骨形态形成蛋白修复大面积关节软骨缺损的实验研究

将骨形态形成骨蛋白用于大面积关节软骨缺损的修复，探讨应用方法，观察修复效果。在５１只成年兔股骨的髌髁关节面上制造５ｍｍ×１０ｍｍ的骨软骨缺损，深３－５ｍｍ。缺损内分别填充骨形态形成蛋白和纤维蛋白粘合剂复合物，ＢＭＰ，ＦＳ，植入物均用自体游离骨膜覆盖，并设单独骨膜覆盖缺损组和空白对照组。     

活动对自体骨膜游离移植修复关节软骨缺损的影响实验研究

为了探索活动对骨膜游离移植修复关节软骨缺损的影响，选用成年健康家兔６０只，随机分三组：笼外活动组，笼内活动组及制动组。手术造成双后肢骨宾股关节股骨关节面０．５ｃｍ×１ｃｍ全层软骨缺损各一处，将自体胫骨骨膜游离移植于左侧软骨缺损处，右侧作为对照。分别于术后第４，８和１２周处死动物，取关节软骨标本。通过大体组织学、组织化学及电镜观察软骨再生情况，并比较活动量的多少对骨膜化生软骨的影响。结果表明，游离骨膜产生类透明软骨能有效地修复全层关节软骨缺损；统计学分析，笼外活动组与笼内活动组差异显著（Ｐ＜０．０５），笼内活动组与制动组差异显著（Ｐ＜０．０５），笼外活动组与制动组差异高度显著（Ｐ＜０．０１）。证明，活动量的大小对骨膜化生软骨有着重要的影响     

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.     

骨膜移植和钻孔术修复关节软骨缺损的实验比较

用中国白兔２４只，在股骨关书面造成６ｍｍ×８ｍｍ全层软骨缺损，分别进行游离骨膜自体移植和钻孔术。术后４、８周取材做组织学及电镜观察并进行比较。结果表明：（１）钻孔、移植骨膜和对照组的优势修复组织分别为类透明软骨、幼稚软骨和纤维组织。（２）修复组织平均数量，移植骨膜明显优于钻孔和对照组。（３）骨膜移植、钻孔和对照组修复组织来源分别为骨膜本身、髓腔和软骨下骨及与缺损毗连的软骨。初步结论：移植骨膜和软骨下骨钻孔均能修复关节软骨缺员，单纯刮除后修复能力最差。     

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。     

自体软骨细胞移植修复猪膝关节软骨缺损的实验研究

目的评价传统和复层高密度培养（复层培养）的自体软骨细胞移植修复关节软骨缺损的效果。方法在8头猪16膝髌切迹上下共建立32个全层软骨缺损，右膝为空白和自体骨膜移植对照组，左膝为传统的自体骨膜覆盖下细胞注射移植组和自体骨膜覆盖下复层培养细胞移植组（上下缺损随机进入各对照和实验组）。术后5-6个月对缺损部位行大体、组织学、免疫组织化学检查。采用O’Driscoll软骨组织形态学评分评价软骨缺损的修复质量。结果四组的O’Driscoll软骨组织形态学评分分别为（3．14±1．95）分、（10．57±3．60）分、（16．29±2．63）分、（20．43±1．81）分，各组之间差异均有统计学意义（P〈0．01）。空白对照组缺损被少量的纤维组织覆盖；骨膜移植组被纤维组织和少量的纤维软骨修复，且与周围组织整合差。自体软骨细胞移植（注射组和复层培养组）缺损被塑形良好的修复组织覆盖，组织学、基质特殊染色示修复组织为纤维软骨和透明软骨，与周边软骨和软骨下骨整合良好。复层培养组修复组织的细胞形态、基质染色比注射细胞组更接近正常软骨。结论自体软骨细胞移植可成功修复全层关节软骨缺损。扩增后的软骨细胞经短期复层培养更有利于软骨缺损的修复。     

骨膜移植修复骨缺损的实验研究及临床应用

为探寻大块骨缺损修复方法的新途径，进行了自体骨膜游离移植修复骨缺损的实验及临床研究。实验用４２只兔，于双侧胫骨作人工骨缺损模型（６ｍｍ×１８ｍｍ×５ｍｍ）。一侧随机植入自体游离骨膜片，另一侧不植入，作为对照。以组织学、Ｘ线和放射性同位素为观察指标，研究成骨过程。结果表明，骨膜植入侧骨缺损的愈合比对照侧缺损的愈合快一倍。原因可能是骨膜提供了大量成骨细胞并直接呈膜内成骨而非软骨内化骨。在此基础上，为２１例骨缺损患者应用自体胫骨骨膜片植入治疗骨缺损，面积最大１０．５ｃｍ×４ｃｍ×４ｃｍ，最小２ｃｍ×２ｃｍ×２ｃｍ；其中１７例为良性骨肿瘤，４例为骨慢性感染。骨缺损均得到修复，关节功能恢复满意。为腔洞性骨缺损的修复提供了一种新方法。     

