缺损修复的比较

目的研究比较自体骨膜移植软骨再生修复不同龄动物大块关节软骨缺损。方法用52只不同龄家兔自体骨膜游离移植修复大块关节软骨缺损,比较移植骨膜生发层朝向关节腔与松质骨时再生软骨的差别。结果经不同时期肉眼和组织学检查证实,幼年兔和成年兔的骨膜移植都能生成软骨,修复大块关节软骨缺损。在成年兔骨膜再生的软骨与成年兔本身周围正常软骨的厚度、组织结构一样。移植骨膜生发层朝向关节腔与松质骨二者间再生软骨结果无明显差别。结论骨膜具有再生软骨的能力,可用来移植修复关节软骨的缺损。骨膜移植生发层不同朝向对软骨再生无明显影响。成年后骨膜移植修复关节软骨缺损能够生成与自身相适应的软骨。     

自体软骨膜、骨膜游离移植修复软骨缺损治疗骨性关节炎

目的：评价自体软骨膜或骨膜游离移植术修复膝关节大面积软骨缺损，治疗膝关节骨性关节炎的疗效。方法：将髌骨及股骨髁，胫骨平台病损软骨清除，游离移植软骨或骨膜修复软骨缺损，治疗骨性关节炎124例，术后不需外固定，4天后持续被动关节活动器作持续动活动。2周后下床活动，结果：术后平均随访6年，治疗效果满意。结论：采用自体软骨膜，骨膜游离移植修复大面积软骨缺损，治疗骨性关节炎，可取得满意效果。     

筋膜蒂骨膜移植修复关节软骨缺损的实验研究

目的：应用筋膜蒂骨膜移植修复关节软骨缺损，观察骨膜在关节腔内再生关节软骨的情况，探讨骨膜再生软骨的机制。方法：用成年家兔２２只，在右侧膝关节造成胫骨关节面软骨缺损，行筋膜蒂骨膜移植；在左侧膝关节行游离骨膜移植，作为对照。观察方法包括形态学观察、Ｘ线和组织学检查。结果：实验组再生关节软骨的质与量均优于对照组。结论：筋膜蒂骨膜移植，由于有血液供应，能够再生关节软骨，修复关节软骨缺损。     

自体骨—骨膜移植修复骨软骨缺损：初步报告

根据骨膜具有再生关节软骨能力的特性，我们将取自于胫骨内侧骺端的骨一骨膜用于修复膝关节表面的骨软骨缺损。临床应用５例，包括剥脱性骨软骨炎、软骨下骨坏死及陈旧性髌骨骨折。经１６～２６个月随访，以膝关节功能评分法评价，移植手术获满意效果。摄片及磁共振影像证实缺损得以修复。此结果表明骨一骨膜移植是修复骨软骨缺损的有效方法。     

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。     

自体软骨细胞团块植入修复兔陈旧性关节软骨缺损的实验研究

目的 观察自体软骨细胞团块植入对兔关节软骨缺损的修复作用. 方法 24只成年新西兰大白兔48侧膝关节,随机分为三组(n=16)并制备双膝关节股骨滑车软骨缺损模型.空白对照组无特殊处理,骨膜移植组将骨膜覆盖缺损并缝合于缺损两侧的股骨髁上,实验组将自体软骨细胞团块植入缺损中.术后3、6个月分别取材(n=8),进行大体和组织学观察,修复组织行Wakitani评分并进行比较. 结果实验组共成功取材11个缺损关节,9个为透明软骨修复,2个因植入细胞生长状态差未修复;骨膜移植组修复组织为纤维软骨或纤维组织,修复组织薄,基质异染弱;空白对照组仅有少量纤维组织填充缺损底部.修复组织Wakitani评分:实验组3.82分,骨膜移植组6.71分,空白对照组9.23分,差异有统计学意义(F=5.96,P=0.00). 结论自体软骨细胞团块植入能较好修复关节软骨缺损,修复的质量与植入细胞的质量有关.     

