异体软骨细胞移植修复猪膝关节软骨缺损的免疫学观察

目的应用同种异体软骨细胞移植修复猪膝关节软骨缺损,评估术后免疫学表现及其修复效果。方法供体为1月龄上海白猪2头,取其膝关节全层软骨片,0.25%胰蛋白酶和0.2%型胶原酶消化,分离培养软骨细胞。受体为2月龄巴马猪8头,在两侧股骨髁关节负重面造成0.5cm×0.5cm软骨缺损,深及软骨下骨。于受体猪双侧膝关节软骨缺损处注入软骨细胞悬液0.2ml,含软骨细胞(1.0～2.0)×106个,密封入口。术前、术后3、5、7、12周分离受体外周血淋巴细胞,检测其与异体软骨细胞混合后的刺激指数(stimulationindex,SI);术后5、7、24周取修复区软骨及软骨下骨观察局部组织学反应。结果术后3、5、7、12周受体外周血淋巴细胞SI分别为1.457±0.062、1.739±0.142、1.548±0.047和1.216±0.028,较术前(1.102±0.034)增高,且差异均有统计学意义(P<0.05);SI术后3周开始升高,5周达高峰,7、12周后开始逐渐下降;3、7、12周SI与5周比较差异均有统计学意义(P<0.05)。组织学观察:术后5周,出现较多的炎性浸润,分散于软骨下骨、修复组织与正常软骨的整合部等;7周,炎性浸润开始减退,仅在软骨下骨可见;24周,缺损处修复软骨与周围软骨整合良好。结论异体软骨细胞移植后,免疫反应一般在移植早期开始出现,并逐渐达到高峰,但随着软骨基质的重新合成,免疫反应也逐渐下降,并最终修复全层关节软骨缺损。     

异体软骨细胞移植修复关节软骨缺损免疫反应及处理

采用异体软骨细胞移植修复关节软骨缺损是当今研究热点。该文综述了异体软骨细胞移植产生免疫反应的原因及解决的方法。     

同种异体兔关节软骨细胞移植实验研究

将幼兔关节软骨细胞复层培养后植入同种成年兔关节软骨面人为缺损部，术后１．５个月－６．５个月采用大体、ＨＥ、甲苯胺兰、番红花精０染色及放射性自显影等方法观察软骨细胞修复缺损情况。结果表明移植术后１．５个月，植入软骨细胞在局部生长，与周围原有软骨组织融合，将缺损填平。２．５个月－６．５个月仍保持透明软骨性质，无纤维化、骨化、血管长入，免疫排斥反应等。     

同种异体组织工程化软骨修复关节软骨缺损

目的　探讨应用同种异体组织工程化软骨修复软骨缺损的可行性。方法　取新西兰大白兔双膝关节软骨细胞 ,经体外培养扩增 ,与PlruonicF12 7混合 ,植入人为造成的异体兔膝关节软骨缺损。结果　空白对照组和材料对照组只见少许纤维组织修复 ,缺损凹陷 ;实验组 8周后关节软骨缺损区由部分白色透明样软骨组织充填 ,Masson三色染色见胶原分布较均匀 ,软骨陷窝多见 ,未见明显炎症现象。 16周后缺损完全修复 ,缺损表面较光滑 ,部分颜色呈淡兰色 ,软骨陷窝清晰 ,细胞与基质分布均匀 ,未见炎症和退变现象。结论　同种异体组织工程化软骨可用于修复关节软骨缺损。     

吻合血管的同种异体骨关节移植的免疫反应

近 3 0年来 ,随着显微外科技术的不断发展 ,吻合血管的骨移植在修复重建外科作用也日益重要 ,特别是在因外伤、恶性骨肿瘤切除以及先天性骨缺损引起的大段骨关节缺损治疗中的作用优势明显。吻合血管的自体骨关节移植因其缺乏抗原性 ,愈合过程类似于骨折而受到外科医师的青睐 ,但自体骨关节来源有限 ,且有供骨区损伤的问题 ;目前多采用同种异体骨关节移植 ,而非血管化的同种异体骨关节移植修复缺损时 ,常因病理骨折、骨不连接、感染、移植骨吸收等并发症而不理想〔1〕。吻合血管的同种异体骨关节移植 ,作为血管化的骨关节移植有供源广、形状…     

兔同种异体异位膝关节移植模型的建立

目的建立兔同种异体异位膝关节移植实验动物模型.方法在观察兔的下肢解剖的基础上,设计兔同种异体异位膝关节移植术;观察吻合血管同种异体膝关节移植组的自然病程.结果兔吻合血管同种异体膝关节移植共9例,移植成功7例,移植物平均存活(13.57±0.79)d,移植物液化坏死,受体生命体征受排斥反应影响大.结论兔同种异体异位膝关节移植模型操作简便,表现稳定,适用于复合组织移植免疫的研究.     

