唑来膦酸钠在抑制钛颗粒诱导骨溶解中作用的体外实验研究

目的：研究唑来膦酸钠（ ZOL）对钛颗粒诱导的骨溶解的影响。方法分离6～8周C57BL/6J小鼠长骨中的前体破骨细胞（ OCP）并分为6组,A组：OCP＋细胞培养液,B组：OCP＋巨噬细胞集落刺激因子（M-CSF）＋NF-κB 受体活化因子配体（RANKL）＋细胞培养液,C组：OCP＋钛颗粒＋细胞培养液,D组：OCP＋上清液（钛颗粒刺激巨噬细胞24 h后上清液）＋细胞培养液,E组：OCP＋M-CSF＋RANKL＋ZOL＋细胞培养液,F组：OCP＋上清液＋ZOL＋细胞培养液。每组细胞分别接种在玻璃盖玻片、皮质骨磨片和含骨检测表面的96孔板上,10 d后检测玻璃盖玻片上细胞抗酒石酸磷酸酶（ TRAP）的表达及皮质骨磨片上骨吸收陷窝的形成,并以骨检测表面的骨吸收面积为指标比较各组破骨细胞的骨吸收活性。结果 B组、D组、E组和F组的OCP均能分化为能被TRAP染色成阳性的破骨细胞并形成骨吸收陷窝,其余组均未发现TRAP染色阳性的破骨细胞和骨陷窝。加入ZOL的F组骨吸收面积（5.54％±1.25％）较D组（10.34％±1.69％）明显减少,差异具有统计学意义（t＝5.61,P＜0.01）。结论在体外实验中钛颗粒并不能直接刺激前体破骨细胞向破骨细胞转化;唑来膦酸钠可以抑制钛颗粒诱导的骨溶解作用。     

阿仑膦酸钠对假体周围骨溶解中OPG及RANKL的影响

目的 探讨阿仑膦酸钠对聚乙烯颗粒诱导假体周围骨溶解中OPG及RANKL影响.方法 将钛金属棒植入新西兰大白兔左侧股骨内,每2周向左膝关节内注射聚乙烯颗粒,使用随机数字表随机分成实验组和对照组,各6只.实验组给予阿仑膦酸钠灌胃,而对照组给予等量的0.9％的氯化钠溶液灌胃.12周后处死动物,取出假体周围界膜组织,用ELIS...     

唑来膦酸钠应用近期安全性观察

目的 了解唑来膦酸钠用药后的近期副反应。方法 自2009年11月至2012年2月对156例骨质疏松症患者应用唑来膦酸钠治疗,观察患者自用药1个月以来的临床表现,重点包括疼痛、发热及其他不良事件。同期观察因脊柱骨折行椎体成形术而未应用唑来膦酸钠治疗的患者107例,比较两组相应事件发生的差异。结果 本组患者全部得到随访,结果显示唑来膦酸钠组疼痛发生率17.3%,发热发生率12.8%,对照组疼痛发生率1.87%,发热发生率0.93%,两组差异有显著性(P < 0.001)。另外唑来膦酸钠组出现1例皮肤过敏,1例心肌缺血、心绞痛。1例多发性骨髓瘤病情加重。1例出现肺炎呼吸衰竭死亡。结论 唑来膦酸钠在骨质疏松症的应用中,有疼痛加重,发热等副反应,甚至可能有致皮肤过敏,心绞痛、多发性骨髓瘤病情加重、肺炎呼吸衰竭等不良事件,建议在用药当天密切监护下使用,并加强用药后2w内的随访。     