自体软骨细胞修复关节软骨缺损的机制探讨

目的:观察团块样自体软骨细胞植入关节软骨缺损后的病理变化,探讨自体软骨细胞移植修复关节软骨缺损的病理生理机制。方法:24只3.0kg以上4～6月龄新西兰大白兔,雌雄不限,随机分为两组:实验组和对照组。实验组12只,20%的速眠新(1mg/kg)肌肉注射麻醉后取肩关节软骨组织,0.2%Ⅱ型胶原酶消化分离软骨细胞,体外单层培养,细胞长成肉眼可见的膜状后收集固体的组织样细胞团,动物再次麻醉制造双膝股骨滑车4.0mm×6.0mm方形缺损,植入细胞团块,骨膜覆盖,缝合骨膜于双股骨髁上。对照组12只,同实验组手术方法进行缺损单纯骨膜移植。1、3、12、24周两组各3只动物空气栓塞处死取材,观察细胞团块变化和缺损修复情况。结果:1周时软骨细胞朝向关节面部分细胞变大变圆,产生大量基质;3周时此种变化更加明显,但骨膜与细胞团块已然不能分开;12周时缺损为类透明软骨组织修复;24周时修复组织为透明软骨样组织,对照组为纤维软骨组织修复。结论:关节软骨细胞体外聚集培养形成的细胞团块内的细胞有迁移生长能力;细胞团块移植方法植入的细胞数量大,表型好,细胞在缺损内不会流失;关节软骨缺损修复是由植入的细胞团块生长分化而来;自体关节软骨细胞团块植入关节缺损内后,在关节应力的影响下,先从朝向关节面的一侧逐渐发生细胞成熟分化和软骨基质产生,逐渐完成缺损的修复。     

关节软骨缺损修复研究进展

关节软骨缺损是临床常见疑难病症之一。滑膜关节表面缺损后难以修复。现就关节软骨缺损的自发修复、自体或异体移植修复、软骨膜或骨膜移植修复、软骨细胞移植修复，以及三维立体细胞培养及组织工程技术修复等五个方面，综述了滑膜关节软骨缺损修复重建的方法学进展     

Reconstruction of Articular Cartilage Defects with Free Periosteal Grafts: An Experimental Study

The chondrogenic potential of free autogenous periosteal grafts was studied histologically in 6-month-old rabbits. The grafts were taken from the tibia and transplanted to 7 × 14 mm large artificial defects of the femoral articular cartilage. The results revealed that the defects were repaired and filled after 4 weeks with a hyaline-like cartilage which was histologically similar to the cartilage adjacent to the transplant. The tissue maintained this morphology after 1 year of observation. In control animals where no periosteum was transplanted to the defect, no real cartilage was found. The tissue which partially filled the defect was a variable mixture of fibrous tissue and fibrocartilage.     

自体骨膜游离移植一期修复髌骨骨折及软骨缺损

目的　探讨自体骨膜游离移植一期修复严重粉碎性髌骨骨折软骨缺损的临床疗效。方法　 1992年 1月～ 1998年 8月 ,用自体骨膜游离移植一期修复严重粉碎性髌骨骨折软骨缺损 17例 ,其中全髌骨粉碎骨折 9例 ,髌骨中上极骨折 3例 ,髌骨中下极骨折 5例。采用膝内侧 S形切口经膝关节腔入路。骨折复位后钢丝内固定 ,清除骨折区残存软骨 ,露出松质骨 ,修理平整 ;胫骨上端前内侧切取骨膜 3cm× 4cm～ 5 cm× 6 cm,游离移植到软骨缺损区并固定 ,术后 1周开始关节被动活动。结果　 17例全部获得随访 ,时间为 8个月～ 6年 2个月 ,平均为 2 .8年。优 12例 ,膝关节功能正常 ;良 4例 ,膝关节功能基本正常 ;可 1例 ,膝关节活动受限 ,但屈曲大于 90°。结论　自体骨膜游离移植一期修复髌骨骨折软骨缺损 ,取材容易 ,操作简便 ,疗效满意 ,有临床应用价值。     

Regeneration of the mandibular head from grafted periosteum

Grafted periosteum has a rich potential to induce heterotopic bone formation. In the current study the authors investigate whether autogenous periosteal grafts can regenerate the mandibular head in a rabbit model. They removed the mandibular head of Japanese white rabbits and grafted tibial periosteum to the cut surface of the mandible. Grafted periosteum was observed histologically and radiographically at day 7, 14, 21, and 45 after surgery. At day 7 after grafting, grafted tissue showed remarkable cell proliferation. By 14 days these cells had differentiated into chondrocytes to form cartilage, and endochondral ossification took place after 21 days. At 45 days after surgery, soft X-ray findings showed a newly formed mandibular head, which was similar histologically to that of a normal mandibular head. The cut mandible without periosteal graft showed no regeneration. These findings indicate that grafted periosteum can regenerate the mandibular head without special procedures such as bone fixation in a rabbit model, and suggest that this technique may be useful clinically.     

