同种异体骨软骨移植修复关节软骨缺损研究现状

关节软骨缺损常因软骨再生能力低而难以自行修复.新鲜的同种异体骨软骨移植修复关节软骨缺损的疗效稳定,成功率逐渐提高.冷冻保存的同种异体骨软骨移植修复关节软骨缺损的成功率可与新鲜的同种异体骨软骨移植媲美,梯度降温法是目前保存软骨细胞存活率最好的冷冻方法.该文就同种异体骨软骨移植的实验和临床研究、免疫排斥问题及其相关研究动态作一综述.     

同种异体骨软骨移植修复软骨损伤的研究进展

关节软骨损伤的治疗是骨科领域一个难题,同种异体骨软骨移植是修复全层软骨损伤的有效方法.文中综述了同种异体骨软骨移植修复软骨损伤的研究进展,包括该方法的适应证和禁忌证,目前在临床的应用情况及治疗效果,并进一步探讨了同种异体骨软骨移植中的3个热点话题,如疾病传播、免疫排斥和移植物的保存.这些文献综述显示同种异体骨软骨移植修复关节软骨损伤具有特定优势,但进一步推广该方法仍有一些需要解决的问题.     

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。     

骨软骨移植联合rhBMP -2/bFGF修复软骨缺损

[目的]探讨采用冷冻同种异体骨软骨移植,配合骨形态发生蛋白(rhBMP-2)/碱性成纤维细胞生长因子(bFGF)修复软骨缺损的效果,为进一步应用于临床提供理论依据。[方法]48只日本大耳白兔,96个关节,随机分A、B、C、D组。A组采用骨软骨移植联合rhBMP-2/bFGF,B组单纯应用骨软骨移植,C组单纯局部注射rh-BMP-2/bFGF混悬液,D组用作空白对照。无菌条件下制作骨软骨缺损模型。术后第4、8、12周作解剖学观察、磁共振检查、组织学检查及软骨细胞记数、免疫组化检查。[结果]A组软骨缺损修复面光滑,呈瓷白色、半透明,B、C组未完全修复,D组无明显修复。A组修复软骨组织学评分与其他组比较差异有统计学意义(P<0.05),图像分析仪软骨细胞记数与其他组比较差异有统计学意义(P<0.05)。修复软骨Ⅱ型胶原免疫组化染色阳性。[结论]联合应用冷冻同种异体骨软骨移植、rhBMP-2/bFGF,能促进新生软骨的形成,提高软骨缺损修复的质量。     

新鲜异体软骨移植修复髋关节损伤

[目的]介绍新鲜同种异体软骨移植修复髋关节损伤的手术技术与初步结果.[方法] 2019年3月～2019年5月,对3例股骨头和(或)髋臼骨缺损采用新鲜同种异体软骨移植技术修复,外科脱位暴露髋关节,清除损伤软骨及软骨下骨组织,将新鲜同种异体骨软骨制备成适合大小,嵌压固定于缺损处.[结果]所有患者术后无严重并发症.术后卧床1...     

慢速梯度降温冷冻保存同种异体骨软骨移植治疗膝关节全层软骨缺损(附三例报告)

目的总结冷冻保存同种异体骨软骨移植物治疗3例膝关节全层骨软骨缺损的手术方法及疗效。方法应用梯度降温冷冻保存的6枚同种异体骨软骨移植物治疗3例膝关节全层骨软骨缺损,2例在关节镜下同种异体骨软骨移植,1例行关节切开移植。膝关节股骨髁关节软骨全层缺损平均面积2.16 cm2。所有患者在手术后第1个月、第3个月时进行膝关节MRI检查,了解移植物与周围骨软骨组织的愈合情况。并于门诊复查时进行Brittberg-Peterson膝关节功能评分,了解功能恢复情况。结果随访4～6个月,平均4.7个月。所有患者术后疼痛消失;无排异反应发生。术后3个月时,MRI检查示术后移植物与宿主软骨下骨整合良好,移植软骨组织结构与内部信号良好。Brittberg-Peterson评分术后6个月比手术前明显降低。结论经梯度降温冷冻保存的同种异体骨软骨移植治疗膝关节软骨缺损早期效果满意。     

自体髌骨外侧软骨移植治疗膝关节软骨缺损的实验研究

自体髌骨外侧软骨移植治疗膝关节软骨缺损的实验研究杨述华李进杜靖远罗怀灿夏志道汪岚关节软骨缺损的修复是至今未能解决的问题，软骨移植治疗软骨缺损仍处于实验阶段，且多数采用同种异体软骨移植，因取材困难以及排异反应仍不能用于临床。作者设计自体髌骨外侧软骨移植...     

