自体与新鲜同种异体骨软骨移植的实验研究

目的:评价自体与新鲜异体骨软骨镶嵌移植两种方法修复全层关节软骨缺损的生物学特性和修复效果。方法:采用完全随机设计,将16只新西兰大白兔的左右后肢制成全层软骨缺损模型,分别进行自体骨软骨镶嵌移植、同种异体骨软骨镶嵌移植修复。术后第12周处死动物取材,分别进行膝关节活动度测定、大体观察、光镜观察及Wakitani组织学评分法,数据行统计学分析。结果:膝关节伸屈活动度、大体观察、组织学光镜及组织修复评分显示自体骨软骨移植在第12周时能以类透明软骨组织修复缺损,而新鲜同种异体骨软骨移植为纤维肉芽组织。结论:自体骨软骨镶嵌移植可以修复兔关节软骨的缺损。新鲜无处理的同种异体骨软骨镶嵌移植修复关节软骨缺损不可行,其排斥反应及吸收破坏严重。     

软骨下骨厚度及固定对兔膝关节软骨下骨移植后软骨的影响

目的 :研究兔膝关节软骨下自体骨移植后软骨下骨厚度及固定对软骨的影响。方法 :健康大耳白兔74只。其中 10只分为I、II两组 ,每组 5只 ,分别建立保留不同软骨下骨厚度的膝关节软骨下大块骨缺损的动物模型 ,术后 1周测量软骨下骨残留厚度。另 6 4只建立动物模型后 ,采用立柱式自体髂骨移植。按保留软骨下骨厚度及术后是否固定随机分为A、B、C、D 4组。A、B组采用I组模型 ,C、D采用II组模型 ,A、C组术后膝关节屈曲角度6 0°位固定 ,4周解除固定 ,B、D组未固定。于术后 12周取材 ,采用光镜、透射电镜 ,对关节软骨进行形态学观察。结果 :I组平均残留软骨下骨的厚度为 1.42mm ,1周时软骨下骨未发生坏死。II组软骨下骨平均残留厚度为 0 .86mm ,1周时全部软骨下骨发生坏死。A、B组 12周时软骨面无 1例塌陷 ,A组 6 .2 5 %的软骨发生退变 ,B组 12 .5 %的软骨发生退变。C、D组 12周时仅 1例关节软骨面塌陷 ,C组 18.7%的软骨发生退变。D组 43.7%的软骨发生退变。经 χ2 检验 ,B组与D组相比 ,有显著性差异 (P<0 .0 5 ) ,C组与D组相比无显著性差异 (P >0 .0 5 )。结论 :关节软骨下骨移植时过多刮除软骨下骨将引起软骨下骨缺血坏死 ,增加软骨退变的发生率。术后固定对关节软骨退变无显著性影响。     

同种异体骨移植修复骨缺损的生物学基础

大块骨缺损的修复一直是骨科领域研究的重要课题,也是骨科界面临的棘手的难题之一［１～３］。长期以来,骨缺损的修复主要采用自体骨移植结合同种异体骨移植的方法。但两种方法各有优缺点。公认的自体骨移植疗效最佳,但骨源有限,且供区有潜在的合并症。同种异体骨是临床上常用的替代自体骨移植材料。但同种异体骨移植可诱发宿主产生免疫排异反应,且目前临床上多采用经冷冻、冻干、脱钙或其它化学处理的同种异体骨,其细胞成分多已坏死,因此同种异体骨移植在成骨机理、愈合过程中的表现、免疫反应等方面与自体骨移植有一定的差异。本文…     

软骨移植及关节软骨组织工程技术研究进展

关节软骨缺损是临床常见的疑难病例,目前的治疗包括自体或异体骨软骨移植修复、软骨膜或骨膜移植修复及软骨细胞移植修复.组织工程技术的发展使软骨移植进入了一个新的发展时期,本文就软骨移植及关节软骨组织工程技术进展进行综述.     

