人工关节无菌性松动界膜中的免疫反应

目的 探讨人工关节无菌性松动界膜的免疫反应状况及其与无菌性松动的关系。方法 对 9例人工关节无菌性松动界膜标本进行组织学观察 ,并行 T淋巴细胞及其亚群 (CD 3、 CD 4、 CD 8)、 IL－ 2受体 (CD 25)、巨噬细胞 (CD 68)及组织相容性抗原 (HLA－ DR)的免疫组织化学染色,观察 T淋巴细胞的表达情况及其与炎症反应的关系。结果 所有界膜标本中均可见肉芽肿性组织,有大量的巨噬细胞和异物巨细胞浸润,其中 5例 T淋巴细胞数量增加, T淋巴细胞及其亚型的分布大致相同,且 3例 T淋巴细胞呈现 IL－ 2R阳性。 T淋巴细胞的增加及 IL－ 2受体阳性的 T淋巴细胞的表达与巨噬细胞、 HLA－ DR的表达和炎症反应程度有一定的相关性。结论 在部分假体周围存在着针对假体材料的超敏反应,并且可能是导致假体无菌性松动的原因之一。     

基质金属蛋白酶及其组织抑制剂与全髋关节置换术后假体无菌性松动中的关系

全髋关节置换术能有效重建髋关节功能 ,减轻疼痛 ,提高生活质量。随着假体设计改进 ,骨水泥技术提高 ,人工关节假体无菌性松动已日益成为影响人工关节使用寿命的主要因素。目前文献[1] 已证实假体周围界膜的存在 ,普遍认为该界膜具有生物学活性并在假体松动中起重要作用。其中基质金属蛋白酶与其组织抑制剂的相互作用关系在假体松动中越来越为人们所重视。1　基质金属蛋白酶 (ＭａｔｒｉｘＭｅｔａｌｌｏｐｒｏｔｅｉｎａｓｅｓ,ＭＭＰｓ)ＭＭＰｓ是一组结构中含Ｚｎ2 + 和Ｃａ2 + 的蛋白水解酶家族 ,目前至少有 16个成员。根据其作用底物不…     

THA术后无菌性松动假体周围界膜组织中IL-34的表达及临床意义

目的通过检测白细胞介素-34(IL-34)在全髋关节置换(THA)术后无菌性松动假体周围界膜组织中的表达情况,探讨其与假体周围骨溶解之间的关系。方法观察组纳入自2015-01—2017-01因THA术后髋关节假体无菌性松动而行翻修手术的患者,标本取自髋臼杯周围的界膜组织。对照组纳入同期因股骨头坏死或股骨颈骨折而行初次THA的患者,标本取自髋臼周围关节囊组织和筋膜组织。采用实时定量聚合酶链式反应(Real-time PCR)、酶联免疫吸附试验(ELISA)检测界膜组织中IL-34的表达水平,并分析其与假体无菌性松动之间的相关性。结果 Real-time PCR与ELISA检测结果均表明,观察组无菌性松动假体周围界膜组织中IL-34的表达明显高于对照组,差异有统计学意义(P 0.05)。结论 IL-34在THA术后无菌性松动假体周围界膜组织中的表达显著升高,其表达水平与无菌性假体松动的发生具有密切关系,IL-34有可能成为诊断无菌性假体松动的一个潜在靶点。     

白介素－1受体相关激酶M在关节假体周围组织中的表达及意义

目的本研究旨在通过检测关节假体周围组织中白介素-1受体相关激酶M(IRAK－M)的表达情况,探讨各种界膜中的表达差异及其临床意义。 方法选取中山大学附属第一医院关节外科人工髋关节置换(THA)患者作为研究对象,排除免疫系统紊乱、免疫功能缺陷及结缔组织病。对初次THA,术中收集滑膜或关节囊组织;对无菌性松动及假体感染患者,翻修术中收集髋臼侧及股骨侧假体周围界膜组织。按照常规免疫组化操作规程进行染色,显微镜下观察IRAK-M的表达情况。多组间采用Kruskal-Wallis H检验,两独立样本计量资料采用的t检验,两组间阳性表达百分比采用Mann-Whitney U检验,P < 0.05表示差异有统计学意义。 结果通过免疫组化检测发现假体感染、无菌性松动的界膜IRAK-M蛋白高表达,且IRAK-M染色阳性细胞几乎仅见于单核巨噬细胞和多核巨细胞,两种类型的松动界膜表达程度无明显统计学差异(P> 0.05),而对照组骨关节炎滑膜IRAK-M表达不明显。 结论IRAK-M在假体周围的界膜中有着高表达,可能参与免疫耐受,导致关节置换术后,局部抗感染免疫下降。     

