阿伦膦酸钠对破骨细胞展平的影响

目的 :以破骨细胞展平面积为功能活性指标 ,探讨不同浓度阿伦膦酸钠对破骨细胞功能的影响。方法 :含破骨细胞的骨髓细胞取自出生 2 4h内的新生大鼠 ,以 1× 10 5/ml的浓度种植于 4个 3 5mm培养碟中 ,其中 1个为对照组 ,3个为含有 10 -11M、10 -10 M、10 -9M 3个浓度阿伦膦酸钠的实验组。每组随机选取 6个破骨细胞为观察对象 ,定时观察拍照 ,并将照片扫描输入计算机计算破骨细胞展平面积。结果 :10 -11M阿伦膦酸钠对破骨细胞展平面积无明显影响 (P>0 .0 5 )。 10 -10 M、10 -9M阿伦膦酸钠降低破骨细胞展平面积 (P <0 .0 1)。 10 -9M比 10 -10 M阿伦膦酸钠降低破骨细胞展平面积的程度大 (P <0 .0 1)。结论 :破骨细胞展平面积是与骨吸收功能密切相关的形态指标。展平面积的测量方法简便、迅速。 10 -10 M以上浓度的阿伦膦酸钠能抑制破骨细胞的功能活性 ,随阿伦膦酸钠浓度增加抑制作用增强     

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。     

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。     

辛伐他汀对PTHrP诱导的破骨细胞性骨吸收及小鼠颅盖骨代谢的影响

[目的]探讨辛伐他汀对体外甲状旁腺素相关肽（PTHrP）诱导小鼠的破骨细胞骨吸收功能的作用及其小鼠骨代谢的影响。[方法]采用PTHrP诱导小鼠骨髓细胞培养破骨细胞和小鼠颅盖骨培养体系，检测辛伐他汀作用8d后破骨细胞骨吸收陷窝和培养上清钙的变化；检测小鼠颅盖骨培养上清碱性磷酸酶和钙含量，组织学观察小鼠颅盖骨形态学变化。[结果]辛伐他汀体外可明显抑制PTHrP诱导小鼠的破骨细胞骨吸收陷窝的形成及培养上清钙的释放，辛伐他汀体外可增强小鼠颅盖骨培养上清碱性磷酸酶的活性，组织学观察到辛伐他汀使小鼠颅盖骨矿化增强。[结论]辛伐他汀体外不仅可促进小鼠颅盖骨的成骨活性，并且可明显抑制PTHrP诱导小鼠的破骨细胞骨吸收功能，对骨吸收性疾病有着重要的防治作用。     

1，25-二羟基维生素D3对小鼠骨髓细胞形成破骨细胞样细胞及其骨吸收效应的影响

目的动态观察1,25-二羟基维生素D3[1,25-(OH)2D3]对NH小鼠骨髓细胞培养在体外形成破骨细胞及其骨吸收的剂量效应和作用时象.方法收集NH小鼠骨髓细胞于含10%胎牛血清的α-MEM培养基中行体外培养,设置不同的1,25-(OH)2D3浓度组和给药时间组,并于培养第3、6、9、12天观察记录抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)阳性多核巨细胞[或破骨细胞样细胞(osteroblast-like cell,OLC)]以及骨磨片上骨吸收陷窝的数目.结果高于10-9mmol/L的1,25-(OH)2D3单一刺激可于培养第6天诱导TRAP阳性OLC形成,并且可在骨磨片上观察到骨吸收陷窝;随1,25-(OH)2D3浓度的增高,OLC和骨吸收陷窝数目均随之增加;各浓度组OLC数目于培养第9天达最高值,随后则趋于减少,而骨吸收陷窝数于培养第9和第12天均有所增加;采用单一1,25-(OH)2D3浓度(10-8mmol/L),TRAP阳性OLC和骨吸收陷窝见于培养3 d后用药和全程用药组,而仅在培养前3 d用药则不能诱导OLC和骨吸收陷窝的形成.结论 1,25-(OH)2D3可在体外诱导骨髓单核细胞分化形成OLC并使其具有破骨活性,诱导作用的强弱与1,25-(OH)2D3的浓度相关,而且其作用时象可能是在培养3 d以后.     

