自体游离骨膜—骨块复合组织移植修复关节骨软骨缺损

用三种类型的自体游离骨膜－骨块复合组织移植修复家兔膝关节的骨软骨缺损。实验结果显示，一个月后三组实验动物的膝关节骨软骨缺损区均被骨块和骨膜再生的新生软骨组织完全充填；不剥离翻转骨膜的骨膜－骨块复合组织移植具有操作简便、不影响骨膜再生软骨之优点；关节滑液能为骨膜再生软骨提供充足的营养。     

骨-关节软骨复合组织块构建的实验研究

目的探讨采用组织工程技术构建骨．关节软骨复合组织块的可行性。方法将分离、培养的第2代软骨细胞和成骨细胞分别接种于磷酸三钙支架上，培养2周后通过物理方法将两者连接成一个整体，然后接种于裸鼠皮下。术后8周取材，行组织学观察。结果复合组织块在体内分别形成软骨组织和成骨组织，而且两者界面整合良好。结论以软骨细胞和成骨细胞为种子细胞、以磷酸三钙为支架体外构建的组织工程软骨和骨，可在体内形成组织工程骨．关节软骨复合组织块，有望用于关节软骨缺损的修复。     

骨髓基质细胞源性软骨细胞修复兔全层关节软骨缺损

目的观察体外诱导骨髓基质细胞(MSCs)源性软骨细胞在兔股骨滑车关节面全层软骨缺损修复中的作用. 方法高密度传代培养第3代诱导MSCs分化为软骨细胞,以酸溶性Ⅰ型胶原为载体,两者混合后形成凝胶样植入物(细胞浓度为5×106/ml).于36只新西兰大耳白兔一侧股骨滑车关节面造成3 mm×5 mm全层关节软骨缺损,凝胶样植入为实验侧;另一侧分别为单纯胶原植入组(18个膝关节)和空白对照组(18个膝关节).术后4、8、12、24、32和48周取材观察缺损修复情况及新生组织的类型.参照Pineda标准对新生组织评分. 结果实验侧术后4周,植入细胞类似软骨细胞,周围有异染基质,形成透明软骨样组织;8周,深层有软骨下骨形成,软骨细胞层较正常关节软骨厚;12周,新生软骨厚度减小,与正常软骨相近,细胞呈柱状排列,结构与正常关节软骨相似,软骨下骨形成,潮线恢复;24周,新生软骨厚度较正常薄,约占55%,表面平整,潮线附近仍有肥大的软骨细胞;32周,潮线附近无肥大软骨细胞;48周,组织结构与32周时基本相同,为类透明软骨.Pineda评分24、32和48周间无差异,与4周比较有统计学意义(P＜0.05).实验组2～48周期间关节功能良好.单纯胶原组与空白对照组缺损无修复,48周时软骨下骨外露,关节退变;关节功能逐渐减退,动度受限. 结论 MSCs源性软骨细胞移植体内可形成透明样软骨组织,24周后新生软骨特性稳定,48周时为透明样软骨,能维持良好的关节功能.     

运动对兔膝退变关节软骨形态学的影响

目的：了解不同运动方式对退变关节软骨的形态学影响。方法：以新西兰白兔为研究对象,用伸膝位石膏固定８周的方法建立ＯＡ动物模型,按运动方式不同,随机分成５组,从光镜、扫描电镜两方面观察运动对退变关节软骨的组织形态学影响。结果：关节制动可诱发关节软骨退变,运动对退变软骨具有保护和促进修复作用。结论：关节适宜运动可促进退变关节软骨的修复和重塑,增加关节稳定性。     

骨髓基质干细胞修复兔关节软骨缺损的实验研究

目的研究以多聚乙醇酸(PGA)为支架的骨髓基质干细胞(BMSCs)复合物修复兔膝关节软骨缺损的情况。方法体外培养扩增的自体BMSCs种植于PGA支架并培养72h,然后将支架-细胞复合物植入兔关节软骨缺损模型。术后12周处死动物,标本行大体观察、组织学检查及Ⅱ型胶原免疫组化染色。结果BMSCs-PGA复合物植入后形成丰富的透明软骨样修复组织,新生软骨无明显退变。对照组主要为纤维组织及软骨下骨修复。结论BMSCs-PGA复合物可修复关节软骨缺损。     