不同剂量阿仑膦酸钠对磨损颗粒诱导假体周围骨溶解的影响

目的观察不同剂量阿仑膦酸钠对磨损颗粒诱导假体周围骨溶解的影响,确定其合适剂量.方法雄性SD大鼠24只,经膝关节将钛合金假体及混合磨损颗粒植入胫骨近端（双侧）,随机分为对照组和实验Ⅰ、Ⅱ、Ⅲ组,每组6只,术后对照组每日空腹生理盐水2ml灌胃;实验Ⅰ、Ⅱ、Ⅲ组每日空腹阿仑膦酸钠灌胃,剂量分别为0．01、0．1、1mg／（kg·d）,持续6周.术后12周处死取材,进行组织学观察及组织形态计量学测定.结果对照组和实验Ⅰ组假体柄周围纤维界膜厚,细胞成分多,与纤维界膜连接处新生骨边缘呈虫蚀状,新生骨对假体的支撑作用较差;实验Ⅱ、Ⅲ组假体柄周围纤维界膜较薄,新生骨与假体间可见有直接接触,对假体有良好的支撑作用.假体周围界膜厚度及面积的形态计量学检测并经统计学分析显示：对照组与实验Ⅰ组、实验Ⅱ与Ⅲ组间差异无显著性（P〉0．05）,对照组和实验Ⅰ组分别与实验Ⅱ组和实验Ⅲ组间差异有显著性（P〈0．05）.结论以阿仑膦酸钠预防磨损颗粒诱导的假体周围骨溶解时,最适合剂量为0．1mg／（kg·d）.     

唑来膦酸、伊班膦酸钠及阿伦膦酸钠防治绝经后骨质疏松症的疗效对比研究

目的对比研究唑来膦酸、伊班膦酸钠及阿伦膦酸钠对绝经后骨质疏松症的疗效。方法 180名绝经后妇女随机分为唑来膦酸治疗组(ZOL组)、伊班膦酸钠治疗组(IBA组)和阿伦膦酸钠治疗组(ALN组);ZOL组给予唑来膦酸治疗,IBA组给予伊班膦酸钠治疗,ALN组予以阿伦膦酸钠治疗。治疗前后分别检测3组受试者腰椎及髋部骨密度、血清骨代谢指标、视觉模拟评分(visual analogue scale,VAS)改变及研究期间药物不良反应和骨折发生率。结果药物治疗12个月后3组腰椎(L1~4)及左侧股骨颈骨密度明显增加,显著高于治疗前(P0.05),而3组间比较差异无统计学意义(P0.05),3组治疗有效率比较差异无统计学意义(P0.05)。药物治疗12个月后3组患者的VAS评分均显著降低,显著低于治疗前(P0.05),而3组间比较差异无统计学意义(P0.05)。干预12个月后两组血清I型胶原交联羧基末端肽和抗酒石酸酸性磷酸酶-5b水平均显著降低,显著低于治疗前(P0.05),而3组间比较差异无统计学意义(P0.05)。3组间药物不良反应发生率比较差异无统计学意义(P0.05)。结论唑来膦酸、伊班膦酸钠及阿伦膦酸钠对绝经后骨质疏松症的治疗安全有效,可以显著改善骨密度及骨代谢异常。     

阿仑膦酸钠抑制破骨细胞体外吸收的研究

本研究利用体外分离培养的兔破骨细胞,观察第三代双膦酸盐阿仑膦酸钠（ａｌｅｎｄｒｏｎａｔｅ）对骨吸收的抑制作用。将ａｌｅｎｄｒｏｎａｔｅ加入培养液中使其最终浓度为０Ｍ,１μＭ,１０μＭ,１００μＭ。同时观察两组用ａｌｅｎｄｒｏｎａｔｅ盐溶液１００μＭ,５０μＭ浸泡的骨片对破骨细胞吸收功能的影响。用倒置光相差显微镜在不同时间点观察计数骨吸收陷窝并拍照。结果随着培养液内ａｌｅｎｄｒｏｎａｔｅ浓度增高,骨吸收陷窝数减少,面积亦减少。而用ａｌｅｎｄｒｏｎａｔｅ浸泡过的骨片上未见骨吸收陷窝。说明ａｌｅｎｄｒｏｎａｔｅ具有抑制破骨细胞体外骨吸收的作用。     

唑来膦酸与伊班膦酸对破骨细胞抑制作用比较

目的:探讨唑来膦酸和伊班膦酸抑制破骨细胞分化及骨吸收功能特征。方法:诱导C57小鼠骨髓单核细胞向破骨细胞分化。依据双膦酸盐种类不同分为3组,对照组采用α-MEM培养基常规孵育,加入磷酸盐缓冲液;唑来膦酸组培养基中分别加入浓度为1×10 -8、1×10 -7、1×10 -6、1×10 ...     

