聚甲基丙烯酸甲酯颗粒对人滑膜细胞核因子κB受体激动剂配体/骨保护蛋白表达的影响

背景：多种因子可以影响骨保护蛋白/核因子κB受体激动剂配体/核因子κB受体激动剂(Osteoprotegerin / receptor activator of nuclear factor-kappaB/ligand of receptor-activator of nuclear factor-kappaB,OPG/RANKL/RANK)系统的表达影响骨代谢,那么松动假体周围的骨水泥颗粒是否也通过影响其代谢参与假体松动过程？ 目的：观察聚甲基丙烯酸甲酯颗粒对人滑膜细胞RANKL/OPG表达的影响。 方法：体外培养人滑膜细胞,在滑膜细胞培养体系中分别加入质量浓度为0(空白对照),0.2,1,10 g/L的聚甲基丙烯酸甲酯骨水泥颗粒,采用荧光定量PCR检测上述各浓度PMMA颗粒作用不同时间后滑膜细胞内RANKL、OPG的表达量及其比值。 结果与结论：与空白对照组相比,各实验组滑膜细胞内RANKL、OPG的表达量均有下降;随着与聚甲基丙烯酸甲酯颗粒共培养时间延长,各实验组滑膜细胞内RANKL、OPG表达均有下降趋势,而RANKL/OPG表达比值无明显变化。说明聚甲基丙烯酸甲酯颗粒在对滑膜细胞产生生物学反应时并不干扰假体周围骨代谢的动态平衡,并未直接参与人工关节假体的无菌性松动。     

人工关节假体植入后无菌性松动的发病机制

背景：对于人工关节无菌性松动的基础和临床研究很多,但其确切机制仍不甚明确。 目的：综述人工关节假体植入后无菌性松动的发病机制。 方法：由第一作者应用计算机检索PubMed和中国期刊全文数据库2006/2011相关文献。在标题、摘要、关键词中以“artificial prosthesis,aseptic loosening”或“人工关节、无菌性松动”为检索词进行检索。共检索到77篇文献,最终纳入符合标准的文献38篇。 结果与结论：无菌性松动已成为人工关节置换后远期失败的主要原因之一。近年来对人工全髋关节置换后翻修病例回顾性分析的统计显示,无菌性松动位于翻修原因第1位。关于人工关节无菌性松动的机制主要分为机械机制、生物机制及其他机制,它们的共同结果是造成骨吸收、骨溶解,最终导致假体松动。随着研究的进一步深入,以及植入方式及假体材料的不断改进,无菌性松动将最终被最小化,使假体使用寿命更长久。     

中药淫羊藿苷干预人工关节磨损微粒诱导的骨溶解

背景：人工关节周围产生骨溶解是关节松动失败的重要原因,各国学者都在寻求一种能有效抑制骨溶解反应的药物,以减少假体松动的发生。 目的：观察不同剂量淫羊藿苷干预人工关节磨损微粒诱导骨溶解的效果。 方法：将钛合金及骨水泥磨损微粒制成的混悬液分别置入清洁小鼠颅盖骨进行骨溶解建模后,空白组滴入生理盐水到颅盖骨中,药物干预组给予不同浓度的淫羊藿苷(剂量分别为30,60,120 mg/kg)灌胃,每日1次。建模2周后处死小鼠,在显微镜下观察小鼠颅盖骨的结构。经苏木精-伊红染色和免疫组化分析计算破骨细胞数量和小鼠颅盖骨溶解面积的变化。 结果与结论：与空白组比较,钛合金微粒和骨水泥微粒建模后的小鼠颅盖骨切片上的骨溶解陷窝数量及破骨细胞数明显增加,图像分析骨溶解陷窝面积也增大(P < 0.05)。淫羊藿苷干预后骨溶解面积减小及破骨细胞数量减少  (P < 0.05),以120 mg/kg灌胃组最为显著,其次是60 mg/kg,30 mg/kg。结果可见钛磨损微粒和骨水泥磨损微粒能促进破骨细胞的增殖,诱导骨溶解;淫羊藿苷能抑制磨损微粒诱导的破骨细胞形成,从而抑制骨溶解。中国组织工程研究杂志出版内容重点：人工关节;骨植入物;脊柱;骨折;内固定;数字化骨科;组织工程全文链接：     

