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。     

Comparative Study of the Reconstruction of Articular Cartilage Defects with Free Costal Perichondrial Grafts and Free Tibial Periosteal Grafts: An Experimental Study on Rabbits

The purpose of this study was to compare the chondrogenic potential of free perichondrial with free periosteal grafts in the resurfacing of full-thickness defects of patellar articular cartilage in rabbits. We used adolescent New Zealand rabbits weighing between 2.4 and 3.6 kg. A 6-mm wide and 3-mm thick defect was created on the patellar articular surface. A total of 30 rabbits were randomly divided into a control group and two test groups. One test group received free perichondrial grafts (PC); the other received free periosteal grafts (PO). All the animals were killed 8 weeks after surgery. All the histological samples were scored from 0 to 17 according to a standard scoring system. Differences in the quality of the regenerated tissue were only found between the control and the test groups. There were no statistically significant histological differences between the grafted defects of the PC and the PO groups that there are not on any of the variables. The results of this study support that there are not significant differences in the quality of the repair tissue when using these two types of biological grafts. Received: 7 July 1998 / Accepted: 8 January 1999     

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.     

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.     

Repair of full-thickness articular cartilage defects using injectable type II collagen gel embedded with cultured chondrocytes in a rabbit model

BACKGROUND: Recently, tissue-engineered chondrocyte transplantation has been tried to treat full-thickness cartilage defects. We developed an injectable type II collagen gel scaffold by chemically reacting type II collagen with polyethylene glycol crosslinker. This type II collagen was prepared from the nasal septa of cattle. In the present study, chondrocytes embedded in type II collagen gel were injected into rabbit full-thickness cartilage defects without a periosteal graft, and the feasibility for clinical application of the gel was evaluated. METHODS: Chondrocytes were isolated from 1-kg New Zealand white rabbits. A full-thickness articular cartilage defect (5 mm diameter, 4 mm depth) was created on the patellar groove of the femur of 16 male 3-kg New Zealand white rabbits. A type II collagen solution of mixed chondrocytes at a density of 1 x 10(7) cells/ml was injected and transplanted into the defect in the right knee. The controls were the defect only in the left knee. At 4, 8, 12, and 24 weeks after operation, four cases from each group were evaluated macroscopically and histologically. RESULTS: After injection into the cartilage defect, the gel bonded to the adjacent cartilage and bone within several minutes. Macroscopic examination revealed that the surface of the transplanted area was smooth and exhibited similar coloration and good integration with the surrounding cartilage at 12 and 24 weeks after transplantation. Histological examination at 8 weeks revealed favorable hyaline cartilage regeneration with good chondrocyte morphology. At 12 and 24 weeks, reparative cartilage remained rich in type II collagen. According to O'Driscoll histological scores, significant differences between the transplanted and control groups were apparent at 12 and 24 weeks. Immunohistochemical staining indicated sufficient type II collagen synthesis in regenerated cartilage 8 weeks after transplantation, and it was maintained until 24 weeks. CONCLUSIONS: These results indicate that type II collagen gel is suitable for injection into cartilage defects without any covering of a graft and offers a useful scaffold during chondrocyte transplantation.     

Biological resurfacing of full-thickness defects in patellar articular cartilage of the rabbit. Investigation of autogenous periosteal grafts subjected to continuous passive motion.

We compared the effects of continuous passive motion with those of intermittent active motion on the results of the resurfacing with autogenous periosteal grafts of full-thickness defects on the articular surface of rabbit patellae. Of 45 rabbits with defects, 30 received grafts. Fifteen of these had continuous passive motion for two weeks and intermittent active motion for four weeks; the other 15 had intermittent active motion for six weeks. In 15 the defects were not grafted (control group) and they had intermittent active motion for six weeks. Ten more rabbits had a sham operation. Six weeks after surgery, the results were assessed by the gross appearance, histology, histochemistry, immunohistochemistry and electron microscopy. By all assessments the quality of neochondrogenesis produced by periosteal grafts was superior to that in ungrafted defects (p less than 0.05) and the results in continuous passive motion treated animals were superior to those in intermittent active motion treated animals (p less than 0.05). The periosteal grafts produced hyaline cartilage containing type II collagen but the organisation of its fibres was irregular.     

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.     