高产量高纯度的破骨细胞分离培养

目的 获得高产量高纯度破骨细胞，为骨吸收的体外研究提供丰富的细胞来源。方法 将经典分离乳兔四肢骨破骨细胞的方法与1，25(OH)2D3诱导骨髓干细胞生成破骨样细胞的方法相结合，并应用0．05％胰蛋白酶／0．02％ EDTA联合消化的方法将其纯化。结果 该方法可诱导生成大量的破骨样细胞，0．05％胰蛋白酶／0．02％ EDTA联合消化可使破骨细胞纯化率达95％。诱导生成的破骨样细胞TRAP染色阳性，体外培养具有噬骨能力。结论 该培养方法可产生大量高纯度的破骨样细胞，可为破骨细胞的体外分子生物学研究提供丰富的细胞来源。     

辛伐他汀抑制甲状旁腺素相关肽诱导的破骨细胞生成和功能

目的:探讨辛伐他汀对骨髓来源的破骨细胞形成和功能的影响。方法:取Balb/C雄性小鼠双侧股骨和胫骨的骨髓,以不含血清的α-MEM培养液洗涤并收集骨髓细胞,再将细胞重新悬浮于含100 m l/L胎牛血清的α-MEM培养液中,细胞计数后,配成1.5×109/L的细胞悬液,同时加入甲状旁腺素相关肽(PTHrP)和不同剂量的辛伐他汀(10-7、10-6、10-5mol/L)于24孔培养板进行培养,并设阳性对照(只加PTHrP)和阴性对照(PTHrP和辛伐他汀都不加)组,每组均有1孔放置骨磨片1片,培养6 d后;去除上清,抗酒石酸(TRAP)染色检测培养板底部破骨细胞形成;骨磨片用甲苯胺蓝染色,电镜检测骨磨片的吸收陷窝。结果:小鼠骨髓细胞在PTHrP的诱导下获得大量的TRAP染色阳性的破骨细胞,骨磨片有吸收陷窝形成;用辛伐他汀(10-7、10-6mol/L)和PTHrP共同培养下TRAP染色阳性的破骨细胞形成数量均明显减少(P<0.01),辛伐他汀在10-5mol/L时则无TRAP染色阳性的破骨细胞形成;辛伐他汀在10-7mol/L时骨磨片有吸收陷窝的形成但少于阳性对照组(P<0.01),在10-6、10-5mol/L时骨磨片则无吸收陷窝的形成。结论:辛伐他汀对小鼠骨髓来源的破骨细胞的形成有着明显的抑制作用,并且呈剂量依赖关系。     

阿司匹林对大鼠破骨细胞分化成熟和骨吸收活性的影响

目的 研究不同浓度阿司匹林对体外培养大鼠破骨细胞(Osteoclast,OC)分化成熟及骨吸收活性的影响.方法 建立由核激活因子受体配体(receptor activator of NF-κB ligand,RANKL)和巨噬细胞集落刺激因子(Macrophage colony stimulating factor,M-CSF)共同作用的大鼠破骨细胞骨髓诱导体系,将雌激素(10-6 mmol/L)和不同浓度的阿司匹林(0.25 mmol/L、0.5 mmol/L、1.0 mmol/L、1.5 mmol/L)分别作用于破骨细胞.诱导培养后分别对破骨细胞进行抗酒石酸酸性磷酸酶(The tartrate-resistant acid phosphatase,TRAP)染色,观察细胞形态,并计数破骨样细胞数量;将各组破骨细胞接种于骨磨片上,建立破骨细胞-骨磨片活性分析模型,于不同时间点对骨磨片进行光镜和扫描电镜观察,分析计算骨吸收陷窝面积.结果 与正常对照组相比,雌激素组破骨细胞数量和骨吸收陷窝面积低于正常对照组,差异有统计学意义(P＜0.05);且随着阿司匹林浓度的增加,阿司匹林组TRAP阳性多核破骨细胞数量、骨吸收陷窝面积逐渐减少直至消失,差异有统计学意义(P＜0.05).与雌激素组相比,低浓度阿司匹林组(0.25mmol/L)没有明显差异;但中、高浓度阿司匹林实验组(0.5mmol/L、1.0mmol/L、1.5mmol/L)破骨细胞数量和骨吸收陷窝面积减少,差异有统计学意义(P＜0.05).结论 阿司匹林对破骨细胞的分化成熟及骨吸收功能有抑制作用,且呈剂量依赖性,从而具有抗骨质疏松的作用.     

