辛伐他汀对钛颗粒刺激人单核细胞形成破骨细胞的影响

目的研究体外辛伐他汀对钛颗粒刺激单核细胞形成破骨细胞的影响,探讨辛伐他汀防治人工关节无菌性松动的可能性。方法体外分离培养人外周血单个核细胞并分成5组,A组为钛颗粒刺激组（单核细胞和磨屑混合培养）,B组为10^-5mol／L辛伐他汀组（单核细胞、磨屑混合培养＋10^-5mol／L辛伐他汀）,C组为10^-6mol／L辛伐他汀组（单核细胞、磨屑混合培养＋10^-6mol／L辛伐他汀）,D组为10^-7mol／L辛伐他汀组（单核细胞、磨屑混合培养＋10^-7mol／L辛伐他汀）,E组为单核细胞组。各组细胞培养24h后取上清液,用ELISA法检测上清液中肿瘤坏死因子（TNF-α）、单核细胞趋化蛋白-1（MCP-1）的含量。分别培养10d、18d后进行TRAP染色阳性细胞计数,采用扫描电镜检测骨磨片的吸收陷窝,观察钛颗粒对破骨细胞形成的影响。结果磨屑刺激单个核细胞分泌溶骨因子,辛伐他汀抑制磨损颗粒刺激单核／巨噬细胞分泌TNF-α及MCP-1;且破骨细胞数明显减少,骨吸收陷窝数减少,与钛颗粒组刺激组比较,差异均有统计学意义（P〈0．05）。结论辛伐他汀通过抑制TNF-α、MCP-1的释放而有效防止磨屑诱导的骨溶解,有望成为防治人工关节无菌性松动的一种有潜力的药物。     

二磷酸盐对关节磨屑刺激单核细胞分泌IL-1β的抑制作用

目的:研究二磷酸盐对关节磨屑刺激单核细胞分泌IL-β的影响。方法:分离培养人外周血单核细胞,加入关节磨屑及不同浓度的阿伦膦酸钠,ELISA检测细胞上清中IL-1β的含量,原位杂交检测IL-1β mRNA的表达。结果:关节磨屑组IL-1β及基因的表达显著高于对照组(P<0.01),不同浓度的阿伦膦酸钠组IL-1β及基因的表达明显低于关节磨屑组(P<0.01)。结论:阿伦膦酸钠能通过下调人外周血单核细胞IL-1β3及mRNA的表达,对人工关节松动可能起防治作用。     

阿伦膦酸钠对破骨细胞凋亡的影响

目的：本研究采用吖啶橙染色方法观察阿伦膦酸钠对破骨细胞凋亡的影响。方法：将阿伦膦酸钠加入培养液配成终浓度为10^-10mol/L、10^-8mol/L、10^-6mol/L。阿伦膦酸钠作用6h，吖啶橙染色，倒置荧光显微镜下观察凋亡破骨细胞所占的比例。结果：吖啶橙染色能展示凋亡破骨细胞的形成。阿伦膦酸钠促进破骨细胞凋亡，随阿伦膦酸钠浓度的增加，破骨细胞凋亡的比例也增加。结论：吖啶橙染色是检测破骨细胞凋亡简便实用的方法，阿伦膦酸钠通过促进凋亡降低破骨细胞数量，降低骨吸收。     

载阿仑膦酸钠骨水泥抑制钛磨屑诱导的假体周围骨溶解

目的 比较载阿仑膦酸钠丙烯酸骨水泥与皮下注射阿仑膦酸钠抑制钛磨眉诱导的骨溶解的效果.方法 48只成年雄性新西兰兔随机均分为无钛磨屑且无阿仑膦酸钠组(A组),有钛磨屑注射且无阿仑膦酸钠组(B组),钛磨屑分别注射0.1%、0.5%、1.0%载阿仑膦酸钠丙烯酸骨水泥组(C、I)、E组),钛磨屑注射且皮下手射阿仑膦酸钠组(F组),每组8只.将载阿仑膦酸钠骨水泥植入兔股骨远端.制备磨屑诱导骨溶解动物模型.术后8周对股骨行组织形态学分析、骨密度(bone mineral density,BMD)测定及界面力学测试结果 B组假体周围可见明显的骨溶解,而C、D、E、F组骨溶解明显少于B组.B组假体周围BMD和骨-骨水泥界面抗剪强度分别较A组下降17%和56%;D组假体周围BMD和界面抗剪强度较B组分别增加29%和62%;E组假体周围BMD和界画抗剪强度较B组分别增加37%和29%;F组假体周围BMD和界面抗剪强度较B组分别增加51%和69%;C组、D组、E组分别与F组比较,假体周围BMD和界面抗剪强度的差异均无统计学意义.结论 载阿仑瞵酸钠丙烯酸骨水泥与皮下注射阿仑瞵酸钠均可在一定程度上抑制磨屑诱导的骨吸收,增强界画抗剪强度.     