脱钙骨植入修复桡骨缺损的动物实验

在54只兔的108例桡骨上分别造成1cm的骨缺损,将其平分为三组,分别植入脱钙同种异体骨、新鲜自体骨和新鲜同种异体骨。并于第1、3、5、7、10和13周,每组杀3只兔,用X线照片及组织学切片观察骨的愈合情况。结果发现新鲜异体骨植入后有排斥反应,而脱钙异体骨植入无此反应。实验证明脱钙同种异体骨是一种较好的植入物。彼优于新鲜异体骨,与新鲜自体骨植入无显著差异。新骨及软骨出现在植入物中而不在邻近宿主骨处,表明新骨形成可能是骨诱导作用的结果。     

关节软骨缺损修复的研究进展

关节软骨缺损的修复一直是骨科临床和实验研究中亟待解决的难题之一 ,人们一直在探索一种有效的修复方式。本文就关节软骨缺损的自身修复、自体或异体骨软骨移植、骨膜或软骨膜移植、软骨细胞移植、三维立体细胞培养组织工程技术及干细胞定向诱导分化的修复等六个方面 ,综述了关节软骨缺损修复的研究进展。1　关节软骨缺损的自身修复软骨细胞的增殖能力极低 ,在正常成熟关节中几乎见不到有丝分裂 ,其分裂周期不是以日、月计 ,而是以年计。一般按损伤的程度可分为部分厚度的软骨缺损和全层关节软骨缺损两类。部分厚度的软骨缺损 ( PTCD) ,…     

自体骨—骨膜转移修复关节软骨缺损的实验研究

目的：通过自体骨－骨膜移植修复兔关节软骨缺损，探讨骨膜间充质细胞对关节透明软骨组织全层缺损修复的作用与机制。方法：以成年兔胫骨内上侧的骨－骨膜组织作为供体，镶嵌式植入兔自体髌股关节之股骨滑车关节软骨等大、相同形状的全层缺损区，术后4－24周分别对缺损修复情况进行大体、组织学及电镜观察。结果：自体骨－骨膜移植组织与缺损周围组织完全愈合，组织学及电镜观察显示修复组织为透明软骨组织。结论：自体骨－骨膜组织移植修补关节软骨缺损可以诱导关节软骨缺损以透明软骨方式进行修复，这为临床应用自体骨－骨膜组织移植修复关节软骨缺损提供了理论依据。     

同种异体脱钙骨移植骨科临床应用研究

<正> 对于各种原因的骨缺损治疗,传统的办法是自体骨移植,但较大的缺损及小儿患者自身取骨量受限,因而寻找骨移植的替代材料为众多的骨科学者所关注。我科自1988年起用同种异体骨盐酸脱钙法制备成脱钙骨,临床应用28例,疗效满意。现报告如下: 材料与方法 脱钙骨取材于新鲜的同种异体骨,即创伤截肢(指、趾)无污染骨,股骨颈骨折行人工关节置换术中取出的股骨头,后路全椎板手术切除     

天然支架在软骨组织工程中应用的研究现状

由于软骨内无血管，缺乏血液供应，细胞代谢缓慢，临床上常见的由创伤和各种疾病（如骨关节炎，剥脱性骨软骨炎、骨坏死等）引起的关节软骨缺损难以自行修复。修复关节软骨缺损的传统的方法（关节灌洗清理术、微骨折、软骨下钻孔、骨膜和软骨膜移植）都存在一些技术上的局限性，均不能实现长期的软骨修复。自体软骨移植在临床上技术较成熟，但也存在供区缺损、来源有限等不足，异体软骨移植因免疫排斥反应而受到限制。组织工程化软骨的出现为软骨缺损修复提出了新的途径。1977年Green将分离、培养的软骨细胞与脱钙骨支架联合培养，开始尝试软骨组织工程的研究，并认识到细胞培养支架的重要性。目前，软骨组织工程中支架分为天然和人工合成支架。     