同种异体骨移植的免疫反应

同种异体骨移植免疫反应过程和减弱其免疫排斥反应的研究是骨科领域的研究重点和难点。本文讨论了同种异体骨移植的抗原性和免疫反应、冷冻和冻干骨的免疫性、组织配型技术和应用免疫抑制剂对同种异体骨移植免疫反应的影响。     

抑制同种异体软骨细胞复合支架免疫反应研究进展

损伤或疾病造成的关节软骨缺损在临床上较常见,许多用于修复关节软骨损伤的方法各有优点,但均存在一定缺陷。组织工程学技术为关节软骨缺损的再生与修复提供了新思路,利用体外培养扩增的同种异体软骨细胞复合支架修复软骨缺损已有大量报道。该文就同种异体软骨细胞复合支架修复关节软骨损伤的免疫反应,在软骨细胞免疫原性、免疫反应发生途径、降低免疫反应机制及方法等方面作一综述。     

用同种异体骨基质明胶修复颅骨大面积缺损

本文报道同种异体骨基质明胶促进颅骨大面积缺损愈合的实验结果。以家兔颅顶骨直径1.0mm圆形骨缺损作为颅骨缺损模型,分别植入同种异体骨基质明胶、脱钙骨基质和不脱钙骨。植入后4周、8周、12周进行X线摄片和组织学位检查,结果表明同种骨基质明胶没有免疫排斥反应,并具有良好的骨诱导作用。术后8周已有大量新骨出现于骨缺损中央部,12周已使骨缺损完全愈合。说明同种骨基质明胶是修复儿童颅骨缺损较理想的植骨材料。     

自体软骨细胞移植修复猪膝关节软骨缺损的实验研究

目的评价传统和复层高密度培养（复层培养）的自体软骨细胞移植修复关节软骨缺损的效果。方法在8头猪16膝髌切迹上下共建立32个全层软骨缺损，右膝为空白和自体骨膜移植对照组，左膝为传统的自体骨膜覆盖下细胞注射移植组和自体骨膜覆盖下复层培养细胞移植组（上下缺损随机进入各对照和实验组）。术后5-6个月对缺损部位行大体、组织学、免疫组织化学检查。采用O’Driscoll软骨组织形态学评分评价软骨缺损的修复质量。结果四组的O’Driscoll软骨组织形态学评分分别为（3．14±1．95）分、（10．57±3．60）分、（16．29±2．63）分、（20．43±1．81）分，各组之间差异均有统计学意义（P〈0．01）。空白对照组缺损被少量的纤维组织覆盖；骨膜移植组被纤维组织和少量的纤维软骨修复，且与周围组织整合差。自体软骨细胞移植（注射组和复层培养组）缺损被塑形良好的修复组织覆盖，组织学、基质特殊染色示修复组织为纤维软骨和透明软骨，与周边软骨和软骨下骨整合良好。复层培养组修复组织的细胞形态、基质染色比注射细胞组更接近正常软骨。结论自体软骨细胞移植可成功修复全层关节软骨缺损。扩增后的软骨细胞经短期复层培养更有利于软骨缺损的修复。     

Pluronic F-127负载同种异体软骨细胞修复兔全厚关节软骨缺损的实验研究

目的:探讨Pluronic F-127作为软骨细胞移植载体的可行性。方法:将培养的软骨细胞与20％的Pluronic F-127的混合,移植到兔膝关节软骨缺损处。在移植后4、8、12周对缺损的修复情况进行大体、光镜组织学评估和电镜观察。结果:移植的软骨细胞在载体中生长良好,形成了透明软骨。扫描电镜下可见多数成熟的透明软骨细胞。Pluronic F-127平均降解时间为6～8周,与正常软骨的再生速度基本一致。根据Wakitani制定的评分标准,采用盲法对修复质量作出评价。Pluronic F-127组和对照组的组织学评分在各个时期无显著的差异(P>0.001),而软骨细胞-载体复合体组和对照组相比在各个时期均存在显著的差异(P<0.001)。结论:Pluronic F-127可作为工程化软骨细胞安全有效的载体。     

