自体骨膜移植修复关节软骨缺损生发层不同朝向的比较研究

为了观察骨膜再生软骨中，骨膜生发层不同的朝向是否影响软骨再生。应用人体纤维蛋白粘合剂粘合自体骨膜修复兔关节软骨缺损，对移植骨膜生发层不同朝向（即朝向关节腔或软骨下骨）的差别进行比较。结果表明：６周前，朝向关节腔组增殖较快，６周后两组无明显差别，故生发层的朝向不是一个重要影响因素。另外，应用藏红Ｏ蛋白多糖染色的电镜观察，证实两组再生软骨均为透明软骨。     

兔自体松质骨颗粒修复关节软骨损伤

目的研究兔自体松质骨颗粒在膝关节软骨损伤处移植后能够诱导软骨组织生成、促进关节软骨损伤修复的现象。方法 12只新西兰大白兔麻醉后在兔的右侧膝关节股骨远端内、外侧髁负重区用电钻分别造成直径、深度均为3 mm的骨软骨缺损创面,取同侧髂骨松质骨,制成直径约为1 mm松质骨颗粒植入股骨内侧髁软骨缺损处,作为实验组,外侧髁软骨缺损不做处理作为对照组。术后12周进行膝关节大体观察、病理切片染色,评估关节软骨损伤的修复情况。结果兔膝关节实验组软骨缺损处被新生软骨填充,软骨面光滑,组织切片染色显示有关节软骨形成。对照组缺损创面仍然凹陷,仅在缺损边缘有少量软骨生长。结论兔自体松质骨颗粒在膝关节软骨损伤处能够诱导软骨生成,促进关节软骨的修复,是一种良好的关节软骨损伤修复方法。     

骨膜诱导再生关节软骨的组织学观察

实验选用成年健康家兔24只,手术造成双侧髌股关节股骨关节面直径4.5mm全层软骨缺损各2处,用纤维蛋白粘合剂将自体骨膜及软骨膜游离移植于软骨缺损处,通过光镜、电镜等方法进行动态观察。结果表明:二者均具有诱导再生成熟关节软骨的能力,其衍化过程一致,说明可利用骨膜代替软骨膜移植治疗关节软骨缺损。     

自体骨软骨移植与含富集骨髓干细胞松质骨移植修复兔关节软骨缺损

目的:评价自体骨软骨移植与含富集骨髓干细胞松质骨镶嵌移植两种方法修复全层关节软骨缺损的生物学特征和效果。方法:采用新西兰大白兔制作左右后肢全层软骨缺损模型,分别进行自体骨软骨镶嵌移植、含富集骨髓干细胞松质骨镶嵌移植修复,对照组不作任何修复,每组12只。术后第4、8、12周处死动物取材,分别进行膝关节活动度测定、大体观察、光镜观察与电镜观察。结果:移植实验组在第12周时均能以类透明软骨组织修复缺损,对照组为纤维肉芽组织。形态学检查表明,两种方法均能以类透明软骨组织覆盖缺损,骨软骨移植组无明显免疫排斥现象,随着时间延长,修复高度逐渐增加。骨软骨移植组同含富集骨髓干细胞松质骨镶嵌移植组效果无显著差别。结论:骨软骨移植、含富集骨髓干细胞松质骨镶嵌移植两种方法均能以类透明软骨组织修复全层关节软骨缺损,含富集骨髓干细胞松质骨镶嵌移植更适用于较大面积软骨缺损的修复。     

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.     