自体肋软骨膜游离移植修复下颌髁状突软骨的组织学研究

为了观察自体肋软骨膜游离移植再生关节软骨的组织学改变，用５０只中国本兔进行了实验。通过组织学、电镜和放射自显影方法，对自体肋软骨膜游离移植于下颌髁状突切骨面后再生软骨的生长发育过程进行了观察。发现：术后１０天移植的软骨膜增厚，间充质样细胞增生。至术后３０天，软骨细胞逐渐发育成熟。６０天后，新生的软骨关节面光滑，细胞形态和排列趋于规则，与正常关节软骨形态相似。结果表明，自体肋软骨膜游离移植于髁状突骨面可以形成新的软骨关节面，其组织学形态与正常关节软骨相同；新生软骨生长发育过程与正常软骨相同；关节运动和负重能促进新生软骨形成并进行生物性改建，逐渐形成正常的关节形态。为临床应用软骨膜移植修复髁状突软骨损伤提供了依据。     

诱导型一氧化氮合酶抑制剂与软骨修复的研究进展

随着社会的老龄化,关节软骨的损伤和退变日益成为骨科最普遍、最重要的问题,而关节软骨的自我修复作用非常有限。因此,怎样促进关节软骨修复重建一直是骨科工作者不断追求,渴望解决的难题。然而,无论是自体或异体软骨移植、软骨膜或骨膜移植、软骨细胞移植,还是三维立体细胞培养及组织工程技术的应用,都仅能在早期生成透明和类透明软骨组织,且随后不可避免会发生退变 [1]。而这些再生的软骨组织,无论是其结构、生物化学或者生物力学特性都与正常透明软骨不同 [2]。因此怎样提高再生软骨质量和防止其发生退变就成为亟待解决的问题。…     

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。     

分别构建膜内成骨和软骨内成骨骨修复模型的研究

[目的]分别构建膜内成骨骨修复模型和软骨内成骨骨修复模型。[方法]取BALB/c小鼠64只,随机分为骨皮质钻孔模型组和骨膜划痕模型组。骨皮质钻孔模型组在小鼠右侧胫骨前内侧实施单纯性骨皮质钻孔损伤用于构建膜内成骨模型;骨膜划痕模型组在小鼠右侧胫骨前内侧实施骨膜划痕损伤用于构建软骨内成骨模型。术后第7、10、14和21 d,每组每个时间点处死8只小鼠,观察损伤部位的组织修复。[结果]骨皮质钻孔模型组小鼠损伤后第7 d在损伤部位出现新生小梁骨构成的骨痂组织。损伤后第10 d新生小梁骨充满损伤骨皮质及骨髓腔。损伤后第14 d新生骨痂组织进入改建过程,至第21 d,多数骨痂组织基本改建完成。在修复过程中无软骨细胞出现。骨膜划痕模型组小鼠损伤后第7 d在损伤部位出现新生软骨组织构成的软骨痂,并有部分软骨细胞已分化为成熟软骨细胞。损伤后第10 d软骨痂中心区域软骨基质溶解,第14 d后新生小梁骨出现,逐渐替代软骨组织。损伤后第21 d,软骨痂已基本被骨小梁替代。在修复过程中,首先出现软骨基质形成的骨痂组织,随后骨化中心形成,骨性骨痂组织逐渐替代软骨,完成骨修复。[结论]成功构建了以膜内成骨方式愈合的骨修复模型和以软骨内成骨方式愈合的骨修复模型,为今后深入研究骨愈合机制提供了基础。     

风湿骨痛药酒药槌外治法防治关节软骨退变的实验研究

目的 探讨风湿骨痛药酒药槌外治法防治关节软骨退变的作用机理。方法 选用日本大耳白兔18只，随机将12只通过结扎右股静脉造成膝关节骨性关节炎模型。未造模兔6只为正常组，12只造模兔随机分为造模组和治疗组。造模8周后，治疗组施以风湿骨痛药酒槌外治法进行治疗。造模11周后，观察软骨病理组织形态学、滑膜中NO浓度、软骨中PA及PAI活性和关节液中HA含量变化。结果 治疗组滑膜中NO浓度、软骨中PA及PAI活性和关节液中HA含量与正常组相比有显著性差异。造模组所测各项指标与治疗组相比有显著性差异。治疗组滑膜中NO浓度及软骨中PA含量明显低于造模组，而软骨中PAI含量及关节液中HA含量明显高于造模组。治疗组关节软骨退变不明显，类似正常组，而造模组关节软骨退行性变明显。结论 风湿骨痛药酒药槌外治法能通过改善骨内及周围组织的微循环，抑制滑膜组织中NO的过度产生，降低软骨中PA的活性，提高PAI的活性，消除导致关节软骨退变的内在因素，提高关节液中HA的含量，从而达到保护关节软骨、抗关节软骨退变的目的。     