阿仑磷酸钠对成骨细胞增殖、分泌和成骨功能的影响

目的　观察阿仑膦酸钠对人成骨细胞功能的影响 ,探讨使用此药防治人工关节无菌性松动的可行性。方法　采用体外培养成骨细胞的方法 ,观察不同浓度的阿仑膦酸钠 (1× 10 -11、1× 10 -9、1× 10 -7、1× 10 -5mol/L)对成骨细胞的增殖、碱性膦酸酶活性、分泌骨钙素及成骨能力的影响。结果　阿仑膦酸钠浓度达 1× 10 -5mol/L时抑制成骨细胞增殖 ;浓度为 1× 10 -7、1× 10 -5mol/L时增强成骨细胞的碱性膦酸酶活性和骨钙素分泌 ;对其成骨能力 ,浓度在 1× 10 -11、1× 10 -9mol/L时具有刺激作用 ,1× 10 -5mol/L有抑制作用 ,中间浓度 (1× 10 -7mol/L)无影响。结论　阿仑膦酸钠低浓度时有利于成骨细胞的成骨 ;局部使用有可能成为防治人工关节无菌性松动的方法之一。     

阿仑膦酸钠局部持续使用防治人工关节无菌性松动可行性的细胞学研究

目的验证不同浓度的阿仑膦酸钠对人成骨细胞和中性粒细胞功能的影响,从而决定局部持续使用此药防治人工关节无菌性松动的可行性。方法用体外细胞培养的方法,测试不同浓度1×10-11mol/L、1×10-9mol/L、1×10-7mol/L、1×10-6mol/L、1×10-5mol/L的阿仑膦酸钠对人成骨细胞的增殖、分泌和成骨能力的影响,以及对人中性粒细胞的运动、呼吸爆发、吞噬和杀灭细菌能力的影响。结果高浓度1×10-6mol/L、1×10-5mol/L的阿仑膦酸钠虽然有利于中性粒细胞的抗感染作用,但浓度为1×10-5mol/L时可抑制成骨细胞的成骨能力;低浓度1×10-11mol/L、1×10-9mol/L、1×10-7mol/L的阿仑膦酸钠,虽然有利于成骨细胞的成骨能力,但可抑制中性粒细胞的吞噬和杀菌作用。浓度为1×10-11mol/L和1×10-9mol/L可促进成骨,而浓度为1×10-7mol/L和1×10-6mol/L的阿仑膦酸钠对成骨无影响。结论局部持续使用阿仑膦酸钠防治人工关节无菌性松动应慎重进行,需保证假体周围的药物浓度可随时间而发生相应变化,即早期浓度不低于1×10-6mol/L,后期浓度则应以1×10-11mol/L或1×10-9mol/L为宜。     

阿伦膦酸钠对骨髓生成破骨细胞及骨吸收作用的影响

[目的]研究阿伦膦酸钠对体外培养的小鼠骨髓生成破骨细胞及其骨吸收作用的影响。[方法]收集小鼠骨髓细胞于含有10^-8mol／L的1，25二羟基维生素D3[1，25-（OH）2D3]的α-MEM完全培养基中体外培养，设置不同浓度的阿伦膦酸钠给药，并于培养的第6、9、12d观察记录抗酒石酸酸性磷酸酶（tartrate-resistant acidphosphatase，TRAP）阳性多核巨细胞[破骨样细胞（osteoclast-like cell，OLC）]生成数，反映破骨细胞生成情况。记数培养12d骨磨片上骨吸收陷窝数及吸收面积，反映骨吸收情况。[结果]随着阿伦膦酸钠浓度的增高，TRAP阳性的细胞数减少，骨吸收陷窝数及面积均减少。[结论]阿伦膦酸钠可抑制骨髓细胞体外培养中破骨样细胞的形成，体外可抑制破骨细胞骨吸收作用。     