孔数不同的软骨下骨钻孔术对兔软骨缺损修复的影响

目的：为了观察软骨缺损的修复过程，比较不同数目钻孔术对软骨缺损的修复效果。方法：用中国白兔２４只，在股骨关节面造成６ｍｍ×８ｍｍ全层软骨缺损，分别施行１０孔及５孔钻孔术，术后４、８周取材，做组织学及电镜观察，并进行评估。结果：（１）１０孔、５孔和对照组的优势修复组织分别以类透明软骨，类透明软骨加纤维软骨和纤维组织为主。（２）修复组织厚度１０孔及５孔无显著差异。（３）修复组织覆盖缺损的面积，１０孔＞５孔＞对照组。初步结论：软骨下骨钻孔可修复关节软骨全层缺损；多孔比少孔修复好；非钻孔的缺损修复效果较差。     

Demineralized Dentin Matrix Acts as a Scaffold for Repair of Articular Cartilage Defects

Articular cartilage repair remains a major obstacle in tissue engineering. In the present study, we investigated the potential for demineralized dentin matrix (DDM; organic material derived from dentin) obtained from extracted teeth to be used as bone graft material. To evaluate the extent to which DDM induces osteochondral regeneration, we implanted DDM from bovine teeth in rabbit knees with full-thickness articular cartilage defects. Thirty-three 13-week-old male rabbits weighing 2.5–3.0 kg were randomly assigned to a control group (n = 11) and two experimental groups (n = 11 for each group). The knees were divided into three groups according to the subsequent treatment: in group I (n = 22), the control group, the defect was left untreated; and in groups II (n = 22) and III (n = 22), 50 and 100 mg of DDM, respectively, was implanted. The rabbits were killed 1, 3, 6, or 9 weeks after the surgical procedure, and the knees were collected. The harvested tissues were examined radiographically and histologically. The 100-mg DDM group (group III) had significantly more new bone forming inside the defect (as measured using the BV/TV value) compared with the other two groups as early as at week 3 postoperatively, but thereafter, the difference gradually decreased. Cartilage repair in the surface region remained significantly better in group III because hyaline-like cartilage appeared in the peripheral area of the defect at week 6 and the surface was covered with hyaline-like cartilage with a thickness similar to that of normal cartilage at week 9. In conclusion, the results of this study suggest that DDM acts as a scaffold for osteochondral regeneration, yielding active new bone formation early in the postoperative period. Thus, DDM may represent an effective bone implant material.     

肢体缺血/再灌注损伤兔膝关节软骨细胞核DNA含量的图像分析

本实验研究通过扫描电镜对缺血及再灌注后关节软骨细胞进行了较全面的观察.对其形态学特征进行了较系统的描述,并运用图像分析手段对细胞核中DNA含量进行定量测定,数量化地描述了关节软骨细胞在此损伤进程各阶段的图像变化特征,为临床进一步研究提供了一些较为可靠的参考指标.     

Sizable Scaffold‐Free Tissue‐Engineered Articular Cartilage Construct for Cartilage Defect Repair

This study introduces an implantable scaffold‐free cartilage tissue construct (SF) that is composed of chondrocytes and their self‐produced extracellular matrix (ECM). Chondrocytes were grown in vitro for up to 5 weeks and subjected to various assays at different time points (1, 7, 21, and 35 days). For in vivo implantation, full‐thickness defects (n = 5) were manually created on the trochlear groove of the both knees of rabbits (16‐week old) and 3 week‐cultured SF construct was implanted as an allograft for a month. The left knee defects were implanted with 1, 7, and 21 days in vitro cultured scaffold‐free engineered cartilages. (group 2, 3, and 4, respectively). The maturity of the engineered cartilages was evaluated by histological, chemical and mechanical assays. The repair of damaged cartilages was also evaluated by gross images and histological observations at 4, 8, and 12 weeks postsurgery. Although defect of groups 1, 2, and 3 were repaired with fibrocartilage tissues, group 4 (21 days) showed hyaline cartilage in the histological observation. In particular, mature matrix and columnar organization of chondrocytes and highly expressed type II collagen were observed only in 21 days in vitro cultured SF cartilage (group 4) at 12 weeks. As a conclusion, cartilage repair with maturation was recapitulated when implanted the 21 day in vitro cultured scaffold‐free engineered cartilage. When implanting tissue‐engineered cartilage, the maturity of the cartilage tissue along with the cultivation period can affect the cartilage repair.     