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。     

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     

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。     

自体骨膜游离移植修复髋膝关节软骨大面积缺损的远期观察

目的观察自体骨膜游离移植修复髋、膝关节软骨大面积缺损的远期临床效果. 方法在动物实验基础上,于1987年2月～1996年8月,采用自体骨膜游离移植修复52例关节软骨大面积缺损的患者,其中有完整随访资料的37例中,先天性髋关节脱位16例,创伤性髋关节炎6例,股骨头低毒性感染1例,强直性脊柱炎2例,膝关节内骨折6例,骨性关节炎4例,创伤感染后膝关节僵直2例.术前有髋关节脱位者行松解牵引术,术中切除病变的软骨组织达出血骨组织;从胫骨前内侧切取大于软骨缺损10%～15%的骨膜,生发层朝向关节腔,缝合固定在软骨缺损表面;双侧移植骨膜的关节间放置硅胶膜;术后进行CPM练习,逐渐增加活动范围,活动时间为4～6周;出院后6个月内避免负重,可主动练习关节活动. 结果术后37例获7～15年随访,平均10.5年.功能评价标准按有无关节疼痛、关节活动角度、日常生活能力和X线片测量关节间隙宽度进行评价,结果:优11例,良18例,差8例. 结论游离骨膜移植修复关节软骨缺损,经远期随访应用于临床是可行的.     

The chondrogenic potential of free autogenous periosteal grafts for biological resurfacing of major full-thickness defects in joint surfaces under the influence of continuous passive motion. An experimental investigation in the rabbit

A rectangular graft of autogenous tibial periosteum was sutured (with its cambium layer facing into the joint) onto the base of a five by ten-millimeter full-thickness defect in the patellar groove of each of 143 adolescent and adult rabbits. The rabbits were managed postoperatively by either immobilization, intermittent active motion, continuous passive motion for two weeks, or continuous passive motion for four weeks. When the animals were killed four weeks postoperatively, the contour of the patellar groove had been restored in all of the rabbits in the group that had had four weeks of continuous passive motion, and the newly formed tissue in all of the defects in this group had the gross, histological, and histochemical appearance of smooth, intact hyaline articular cartilage. Histologically, the nature of the tissue that had formed, as well as its surface regularity, structural integrity, and bonding to the adjacent cartilage, were significantly better in the group that had had four weeks of continuous passive motion than in any of the other groups. The results were significantly worse when the orientation of the periosteal graft was reversed (that is, when it had been sutured into the defect with the cambium layer of the graft facing the subchondral bone rather than into the joint) or when no periosteal graft was used. Biochemical analyses revealed that, in the group that had had four weeks of continuous passive motion, the total hexosamine content, the levels of chondroitin sulphate and keratan sulphate, and the ratio of galactosamine to glucosamine were all comparable with the values for normal articular cartilage. In contrast, in the groups that were treated by immobilization, intermittent active motion, or two weeks of continuous passive motion, as well as in the adult rabbits, the content of the first three of these substances was significantly less than normal. In the groups that were treated by immobilization, intermittent active motion, or two weeks of continuous passive motion, 32 to 47 per cent of the total collagen was type II, while in the group that had had four weeks of continuous passive motion, 93 per cent of the total collagen was type II. These results demonstrate that, under the influence of continuous passive motion, free autogenous periosteal grafts can repair a large full-thickness defect in a joint surface by producing tissue that resembles articular cartilage grossly, histologically, and biochemically, and that contains predominantly type-II collagen.     

Differentiation of the Osteochondrogenic Cells of the Periosteum in Chondrotrophic Environment

The behaviour of the cells in free periosteal grafts was studied in growing rabbits in three chondrotrophic recipient milieus: costal cartilage, ear cartilage and synovial fluid of the knee joint. The periosteal grafts first formed cartilage, which was then quite rapidly transformed into bone on the costal cartilage, quite slowly and in smaller amounts in the ear cartilage, whereas no bone was found in the cartilaginous loose bodies formed in the knee joint. In bone formation vascularization plays a major role, but other factors are also involved.     