培养软骨移植修复关节软骨缺损的实验研究

目的：为探讨一种新的关节软骨缺损修复方法。方法：将体外培养２周形成软骨样组织,移植修复兔关节软骨全层缺损。于移植术后２、４、８周分别行功能评价、大体形态及组织学检查。结果：全部实验兔于术后２周内恢复正常活动。２周时移植修复组织由非成熟透明软骨组成。４周时部分移植组出现成熟透明软骨。８周时移植组关节软骨缺损全部由成熟透明软骨充填修复,修复组织与邻近关节软骨融合。培养软骨移植修复关节软骨全层缺损明显优于自身修复（Ｐ＜００１）。结论：本实验提示使用具有高有丝分裂率的软骨细胞,经离心管培养形成骺软骨样组织,植入关节软骨全层缺损后,软骨细胞生长良好,逐渐成熟和转化,能发挥良好的修复作用。     

关节游离体软骨细胞生物学特性的初步观察

目的观察关节游离体软骨细胞的生物学特性，寻找组织工程软骨种子细胞来源。方法分别取游离体软骨标本5例，正常软骨标本2例和骨关节炎软骨标本6例，组织学观察其细胞分布、尿嘧啶脱氧核苷末端转移酶介导的三磷酸脱氧核苷缺口标记法观察细胞凋亡，细胞分离培养观察原代软骨细胞形态，并进行比较。结果与正常软骨、骨关节炎软骨比较，游离体软骨细胞分布密度较高，细胞凋亡明显．体外培养时原代细胞已呈成纤维细胞样形态。结论游离体软骨细胞已具有成纤维细胞特性，不宜直接作为组织工程的种子细胞来源。     

髋臼后壁重建后关节软骨修复的实验研究

目的 研究髋臼后壁重建区臼面软骨修复的组织学特点,初步探讨软骨柱播种在其修复中的作用。方法 选取12只家犬共24髋,随机分为3组,均采用后壁截骨法建立髋臼后壁60°弧1/2缺损的动物模型,两侧髋臼后壁缺损区分别选用不同的重建方法：A组：解剖重建+软骨柱播种,B组：解剖重建,C组：普通重建。术后12周取材进行大体形态、光镜观察,参照Pineda标准对新生组织进行评分。结果 A组中以成熟、新生类透明软骨修复缺损,B组为类纤维软骨修复,C组为纤维肉芽组织或板层骨修复。A、B、C组关节软骨修复Pineda总评分平均分别为(5.1±0.1)、(6.5±0.2)、(12.3±0.2)分,差异有统计学意义(F=3.157,P=0.000),3组间两两比较差异均有统计学意义(P＜0.05)。结论 解剖重建促使部分软骨柱集中在缺损区,其修复组织具备部分关节软骨的组织特性,能够较好地替代关节软骨。     

The distribution of S-100 protein positive chondrocytes in the human articular cartilages under aging or diseased conditions]

There have been few reports on the localization of S-100 protein positive chondrocytes in the human articular cartilages. We studied 59 articular cartilages of the aged subjects, 65 osteoarthritic (OA) and 39 rheumatoid arthritic (RA) articular cartilages, to detect the histological localization of S-100 protein using immunoperoxidase method (ABC). The results obtained from normal cartilages demonstrated strongly positive cells representing hypertrophic chondrocytes in the perivascular areas of the neonatal articular cartilage and in the deep zone of the infant articular cartilage. The moderately positive cells were found in the intermediate zone of infant and adult articular cartilages. In mild OA, there were many positive chondrocytes in the intermediate zone with erosion of the surface layer, while in moderate or severe OA many strongly positive cells were found in clusters. The hypertrophic cells in the metaplastic cartilage arising from bone marrow in subjects with severe OA, or from pannus after RA were also positive. It is therefore, suggested that S-100 protein may be correlated with the metabolic activity of the cartilage matrix such as collagen and proteoglycan, as reported in the literature. S-100 protein further, appears to be useful for evaluating histologically the activity of cartilage repair in the pathologic human articular cartilages.     

组织工程化软骨修复兔膝关节软骨缺损的实验研究

目的 评估同种异体组织工程软骨修复兔膝关节全层软骨缺损的有效性。方法分离收集成年新西兰大白兔软骨细胞进行体外培养。建立双侧兔膝关节软骨缺损模型，用去端肽胶原（atelocollagen）凝胶与所培养的异体兔关节软骨细胞共同植入兔膝关节软骨缺损处，并设对照组。分别于手术后4周、8周观察大体标本以及组织学修复结果，并进行Wakitan的评分，评估此方法的有效性。结果大体观察结果表明，与对照组相比，实验组缺损处由软骨组织修复而对照组缺损处由纤维样组织填充。组织学观察可以见到实验组关节软骨缺损处有密集的软骨细胞而对照组关节缺损处只有纤维细胞无软骨细胞。结论通过短期观察表明以同种异体软骨细胞-去端肽胶原复合物修复全层软骨缺损的方法是有效可行的，为其进一步临床应用提供了参考。     

Application of microscopic MR for evaluation of cartilage repair]

Magnetic resonance imaging is gold standard for noninvasive evaluation of articular cartilage damage and has been also used for monitoring cartilage repair. The aim of this study was to find correlation between histological microscopy and microscopic MR in evaluation of the repair of osteochondral defects in articular cartilage. Study was based animal model (rabbit). The cartilage repair process was evaluated histology and micro MR. Most of the defects were filled with fibrocartilage and fibrous tissue formed. Both methods were equally efficient to show repair tissue thickness, subchondral bone reconstruction and disintegration. Result of observation by both histological and MR microscopy and showed good correlation. Micro MR is promising evaluation tool for cartilage repair monitoring. Results of micro MR correlate well with standard microscopy.     