关节软骨损伤细胞修复研究进展

关节软骨损伤主要由创伤和各种疾病如骨关节炎、剥脱性骨软骨炎、骨坏死等引起，容易导致关节功能障碍。关节软骨损伤目前治疗方法很多，包括微骨折、软骨下骨打孔术、骨膜和软骨膜移植、自体和异体软骨移植、组织工程软骨移植、细胞移植、基因治疗等方法，但效果各异。他们都旨在解决两个主要问题：（1）利用与关节软骨相似的组织或材料充填缺损；（2）促使修复组织和原先组织在形态上相似，在结构和功能上相近。     

不同营养来源的骨膜移植修复关节软骨缺损的对比研究

在39侧狗骨关节面上造成20×25mm的软骨缺损,用三种不同营养来源的骨膜瓣移植修复作对比研究结果表明。依靠静脉血营养的骨膜瓣能够成活并化生为透明软骨,可修复大范围软骨缺损游离骨膜瓣只能部分存活、化生软骨,且新生软骨质量较差。动脉血营养骨膜瓣只能成骨,不能用干修复关节软骨缺损。     

关节软骨损伤和缺损修复策略

关节软骨的损伤是临床中常见的疾病。由于关节软骨自身修复能力有限,采用关节镜下清创术、软骨移植、软骨细胞移植、组织工程技术及凝胶类关节软骨修复材料是目前对关节软骨损伤进行修复的主要手段。本文对目前用于关节软骨损伤和缺损修复的主要策略及各类修复技术的优缺点进行了综合评述。关节镜下清创术对早期骨性关节炎疗效显著;软骨及软骨细胞移植对小面积软骨缺损修复效果较为理想;组织工程技术是目前对关节软骨损伤和缺损修复的一个热点方向,但存在支架材料与软骨缺损区整合不紧密等问题;凝胶类关节软骨修复材料具有与自然关节软骨相似的力学和生物摩擦学特性,但其生物活性及与自然关节软骨间的结合强度有待进一步提高,如何实现材料生物活性、生物力学性能和生物摩擦学性能功能一体化是凝胶类关节软骨修复材料亟待解决的焦点问题。     

生长板损伤修复的实验与临床研究

目的 综述生长板损伤修复的研究进展。方法 广泛查阅近期有关生长板损伤修复文献，介绍生长板损伤修复的主要方法，重点阐述应用组织工程方法修复生长板损伤。结果 生长板损伤修复是小儿矫形外科实验研究和临床治疗面临的一大难题。游离生长板软骨移植，因缺乏血供而应用困难；血管化生长板移植解决了血循环问题，但是自体移植供体来源有限，异体移植存在免疫排斥；非软骨组织材料只能阻止小面积生长板缺损内骨桥形成，无再生修复能力。软骨细胞及组织工程软骨移植可能是生长板损伤修复的最佳选择。结论 由于缺乏安全有效的生长板损伤修复方法，应进一步开展软骨细胞及组织工程软骨研究。     

Rabbit articular cartilage defects treated by allogenic chondrocyte transplantation

Articular cartilage defects have a poor capacity for repair. Most of the current treatment options result in the formation of fibro-cartilage, which is functionally inferior to normal hyaline articular cartilage. We studied the effectiveness of allogenic chondrocyte transplantation for focal articular cartilage defects in rabbits. Chondrocytes were cultured in vitro from cartilage harvested from the knee joints of a New Zealand White rabbit. A 3 mm defect was created in the articular cartilage of both knees in other rabbits. The cultured allogenic chondrocytes were transplanted into the defect in the right knees and closed with a periosteal flap, while the defects in the left knees served as controls and were closed with a periosteal flap alone, without chondrocytes. Healing of the defects was assessed at 12 weeks by histological studies. Allogenic chondrocyte transplantation significantly increased the amount of newly formed repair tissue (P=0.04) compared with that found in the control knees. The histological quality score of the repair tissue was significantly better (P=0.05), with more hyaline characteristics in the knees treated with allogenic chondrocytes than in the control knees. Articular cartilage defects treated with allogenic chondrocyte transplantation result in better repair tissue formation with hyaline characteristics than those in control knees.     

Articular cartilage repair using tissue engineering technique--novel approach with minimally invasive procedure

Articular cartilage has very limited potential to spontaneously heal, because it lacks vessels and is isolated from systemic regulation. Although there have been many attempts to treat articular cartilage defects, such as drilling, microfracture techniques, soft tissue grafts or osteochondral grafts, no treatment has managed to repair the defects with long-lasting hyaline cartilage. Recently, a regenerative medicine using a tissue engineering technique for cartilage repair has been given much attention in the orthopedic field. In 1994, Brittberg et al. introduced a new cell technology in which chondrocytes expanded in monolayer culture were transplanted into the cartilage defect of the knee. As a second generation of chondrocyte transplantation, since 1996 we have been performing transplantation of tissue-engineered cartilage made ex vivo for the treatment of osteochondral defects of the joints. This signifies a concept shift from cell transplantation to tissue transplantation made ex vivo using tissue engineering techniques. We have reported good clinical results with this surgical treatment. However, extensive basic research is vital to achieve better clinical results with this tissue engineering technique. This article describes our recent research using a minimally invasive tissue engineering technique to promote cartilage regeneration.     