人工关节无菌性松动界膜中的溶骨因子

背景:研究证实人工关节无菌性松动是一个溶骨的过程,溶骨因子在其中具有重要作用,但目前只有肿瘤坏死因子、前列腺素E2的相关报道,对于其他溶骨因子在这一过程中的作用尚未见相关研究报道。目的:检测人工关节无菌性松动患者界膜组织中溶骨因子的表达。方法:将2006-03/2008-12武警医学院附属医院及协作医院收治的发生人工关节无菌性松动并行关节翻修的54例患者设为观察组,翻修术中收集界膜。同时收集武警医学院内固定取出术患者56例设为对照组,内固定取出术中收集内固定物与骨之间组织。两组标本均以免疫组化方法检测白细胞介素1、白细胞介素6、白细胞介素11、单核细胞集落刺激因子等促进溶骨细胞因子的染色指数,同时以反转录-聚合酶链反应方法检测细胞因子mRNA表达情况。结果与结论:白细胞介素1、白细胞介素6、白细胞介素11、单核细胞集落刺激因子在人工关节无菌性松动患者界膜组织中的表达水平明显高于对照组(P0.05)。提示白细胞介素1、白细胞介素6、白细胞介素11、单核细胞集落刺激因子等溶骨因子在人工关节无菌性松动病理过程中发挥了重要作用,如能阻断其病理过程将有效减缓和预防人工关节无菌性松动的发生。     

骨替代材料引导骨组织再生过程中白细胞介素4对破骨细胞分化的调控

背景：研究显示,一定剂量的白细胞介素4干预可以得到适宜比例的M1/M2巨噬细胞谱,产生利于骨愈合的微环境,促进骨再生。目的：探讨在骨替代材料引导骨组织再生的过程中,白细胞介素4对NLRP3炎性小体及破骨细胞分化的影响。方法：选取6-8周龄雄性SD大鼠48只,在左侧颅骨制备直径5 mm的骨缺损并同期植入Bio-Oss骨替代材料,缝合骨膜。术后第3天,采用随机数字表法分为实验组与对照组,每组24只,实验组、对照组分别于骨缺损区局部注射白细胞介素4或PBS,1次/d,连续注射5 d。注射后1,2周,取颅骨样本,免疫组织化学染色检测裂解caspase-1和白细胞介素1β蛋白的表达,免疫荧光染色观察M1表面标志物(诱导型一氧化氮合酶)、NLRP3指标(裂解caspase-1)的表达,RT-qPCR检测caspase-1、白细胞介素1β及组织蛋白酶K基因的表达,抗酒石酸酸性磷酸酶染色观察破骨细胞分化及数量。注射后6,12周,取颅骨样本,行Micro-CT检测及苏木精-伊红染色。结果与结论：(1)免疫荧光染色显示,实验组注射后1,2周的诱导型一氧化氮合酶、裂解caspase-1双染细胞数量显著少于对...     

内质网应激介导成骨细胞凋亡在骨溶解骨组织中的作用及机制

背景：磨损微粒能够在体外诱导成骨细胞凋亡,但是发生骨溶解的骨组织中是否也存在成骨细胞的凋亡以及骨组织中的成骨细胞凋亡信号通过何种途径进行传导目前尚不清楚。 目的：分析内质网应激反应在骨溶解骨组织中成骨细胞凋亡和骨溶解发生发展中的作用。 方法：制备磨损微粒诱导骨溶解动物模型。实验分为4组：空白对照组只接受PBS的刺激;磨损微粒组只接受纳米合金粉末悬液的刺激;内质网应激阳性对照组接受纳米合金粉末+毒胡萝卜素的刺激;内质网应激抑制组接受纳米合金粉末悬液及造模后当时、造模后1,2,3和5 d分别腹腔注射4-苯基丁酸。通过甲苯胺蓝染色、苏木精-伊红染色和碱性磷酸酶染色观察骨溶解的病理变化;分析骨溶解颅骨组织中成骨细胞分化成熟情况;Western Blot方法检测骨溶解颅骨组织内内质网应激反应标志蛋白的表达变化;TUNEL和Caspase-3免疫组织化学方法检测骨溶解颅骨组织内成骨细胞的凋亡情况。 结果与结论：磨损微粒能够在体外诱导小鼠颅骨骨溶解的发生、加重炎症细胞的浸润以及抑制成骨细胞分化成熟,同时磨损微粒还可以上调成骨细胞内质网应激反应标志蛋白以及促进骨溶解骨组织中成骨细胞的凋亡。经内质网应激抑制剂(4-苯基丁酸)的治疗后,骨溶解症状明显缓解,骨侵蚀和炎症浸润显著降低,成骨细胞的分化成熟得到改善,凋亡的成骨细胞急剧减少,内质网应激标志蛋白的表达逐渐减弱。表明内质网应激反应参与骨溶解的形成并在骨溶解的发生发展中发挥重要作用。提示内质网应激可作为一种新的治疗靶点,为临床逆转或治疗骨溶解和无菌性松动提供新的思路和方法。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程全文链接：     