膝关节软骨损伤的外科治疗进展

关节软骨损伤后,软骨缺损通常缺乏自行修复能力,要求外科修复。传统外科治疗软骨损伤包括关节镜下冲洗清理术、微骨折术、自体骨软骨移植术、异体骨软骨移植术和自体软骨细胞移植等方法。关节冲洗清理术去除了关节内致痛因素,操作简单,应用广泛,早期疗效确切。微骨折术及自体骨软骨移植对小面积的软骨缺损修复较为理想,然而远期临床观察发现钻孔渗透修复的纤维软骨会降低微骨折术后疗效,相对于重建负重区关节面完整性自体骨软骨移植更具有优势。自体软骨细胞移植及异体骨软骨移植适用于更大面积的软骨缺损,异体骨软骨移植术后存活率受到局部排斥反应影响,从而降低了远期疗效。软骨组织工程技术可最大限度地提高自体软骨细胞移植的修复质量,实现修复组织接近透明软骨,但对于累及软骨下骨板、反应性骨水肿、严重骨量丢失或下肢轴线不良具有局限性。近年来许多新技术陆续应用于软骨损伤治疗领域,创伤小、操作简便、恢复快、疗效好、花费低、多技术联合应用的外科修复技术将会成为未来的治疗软骨损伤的重要手段。目前如何提高软骨修复质量,更具抗压、耐磨性,仍亟待解决。     

关节镜辅助下同种异体骨软骨移植修复膝关节剥脱性骨软骨炎

目的 评价关节镜辅助下采用同种异体骨软骨移植修复膝关节剥脱性骨软骨炎(osteochondritis dis-secans,OCD)伴大面积骨软骨缺损的可行性和有效性.方法 2004年1月-2007年5月,收治13例膝关节股骨髁OCD伴大面积骨软骨缺损患者.男7例,女6例;年龄18～59岁.左膝8例,右膝5例;病程7 d～20年,中位病程为42个月.4例有明确膝关节扭伤史.受累部位:股骨内侧髁外侧份5例,股骨内侧髁滑车面2例,股骨外侧髁6例.主动关节活动度为(95.0±13.5)°.Lysholm膝关节功能评分为(62.23±7.79)分.软骨损伤根据国际软骨修复协会分型和关节镜下的Guh1分型,均为Ⅳ型.关节镜下见骨软骨缺损面积为3～7 cm2,平均4.32 cm2;缺损深度0.8～2.0 cm,平均1.55cm.采用同种异体骨软骨块修复缺损,并联合可吸收钉固定骨软骨块.术后指导功能锻炼.结果 术后伤口均Ⅰ期愈合.13例均获随访,随访时间1年2个月～4年,平均2.2年.术后3个月2例持续活动1 h以上后膝关节疼痛,指导其功能锻炼后缓解.术后6个月2例关节摩擦感明显,关节稳定性尚好,予药物保守治疗.末次随访时主动关节活动度为(137.0±9.8)°,与术前比较差异有统计学意义(P<0.05).X线片检查示术后4～6个月移植骨软骨与受区松质骨融合,关节面平整,关节间隙正常.末次随访EMRI检查示骨软骨愈合,关节面完整;骨软骨移植物无缺损、退行性变.术后1年Lysholm膝关节功能评分为(92.08±7.64)分,与术前比较差异有统计学意义(P<0.05).结论 对于膝关节股骨髁OCD伴大面积骨软骨缺损,在关节镜辅助下采用同种异体骨软骨移植修复缺损能重建关节面的完整性和恢复关节稳定性,可取得良好的治疗效果.     