Connective tissue growth factor mRNA expression pattern in cartilages is associated with their type I collagen expression

Connective tissue growth factor (CTGF) has been identified as a secretory protein encoded by an immediate early gene and is a member of the CCN family. In vitro CTGF directly regulates the proliferation and differentiation of chondrocytes; however, a previous study showed that it was localized only in the hypertrophic chondrocytes in the costal cartilages of E 18 mouse embryos. We described the expression of CTGF mRNA and protein in chondrocytes of different types of cartilages, including femoral growth plate cartilage, costal cartilage, femoral articular cartilage, mandibular condylar cartilage, and cartilage formed during the healing of mandibular ramus fractures revealed by in situ hybridization and immunohistochemistry. To characterize the CTGF-expressing cells, we also analyzed the distribution of the type I, type II, and type X collagen mRNA expression. Among these different types of cartilages we found distinct patterns of CTGF mRNA and protein expression. Growth plate cartilage and the costal cartilage showed localization of CTGF mRNA and protein in the hypertrophic chondrocytes that expressed type X collagen mRNA with less expression in proliferating chondrocytes that expressed type II collagen mRNA, whereas it was also expressed in the proliferating chondrocytes that expressed type I collagen mRNA in the condylar cartilage, the articular cartilage, and the cartilage appearing during fracture healing. In contrast, the growth plate cartilages or the costal cartilages were negative for type I collagen and showed sparse expression of CTGF mRNA in the proliferating chondrocytes. We found for the first time that CTGF mRNA could be differentially expressed in five different types of cartilage associated with those expressing type I collagen. Moreover, the spatial distribution of CTGF mRNA in the cartilages with type I collagen mRNA suggested its roles in the early differentiation, as well as in the proliferation and the terminal differentiation, of those cartilages.     

Chondroitin sulphate impedes the migration of a sub-population of articular cartilage chondrocytes

OBJECTIVE: To determine whether chondroitin sulphate (CS) impedes the migration of primary articular chondrocytes. DESIGN: Articular chondrocytes were isolated from young and skeletally mature bovine animals. Boyden chambers were used to quantify chondrocyte migration on aggrecan in the presence and absence of CS chains. A novel in vitro model of cell migration into articular cartilage explants was designed to visualise and quantify the migration of labelled chondrocytes into cartilage matrix which had been treated with chondroitinase ABC to remove CS chains present. RESULTS: A consistent trend of increased migration with both age groups of a sub-population of chondrocytes was demonstrated on aggrecan in the absence of CS. These data were supported by results from the in vitro model of chondrocyte migration which demonstrated increasing numbers of a chondrocyte sub-population from both age groups of cartilage migrating into the chondroitinase ABC digested cartilage explants with time in culture. Minimal migration of these chondrocytes was demonstrated into phosphate buffered saline (PBS) treated control explants. CONCLUSIONS: We confirm that a sub-population of chondrocytes isolated from both young and skeletally mature articular cartilages have the ability to migrate. We also demonstrate that CS chains inhibit the migration of these articular chondrocytes and that their removal by chondroitinase ABC digestion enhances the migration of these chondrocytes. Such findings may provide a clinical application for improving cell-based cartilage repair strategies by enhancing integration between endogenous and repair tissue.     

培养软骨移植修复兔生长板缺损

目的 将体外培养软骨移植修复兔生长板缺损,防止生长板缺损早闭造成的肢体发育畸形。方法 分离收集1月龄兔关节软骨细胞,经离心管内培养形成软骨。将培养2周软骨植入6周龄兔胫骨上端生长板缺损区,于4、16周时对术肢行X线摄片、组织学及免疫组织化学等检查。结果 兔胫骨上端生长板缺损区移植培养软骨4周时,胫骨无明显畸形发生,组织学检查显示生长板缺损由软骨充填,Ⅱ型胶原免疫组织化学染色阳性。16周时移植侧胫骨仍无明显畸形,组织学检查显示生长板已近闭合。而对照侧胫骨发生严重畸形,生长板闭合。结论 培养软骨异体植入兔生长板缺损,可替代缺损的生长板软骨组织,维持肢体正常生长,防止肢体发育畸形。