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。     

应用骨形态发生蛋白(BMP)修复关节软骨缺损的实验研究

目的探讨关节软骨全层缺损应用骨形态发生蛋白修复的效果。方法于2004年5月至2005年12月,30只新西兰种成年兔随机分为A,B,C三组,每只兔子左膝股骨髁间凹做一大小为4mm×5mm×2.5mm的全层关节软骨缺损。A,B组缺损内分别填充骨形态发生蛋白/纤维蛋白胶(BMP/FG)及FG,C组为空白。术后28周对缺损修复情况行大体形态、组织学和电镜观察。结果BMP/FG组,缺损组织以透明软骨修复,接近正常组织,而FG组和空白组则以纤维组织修复为主。结论BMP/FG能较好的完成关节骨软骨全层缺损的修复,并随着时间的延长修复的软骨越接近正常软骨,但修复软骨缺损的组织与邻近正常软骨组织连接性仍不是十分理想。     

Effects of low-intensity pulsed ultrasound in repairing injured articular cartilage

Articularcartilageisakindofavascularmusculoskeletaltissueswitharelativelylowcapacityforintrinsicrepairorregeneration.Intheabsenceofavailablestemcells,thereislittlehopeforcartilagerepairafterinjuryandthetissuesprogress alongadegenerativepathwaythatdestroysthe biochemicalandbiomechanicalproperties.Sorepairof damagedarticularcartilageisoneofthemostdifficult problemsinorthopaedicsandnosatisfactorytreatment methodhasbeenfoundinthisfield.Ultrasoundhas beenwidelyusedinclinicaldiagnosisandtreatment.It…     

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.     

Current treatments of isolated articular cartilage lesions of the knee achieve similar outcomes

Background  

Many surgical techniques, including microfracture, periosteal and perichondral grafts, chondrocyte transplantation, and osteochondral grafts, have been studied in an attempt to restore damaged articular cartilage. However, there is no consensus regarding the best method to repair isolated articular cartilage defects of the knee.     

The biologic concept of continuous passive motion of synovial joints. The first 18 years of basic research and its clinical application

The notoriously limited capacity of articular cartilage to heal or to regenerate plus the author's clinical observations and research on the deleterious effects of immobilization on joints led him to the biologic concept of continuous passive motion (CPM) of synovial joints in 1970. The hypothesis that CPM should stimulate pluripotential mesenchymal cells to differentiate into articular cartilage and should accelerate the healing of articular tissues has been validated by numerous scientific investigations of a variety of experimental models of the knee joint. These models have included full-thickness defects, intraarticular fractures, acute septic arthritis, partial thickness lacerations of the patellar tendon, semitendinosus tenodesis to replace the medial collateral ligament, autogeneic osteoperiosteal grafts in major defects, free autogeneic periosteal grafts, and periosteal allografts. In 1978, the author collaborated with Saringer, an engineer, to develop CPM devices for humans. CPM is clinically indicated following such procedures as open reduction of fractures, arthrolysis for posttraumatic arthritis, synovectomy, drainage of septic arthritis, release of joint contractures, total arthroplasty, tendon repair, and ligament reconstruction. Clinically, CPM is an important stimulus to joint regeneration processes.     

微粒骨膜-三维支架修复大面积关节软骨缺损

目的 探讨微粒骨膜-三维支架修复大面积关节软骨缺损的有效性和可行性.方法 于兔股骨滑车关节面制作直径4.5 mm深达软骨下骨板的全层软骨缺损模型,缺损处随机行自体微粒骨膜-纤维蛋白混凝物、单纯纤维蛋白"浇铸"移植.分别于术后3 h、4 d及1、2、4、8、12、24周取材,行大体观察、苏木素.伊红(HE)、Masson及藏红花(safranin-0)染色组织学检查,并进行组织学评分半定量分析.结果 微粒骨膜.三维支架制备简便.微粒骨膜被均匀种植于纤维蛋白三维支架中,可随意"浇铸"充填骨软骨缺损,移植物不易脱落,手术1次完成.术后微粒骨膜在缺损空间内全方位迅速增殖、分化、分泌基质完成缺损骨软骨修复.新生软骨具有与周围正常软骨基本一致的厚度、细胞形态及排列、基质胶原及蛋白多糖染色,且与周边软骨及软骨下骨结合良好.术后4、8、12及24周,两组组织学评分差异有统计学意义(P<0.05).结论 该方法能简单高效地构建工程化组织复合体,随意浇铸充填软骨缺损,完成较大面积关节软骨缺损的生物性修复.     