冷冻保存胎兔颅骨骨膜修复关节软骨缺损的实验研究

为探讨关节软骨缺损的修复方法，采用家兔３０只，平均分成两组。在每个动物的双侧髌股关节股骨关节面作４ｍｍ×７ｍｍ全层软骨缺损。将经二步冷冻法保存的胎兔颅骨骨膜移植于一组动物一侧后肢作为实验组，另一侧用自体骨膜移植作为自体对照组。在另一组动物的一侧后肢用新鲜胎兔颅骨骨膜移植作为新鲜对照组，另一侧关节软骨缺损不修复作为空白对照组。术后肢体不作外固定。１６周后取材，经大体观察、组织学观察及氨基酸成分分析。结果发现，经冷冻保存的胎兔颅骨骨膜移植后能形成透明样软骨，与自体对照组无显著性差异（Ｐ＞０．０５），与新鲜对照组及空白对照组主要指标有显著性差异（Ｐ＜０．０５）。氨基酸分析示实验组新生物接近纤维软骨（Ｐ＞０．０５）。认为，冷冻胎兔颅骨骨膜移植为关节软骨缺损的修复提供了实验依据。     

Xeno-implantation of pig chondrocytes into rabbit to treat localized articular cartilage defects: an animal model

Articular cartilage has only a limited ability to regenerate. The transplantation of autologous chondrocytes is currently used to treat focal defects in human articular cartilage, although use of organs, tissues, or cells from different species is being investigated as an alternative treatment. The object of this study was to use xeno-transplantation of cultured pig chondrocytes for the repair of rabbit chondral defects, and to analyze the significance of tissue rejection in this animal model. Partial chondral defects, including removal of cartilage tissue and a part of the subchondral bone, were created in the lateral femoral condyles of 30 adult New Zealand White rabbits. A periosteal flap was sutured to the native cartilage with the cambium layer facing the defect. As a control, culture medium was injected into the defect void of one group of rabbits while in a treatment group, chondrocytes, isolated from normal femoral pig cartilage, were injected into the defect void. All rabbits were killed by 24 weeks. Macroscopic changes of the cartilage were analyzed using Mankin's score. The distal femoral portion was studied histologically using hematoxylin and eosin, alcian blue, toluidine blue, and Mason's trichrome. Pig cells and pig genetic material were detected in the neo-synthesized tissue by immunohistochemical detection of SLA-II-DQ and polymerase chain reaction analysis of the gene SLA-II-DQB. The synovial membrane was studied histologically by hematoxylin and eosin staining. In the control group, on average, less than 25 percent of the chondral defect was filled. The repair tissue had an irregular surface with few cells similar to chondrocytes or fibroblasts and a minimal formation of extracellular matrix. In the treatment group, the chondral defect was approximately 90 percent filled with good integration between the neo-synthesized cartilage and the native cartilage. The repair tissue had a smooth surface with cells similar to chondrocytes and a hyaline-like extracellular matrix. The neo-synthesized cartilage was morphologically similar to hyaline cartilage. Importantly, there were no signs of graft-vs.-host rejections or infiltration by immune cells. In the neo-synthesized tissue, pig genetic material was detected in 27 +/- 5 percent of all cells. These cells containing pig genetic material were distributed throughout the neo-synthesized cartilage. We conclude that the xeno-transplantation of chondrocytes could be an alternative method for the repair of articular cartilage defects.     

Value of autologous chondrocyte transplantation in the reconstruction of experimental cartilage defects. Part I. Extent of defect, macroscopic appearance of reconstructed articular surface and microscopic analysis of predominant tissue]

Articular cartilage defect is one of the main reasons of osteoarthritis. Currently, tissue engineering techniques are the methods concerning better cartilage reconstruction. The aim of this part of the study was macroscopic evaluation of degree of defect feeling, macroscopic appearance of repair tissue and microscopic analysis of predominant tissue after autologous chondrocytes transplantation. Repair of partial thickness cartilage defect on distal part of femur was evaluated (25 adolescent rabbits). Procedures were performed in II groups: I--autologous chondrocytes transplantation under periosteal flap, II--periosteal graft. Chondrocytes were isolated from the cartilage specimens by enzymatic digestion and cultured in vitro. The regenerates were inspected 4, 8 and 12 weeks after the operation. Macroscopic analysis in group I, in most cases revealed filling of the defect with tissue resembling surrounding cartilage. In group II the defect was partially filled, and there was many fissures and cracks in all regenerates. In microscopic analysis in group I, after 4 and 8 weeks following the transplantation the tissue similar to juvenile hyaline cartilage predominated. After 12 weeks it resembled mature hyaline cartilage. In group II, in all cases fibrous cartilage was observed after 4, 8, 12 weeks. Obtained results indicate, that macroscopic and microscopic characteristics of repair tissue after autologous chondrocytes transplantation more closely resembled hyaline cartilage, than in periosteal graft group. 12 weeks after autologous chondrocytes transplantation the repair tissue reached maturity, and demonstrated microscopic characteristics of hyaline-like cartilage. The method of autologous chondrocytes transplantation provides potential for clinical application.     