伊班膦酸钠对钛颗粒诱导的骨溶解作用

[目的]通过建立钛颗粒诱导的小鼠颅骨骨溶解动物模型,研究伊班膦酸钠对骨溶解的影响,并探讨其作用机制.[方法]24只昆明系小鼠随机分成4组,每组6只,建立钛颗粒诱导的小鼠颅骨骨溶解模型.空白对照组进行假手术;钛颗粒组在小鼠颅顶植入30 mg钛颗粒;1单位伊班膦酸钠组和10单位伊班膦酸钠组分别于植入钛颗粒第2 d腹腔注射伊班膦酸钠10 mg、100 mg.手术后第10 d,取出颅骨进行病理学分析.[结果]空白对照组骨溶解面积为(0.070±0.009)mm2,没有发生明显骨溶解.钛颗粒组骨溶解面积为(0.369±0.050)mm2,和空白对照组相比发生了更大面积的骨溶解(P<0.05).1单位伊班膦酸钠组和10单位伊班膦酸钠组骨溶解面积分别为(0.164±0.018)mm2、(0.018±0.075)mm2,均小于钛颗粒组骨溶解面积(P<0.05).而且10单位伊班膦酸钠组比1单位伊班膦酸钠组骨溶解面积更小(P<0.05).骨溶解区域破骨细胞数量各组比较结果和骨溶解面积结果类似.[结论]伊班膦酸钠能够有效抑制钛颗粒诱导的小鼠颅骨骨溶解.     

阿仑膦酸钠对人工关节假体周围骨长入的影响

目的观察阿仑膦酸钠对人工关节假体周围骨长入的影响。方法SD雄性大鼠16只,双侧胫骨上端经膝关节植入定制钛合金假体,随机分成对照组和实验组,对照组术后每日空腹生理盐水灌胃;实验组术后每日空腹阿仑膦酸钠灌胃,剂量0.1 mg/(kg.d),持续6周。术后12周处死取材,进行组织学观察及组织形态计量学测定。结果组织学观察发现,对照组假体周围由新生骨、类骨质和纤维界膜构成。纤维界膜较厚,与新生骨或类骨质间界限清晰。实验组假体周围纤维界膜薄且稀少,新生骨与假体界面多为直接接触,有些部位新生骨与假体界面完全整合。组织形态计量学测定发现,对照组假体周围界膜的厚度和面积均明显大于实验组,差异有显著性(P<0.05)。结论阿仑膦酸钠经胃肠给药可能对钛合金人工关节假体周围骨长入有一定的促进作用。     

局部应用唑来膦酸预防股骨头坏死塌陷

[目的]建立大鼠股骨头骨坏死动物模型,观察以聚乳酸-羟基乙酸共聚物(polylactic-co-glycolic acid,PLGA)为载体,局部应用唑来膦酸(zoledronate acid,ZOL)防止大鼠股骨头骨坏死塌陷的效果。[方法]选用PLGA作为局部应用ZOL的载体,制备含有30μg的ZOL的PLGA棒。取SD雄性大鼠30只,随机分为三组,建立创伤性股骨头骨坏死模型。股骨头钻孔,分别植入含有ZOL的PLGA棒(PLGA-ZOL组),以不含ZOL的PLGA棒(PLGA组)和空置(单纯打孔组)为对照组。6周后将大鼠处死取材,进行X线检查计量股骨头高度比,进行MicroCT扫描,分别计算比较各组的骨矿密度(bone mineral density,BMD)、骨体积分数(bone volume fraction,BVF)、骨小梁厚度(trabecular plate thickness,Tb.Th)和骨小梁间隙(trabecular spacing,Tb.Sp)。扫描后进行HE和Masson三色染色了解各组间组织学改变情况。[结果]与打孔组和PLGA组相比,PLGA-ZOL组中BMD(535.95±15.08,P<0.05),BVF(0.77±0.04,P<0.05)和Tb.Th(0.64±0.05,P<0.05)显著增加,而Tb.sp(0.37±0.06,P<0.05)显著降低,而打孔组和PLGA组并无统计学差异。而且PLGA-ZOL组股骨头有更好的形态,骨小梁保留更完整,新生骨更多。[结论]使用ZOL能够抑制局部骨吸收活动,促进局部骨生成,预防股骨头塌陷。     