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     

磨屑在人工关节无菌性松动中作用的实验研究

目的：观察磨屑在动物体内引起的组织学反应，比较了不同磨屑所致反应差别，比较磨屑在羟基磷灰石涂层钛合金棒－骨界面和光滑钛合金棒－骨界面间移动差别，探讨人工关节无菌性松动机制。方法：６４只家兔分为８组（ｎ＝８），分别将羟基磷灰石涂层钛合金棒和光滑钛合金棒经膝关节置入股骨远端，定期膝关节注入聚乙烯，钛合金及两者的混合磨屑。光镜、偏振光显微镜和电镜观察关节滑膜、两种钛合金棒－骨界面的组织学和超微结构。结果     

Effects of alendronate on particle-induced osteolysis in a rat model

BACKGROUND: Particle-induced osteolysis is currently a major problem affecting the long-term survivorship of total joint replacements. Alendronate is a third-generation bisphosphonate that blocks osteoclastic bone resorption. The objective of this study was to determine whether alendronate could prevent particle-induced osteolysis or restore (reverse) bone loss in established osteolysis. METHODS: A rat model of particle-induced osteolysis was used. A specially designed polyethylene implant was placed in the proximal part of the right tibia of seventy-two animals. Following four weeks of healing, the animals were randomized into control groups, a prevention group, or a treatment group. In the prevention group, animals received intra-articular injections of high-density polyethylene particles (mean size, 2 m; all <10 m) at four, six, and eight weeks postoperatively. Alendronate (0.01 mg/kg/day) was administered concomitantly through an implantable pump from the fourth week through the tenth week. In the treatment group, animals were also exposed to polyethylene particles at four, six, and eight weeks, to establish bone loss, but they received alendronate subsequently, from the tenth week through the sixteenth week, to treat the bone loss. Positive (particle-only) and negative (saline-solution-only) control groups were assessed as well. Tissues were harvested at ten weeks in the prevention group and at sixteen weeks in the treatment group. Histological analyses and histomorphometric determinations of the periprosthetic bone volume were carried out. RESULTS: Histological examination showed a rim of new bone (neocortex) around the implant in the untreated and saline-solution-treated control animals (no polyethylene particles). Treatment with saline solution (no polyethylene particles) did not affect periprosthetic bone. Animals exposed to polyethylene particles had bone loss. In those that received alendronate, the bone loss was either prevented or reversed, and the quantity of neocortical and trabecular bone was increased compared with that of the controls. Alendronate effectively preserved periprosthetic bone in both the prevention and treatment groups. In the prevention arm, the mean periprosthetic bone volume of the neocortex and the surrounding trabecular bone, as determined with histomorphometry, was 21.5% +/- 6.5% in the saline-solution-treated controls (no particles), 13.1% +/- 5.9% in the particle-treated animals, and 32.6% +/- 6.4% in the alendronate-treated animals (p < 0.001). In the treatment arm, the mean periprosthetic bone volume was 27.2% +/- 5.6% in the saline-solution-treated controls, 17.7% +/- 6.2% in the particle-treated animals, and 30.2% +/- 5.9% in the alendronate-treated animals (p = 0.002). CONCLUSIONS: In our model, the intra-articular injection of polyethylene particles caused substantial bone loss around a loaded implant. Alendronate effectively prevented and treated the particle-induced periprosthetic bone loss.     