磨损颗粒与人工关节无菌性松动

人工关节无菌性松动是影响人工关节长期使用和妨碍人工关节发展的最重要的并发症。许多研究表明,人工关节无菌性松动与假体磨损有着密切关系。磨损颗粒的产生及其所诱发的一系列生物反应是导致假体周围骨溶解及假体无菌性松动的重要因素,其中巨噬细胞、成纤维细胞释放的骨吸收介质和成骨细胞的功能抑制可能是假体无菌性松动发生的重要机制。本文就近年来有关磨损颗粒与人工关节无菌性松动机制的一些研究进展进行综述     

人工关节无菌性松动的早期诊断

背景：人工关节无菌性松动是影响关节假体寿命最主要的因素。 目的：旨在探讨人工关节无菌性松动的早期诊断方法,以利于开展积极的早期治疗。 方法：由第一作者应用计算机检索PubMed、中国期刊全文数据库(CNKI)、维普数据库和万方数据库 1997-05/2010-10相关文献。在标题、摘要、关键词中以“aseptic loosening,artificial joint,prosthesis,osteolysis,early diagnosis”或“无菌性松动,人工关节,假体,骨溶解,早期诊断”为检索词进行检索。选择文章内容与假体无菌性松动有关者,同一领域文献则选择近期发表在权威杂志文章。初检得到182篇文献,根据纳入标准选择关于假体无菌性松动分析及处理的39篇文献进行综述。 结果与结论：假体无菌性松动和骨溶解与骨代谢异常和炎症反应之间的关系密切,所以骨代谢相关指标和磨损颗粒诱导炎症反应的相关细胞因子都可用于早期诊断;核素示踪剂不仅能显示组织器官形态,而且通过显示器官或组织的生理与生化过程来反映组织器官的功能,可早于影像学数周甚至数月发现病变。     

他汀类药物用于人工关节无菌性松动的防治

背景：人工关节无菌性松动是影响关节假体寿命最主要的因素。他汀类药物被发现具有稳定人工关节无菌性松动的新作用。 目的：从药理机制、临床实验、不良反应的最新研究进展3方面进行综述,为药物预防人工关节无菌性松动提供参考。 方法：由第一作者应用计算机检索PubMed和中国期刊全文数据库(CNKI)2007至2011年相关文献。在标题摘要关键词中以“statins,artificial joint,aseptic loosening,prosthesis,osteolysis”或“他汀类药物;人工关节;无菌性松动;假体;骨溶解”为检索词进行检索。选择文章内容与他汀类药物预防人工关节无菌性松动有关者,同一领域文献则选择近期发表在权威杂志文章。初检得到167篇文献,根据纳入标准选择筛选出33篇文献进行综述。 结果与结论：他汀类具有促进假体周围新骨形成、增强的三点弯曲力量进而稳定人工关节周围骨结构的新作用,同时也有实验表明高剂量的辛伐他汀可能对多发性骨髓瘤患者有害、在一定浓度可能引起炎症反应等。作为预防人工关节无菌性松动的新药物,还需要进行进一步的研究以确定最佳的治疗阈值、给药方式和对人类骨再生的有效性。     