Repair of articular cartilage defects with cultured chondrocytes on polysulphonic membrane: experimental studies in rabbits

INTRODUCTION: Autologous osteochondral transplantation is one method that can be used to create hyaline or hyaline-like repair in a defect area. The purpose of the present study was to repair full-thickness articular cartilage defects in 9 rabbit knee joints with autologous cultured chondrocytes. METHODS: An articular cartilage defect was created on the patellar groove of the femur. The defect was filled with chondrocytes cultured in vitro and placed into the knee on a polysulphonic membrane. At 8 weeks after the operation, the reparative tissue was analyzed macroscopically and histologically. RESULTS: At 8 weeks after the operation, the surfaces of the reparative tissue were smooth, and the defects were filled with mature hyaline cartilage in 5 cases. In 2 cases, the reparative hyaline cartilage was immature and there was worse integration of grafted tissue into the adjacent normal cartilage. In 2 cases, the surface of the grafted area was irregular, and the reparative tissue was disintegrated and incompletely differentiated. CONCLUSION: The results suggest that transplantation of autologous chondrocytes cultured in vitro and placed into the knee on polysulphonic membrane is effective in repairing an articular cartilage defect.     

同种脱细胞软骨基质与软骨细胞复合物修复兔甲状软骨缺损的实验研究

目的 探讨脱细胞软骨基质(acelluar cartilaginous matrix，ACM)与软骨细胞复合后植入兔体内，能否形成软骨修复甲状软骨缺损。方法 分离培养免甲状软骨细胞，与ACM体外复合培养，形成ACM-软骨细胞复合物，进行组织学分析及用于修复兔甲状软骨缺损。新西兰大白兔18只，制成两侧甲状软骨缺损模型，随机分为3组，每组6只。对照组：只制备缺损，不作修复；ACM修复组：采用单纯ACM修复缺损；实验组：用ACM-软骨细胞复合物修复。术后8周处死动物，取出标本，进行大体和组织学观察。结果 体外培养时，软骨细胞能在ACM表面生长，未长入基质内。动物实验结果，对照组：甲状软骨缺损处被肌肉、结缔组织充填；ACM修复组：ACM内炎性细胞浸润，轻度吸收变形；实验组：复合物植人体内后8周未形成软骨，甲状软骨缺损修复不理想。结论 ACM体外培养和体内植入均未能为软骨细胞生长提供支持，作为一种天然细胞培养支架，尚有待进一步改进与完善。     

Integration of periosteum covered autogenous bone grafts with and without autologous chondrocytes. An animal experiment using the Göttinger minipig

Autologous osteochondral transplantation has the major disadvantage of significant damage to a healthy joint surface at the donor site.The purpose of this study was to examine the effect of autogenous chondrocytes injected into the periosteum of autologous bone grafts in order to provide an alternative method for cartilagerepair. A total of 22 Göttinger minipigs were operated twice on both knees.The first operation served for cartilage biopsy for the chondrocyte culture.During the second operation an osteochondral defect was created in the medial facet of the trochlear groove.The defect was treated differently with an autologous cortico-cancellous bone cylinder,harvested from the proximal tibia.Group A: untreated defect (control);B: bone-graft;C: bone-graft covered with periosteum; D: bone-graft with periosteum and injected autologous chondrocytes.The animals were killed after 6,12,26 and 52weeks.The regenerated areas were evaluated macroscopically, tested biomechanically (long-term specimens; indentation-test) and a histological, blind evaluation was carried out according to a semi-quantitative scoring system. The periosteum covered bone cylinders in Groups C and D showed good repair of the bone and cartilage defect.The repaired tissue consisted predominantly of fibrocartilage with the partial formation of hyalin like tissue.The regenerated areas were integrated with the adjacent cartilage and were biomechanically superior when compared with the other groups. The additional injection of chondrocytes did not produce significantly better results. Our findings suggest that the transplantation of periosteum-covered bone cylinders may provide an alternative method for treating chondral and osteochondral defects and can be recommended for filling large donor site defects in joint surgery.The additional transplantation of chondrocytes does not seem to be justified.