关节镜下微骨折术修复膝关节软骨缺损

目的探讨关节镜下微骨折术修复膝关节软骨缺损的临床效果。方法选取本院2010年10月至2012年10月收治的膝关节软骨缺损患者68例,根据术式分为两组,34例患者在关节镜下采用关节清理术治疗为对照组,34例患者在关节镜下采用微骨折术治疗为观察组,术后随访6个月。两组患者治疗前后均行Tegner运动能力评分和美国特种外科医院（the hospital special surgery,HSS）膝关节功能评分,分析两组患者的临床症状改善情况、手术治疗的临床效果及术后并发症情况。结果治疗后,两组患者Tegner运动能力评分、HSS膝关节功能评分均显著升高。观察组患者Tegner运动能力评分（4.3±1.2）分,HSS膝关节功能评分（84.3±15.2）分,均明显高于对照组（2.7±0.8）分、（61.5±14.8）分,观察组患者术后疼痛病症消失情况（97.1%）,肿胀病症消失情况（100.0%）均明显好于对照组（76.5%）,（82.4%）,观察组患者手术治疗的总有效率（94.1%）明显高于对照组（73.5%）,差异均有统计学意义（P〈0.05）。观察组患者的术后并发症发生率（0）低于对照组（5.9%）,差异无统计学意义（P〉0.05）。结论关节镜下微骨折术是修复膝关节软骨缺损的有效方法,可明显改善患者的临床症状,改善膝关节功能和活动能力,提高治愈率,安全性高。     

异体骨基质海绵吸附自体骨髓细胞修复关节软骨的实验研究

在兔股骨髌股关节面上造成软骨缺损,采用同种异体骨基质海绵吸附自体骨髓细胞移植,分期取材.通过肉眼、光镜、电镜观察软骨修复过程中的组织学形态.结果显示骨髓细胞在关节腔内血供差、低氧压及滑液的环境中具有较好的成软骨作用.认为自体骨髓细胞来源充足、易采取,创伤小,无不良并发症,无排斥反应.有可能成为一种修复关节软骨缺损的新途径.     

Long-Term Evaluation of Allogenic Chondrocyte-Loaded PVA–PCL IPN Scaffolds for Articular Cartilage Repair in Rabbits

ObjectiveThis study tested the long-term efficacy of two synthetic scaffolds for osteochondral defects and compare the outcomes with that of an established technique that uses monolayer cultured chondrocytes in a rabbit model.MethodsArticular cartilage defect was created in both knees of 18 rabbits and divided into three groups of six in each. The defects in first group receiving cells loaded on Scaffold A (polyvinyl alcohol–polycaprolactone semi-interpenetrating polymer network (Monophasic, PVA–PCL semi-IPN), the second on Scaffold B (biphasic, PVA–PCL incorporated with bioglass as the lower layer), and the third group received chondrocytes alone. One animal from each group was sacrificed at 2 months and the rest at 1 year. O’Driscoll’s score measured the quality of cartilage repair.ResultsThe histological outcome had good scores (22, 20, and 19) for all three groups at 2 months. At 1-year follow-up, the chondrocyte alone group had the best scores (mean 20.0 ± 1.4), while the group treated by PVA–PCL semi-IPN scaffolds fared better (mean 15 ± 4.2) than the group that received biphasic scaffolds (mean 11.8 ± 5.9). In all three groups, defects treated without cells scored less than the transplant.ConclusionThese results indicate that while these scaffolds with chondrocytes perform well initially, their late outcome is disappointing. We propose that for all scaffold-based tissue repairs, a long-term evaluation should be mandatory. The slow degrading scaffolds need further modifications to improve the milieu for long-term growth of chondrocytes and their hyaline phenotype for the better incorporation of tissue-engineered constructs.