磨损颗粒引起人工关节无菌性松动的研究与进展

背景：研究显示磨损颗粒诱导的假体周围骨溶解是导致人工假体无菌性松动的最主要原因。 目的：对磨损颗粒引起人工关节假体无菌性松动在分子生物学方面的研究现状及新进展作一综述。 方法：应用计算机检索CNKI和Pubmed数据库中1991年1月至2012年3月关于人工关节磨损颗粒的文章,在标题和摘要中以“关节置换,磨损颗粒,假体松动”或“arthroplasty,wear particles,loosening of the prosthesis”为检索词进行检索。最终选择30篇文献进行综述。 结果与结论：假体无菌性松动是人工关节置换后手术失败的最主要原因,在此过程中磨损颗粒起到关键性作用。通过对细胞因子及信号传导途径研究的综述,为预防及药物治疗假体无菌性松动提供理论依据。然而,假体的无菌性松动受多种因素的影响,所以预防应从整体着眼,全面深入研究,为假体的松动防治提供方向。     

Protection Against Titanium Particle-Induced Inflammatory Osteolysis by the Proteasome Inhibitor Bortezomib In Vivo

Wear particle-induced vascularized granulomatous inflammation and subsequent inflammatory osteolysis is the most common cause of aseptic loosening after total joint replacement (TJR); however, the precise mechanism by which this occurs is unclear. This study investigates the effects of the proteasome inhibitor bortezomib (Bzb) on the expression of key biochemical markers of bone metabolism and vascularised granulomatous tissues, such as receptor activator of nuclear factor-κB ligand (RANKL), osteoprotegerin (OPG), vascular endothelial growth factor (VEGF) and tumor necrosis factor receptor-associated factor 6 (TRAF6). In addition, the effect of Bzb on apoptosis of CD68+ cells was examined. A total of 32 female BALB/C mice were randomly divided into four groups. After implantation of calvaria bone from syngeneic littermates, titanium (Ti) particles were injected into established air pouches for all mice (excluding negative controls) to provoke inflammatory osteolysis. Subsequently, Bzb was administered at a ratio of 0, 0.1, or 0.5 mg/kg on day 1, 4, 8, and 11 post-surgery to alleviate this response. All of the air pouches were harvested 14 days after the surgical procedure and were processed for molecular and histological analysis. The results demonstrated that Ti injection elevated the expression of RANKL, OPG, VEGF, and TRAF6 at both the gene and protein levels, increased counts of infiltrated cells and thickness of air pouch membranes, and elevated the apoptosis index (AI) of CD68+ cells. Bzb treatment significantly improved Ti particle-induced implanted bone osteolysis, attenuated vascularised granulomatous tissues and elevated AI of CD68+ cells. Therefore, the proteasome pathway may represent an effective therapeutic target for the prevention and treatment of aseptic loosening.     

Distinct gene expression of receptor activator of nuclear factor-kappaB and rank ligand in the inflammatory response to variant morphologies of UHMWPE particles

Recent studies have examined the role of wear debris-induced bone resorption in the aseptic loosening of orthopedic prostheses. Research has shown that inflammation depends not only on the amount of particulate debris, but also the shape and size of the accumulated wear particles. Our previous studies have demonstrated that variant shapes of ultra-high molecular weight polyethylene (UHMWPE) particles induce diverse cellular and apoptotic responses in a murine inflammation model. Since enhanced osteoclastogenesis is recognized as a hallmark of bone loss in prosthetic loosening, we have now investigated the gene expression of receptor activator of nuclear factor-kappaB (RANK) and receptor activator of nuclear factor-kappaB ligand (RANKL) during the inflammatory response to different shapes of UHMWPE particles. Two shapes of UHMWPE particles (globular or elongated) were implanted in established air pouches on BALB/c mice, and pouches harvested 7 days after stimulation with UHMWPE particles. Gene levels of RANK, RANKL, TNFalpha, IL-1beta, and cathepsin K (CK) were quantified by real time RT-PCR, and TRAP staining of pouch membrane was used to evaluate osteoclastogenesis. We found that (i) elongated particles generated significantly higher RANK and RANKL gene expression than globular particles in pouch tissue; (ii) elongated particles provoked significantly higher IL-1beta and TNFalpha gene expression; (iii) a positive association was found between tissue inflammation status and the gene level of RANK/RANKL; and (iv) elongated particles stimulated significantly higher CK gene expression in comparison with globular particles. Histology revealed that clusters of TRAP+ cells were located in regions in contact with elongated particles. Overall, these data suggest that the morphology of wear debris may be a critical factor in the pathogenesis of prosthetic loosening.     