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。     

The chondrogenic repair response of undifferentiated mesenchymal cells in rat full-thickness articular cartilage defects

OBJECTIVE: The aim of this study is to develop a rat model of full-thickness articular cartilage defects that is suitable for detailed molecular analyses of the regenerative repair of cartilage. MATERIALS AND METHODS: The V-shaped full-thickness defects (width: 0.7 mm; depth: 0.8 mm; and length: 4mm) were created in the femoral patellar groove of 6 weeks old male rats using a custom-built twin-blade device. Prior to starting the repair experiments, our device was examined for its accuracy and reliability in generating defects. Then, the time course of the repair response in these cartilage defects was examined using a semi-quantitative histological grading scale. The expression of chondrogenic differentiation markers in the reparative regions was examined with immunohistochemistry and in situ hybridization. RESULTS: Our device creates full-thickness articular cartilage defects uniformly. In these defects, undifferentiated mesenchymal cells filled the defect cavities (4 days) and initiated chondrogenic differentiation at the center of the defect (7 days). Cartilage formation was observed in the same region (2 weeks). Finally, hyaline-like articular cartilage and subchondral bone layers were reconstituted in their appropriate locations (4 weeks). CONCLUSIONS: We have successfully developed a rat model containing identically sized full-thickness defects of articular cartilage that can undergo chondrogenic repair in a reproducible fashion.     

Repair of large full-thickness articular cartilage defects in the rabbit: the effects of joint distraction and autologous bone-marrow-derived mesenchymal cell transplantation

We produced large full-thickness articular cartilage defects in 33 rabbits in order to evaluate the effect of joint distraction and autologous culture-expanded bone-marrow-derived mesenchymal cell transplantation (ACBMT) at 12 weeks. After fixing the knee on a hinged external fixator, we resected the entire surface of the tibial plateau. We studied three groups: 1) with and without joint distraction; 2) with joint distraction and collagen gel, and 3) with joint distraction and ACBMT and collagen gel. The histological scores were significantly higher in the groups with ACBMT collagen gel (p < 0.05). The area of regenerated soft tissue was smaller in the group allowed to bear weight (p < 0.05). These findings suggest that the repair of large defects of cartilage can be enhanced by joint distraction, collagen gel and ACBMT.     

The effect of hyaluronic acid on a rabbit model of full-thickness cartilage repair

The current study investigated whether hyaluronate exerts a beneficial effect on articular cartilage repair. Nineteen rabbits had bilateral knee arthrotomies, and 2-mm full-thickness cartilage defects were created on each medial femoral condyle. Rabbits received intraarticular injections (0.5 mL) of hyaluronic acid once a week for 3 weeks in the right knee, started at either 1 or 3 weeks after injury. The left knees, which served as controls, were injected with 0.5 mL normal saline. Cohorts of each group were euthanized at 2 and 6 months, and histologic sections of the injury sites were evaluated for repair tissue. No significant differences were seen in the quantity or quality of the repair tissue at either 2 or 6 months. Hyaluronate and saline-treated defects showed persistent fibrillation, poor matrix staining, and incomplete void filling, irrespective of the injection timing. Hyaluronate did not provide protection to zones peripheral to the injury site, and did not significantly alter the healing process in this model of acute full-thickness cartilage injuries.     

人重组骨形态发生蛋白-2修复全层关节软骨缺损的实验研究

目的 :研究人重组骨形态发生蛋白 2 (rhBMP 2 )对全层关节软骨缺损的修复 ,观察修复效果。方法 :家犬 8只 (16膝 ) ,每个膝内外髁均做全层软骨缺损 ,内外髁共 3 2个缺损。随机分为 4组 ,每组 2只。每只犬一侧关节行胶原海绵吸附rh BMP填充内外髁缺损 ,另一侧以单纯胶原海绵填充作对照 ,不处理组为空白对照。术后 2、4、 8、 12周取材作大体、光镜、透射电镜观察。结果 :rh BMP组为类软骨细胞修复 ,而单纯胶原海绵组和空白组均为纤维性修复。结论 :rhBMP 2有效地促进关节软骨缺损的修复 ,可以作为临床上治疗关节软骨缺损的方法     

自体游离骨膜再造膝关节半月板的实验研究

目的：对于严重损伤的半月板全切除术后行自体游离骨膜重建半月板，以维持膝关节结构、生理及功能特性。方法：使用１７只成年犬，５只幼犬。外侧半月板全切除以后，胫骨内侧近端的骨膜游离并移植到外侧半月板部位。于２、４、８、１２、２４、４８周取下重建半月板及不同部位的关节面软骨进行大体、光镜及电镜观察。结果：术后２个月，重建半月板的大体形态、组织结构与正常半月板近似。外侧股骨软骨面及没有被覆盖的胫骨软骨面表现退行性改变，被移植物覆盖的软骨面退变较轻。幼犬的退变更早、更严重。结论：本研究说明了自体游离骨膜在滑液环境及受到合适的应力刺激会向纤维软骨演化，在半月板全切除后，应用游离骨膜重建半月板是可行的