Role of polyethylene particles in peri-prosthetic osteolysis: A review

There is convincing evidence that particles produced by the wear of joint prostheses are causal in the peri-prosthetic loss of bone, or osteolysis, which, if it progresses, leads to the phenomenon of aseptic loosening. It is important to fully understand the biology of this bone loss because it threatens prosthesis survival, and loosened implants can result in peri-prosthetic fracture, which is disastrous for the patient and presents a difficult surgical scenario. The focus of this review is the bioactivity of polyethylene (PE) particles, since there is evidence that these are major players in the development and progression of osteolysis around prostheses which use PE as the bearing surface. The review describes the biological consequences of interaction of PE particles with macrophages, osteoclasts and cells of the osteoblast lineage, including osteocytes. It explores the possible cellular mechanisms of action of PE and seeks to use the findings to date to propose potential non-surgical treatments for osteolysis. In particular, a non-surgical approach is likely to be applicable to implants containing newer, highly cross-linked PEs (HXLPEs), for which osteolysis seems to occur with much reduced PE wear compared with conventional PEs. The caveat here is that we know little as yet about the bioactivity of HXLPE particles and addressing this constitutes our next challenge.     

Micro-CT评价唑来膦酸对磨损颗粒诱导假体周围骨溶解模型的抑制作用

目的 :研究唑来膦酸对磨损颗粒诱导假体周围骨溶解大鼠模型的骨微结构的影响。方法 :选取30只成年雄性SPF级SD大鼠(重250~300 g),随机分成假手术组、模型组和唑来膦酸组,每组10只。通过往大鼠右侧股骨置入钛钉和聚乙烯颗粒进行造模,术后3 d假手术组、模型组皮下注射0.9%生理盐水2 ml/kg,唑来膦酸组皮下注射唑来磷酸0.1 mg/kg,每周1次,共8周。8周后取各组大鼠右侧股骨标本采用Micro-CT对大鼠股骨松质骨微结构进行扫描,应用三维重建处理和分析软件进行处理得到图像及BMD、BV/TV、Tb.N、Tb.Th、SMI、BS/BV、Tb.Sp、Tb.Pf等参数进行分析。结果:Micro-CT三维成像显示,模型组大鼠股骨骨密度较假手术组明显下降,骨微结构破坏严重,骨小梁稀疏变细,连续性降低;唑来膦酸组与模型组比较,骨微结构又出现明显改善。对大鼠股骨微结构的Micro-CT参数进行分析,模型组大鼠股骨BMD(0.081±0.020)明显低于假手术组(0.160±0.018)及唑来膦酸组(0.125±0.012),差异有统计学意义;模型组大鼠股骨BV/TV(10.563±1.070)低于假手术组(27.935±1.834)及唑来膦酸组(14.559±1.258),差异有统计学意义;模型组大鼠股骨Tb.N(1.005±0.165)低于假手术组(2.058±0.108)及唑来膦酸组(1.515±0.126),差异均有统计学意义;模型组大鼠股骨Tb.Th(0.075±0.016)明显低于假手术组(0.158±0.016)及唑来膦酸组(0.124±0.011),差异均有统计学意义。同时,模型组大鼠股骨SMI(1.817±0.127)明显高于假手术组(1.104±0.120)及唑来膦酸组(1.547±0.122),差异均有统计学意义;模型组大鼠股骨BS/BV(35.784±1.650)明显高于假手术组(21.506±2.771)及唑来膦酸组(30.399±2.730);模型组大鼠股骨Tb.Sp(0.735±0.107)高于假手术组(0.423±0.057)及唑来膦酸组(0.577±0.082),差异均有统计学意义;模型组大鼠股骨Tb.Pf(9.088±1.283)明显高于假手术组(2.447±0.703)及唑来膦酸组(5.862±1.042),差异有统计学意义。结论:唑来膦酸可以通过改变大鼠骨微结构达到抑制磨损颗粒诱发的骨溶解的作用,为使用唑类酸作为一种治疗性干预手段来防止假骨溶解提供了科学依据。     

The effectiveness of polyethylene versus titanium particles in inducing osteolysis in vivo.