新鲜和深低温冷冻同种异体骨软骨移植长期转归实验研究

[目的]观察和比较新鲜和深低温冷冻同种异体骨软骨移植后的长期转归情况,为同种异体骨软骨移植治疗关节软骨缺损进一步提供理论基础。[方法]建立兔膝关节软骨缺损模型,行新鲜和深低温冷冻同种异体骨软骨移植。在术后12个月和18个月,骨软骨移植物取材进行关节软骨蛋白多糖、软骨细胞活性检测和软骨细胞超微结构观察。[结果]新鲜和冷冻骨软骨移植物中蛋白多糖阿尔新蓝染色强度和存活软骨细胞比率术后均下降,且在各时间点新鲜移植组的结果优于冷冻移植组,有统计学差异(P0.05)。超微结构显示软骨细胞退行性改变。[结论]新鲜和深低温冷冻同种异体骨软骨在移植术后较长时间均出现严重退变,冷冻骨软骨移植物退变较严重,其作为关节软骨缺损治疗方法目前是不可行的。     

Repairing articular cartilage defects with tissueengineering cartilage in rabbits

Objective: To investigate the effect of cancellous bone matrix gelatin ( BMG ) engineered with allogeneic chondrocytes in repairing articular cartilage defects in rabbits. Methods: Chondrocytes were seeded onto three-dimensional cancellous BMG and cultured in vitro for 12 days to prepare BMG-chondrocyte complexes. Under anesthesia with 2.5% pentobarbital sodium (1ml/kg body weight), articular cartilage defects were made on the right knee joints of 38 healthy New Zealand white rabbits (regardless of sex, aged 4-5 months and weighing 2. 5-3 kg) and the defects were then treated with 2. 5% trypsin. Then BMG-chondrocyte complex ( Group A, n = 18 ), BMG (Group B, n = 10), and nothing (Group C, n = 10) were implanted into the cartilage defects, respectively. The repairing effects were assessed by macroscopic, histologic, transmission electron microscopic ( TEM ) observation, immunohistochemical examination and in situ hybridization detection, respectively, at 2, 4, 8, 12 and 24 weeks after operation. Results: Cancellous BMG was degraded within 8 weeks after operation. In Group A, lymphocyte infiltration was observed around the graft. At 24 weeks after operation, the cartilage defects were repaired by cartilage tissues and the articular cartilage and subchondral bone were soundly healed. Proteoglycan and type II collagen were detected in the matrix of the repaired tissues by Safranin-O staining and immunohistochemical staining, respectively. In situ hybridization proved gene expression of type II collagen in the cytoplasm of chondrocytes in the repaired tissues. TEM observation showed that chondrocytes and cartilage matrix in repaired tissues were almost same as those in the normal articular cartilage. In Group B, the defects were repaired by cartilage-fibrous tissues. In Group C, the defects were repaired only by fibrous tissues. Conclusions: Cancellous BMG can be regarded as the natural cell scaffolds for cartilage tissue engineering. Articular cartilage defects can be repaired by cancellous BMG engineered with allogeneic chondrocytes. The nature of repaired tissues is closest to the normal cartilage. Local administration of trypsin can promote the adherence of repaired tissues to host tissues. Transplantation of allogeneic chondrocytes has immunogenicity, but the immune reaction is weak.     

Articular cartilage restoration with costal cartilage previously fused with bone

A novel procedure was developed for restoration of an articular cartilage defect using an autologous costal cartilage prepared with iliac bone, and the durability in vivo of this biologic construct was examined. First, an osteochondral complex was prepared (successful preparation, 67 of 80). Cancellous bone blocks isolated from the ilium of male Japanese White rabbits aged 5 months were implanted onto the surface of the costal cartilage before being tied by a pair of 3-0 silk thread sutures that were looped around the costal cartilage from behind. Second, 3 months later, the bone-attached costal cartilage was harvested and implanted into a full-thickness cartilage defect induced in a trochlear groove of the femur. All of the grafts were fixed to the recipient, maintaining its cartilage structure until 6 months (n = 28) and 12 months (n = 12) after implantation. However, when the costal cartilage without any bony portion was implanted into a similarly induced defect, 42% (10 of 24) were detached from the recipient before 12 months after implantation. The nontreated defect did not heal spontaneously to a satisfactory level (n = 12). These findings suggest that an osteochondral fragment, prepared by grafting cancellous bone onto costal cartilage, can be used for articular cartilage restoration.     