人工关节无菌松动界膜中破骨细胞分化因子受体免疫阳性的多核巨细胞

人工关节置换术后最重要的并发症是假体周围骨溶解导致的假体无菌松动 ,松动的机理还不完全清楚。随着骨代谢研究的进展 ,调节破骨细胞形成和功能的肿瘤坏死因子家族 (ＴＮＦｓ)新成员 :骨保护蛋白 (ＯＰＧ)、破骨细胞分化因子 (ＲＡＮＫＬ)及其受体 (ＲＡＮＫ)相继被发现 ,它们在人工关节置换术后无菌松动中的作用成为人工关节研究的新热点[1] 。本研究拟建立界膜多核巨细胞分离培养法 ,并初步探讨ＲＡＮＫ在多核巨细胞上的表达。1.资料和方法 :(1)界膜多核巨细胞分离培养 :人工髋关节置换术后无菌松动病人的界膜经手术翻修取得。新鲜界膜…     

基质金属蛋白酶-9与假体周围骨溶解引起无菌性松动的研究进展

人工关节置换术能够减少关节疼痛并改善关节的功能,有效地提高患者的生活质量,是近年来临床上治疗严重关节疾病比较成功的治疗方法之一.而假体周围骨溶解引起的无菌性松动是人工关节置换术后并发的主要问题,其中破骨细胞增殖分化活跃造成的溶骨性骨破坏是关键因素,在假体周围骨溶解引起的无菌性松动中起至关重要的作用,而基质金属蛋白酶-9(matrix metalloproteinase -9,MMP -9)在破骨细胞中高表达,在细胞外基质的降解中发挥重要作用.破骨细胞分化过程中,MMP -9基因表达受到多种因素的调控.本文将近年来破骨细胞表达的MMP -9作用和调控机制以及与假体周围骨溶解的关系予以综述.     

成纤维细胞在全髋关节置换术后无菌性松动中的潜在作用

全髋关节置换术可明显减轻病人的疼痛，改善患者的关节功能，提高病人的生活质量。然而，置换术后晚期假体无菌性松动已日益成为影响人工关节使用寿命的主要因素。许多文献已证实，假体周围往往存在一层膜性组织，这层膜性组织具有生物活性并在假体松动中起着重要的作用。...     

磨损微粒诱导细胞凋亡与无菌性松动的研究进展

无菌性松动是关节置换术后最常见的远期并发症之一,限制了关节假体的使用寿命。近年来人们对假体周围界膜组织内的巨噬细胞、成骨细胞、破骨细胞及成纤维细胞作了大量的研究发现,人工关节置换产生的磨损颗粒在与界膜组织周围的细胞接触或被吞噬后可诱导假体周围界膜的慢性炎症反应并引起细胞凋亡,最终导致假体周围骨溶解以及无菌性松动的发生,提示细胞凋亡在无菌性松动中起着重要作用,细胞凋亡有可能成为无菌性松动新的治疗方法。本文就磨损颗粒引起细胞凋亡与无菌松动之间关系做一综述。     

界膜

1983年Goldring等发现了松动人工关节与骨之间有一层滑膜样组织,该滑膜样组织能产生大量被称为骨吸收因子的前列腺2（PGE2）和胶原酶。此后越来越多的研究表明界膜是在人工关节中骨-骨水泥或骨-假体界面,特别是在骨溶解部位存在的一层滑膜样纤维结缔组织。松动假体周围的结缔组织膜具有异物肉芽肿的特征并含有假体磨损颗粒。     