Bearing surface wear and periprosthetic osteolysis due to wear particles are among the most common reasons for joint replacement failure. A murine calvarial model of wear particle-induced osteolysis has been used to identify different biologic factors associated with this problem and to test nonsurgical methods of modulating the host response to particulate debris. This model has utilized titanium particles, however, in clinical practice the most common source of particulate debris is polyethylene particles from bearing surface wear. We now report a calvarial model of wear particle-induced osteolysis based on commercially available polyethylene particles. We found that compared to sham surgery osteoclast recruitment and bone resorption can be induced by introduction of the titanium particles or polyethylene particles. However, bone resorption was significantly higher with polyethylene particles compared to titanium particles (p=0.02). We consider the polyethylene based murine calvarial model of wear particle-induced osteolysis a reliable and clinically relevant tool to understand the host factors and potential pharmacologic interventions that can influence wear debris generated osteolysis. This model might serve as an extension of the well-established titanium based bone resorption model.     

人工关节置换术后磨损颗粒与假体周围骨溶解的研究进展

人工关节置换术后磨损颗粒的产生是导致假体周围骨溶解的重要影响因素。目前研究认为磨损颗粒导致假体周围骨溶解的主要原因与植入假体的材料以及其产生的磨损颗粒通过各种作用对相关细胞如巨噬细胞、成骨细胞、破骨细胞等的刺激有关,诱导产生多种炎性细胞因子,激活和开放细胞信号和通路,产生长期慢性的炎症,导致假体周围骨溶解。为此,本文主要研究不同类型的磨损颗粒对假体周围骨溶解的影响,以及磨损颗粒在假体周围骨溶解进展过程中的机制,并且由此探讨如何减少磨损颗粒的产生以及阻断其在骨溶解进程的作用,以此来达到预防和治疗假体周围骨溶解的目的。     

Polyethylene and titanium particles induce osteolysis by similar, lymphocyte-independent, mechanisms.

Periprosthetic osteolysis is a major clinical problem that limits the long-term survival of total joint arthroplasties. Osteolysis is induced by implant-derived wear particles, primarily from the polyethylene bearing surfaces. This study examined two hypotheses. First, that similar mechanisms are responsible for osteolysis induced by polyethylene and titanium particles. Second, that lymphocytes do not play a major role in particle-induced osteolysis. To test these hypotheses, we used the murine calvarial model that we have previously used to examine titanium-induced osteolysis. Polyethylene particles rapidly induced osteolysis in the murine calvaria 5-7 days after implantation. The polyethylene-induced osteolysis was associated with large numbers of osteoclasts as well as the formation of a thick periosteal fibrous tissue layer with numerous macrophages containing phagocytosed polyethylene particles. Polyethylene-induced osteolysis was rapidly repaired and was undetectable by day 21 after implantation. Lymphocytes were noted in the fibrous layer of wild-type mice. However, the amount of osteolysis and cytokine production induced by polyethylene particles was not substantially affected by the lack of lymphocytes in Pfp/Rag2 double knock out mice. All of these findings are similar to our observations of osteolysis induced by titanium particles. These results provide strong support for both of our hypotheses: that similar mechanisms are responsible for osteolysis induced by polyethylene and titanium particles and that lymphocytes do not play a major role in particle-induced osteolysis.     

A novel bisphosphonate inhibits inflammatory bone resorption in a rat osteolysis model with continuous infusion of polyethylene particles.