静脉血骨膜瓣生发层朝向对再生关节软骨的影响

在２４条犬髌股关节面上造成２０ｍｍ×２５ｍｍ的软骨缺损，用静脉血营养骨膜瓣移植修复，根据生发层不同朝向，分两组对比研究。结果表明：生发层朝关节腔组新生软骨组织学结构，超微结构，胶原类型及软骨细胞的营养代谢均明显优于生发层朝骨面组。对于临床应用具有指导意义。     

Kaoko O. Kettunen: Skin Arthroplasty in the light of Animal Experiments with special reference to Functional Metaplasia of Connective Tissue

The behaviour of the cells in free periosteal grafts was studied in growing rabbits in three chondrotrophic recipient milieus: costal cartilage, ear cartilage and synovial fluid of the knee joint. The periosteal grafts first formed cartilage, which was then quite rapidly transformed into bone on the costal cartilage, quite slowly and in smaller amounts in the ear cartilage, whereas no bone was found in the cartilaginous loose bodies formed in the knee joint. In bone formation vascularization plays a major role, but other factors are also involved.     

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.     

Periosteal neochondrogenesis for biologically resurfacing joints: its cellular origin

The purpose of this study was to determine if the hyaline-like cartilage produced in major full-thickness defects of a joint surface, treated by a free periosteal graft and subjected to continuous passive motion (CPM), originated exclusively from the progenitor cells of the cambium of the graft. Free periosteal grafts were raised from the tibia of both hind legs of eight male New Zealand rabbits and transplanted into full-thickness defects across the entire width of the patellar groove of 15 female rabbits. Postoperatively, CPM was instituted and the animals were sacrificed after 3 weeks. Cells from the regenerated tissue were grown from tissue explants and their karyotypes determined. In 33% of the rabbits, all cells contained a Y (male) chromosome, indicating that regenerated tissue originated exclusively from the progenitor cell of the periosteal graft. Karyotypes of the cells from the other 67% were mosaics (both female and male); thus, their cellular origin was from both the periosteal allograft and the pluripotential mesenchymal cells in the subchondral tissues.     

Reconstruction of patellar cartilage defects with free periosteal grafts. An experimental study

The osteo-chondrogenic potential of free periosteal grafts was investigated within the knee joint in 26 rabbits aged four to six weeks. A total of 36 knee joints were operated on. The grafts were stripped from the medial side of the right tibia and sutured on the articular surface of the patella, from which the cartilage had been totally excised to the subchondral bone. In 16 knees the graft was sutured with the cambium layer towards the subchondral bone and in eight knees the fibrous layer faced the bone. In the control group of twelve knees the patellar articular cartilages were excised and no periosteal transplant was grafted to the patellar articular surface. In the transplantation group cartilage formation could be seen already one week after the operation. There were no marked differences between the series with the cambium layer facing the subchondral bone or the group with the fibrous layer facing the bone. At 20 weeks the hypertrophied cartilage had thinned and resembled normal joint cartilage. In the control group the histological picture resembled osteoarthritis.     

Repair of cartilage defects with periosteal grafts.

Alternative sources for repair of cartilage defects are limited and donor sites are associated with morbidity. It is known that cartilage development from periosteal grafts is possible. Various factors have been found positively to affect this process in experimental settings. However, all of these studies were limited to joint cartilage. We conducted an experimental study in rabbits for the investigation of the elastic cartilage regeneration from perichondrial and periosteal grafts together with the effects of hyaluronan on this process. 1 x 1 cm(2) cartilage defects were created on the elastic ear cartilage of rabbits. Four experimental groups with 18 ears in each group were created: Group 1 (repair with perichondrium graft), group 2 (repair with periosteum graft), group 3 (repair with periosteum graft+hyaluronan), group 4 (defect-only control group). Macroscopic and microscopic evaluations were done on the 4th, 8th and 12th weeks. Cellular morphology of the regenerated cartilage and its integration and similarity with adjacent cartilage were evaluated. Cartilage regeneration groups 1, 2 and 3 were found to be statistically different from the control group. There was not a significant difference between groups 1 and 2 or 2 and 3. There is no significant difference between perichondrial and periosteal grafts in cartilage regeneration, and hyaluronan has no beneficial effect on this process.