脱细胞软骨支架材料修复兔关节软骨缺损

目的 观察异种异体脱细胞软骨支架材料(ACM)复合同种异体兔骨髓间充质干细胞(rBMSCs)修复兔股骨内髁关节软骨缺损的效果.方法 (1)密度梯度离心和差速贴壁法获得原代兔BMSCs,选择第3代BMSCs作为种子细胞;(2)利用冷冻干燥、胰酶消化和化学去垢剂等方法制备脱细胞软骨支架材料;(3)3个月龄新西兰兔股骨内髁制备直径4 mm,深3 mm砌关节软骨缺损模型,24只新西兰兔以2个时间段随机分为3组,Ⅰ ACM-BMSCs组:第3代BMSCs 1×106个/ml与ACM于37℃5%CO2饱和湿度复合48 h;Ⅱ ACM组;Ⅲ空白对照组.(4)移植6、12周后大体及组织学观察,免疫组织化学染色观察修复组织Ⅱ型胶原,Wakitani评分评估修复效果.结果 (1)大体观察及组织学观察:6和12周Ⅰ组再生组织与正常关节软骨面平齐,修复部位表面较平整,界限模糊,接近正常软骨.Ⅱ组修复组织表面不平整并有明显下陷,修复组织全层可见成纤维样细胞,深层可见极少数透明软骨样细胞.Ⅲ组未见明显修复,肉芽组织形成伴成纤维样细胞增生;(2)Wakitani组织学评分可见在不同的时间段I组和Ⅱ组均低于Ⅲ组,差异有统计学意义(P<0.05),Ⅰ组和Ⅱ组间组织学评分差异无统计学意义(P>0.05).(3)免疫组织化学:ACM-BMSCs组修复组织的细胞为软骨样细胞,可见柱状排列,周围软骨基质Ⅱ型胶原染色阳性.结论 以ACM为支架材料,同种异体BMSCs为种子细胞制备的组织工程化软骨对兔股骨内髁关节软骨缺损有修复作用,形成的新生软骨为透明软骨样组织.     

组织工程软骨对兔膝关节胫骨平台浅层全关节软骨缺损的修复作用

目的探讨组织工程软骨对兔膝关节胫骨平台外侧髁浅层全关节软骨缺损的修复作用，并检测其修复组织的软骨类型。方法取2周龄乳兔软骨细胞体外培养传代至第3代后，与人胎盘Ⅰ型胶原蛋白海绵复合后植入成年兔的胫骨平台外侧髁完全软骨缺损区，并设立空白对照组，分别于术后4、12、24周取材。观察修复效果。运用天狼星红染色检测Ⅰ型胶原，Ⅱ型胶原单克隆抗体检测Ⅱ型胶原的表达。结果实验组术后4周缺损表面未见明显的新生软骨形成，组织学切片上少数几个呈上皮样生长的软骨细胞，苏木素-伊红（HE）染色胞浆呈深蓝色，周围软骨基质染色成浅蓝色；12周，缺损表面有少量的新生软骨形成，组织学切片上可见软骨细胞呈边缘不规则的，小蜂窝状结构，软骨细胞周围有软骨陷窝形成；24周，缺损表面的新生软骨较4、12周的新生软骨明显，表面光滑，且与周围组织结合紧密，但仍存在部分缺损尚未修复，组织学切片上可见软骨细胞呈边缘不规则的，多层细胞的蜂窝状结构，软骨细胞周围有软骨陷窝形成，并分泌甲苯胺蓝异染的软骨基质。而对照组则均未见明显的修复；随着术后时间的延长，Ⅰ型胶原的表达呈逐渐减弱的趋势，而Ⅱ型胶原的表达呈逐渐增强的趋势。结论该方法制成的组织工程软骨对兔胫骨平台外侧髁全层软骨缺损有修复作用，运用该方法不能完全修复兔胫骨平台外侧髁软骨完全缺损；形成的新生软骨为透明软骨样组织。     