无菌性松动全髋关节假体周围界膜组织中Caspase-3表达和细胞凋亡的研究

目的观察无菌性松动全髋关节假体周围界膜中凋亡调控蛋白Caspase-3表达和细胞凋亡分布的变化,探讨其与假体周围骨溶解之间的关系。方法2001年4月～2006年8月临床上选取16例高分子聚乙烯-金属配伍全髋关节翻修术的患者,其中男10例,女6例;初装年龄45～67岁,翻修年龄55～78岁,使用时间7～13年。根据术前X线片和术中所见,分为松动/非骨溶解组和松动/骨溶解组,每组各8例,取出假体周围各区的假体-骨间界膜组织。另取6例初装人工全髋关节的骨性关节炎患者作为对照组,其中男2例,女4例;年龄54～68岁,病程9～15年;采用免疫组织化学法检测界膜组织中Caspase-3的表达,末端标记法原位检测细胞凋亡,分析并比较Caspase-3和细胞凋亡的阳性表达和分布,与磨损颗粒的局部聚积和骨溶解程度的关系。结果高分子聚乙烯磨损颗粒局部聚积区的Caspase-3阳性细胞率和细胞凋亡指数明显高于金属磨损颗粒;重度磨损细胞凋亡指数高于轻度磨损,差异有统计学意义(P<0.05);松动/骨溶解组Caspase-3阳性细胞率和细胞凋亡指数明显高于松动/非骨溶解组和对照组,差异有统计学意义(P<0.01)。结论假体周围界膜组织Caspase-3的表达和细胞凋亡具有一定规律性,并与磨损微粒的局部聚积和骨溶解程度密切相关,可能是磨损颗粒造成界面骨重建受阻而溶骨大量发生的关键环节之一;细胞凋亡参与了假体无菌性松动的病理过程,且与Caspase-3信号激活有关。     

Increased expression of EMMPRIN in the tissue around loosened hip prostheses

Matrix metalloproteinases (MMPs) have been shown to play a role in aseptic loosening of total hip replacement (THR). Extracellular matrix metallo-proteinase inducer (EMMPRIN) can upregulate expression of several MMPs but has little effect on their tissue inhibitor (TIMP)- Using the avidin-biotin-per-oxidase complex immunostaining method, we detected strong immunoreactivity of EMMPRIN in the lining-like layers, sublining area and vascular endothelium of synovial membrane-like interface tissue around loosened prostheses. In contrast, EMMPRIN staining was very weak in the synovial samples from patients with hip arthrosis. Double immunofluorescence labeling revealed EMMPRIN/MMP-1 double-positive cells in iining-iike layers and the sublining area of interface tissue.

Our findings indicate that EMMPRIN expression is upregulated in interface tissue, and that locally accumulated EMMPRIN may modulate MMP-1 expression. An imbalance in the activity of MMPs and TIMP may lead to tissue destruction and periprosthetic osteolysis. These biological responses, combined with mechanical stress caused by micromotion and oscillating fluid pressure, may eventually cause aseptic loosening of THR.     

Increased expression of EMMPRIN in the tissue around loosened hip prostheses

Matrix metalloproteinases (MMPs) have been shown to play a role in aseptic loosening of total hip replacement (THR). Extracellular matrix metallo-proteinase inducer (EMMPRIN) can upregulate expression of several MMPs but has little effect on their tissue inhibitor (TIMP)- Using the avidin-biotin-per-oxidase complex immunostaining method, we detected strong immunoreactivity of EMMPRIN in the lining-like layers, sublining area and vascular endothelium of synovial membrane-like interface tissue around loosened prostheses. In contrast, EMMPRIN staining was very weak in the synovial samples from patients with hip arthrosis. Double immunofluorescence labeling revealed EMMPRIN/MMP-1 double-positive cells in iining-iike layers and the sublining area of interface tissue.

Our findings indicate that EMMPRIN expression is upregulated in interface tissue, and that locally accumulated EMMPRIN may modulate MMP-1 expression. An imbalance in the activity of MMPs and TIMP may lead to tissue destruction and periprosthetic osteolysis. These biological responses, combined with mechanical stress caused by micromotion and oscillating fluid pressure, may eventually cause aseptic loosening of THR.     

Increased expression of EMMPRIN in the tissue around loosened hip prostheses

Matrix metalloproteinases (MMPs) have been shown to play a role in aseptic loosening of total hip replacement (THR). Extracellular matrix metalloproteinase inducer (EMMPRIN) can upregulate expression of several MMPs but has little effect on their tissue inhibitor (TIMP). Using the avidin-biotin-peroxidase complex immunostaining method, we detected strong immunoreactivity of EMMPRIN in the lining-like layers, sublining area and vascular endothelium of synovial membrane-like interface tissue around loosened prostheses. In contrast, EMMPRIN staining was very weak in the synovial samples from patients with hip arthrosis. Double immunofluorescence labeling revealed EMMPRIN/MMP-1 double-positive cells in lining-like layers and the sublining area of interface tissue. Our findings indicate that EMMPRIN expression is upregulated in interface tissue, and that locally accumulated EMMPRIN may modulate MMP-1 expression. An imbalance in the activity of MMPs and TIMP may lead to tissue destruction and periprosthetic osteolysis. These biological responses, combined with mechanical stress caused by micromotion and oscillating fluid pressure, may eventually cause aseptic loosening of THR.     