Inhibitory effects of erythromycin on wear debris-induced VEGF/Flt-1 gene production and osteolysis

Objectives  A highly vascularized and inflammatory periprosthetic tissue augments the progress of aseptic loosening, a major clinical problem after total joint replacement. The purpose of this study is to investigate the effect of erythromycin (EM) on ultra high molecular weight polyethylene (UHMWPE) particle-induced VEGF/VEGF receptor 1 (Flt-1) gene production and inflammatory osteolysis in a mouse model. Methods  UHMWPE particles were introduced into established air pouches on BALB/c mice, followed by implantation of calvaria bone from syngeneic littermates. EM treatment started 2 weeks after bone implantation (5 mg/kg day, i.p. injection). Mice without drug treatment as well as mice injected with saline alone were included. Pouch tissues were harvested 2 weeks after bone implantation. Expression of VEGF, Flt-1, RANKL, IL-1, TNF and CD68 was measured by immunostain and RT-PCR, and implanted bone resorption was analyzed by micro-CT (μCT). Results  Exposure to UHMWPE induced pouch tissue inflammation, increase of VEGF/Flt-1 proteins, and increased bone resorption. EM treatment significantly improved UHMWPE particle-induced tissue inflammation, reduced VEGF/Flt-1 protein expression, and diminished the number of TRAP⁺ cells, as well as the implanted bone resorption. Conclusion  This study demonstrated that EM inhibited VEGF and Flt-1 gene expression. The molecular mechanism of EM action on VEGF/Flt-1 signaling-mediated osteoclastogenesis warrants further investigation.     

OPG／RANKL在大鼠骨关节炎模型中的表达及意义

目的：检测骨保护素（OPG）和核因子Kβ受体活化因子配体（RANKL）在大鼠骨关节炎（OA）模型软骨下骨中的表达,探讨其与OA软骨下骨病变的关系及意义。方法：36只大鼠分两组。模型组行右膝前交叉韧带＋内侧副韧带横断术造模,对照组仅切开关节囊。术后2,6,8周取两组膝关节标本,观察外观、Mankin评分和骨形态计量参数变化,并检测软骨下骨中OPG和RANKL含量及其比值。结果：模型组软骨Mankin评分随时间逐渐增加（P〈0．01）;骨形态计量学显示造模后2周骨量降低,6、8周增加（P〈0．05）;模型组OPG含量由低变高,RANKL含量则由高变低,其比值逐渐增加（P〈0．01）。结论：OPG、RANKL在OA软骨下骨异常重建中起着重要作用。由此推测,在OA早期对其调控可能抑制异常骨重建,将是治疗OA的一种潜在有效方法。     

Protective effects of COX-2 inhibitor on titanium-particle-induced inflammatory osteolysis via the down-regulation of RANK/RANKL

Particle-wear-induced inflammatory osteolysis remains a major problem for the long-term success of total joint arthroplasty. Previous studies have demonstrated that cyclooxygenase-2 (COX-2) is expressed abundantly in the tissue around a failed implant. However, the role of COX-2 in the development of particle-wear-induced osteoclastogenesis remains unclear. The aim of the study was to test the hypothesis that Dynastat, a COX-2 inhibitor, ameliorates particle-wear-induced inflammatory osteoclastogenesis through the down-regulation of the receptor activators of nuclear factor-κB (RANK) and nuclear factor-κB ligand (RANKL) expression in a murine osteolysis model. Titanium (Ti) particles were introduced into established air pouches in BALB/c mice, followed by the implantation of calvaria bone from syngeneic littermates. Dynastat was given to mice intraperitoneally 2 days before the introduction of Ti particles and maintained until the mice were sacrificed. Pouch tissues were collected 14 days after Ti inoculation for molecular and histological analysis. The results showed that Dynastat has more impact on Ti-particle-induced prostaglandin E(2) expression and less on the expression of interleukin-1β and tumor necrosis factor-α. Dynastat inhibited Ti-particle-induced osteoclastogenesis by reducing the gene activation of RANK and RANKL, and diminishing the RANKL expression in Ti-particle-charged pouches. Dynastat markedly reduced the number of tartrate-resistant acid-phosphatase-positive cells in pouch tissues stimulated by Ti particles. In conclusion, this study provides evidence that Dynastat can markedly inhibit Ti-particle-induced osteoclastogenesis by the down-regulation of RANK/RANKL in a murine air pouch model, and is a promising therapeutic candidate for the treatment of inflammatory osteolysis induced by wear particles.     