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.     

Transplantation of free tibial periosteal grafts for the repair of articular cartilage defect: An experimental study

Background:

Articular chondrocytes have got a long lifespan but rarely divides after maturity. Thus, an articular cartilage has a limited capacity for repair. Periosteal grafts have chondrogenic potential and have been used to repair defects in the articular cartilage. The purpose of the present study is to investigate the differentiation of free periosteal grafts in the patellofemoral joint where the cambium layer faces the subchondral bone and to investigate the applicability of periosteal grafts in the reconstruction of articular surfaces.

Materials and Methods:

The study was carried out over a period of 1 year on 25 adult, male Indian rabbits after obtaining permission from the institutional animal ethical committee. A full-thickness osteochondral defect was created by shaving off the whole articular cartilage of the patella of the left knee. The defect thus created was grafted with free periosteal graft. The patella of the right knee was taken as a control where no grafting was done after shaving off the articular cartilage. The first animal was used to study the normal histology of the patellar articular cartilage and periosteum obtained from the medial surface of tibial condyle. Rest 24 animals were subjected to patellectomy, 4 each at serial intervals of 2, 4, 8, 16, 32 and 48 weeks and the patellar articular surfaces were examined macroscopically and histologically.

Results:

The grafts got adherent to the underlying patellar articular surface at the end of 4 weeks. Microscopically, graft incorporation could be appreciated at 4 weeks. Mesenchymal cells of the cambium layer were seen differentiating into chondrocytes by the end of 4 weeks in four grafts (100%) and they were arranged in a haphazard manner. Till the end of 8 weeks, the cellular arrangement was mostly wooly. At 16 weeks, one graft (25%) had wooly arrangement of chondrocytes and three grafts (75%) had columnar formation of cells. Same percentage was maintained at 32 weeks. Four grafts (100%) at 48 weeks showed columnar orientation. The control side showed no changes over the shaved off articular surface in all the rabbits. One rabbit at 4 weeks had a dislocation of the patella on the control side. None of the rabbits developed any infection or wound dehiscence.

Conclusion:

Autologous periosteal graft transplantation can be a promising substitute for articular cartilaginous defects.     

Cellular origin and evolution of neochondrogenesis in major full-thickness defects of a joint surface treated by free autogenous periosteal grafts and subjected to continuous passive motion in rabbits

A graft of periosteum from the tibia of 27 rabbits was incubated in vitro with tritiated thymidine for 24 hours and then transplanted into a full-thickness defect in the patellar groove. The rabbits were managed after the operation on continuous passive motion (CPM), and the joints excised at intervals of two to 21 days. After one week the cells had begun to synthesize glycosaminoglycan and by two weeks the tissue resembled immature hyaline cartilage. Thymidine-labeled cells were seen throughout the entire regenerated tissue. The cellular origin of the hyaline-like tissue that filled the defects was the progenitor cells of the periosteal graft.     

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。     

Repair of articular cartilage defects with cultured chondrocytes in Atelocollagen gel

We attempted to repair full-thickness articular cartilage defects in rabbit knee joints with allogeneic cultured chondrocytes embedded in Atelocollagen gel. An articular cartilage defect was created on the patellar groove of the femur. The defect was filled with chondrocytes cultured in the collagen gel and covered with periosteal flap (G group). In three other experimental groups, the same defects were transplanted with chondrocytes in monolayer culture with periosteal flap (M group), periosteal graft only (P group), or left empty (E group). At 4, 12, and 24 weeks after operation, the reparative tissue was analyzed macroscopically and histologically. At 4 weeks after operation, the surfaces of the reparative tissue were smooth, and the defects were filled with reparative tissues that resembled hyaline cartilage in all four groups. However, the reparative tissues degenerated gradually with time in the M, P, and E groups. In contrast, in the G group, the reparative tissue retained its thickness, and there was a steady integration of the grafted tissue into the adjacent normal cartilage at 24 weeks after operation. The results suggest that transplantation of allogeneic chondrocytes cultured in Atelocollagen gel is effective in repairing an articular cartilage defect.     

骨膜游离移植修复膝关节软骨缺损及重建半月板

目的　对自体骨膜游离移植修复软骨缺损 ,重建半月板治疗膝骨性关节炎的方法、疗效进行评价。方法　 1996年～ 1999年 ,在膝关节清理术中清除病损软骨及退变碎裂的半月板 ,取胫骨骨膜游离移植修复软骨缺损 2 8例 36膝 ,其中重建半月板 8膝。术后早期在 CPM器上作连续被动功能锻炼 4周 ;半月板重建者术后石膏固定 7天。结果　术后全部病例均获 1～ 4年随访 ,采用主观症状、客观体征及 X线片三方面综合评价。术后膝关节功能优 2 2膝 ,良9膝 ,可 5膝 ,优良率为 86 .1%。结论　采用自体骨膜游离移植修复软骨缺损 ,重建半月板治疗膝骨性关节炎 ,近期可取得较好效果 ,远期效果尚需进一步研究。