High cathepsin B activity in arthroplasty interface membranes: A histochemical study of 9 loose cemented total hip prostheses

We studied biopsies of interface membranes of 9 aseptically loosened total hip prostheses. The morphologic resemblance of the cement-facing surface of the membranes to synovial tissue of arthritic joints, as noticed by others, was confirmed by histochemical techniques. High cathepsin B activity was found in the bone-facing surface of the membranes. Since this enzyme also plays an important role in tissue destruction of arthritic joints, further similarities in the mechanisms of tissue breakdown in arthritis and aseptic loosening of cemented hip prostheses may be conjectured.     

Autonomic neuropeptides in the interface membrane of aseptic loose hip prostheses.

We analyzed the presence of autonomic nerve fibers in the interface membranes (n = 9) surrounding aseptic loosened hip prostheses by immunohistochemistry. The study focused on the autonomic messengers neuropeptide Y (NPY), tyrosine hydroxylase (TH), the rate-limiting enzyme in the synthesis of noradrenaline (NA), and vasoactive intestinal polypeptide (VIP). Protein gene product (PGP) 9.5, a general marker of peripheral nerve fibers, was also analyzed to establish the neuronal character of the immunoreactive structures. PGP 9.5-positive and NPY-positive nerve fibers were identified in all 9 samples, and VIP-immunoreactive and TH-immunoreactive fibers were found in 7. There was a difference in the distribution of nerve fibers both between and within the samples. Among the neuropeptides analyzed, NPY was most abundant. NPY-positive and TH-positive fibers were predominantly found around the blood vessel walls forming varicose nerve terminals. VIP-positive fibers were mainly observed as thin varicose nerve terminals with no relationship to blood vessels. Autonomic neuropeptides exert not only vasoactive and immunoregulatory effects, but also have been found to have direct effects on bone tissue. Moreover, the autonomic nervous system has been strongly implicated in nociception and inflammation. Neuronal NPY, TH, and VIP in the interface membrane may prove to contribute to the pathologic mechanisms leading to aseptic loosening of hip prostheses.     

人工关节松动病因的研究

目的：探讨人工关节松动的病因。方法：选择７例松动人工髋关节，翻修手术时取松动关节周围的界膜组织；同时选择１０例骨折内固定患者，拆除内固定物时取内固定物周围瘢痕组织。标本做组织学检查和肿瘤坏死因子（ＴＮＦ）测定。选择１０只成年兔，将２０只模拟假体分别置入双侧股骨远端。分别于术后第６、８、１０、１２、１４周向右侧膝关节腔注射聚乙烯微粒悬液，作为实验侧；左侧膝关节腔注射生理盐水，作为对照侧。第１６周取股骨远端标本，做组织学检查。结果：松动人工髋关节周围的界膜组织主要含大量的组织细胞和聚乙烯微粒，而骨折内固定物周围的瘢痕组织主要为纤维成分，无聚乙烯微粒。松动关节周围界膜组织中的ＴＮＦ浓度明显高于骨折内固定物周围的瘢痕组织（Ｐ＜０．０１）。动物实验发现，实验侧模拟假体周围有一层充满组织细胞的纤维结缔组织界膜，并有明显骨吸收和骨溶解现象，而对照侧无明显纤维结缔组织界膜，也无骨破坏现象。结论：人工关节磨损后，产生大量的磨损微粒，微粒刺激组织细胞分泌ＴＮＦ等溶骨性因子，这些溶骨性因子直接或间接地激活破骨细胞，从而引起假体周围骨吸收、骨溶解，最终导致假体松动。假体松动后又可加重磨损，产生更多的微粒，形成恶性循环     

Histological analysis and biological effects of granulation tissue around loosened hip prostheses in the development of osteolysis