Inhibition of p38 Mitogen-Activated Protein Kinase Down-regulates the Inflammatory Osteolysis Response to Titanium Particles in a Murine Osteolysis Model

The p38 mitogen-activated protein kinase (p38 MAPK) pathway is involved in the osteoclast differentiation. The aim of the study was to investigate whether SB203580, a p38 MAPK inhibitor, inhibits wear-debris-induced inflammatory osteolysis in mice. Forty-five mice were implanted with calvaria bone from syngeneic littermates; then, titanium (Ti) particles were injected into established air pouches to provoke inflammatory osteolysis. At 14?days after bone/Ti implantation, pouch membranes with intact bone implants underwent histological and molecular analysis. SB203580 had less effect on MMP-9 and TNF-?? expression under wear-debris-induced conditions. SB203580, by inhibiting the expression of p38 MAPK and phospho-p38 MAPK, inhibited Ti particle wear-debris-induced inflammatory osteolysis. It also remarkably decreased the number of tartrate-resistant acid phosphatase positive cells in Ti-particle-induced pouch tissues. Results suggest that p38 MAPK may be critical in a murine osteolysis model. SB203580 may notably inhibit wear-debris-induced inflammatory osteolysis by down-regulating expression of MMP-9 and TNF-?? via the p38 MAPK pathway.     

Therapeutic Potential of the Proteasome Inhibitor Bortezomib on Titanium Particle-Induced Inflammation in a Murine Model

Wear particle-induced aseptic loosening has been recognized as a harmful inflammatory process that jeopardizes the longevity of total joint replacement. The proteasome controls the activation of NF-κB and subsequent inflammatory mediators, such as TNF-α and IL-1β; thus, we investigated whether proteasome inhibition can ameliorate wear particle-induced inflammation in a murine model. A total of 48 BALB/C mice were divided into four groups. Titanium (Ti) particles were injected into the established air pouches of all mice (except negative controls) to provoke inflammation, and then 0.1 or 0.5 mg/kg of Bortezomib (Bzb, a proteasome inhibitor) was administered to ameliorate the inflammation response, while air pouches without Bzb administration were used as loading controls. The air pouches were harvested 2 or 7 days after Bzb injection for molecular and histological analyses. Inflammation responses in the air pouch tissues of Bzb treatment groups are lower than those in the Ti-stimulated group, and this occurs in a dose-dependent manner. Bzb can significantly attenuate the severity of Ti-induced inflammation in air pouches.     

Inhibitory effects of luteolin on titanium particle-induced osteolysis in a mouse model

Wear particles liberated from the surfaces of an implanted prosthesis are associated with peri-implant osteolysis and subsequent aseptic loosening. In the latter wear particle-induced inflammation and osteoclastogenesis have been identified as critical factors, and their inhibition as important steps in the treatment of affected patients, such as those undergoing total hip replacement. In this study the ability of luteolin to inhibit both titanium (Ti) particle-induced osteoclastogenesis in vitro and osteolysis in a murine calvaria Ti particle-induced model of osteolysis was examined. The results showed that luteolin, a highly potent and efficient inhibitor of tumor necrosis factor α (TNF-α) and interleukin-6 expression, inhibited Ti particle-induced inflammatory cytokine release, osteoclastogenesis, and bone resorption in bone marrow macrophages. Microcomputed tomography and histological analyses showed that the Ti particles caused significant bone resorption and increased TRAP(+) multinuclear osteoclasts in the murine calvarial model of osteolysis, whereas this was not the case in the luteolin treatment group, in which osteolytic suppression was accompanied by a decrease in both TNF-α production and serum levels of the osteoclast marker the C-terminal telopeptide fragment of type I collagen. These results support the use of luteolin as a natural compound in the prevention and treatment of aseptic loosening after total replacement arthroplasty.