This study examined the inhibitory effect of a new bisphosphonate (TRK-530) on wear debris-mediated bone resorption in a rat osteolysis model involving continuous infusion of high density polyethylene (HDPE) particles. TRK-530 (TRK) is a novel synthetic bisphosphonate that has been shown to decrease the level of tumor necrosis factor alpha (TNF-alpha) in the bone marrow of rats with adjuvant arthritis. Forty Wistar rats were randomized to two groups (n = 20 each). In each rat, a Kirshner (K) wire was inserted into the femur and HDPE particles were continuously infused into the knee joint. Thereafter, the animals were subcutaneously injected with saline (control group) or 1 mg/kg of TRK (TRK group) every second day, and were sacrificed at 4 or 8 weeks after surgery. Radiographs obtained at the time of sacrifice were evaluated for periprosthetic osteolysis. We also examined the thickness of the reactive membrane as well as the number of osteoclast-like cells around the K-wire. In addition, we examined the expression of genes for bone-resorbing cytokines in the reactive membrane. Radiographic peri-implant osteolysis was more frequent in the control group compared with the TRK group at each time of assessment (p < 0.01). The interfacial membrane was significantly thinner in the TRK group compared with the control group (p < 0.01) and the average number of osteoclast-like cells around the K-wire was significantly fewer in the TRK group (p < 0.01). In addition, the expression of interleukin 1-alpha messenger ribonucleic acid (IL-1alpha mRNA) and TNF-alpha mRNA was suppressed in the TRK group at each time of assessment. We conclude that the TRK can inhibit the formation of inflammatory peri-implant osteolysis induced by HDPE particles.     

阿仑膦酸钠局部释放对假体周围骨溶解的抑制作用

目的 通过阿仑膦酸钠(ALN)复合羟基磷灰石(HA)涂层假体,检测局部释放的ALN对骨溶解的抑制作用. 方法 选择18只雌性新西兰大白兔,随机均分为实验组、阳性对照组和空白对照组,三组均于左侧胫骨平台处向胫骨髓腔竖直植入HA涂层假体.在植入前,实验组于HA涂层表面复合ALN 5 mg,且实验组和阳性对照组涂层表面均匀涂抹25μg超高相对分子质量聚乙烯颗粒,空白对照组不做处理.术后12周取左侧胫骨行骨组织形态学、生物力学以及影像学测馈,检测ALN体外释放周期,并评估抗骨溶解效果. 结果 与对照组相比,实验组的骨皮质厚度(+9.26%)、假体生物力学固定强度(极限抗剪强度:+36.42%、表观抗剪刚度:+37.79%、总能量吸收值:+30.00%)、骨-假体接触率(+44.40%)、假体周围骨体积分数(+16.07%)、骨矿化水平(钙盐沉积岛平均而积:+598.86%、钙盐沉积岛面积分数:+650.11%)和假体周围骨密度(+62.05%)均明显增高,差异有统计学意义(P<0.05);反映成骨细胞活性的各项参数(类骨质表面:+33.53%、类骨质厚度:+49.60%、骨形成速率:+91.62%、松质骨骨矿沉积率:+48.04%、皮质骨骨矿沉积率:+60.42%)也有不同程度的增高,差异有统计学意义(P<0.05). 结论 非多孔HA涂层假体复合ALN的局部释放可以明显抑制超高相对分子质量聚乙烯颗粒诱导的假体周围骨溶解,ALN在预防磨屑诱导的假体周围骨溶解过程中能够增强成骨细胞活性.     

SCID小鼠单次唑来磷酸头皮下注射抑制人假体周围骨溶解

[目的]评价唑来磷酸在新型动物模型中对人关节假体周围界膜组织诱导的骨溶解的抑制效应。[方法]将髋关节翻修病人体内获得的界膜组织移植到SCIDbeige小鼠颅骨上。术后2d将唑来磷酸直接注射至小鼠头皮下,并在术后2、4周处死动物取出带移植组织的颅骨进一步检测。[结果]移植组织在动物体内存活至少4周,HE染色可见界膜组织引起大量的骨胶原纤维碎裂溶解。界膜组织中TNF-α、IL-6、RANKL和CPK的基因表达和蛋白水平均显著高于对照。单次局部注射唑来磷酸显著降低RANKL和CPK的表达但未影响TNF-α和IL-6的水平。[结论]单次局部注射唑来磷酸可有效抑制人界膜组织诱导的骨溶解,且效应持续。该新型动物模型模拟临床程度高,可作为评价假体周围骨溶解药物抑制效应的平台。