Biological responses of human mesenchymal stem cells to titanium wear debris particles

Wear debris-induced osteolysis is a major cause of orthopedic implant aseptic loosening, and various cell types, including macrophages, monocytes, osteoblasts, and osteoclasts, are involved. We recently showed that mesenchymal stem/osteoprogenitor cells (MSCs) are another target, and that endocytosis of titanium (Ti) particles causes reduced MSC proliferation and osteogenic differentiation. Here we investigated the mechanistic aspects of the endocytosis-mediated responses of MSCs to Ti particulates. Dose-dependent effects were observed on cell viability, with doses >300 Ti particles/cell resulting in drastic cell death. To maintain cell viability and analyze particle-induced effects, doses <300 particles/cell were used. Increased production of interleukin-8 (IL-8), but not IL-6, was observed in treated MSCs, while levels of TGF-β, IL-1β, and TNF-α were undetectable in treated or control cells, suggesting MSCs as a likely major producer of IL-8 in the periprosthetic zone. Disruptions in cytoskeletal and adherens junction organization were also observed in Ti particles-treated MSCs. However, neither IL-8 and IL-6 treatment nor conditioned medium from Ti particle-treated MSCs failed to affect MSC osteogenic differentiation. Among other Ti particle-induced cytokines, only GM-CSF appeared to mimic the effects of reduced cell viability and osteogenesis. Taken together, these results strongly suggest that MSCs play both responder and initiator roles in mediating the osteolytic effects of the presence of wear debris particles in periprosthetic zones.     

雷公藤多甙对糖尿病肾病患者微炎症反应的影响

目的探讨雷公藤多甙对糖尿病肾病患者微炎症反应的影响。方法将58例糖尿病肾病患者随机分为普通治疗组（DN组）和雷公藤多甙治疗组（TL组）,共治疗3个月。Ficoll法分离外周血单个核细胞（PBMC）并于体外培养,观察雷公藤多甙对培养的PBMC的作用。用放射免疫法测定患者血清和PBMC上清液肿瘤坏死因子-α（TNF-α）和白细胞介素-1（IL-1）的浓度,EMSA测定PBMC核因子-κKB（NF-κB）的活性。选择20名性别、年龄匹配的健康者作为对照组（NO组）。结果①与NO组治疗前相比,DN组和TL组治疗前血自蛋白降低,24h尿蛋白定量升高,血TNF-α和IL-1升高（P〈0．05）。②治疗后,与DN组相比,TL组患者血白蛋白升高,24h尿蛋白定量降低,血TNF-α和IL-1降低（P〈0．05）。③与NO组治疗前相比,DN组和TL组治疗前PBMC基础分泌TNF-α和IL-1增加,PBMC NF-κB活性升高（P〈0．05）。雷公藤多甙处理后,PBMC分泌TNF-α和IL-1明显减少,NF-κB活性降低（P〈0．05）。结论雷公藤多甙通过抑制PBMC活性及循环微炎症反应发挥治疗糖尿病肾病作用。     

Inhibition of particulate debris-induced osteolysis by alendronate in a rat model.

A rat model was used to study the efficacy of alendronate therapy in inhibition of particle-induced periprosthetic osteolysis. A prosthesis was simulated by inserting a cylindrical polymethylmethacrylate plug into the distal femur of 24 rats allowing the plug to communicate with the joint space. Intra-articular injections of irregularly-shaped ultra-high molecular weight polyethylene particles of 20-200 pm in diameter were administered at 2-week intervals. The rats were randomized into two groups (n=12 each). Group A rats received twice weekly subcutaneous injections of alendronate sodium while group B rats received injections of saline vehicle only. At 10 weeks all rats were sacrificed. The distal femurs were harvested and axial sections were prepared for histologic analysis. Each section was graded on a scale of 1-4, quantifying the degree of osteolysis surrounding the polymethylmethacrylate plug. Microscopic examination showed a significant (P<.0001) difference in the amount of periprosthetic bone. Femurs from group A treated with alendronate demonstrated mostly normal or near-normal periprosthetic trabeculations, whereas femurs from group B treated with saline showed extensive bone resorption. There was no qualitative difference in the inflammatory cellular response between the groups. This study established the ability of alendronate to inhibit the osteoclastic-mediated osteolysis around joint implants.     