耳软骨整体再造鼻侧软骨的可行性分析

目的：以耳软骨为供区,探索整体再造鼻侧软骨的方法。方法：20具（40侧）尸体标本,取下耳软骨40枚,鼻侧软骨40枚。CT扫描后重建三维图像,测量鼻侧软骨各解剖分区的相关数据。设计耳软骨3个供区可整体移植再造同侧鼻侧软骨,测量相关的形态数据。结果：耳软骨3个供区的相关形态数据大于同侧鼻侧软骨的相应数据。结论：耳甲腔、耳屏及两者之间连接的峡部（CVIT区域）,三角窝、耳轮及两者的连接部（TFH区域）,耳甲艇、耳轮及两者之间的连接部分（CBH区域）可整体取下,修整后整体再造同侧鼻侧软骨。     

软骨代谢标志物对骨关节炎软骨改变的反映

目的研究血清中蛋白聚糖和Ⅱ型胶原水平与骨关节炎严重程度间的关系,及对手术后软骨代谢改变的反映。方法研究对象包括65例膝骨关节炎患者及22名正常人。45例患者分别行不同方式手术治疗,术后半年复查。正常人及患者术前和复查时抽取静脉血,摄下肢负重位X线片。应用酶联免疫吸附方法检测血清样本中蛋白聚糖及Ⅱ型胶原的水平。结果骨关节炎患者血清中蛋白聚糖及Ⅱ型胶原水平均显著高于正常对照组,差异有显著性意义(P<0.05)。血清中蛋白聚糖和Ⅱ型胶原水平在轻型骨关节炎组升高,在关节明显狭窄组水平最高,而在严重狭窄组明显降低。术后半年,全膝关节置换术组血清中蛋白聚糖水平较术前明显下降(P<0.05);截骨术组血清中Ⅱ型胶原水平较术前明显下降(P<0.05)。结论血清中蛋白聚糖和Ⅱ型胶原的水平与骨关节炎软骨的破坏程度、软骨细胞的合成反应及软骨的总量有关。蛋白聚糖和Ⅱ型胶原水平是反映软骨代谢改变的较敏感的指标。     

Articular cartilage injuries

The acute and repetitive impact and torsional joint loading that occurs during participation in sports can damage articular surfaces causing pain, joint dysfunction, and effusions. In some instances, this articular surface damage leads to progressive joint degeneration. Three classes of chondral and osteochondral injuries can be identified based on the type of tissue damage and the repair response: (1) damage to the joint surface that does not cause visible mechanical disruption of the articular surface, but does cause chondral damage and may cause subchondral bone injury; (2) mechanical disruption of the articular surface limited to articular cartilage; and (3) mechanical disruption of articular cartilage and subchondral bone. In most instances, joints can repair damage that does not disrupt the articular surface if they are protected from additional injury. Mechanical disruption of articular cartilage stimulates chondrocyte synthetic activity, but it rarely results in repair of the injury. Disruption of subchondral bone stimulates chondral and bony repair, but it rarely restores an articular surface that duplicates the biologic and mechanical properties of normal articular cartilage. In selected patients, surgeons have used operative treatments including penetrating subchondral bone, soft tissue grafts, and cell transplants and osteochondral autografts and allografts to restore articular surfaces after chondral injuries. Experimental studies indicate that use of artificial matrices and growth factors also may promote formation of a new joint surface. However, an operative treatment of an articular surface injury that will benefit patients must not just provide a new joint surface, it must produce better long-term joint function than would be expected if the injury was left untreated or treated by irrigation and debridement alone. Therefore, before selecting a treatment for a patient with an articular cartilage injury, the surgeon should define the type of injury and understand its likely natural history.