冷冻同种异体骨软骨移植修复大面积关节软骨缺损的可行性研究

目的观察冷冻同种异体骨软骨移植修复大面积关节软骨缺损的可行性，为临床应用提供实验依据。方法共选取36只新西兰兔，随机数字表法分成供体组、实验组和对照组，每组12只。供体组制备冷冻同种异体胫骨内侧髁骨软骨移植物后处死；实验组将经程序深低温冷冻保存的同种异体胫骨内侧髁移植；对照组将取下的胫骨内侧髁原位植入。所有实验动物在术后12周处死。应用苏木精-伊红染色观察关节软骨组织的病理学变化，软骨缺损的组织学评分以半定量的改良Wakitani score法评估，内容包括：细胞种类、髓质染色、表面完整性、软骨的厚度、移植软骨在受体周围组织的完整性5个方面，采用0、1、2、3、4评分，最高总分14分。观察两组动物的大体标本、X线片、组织学检查、Waki-tani score评分及免疫组织化学检查结果。结果①移植软骨的大体观察：两组动物关节活动度正常，关节腔无粘连、无积液，实验组移植软骨颜色及高度与周围关节软骨大致相同，关节面较平整，基底部界限模糊。对照组关节软骨面平整、无塌陷，厚度不变。②两组动物关节X线片均见骨折线已模糊，对位、对线良好。③组织学检查：两组动物关节软骨均已被透明软骨覆盖，细胞排列有序，软骨基质大量分泌，未见淋巴细胞和浆细胞浸润。④两组动物关节软骨Wakitani score评分结果无显著性差异，总评分相近（P〉0.05）。⑤两组修复软骨Ⅱ型胶原免疫组织化学染色强阳性。结论冻存同种异体骨软骨移植后可完全修复大面积关节软骨缺损，并与自体骨软骨块修复兔的关节软骨缺损在组织学上相近，未见免疫排斥反应。     

Effects of Structural Changes in Subchondral Bone on Articular Cartilage in a Beagle Dog Model

Objective Using MR T2-mapping and histopathologic score for articular cartilage to evaluate the effect of structural changes in subchondral bone on articular cartilage. Methods Twenty-four male Beagle dogs were randomly divided into a subchondral bone defect group(n = 12) and a bone cement group(n = 12). Models of subchondral bone defectin the medial tibial plateau and subchondral bone filled with bone cement were constructed. In all dogs, the left knee joint was used as the experimental sideand the right knee as the sham side. The T2 value for articular cartilage at the medial tibial plateau was measured at postoperative weeks 4, 8, 16, and 24. The articular cartilage specimens were stained with hematoxylin and eosin, and evaluated using the Mankin score. Results There was a statistically significant difference(P 0.05) in Mankin score between the bone defect group and the cement group at postoperative weeks 16 and 24. There was a statistically significant difference in the T2 values between the bone defect group and its sham group(P 0.05) from week 8, and between the cement group and its sham group(P 0.05) from week 16. There was significant difference in T2 values between the two experimental groups at postoperative week 24(P 0.01). The T2 value for articular cartilage was positively correlated with the Mankin score(ρ = 0.758, P 0.01). Conclusion Structural changes in subchondral bone can lead to degeneration of the adjacent articular cartilage. Defects in subchondral bone cause more severe degeneration of cartilage than subchondral bone filled with cement. The T2 value for articular cartilage increases with the extent of degeneration. MR T2-mapping images and the T2 value for articular cartilage can indicate earlycartilage degeneration.     

Subchondral thickness does not vary with cartilage degeneration on the metatarsal

Osteoarthritis is a disease of synovial joints that involves articular cartilage breakdown with accompanying bone changes, including subchondral sclerosis and osteophytosis. However, conflicting data have been reported concerning the cause-and-effect relationship, if any, between these changes. The authors studied the subchondral plate (subchondral bone plus calcified cartilage) in relation to the degree of articular cartilage degeneration on the distal articular surface of the first metatarsal, a region prone to osteoarthritis. No correlation was found between subchondral plate thickness or porosity and the degree of cartilage degeneration in the study sample of 96 metatarsals. Owing to the suggestion that initiation of cartilage fibrillation may be a result of steep stiffness gradients in the subchondral bone, the ratios of subchondral plate thickness in adjacent regions of the metatarsal head were examined in detail, but no correlation was found with subchondral degeneration. Thus increases in subchondral bone thickness are not associated with increases in cartilage degeneration on the first metatarsal, which may imply that subchondral bone changes do not cause osteoarthritis in this joint.     

Dynamic Alterations in Microarchitecture,Mineralization and Mechanical Property of Subchondral Bone in Rat Medial Meniscal Tear Model of Osteoarthritis

Background:

The properties of subchondral bone influence the integrity of articular cartilage in the pathogenesis of osteoarthritis (OA). However, the characteristics of subchondral bone alterations remain unresolved. The present study aimed to observe the dynamic alterations in the microarchitecture, mineralization, and mechanical properties of subchondral bone during the progression of OA.