雷尼酸锶对钛颗粒诱导炎性骨溶解的抑制作用

目的观察雷尼酸锶（strontium ranelate,SR）对磨损颗粒诱导炎性骨溶解的影响。方法 30只雄性C57BL/J6小鼠,随机分为空白组、对照组和药物组,每组10只。采用钛（Ti）颗粒诱导的小鼠颅骨溶解模型,药物组建模当日经灌胃予SR[600mg/（kg·d）],空白组和对照组不予处理;持续至建模后10d,处死取材。HE染色观察颅骨溶解程度及骨膜厚度;抗酒石酸酸性磷酸酶（tartrate resistant acid phosphatase,TRAP）染色检测成熟破骨细胞;酶联免疫吸附试验（ELISA）检测肿瘤坏死因子（TNF-α）、白细胞介素（IL）-1β和IL-6表达水平。结果HE染色结果,对照组骨膜明显增厚,颅骨溶解区域广;图像分析软件测量结果,对照组骨膜厚度（0.27±0.04）mm,骨溶解率为0.47±0.11,与药物组[（0.11±0.02）mm,0.18±0.05]比较,差异有统计学意义（P〈0.05）;TRAP染色结果,对照组颅骨大片紫红色区域,SR治疗后明显减少;ELISA检测结果,SR加入后,TNF-α、IL-1β和IL-6的表达量分别为[（145.6±14.2）ng/L、（130.2±8.2）ng/L和（137.6±8.2）μg/L],与对照组[（210.2±8.9）ng/L、（159.6±9.7）ng/L、（170.8±9.5）μg/L]比较,差异有统计学意义（P〈0.05）。结论 SR能够减轻Ti颗粒引起的炎症反应、减少炎症因子分泌,抑制骨溶解。     

Comparison of the roles of IL-1, IL-6, and TNFalpha in cell culture and murine models of aseptic loosening

Pro-inflammatory cytokines, such as IL-1, IL-6, and TNF, are considered to be major mediators of osteolysis and ultimately aseptic loosening. This study demonstrated that synergistic interactions among these cytokines are required for the in vitro stimulation of osteoclast differentiation by titanium particles. In contrast, genetic knock out of these cytokines or their receptors does not protect murine calvaria from osteolysis induced by titanium particles. Thus, the extent of osteolysis was not substantially altered in single knock out mice lacking either the IL-1 receptor or IL-6. Osteolysis also was not substantially altered in double knock out mice lacking both the IL-1 receptor and IL-6 or in double knock out mice lacking both TNF receptor-1 and TNF receptor-2. The differences between the in vivo and the cell culture results make it difficult to conclude whether the pro-inflammatory cytokines contribute to aseptic loosening. One alternative is that in vivo experiments are more physiological and that therefore the current results do not support a role for the pro-inflammatory cytokines in aseptic loosening. We however favor the alternative that, in this case, the cell culture experiments can be more informative. We favor this alternative because the role of the pro-inflammatory cytokines may be obscured in vivo by compensation by other cytokines or by the low signal to noise ratio found in measurements of particle-induced osteolysis.     

老年患者炎症因子水平与肾性贫血的相关分析

目的 探讨老年患者贫血的发生与炎症因子水平及。肾功能的相关性。方法 选择老年患者（年龄≥60岁）200例为研究对象,另设30例健康体检者为正常对照组。根据血红蛋白水平将200例老年患者分为贫血组和非贫血组;根据估算肾小球滤过率（eGFR）又分为A组[eGFR〉50ml·min^-1·（1．73m^2）^-1]、B组[eGFR30～50ml min^-1·（1．73m^2）^-1和C组[eGFR〈30mlmin^-1·（1．73m^2）^-1],测定血清超敏C反应蛋白（hs-CRP）、白细胞介素6（IL-6）、肿瘤坏死因子α（TNF-α）水平。结果随着肾功能水平减退,贫血患病率逐渐增高,血清hs-CRP、IL-6、TNF-α水平亦逐渐增高,但仅hs-CRP组间有显著差异（P〈0．05）;血清hs-CRP、IL-6、TNF-α与血红蛋白均呈负相关（r=-0．271、-0．148、-0．155,P〈0．05）;hs-CRP、TNF-α与eGFR亦呈负相关（r=-0．140、-0．142,P〈0．05）,IL-6与eGFR无相关性。结论老年人肾功能减退时的高贫血患病率与炎症因子参与的致病机制有关;hs-CRP在评价。肾功能减退者的机体炎症状态时优于TNF-α和IL-6。