Although aseptic loosening of the prosthesis is a long-term complication after total joint replacement, the detailed mechanism of osteolysis remains unknown. We examined 82 samples from 40 patients with aseptic loosened hip prostheses histologically, and compared the distribution of particles, macrophages/histiocytes, and foreign body giant cells in the retrieved tissue from capsules and around prostheses. Furthermore, to investigate the mechanism of osteolysis, we cultured tissue from a patient with massive osteolysis and examined the effects of the conditioned medium on osteoblasts in vitro. Numerous multinucleated giant cells and histiocytes were present, and polyethylene particles ranging from medium to large were identified in the polarized light. However, the distribution was heterogeneous, and no particles were found microscopically in about 30%–40% of periprosthetic tissues, and in 60% of capsules. The amount of particles correlated with giant cells, but not with histiocytes. The conditioned medium of the granulation tissue culture stimulated osteoblasts to produce interleukin-6 in both protein and mRNA, and this was in part inhibited by anti-tumor necrosis factor- or the interleukin-1 antibody, suggesting that interleukin-6 production is mediated by several cytokines. These findings suggest that interleukin-6, which is produced not only by macrophages but also by osteoblasts, is a contributing factor to aseptic loosening.     

The microscopic anatomy of the bone-cement interface in failed total hip arthroplasties

Thirty specimens were removed from the bone-cement interface in 25 hips revised for aseptic loosening of total hip arthroplasty. Histologic examination of the membranous tissues revealed that histiocytosis, fibrosis, and necrosis were present in every specimen. Other prominent features included particles of acrylic cement, polyethylene debris, and fragments of necrotic bone. Mechanical failure was characterized by cement fractures, and microfracture of bone. The presence of cement debris and bone detritus in the membranes, and smooth appearance of the removed cement mantles substantiated the presence of mechanical failure. The process of loosening was characterized by the recruitment of histiocytes into the interface and the subsequent resorption of bone around the prosthesis. This may be a manifestation of the rate of cement wear and tear, particle size, and the immunologic predispositions of the host. These observations on interfaces of loosened prostheses are reexamined and reinterpreted in the light of radiologic observations on interface radiolucent zones and well-functioning prostheses.     

Presence of interleukin-17C in the tissue around aseptic loosened implants

Purpose

The most common long-term complication of joint arthroplasty is aseptic loosening. The proinflammatory cytokines secreted by macrophages are involved in aseptic loosening. Recently, a novel proinflammatory cytokine IL-17C was reported to participate in inflammatory diseases by synergising with proinflammatory cytokines. However, the relationship between IL-17C and the aseptic loosening is unclear.

Methods

The tissues around aseptic loosened implants were collected during revision surgery and handled by formalin fixation and embedded in paraffin. The presence of IL-17C in the tissues around the aseptic loosened implants was investigated in 12 aseptic loosening patients using immunofluorescence.

Results

The presence of IL-17C protein in the tissues around aseptic loosened implants was detected by immunofluorescence. There are no statistical differences between optical density of IL-17C in aseptic loosening samples and in rheumatoid arthritis samples (positive control).

Conclusions

These results suggest the presence of IL-17C in aseptic loosening. Interleukin-17C was related to the inflammation of aseptic loosening, possibly by contributing to the inflammation and osteolysis in the tissues surrounding aseptic loosened implants.     

Toll‐like receptors and aseptic loosening of hip endoprosthesis—a potential to respond against danger signals?

Bacterial remnants and subclinical biofilms residing on prosthesis surfaces have been speculated to play a role in hip implant loosening by opsonizing otherwise relatively inert wear particles. The innate immune system recognizes these microbial pathogen‐associated molecular patterns (PAMPs) using Toll‐like receptors (TLRs). Our objective was to evaluate the possible presence of TLRs in aseptic synovial membrane‐like interface tissue. Bacterial culture‐negative, aseptic (n = 4) periprosthetic synovial membrane‐like tissue was compared to osteoarthritis synovial membrane (n = 5) for the presence of cells positive for all known human functional TLRs, stained using specific antibodies by immunohistochemistry, and evaluated using morphometry. In comparison to osteoarthtritic synovium, the number of TLR‐positive cells was found to be increased in the aseptic setting, reflecting the considerable macrophage infiltration to the tissues investigated. Thus aseptic periprosthetic tissue seems to be very reactive to PAMPs. It has been recently recognized that TLR do not only respond to traditional PAMPs, but also to endogenous alarmings or danger signals released from necrotic and activated cells. Alarming‐TLR interaction in the periprosthetic tissue might be a novel mechanism of aseptic loosening of endoprosthesis. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:184–190, 2010