Methods:

A medial meniscal tear (MMT) operation was performed in 128 adult Sprague Dawley rats to induce OA. At 2, 4, 8, and 12 weeks following the MMT operation, cartilage degeneration was evaluated using toluidine blue O staining, whereas changes in the microarchitecture indices and tissue mineral density (TMD), mineral-to-collagen ratio, and intrinsic mechanical properties of subchondral bone plates (BPs) and trabecular bones (Tbs) were measured using micro-computed tomography scanning, confocal Raman microspectroscopy and nanoindentation testing, respectively.

Results:

Cartilage degeneration occurred and worsened progressively from 2 to 12 weeks after OA induction. Microarchitecture analysis revealed that the subchondral bone shifted from bone resorption early (reduced trabecular BV/TV, trabecular number, connectivity density and trabecular thickness [Tb.Th], and increased trabecular spacing (Tb.Sp) at 2 and 4 weeks) to bone accretion late (increased BV/TV, Tb.Th and thickness of subchondral bone plate, and reduced Tb.Sp at 8 and 12 weeks). The TMD of both the BP and Tb displayed no significant changes at 2 and 4 weeks but decreased at 8 and 12 weeks. The mineral-to-collagen ratio showed a significant decrease from 4 weeks for the Tb and from 8 weeks for the BP after OA induction. Both the elastic modulus and hardness of the Tb showed a significant decrease from 4 weeks after OA induction. The BP showed a significant decrease in its elastic modulus from 8 weeks and its hardness from 4 weeks.

Conclusion:

The microarchitecture, mineralization and mechanical properties of subchondral bone changed in a time-dependent manner as OA progressed.     

EFFECT OF INTRA-ARTICULAR FRACTURE ON ARTICULAR CHONDROCYTES A TRANSMISSION ELECTRON MICROSCOPIC STUDY

Intra-articular fractures involving the subchondral bone were produced surgically in both left and right medial femoral condyles of twenty rabbits. The edges of the fractures on the left side were well approximated, while those on the right side maintained with a gap of 2 mm. Transmission electron microscopic study of the nearby articular cartilages conducted 1, 2, 3, 4 and 5 months after the fracture revealed the following results: 1) The neighbouring articular chondrocytes tended once to assume the configuration of fibroblasts; 2) Those remote from the fracture showed both degeneration and hyperplasia to form clones; 3) Even after the intra-articular fractures were well-healed, degeneration and hyperplasia of the articular chondrocytes nearby the fracture would persist for some time.     

膝关节软骨损伤的MR与关节镜对比研究

目的:分析MR扫描关节软骨损伤的诊断能力,为关节软骨损伤的临床诊断和治疗提供可靠的影像学依据。方法:对临床行膝关节镜检查的膝关节疼痛患者进行术前MR成像,对MR图像进行三维重建处理。结果:与关节镜对照,关节软骨损伤病例中在病变部位出现与损伤软骨区相对应的软骨下骨及骨髓内片状T1WI低信号影,.T2WI呈高信号影。结论:MRI对关节软骨损伤病变的准确性与关节镜诊断结果之间具有良好的一致性。     

关节软骨微环境的物质输运研究进展

关节软骨是最常见且研究最多的一种透明软骨;其软骨细胞的物质输运主要由扩散和对流2种方式完成.在关节软骨的胞外基质内,生物活性分子的扩散和对流输运在组织生理调节和细胞生物学响应中起到极为重要的作用.近年来,关节软骨与软骨下骨的交互作用备受关注;其交互作用在骨关节炎的发生发展中不容忽视.本文主要对关节软骨内、关节软骨与软骨下骨间物质输运的实验研究进展进行综述,旨在为研究关节软骨微环境的生理病理作用提供一定参考,并为临床治疗骨关节炎提供一定的新思路.     

Repair of articular cartilage on the surface of heat-treated bone by transplantation of cultured chondrocytes

The present study addresses clinical problems associated with the degeneration of articular cartilage, which occurs when heat-treated bone with articular cartilage is used for re-implantation after resection of malignant bone tumors adjacent to the joints. We therefore evaluated the effect of transplantation of chondrocytes embedded in collagen gel on the surface of heat-treated bone. A cylindrical complex of bone and articular cartilage 6 mm in diameter was resected from rabbits' patellar grooves and treated in saline at 60 degrees C for 30 min. In Group A, articular cartilage was resected from the complex and the remaining bone was returned to the patellar groove. Then, autologous chondrocytes cultured in collagen gel were transplanted and covered with periosteum. As controls, the original complex of heat-treated bone and articular cartilage (Group B) and heat-treated bone directly covered with periosteum (Group C) was returned to the patellar groove. In Group A, histological study showed that round cells were mainly observed and the matrix was well stained with Safranin O in the repair tissue after 24 weeks. The repair tissue was as thick as the adjacent normal cartilage. Immunohistological study detected type-II collagen and chondroitin-6-sulphate (3B3+) in the matrix of the repair tissue, but not type-I collagen. The repair tissue was consequently cartilaginous in Group A. The repair tissue was not cartilaginous or was degenerative in the control groups. We believe that this modality of heat-treated joints will contribute to limb salvage reconstruction after resection of malignant bone tumors adjacent to the joints.     

自体骨-骨膜移植修复关节软骨缺损的实验研究

目的　通过自体骨 -骨膜移植修复兔关节软骨缺损 ,探讨骨膜间充质细胞对关节透明软骨组织全层缺损修复的作用与机制。方法　以成年兔胫骨内上侧的骨 -骨膜组织作为供体 ,镶嵌式植入兔自体髌股关节之股骨滑车关节软骨等大、相同形状的全层缺损区 ,术后 4～ 2 4周分别对缺损修复情况进行大体、组织学及电镜观察。结果　自体骨 -骨膜移植组织与缺损周围组织完全愈合 ,组织学及电镜观察显示修复组织为透明软骨组织。结论　自体骨 -骨膜组织移植修补关节软骨缺损可以诱导关节软骨缺损以透明软骨方式进行修复 ,这为临床应用自体骨 -骨膜组织移植修复关节软骨缺损提供了理论依据     

脱钙松质骨复合同种异体软骨细胞修复兔关节骨软骨缺损的实验研究

目的评价脱钙松质骨（DCB）复合同种异体软骨细胞构建组织工程软骨修复兔关节骨软骨缺损的效果。方法分离1月 龄雄性新西兰兔关节软骨细胞,原代培养后复合制备的DCB体外培养2周构建组织工程软骨。4~5月龄新西兰兔30只双侧股 骨内髁制作直径3 mm、深3 mm,穿透软骨下骨板的骨软骨缺损模型,20只右侧关节缺损处植入构建的组织工程软骨（A组）,左 侧缺损处植入DCB（B组）,10只双侧骨软骨缺损未予处理作为空白对照（C组）。分别于术后1、3、6月取修复组织标本,进行大 体形态、组织学及Ⅱ型胶原染色;并对6月修复组织进行组织学评分,比较各组修复效果差异。结果制备的DCB为三维多孔的 海绵结构,孔隙大小约为100~500 μm,相互交通。DCB植入体内后1月开始降解,3月完全吸收。术后6月A组缺损处修复组织 主要为透明样软骨,与周围正常软骨厚度基本一致,修复交界区整合良好,不易辨认。修复组织深层细胞在软骨陷窝内,呈柱状 排列,基质蛋白多糖和Ⅱ型胶原染色接近正常软骨,软骨下骨板完整。B组缺损处以纤维软骨样组织修复为主。C组以纤维组 织填充。组织学评分显示术后6月A组除软骨下骨板重建与B组比较无统计学差异外, 其它各项评分均优于B组和C组,差异 有统计学意义（P<0.05）。结论DCB是一种较好的软骨组织工程支架材料,复合同种异体软骨细胞能修复关节骨软骨缺损,修 复组织为透明样软骨。     

兔膝关节软骨下自体骨移植后不同固定时间对软骨的影响

目的：研究兔膝关节软骨下自体骨转移后不同固定时间对关节软骨的影响。方法：健康大耳白兔５４只按术后膝关节不同固定时间随机分为Ａ、Ｂ、Ｃ３组。建立膝关节软骨下自体骨移植的动物模型,于术后８周取材,采用光镜、透射电镜对关节软骨进行形态学观察。结果：Ａ组２４％的关节软骨出现退变;Ｂ组１３％的关节软骨出现退变;Ｃ组２０．３％关节软骨出现退变。Ｂ组与Ａ组、Ｃ组相比,有显著性差异。结论：术后合理负重可降低关节软