淫羊藿苷对小鼠骨髓源性破骨细胞诱导生成及骨吸收功能的影响

目的探讨淫羊藿苷对破骨细胞诱导产生及骨吸收功能的影响。方法用终浓度分别为25ng·mL^-1、30ng·mL^-1、10^-8mol·L^-1的M—CSF、RANKL、1，25（OH）2VitD3体外诱导培养小鼠骨髓源性破骨细胞，在此过程中加入终浓度分别为0、10^-7mol·L^-1、10^-6mol·L^-1、10^-5mol·L^-1的淫羊藿苷。倒置相差显微镜下观察活体细胞、HE染色、TRAP染色及降钙素受体染色鉴定破骨细胞，计数骨片上骨吸收陷窝数及面积，玻片上TRAP阳性多核细胞数。结果加药组随淫羊藿苷浓度的增加，骨片上形成的骨吸收陷窝数及面积，玻片上的TRAP阳性多核细胞数呈量的依赖性的减少，与非加药组比较，10^-5mol·L^-1、10^-5mol·L^-1浓度的淫羊藿苷组，差异有显著性（P〈0．05）。结论淫羊藿苷具有抑制破骨细胞诱导产生及骨吸收功能的作用，并随浓度增加抑制作用增强。     

Bone cement with reduced proportion of monomer in total hip arthroplasty: Preclinical evaluation and randomized study of 47 cases with 5 years' follow-up

Bone cement with reduced amount of monomer and low curing temperature may improve implant fixation due to reduced toxicity. We analyzed the mechanical, chemical and thermal properties of such a cement (Cemex Rx) using Palacos R as control. The in vivo performance of the 2 cements was also evaluated in a prospective randomized study of 47 hips, where either of the cement types was used to fixate Lubinus SP2 prostheses with the stem made of titanium alloy. Cemex Rx had a reduced tensile strength, probably because this cement was manually mixed, as recommended by the manufacturer. A standardized laboratory test showed lower curing temperature for Cemex, but measurements at 37° and with prechilled Palacos R and Cemex Rx, as in clinical work, showed no difference. In the clinical study radiostereometric measurements of cup and stem migration showed similar values in the 2 groups up to 5 years after the operation. The cement mantle was stable in both groups, but the stems migrated similarly inside the cement mantle regardless of the type of cement used. Proximal wear was low (0.04-0.05 mm/year) and tended to be lower in the Cemex group (p = 0.02). Aluminum and vanadium levels in serum increased 5 years after the operation, but no difference was noted between the 2 groups. Collagen markers (PICP, ICTP) showed similar increases in bone turnover 6 weeks and 6 months after operation in both groups.     

Osteogenesis after Bone and Bone Marrow Transplantation: II. The Initial Cellular Events Following Transplantation of Decalcified Allografis of Cancellous Bone

An experimental study was done in rabbits to investigate the fate of allogeneic iliac cancellous bone, both non-decalcified and decalcified with hydrochloric acid, transplanted to a muscular site for up to 14 days. Some of the treated allografts were impregnated with autologous bone marrow cells, obtained from the femoral medulla by aspiration, and each was compared with allografts alone. Combined myelo-osseous grafts produced bone after 7 to 8 days implantation, as did marrow autografts alone. In addition non-decalcified implants stimulated the production of multinucleated giant cells. Three different types of wash solution were used but these did not influence the cell population seen, nor the new bone formation. It is concluded that the critical events in bone formation after transplantation occur less than 8 days after the transplantation and that marrow cells have osteogenic capacity. This has relevance to the clinical aspects of bone grafting.     

Bone microstructure at the distal tibia provides a strength advantage to males in late puberty: An HR‐pQCT study

Bone is a complex structure with many levels of organization. Advanced imaging tools such as high‐resolution (HR) peripheral quantitative computed tomography (pQCT) provide the opportunity to investigate how components of bone microstructure differ between the sexes and across developmental periods. The aim of this study was to quantify the age‐ and sex‐related differences in bone microstructure and bone strength in adolescent males and females. We used HR‐pQCT (XtremeCT, Scanco Medical, Geneva, Switzerland) to assess total bone area (ToA), total bone density (ToD), trabecular bone density (TrD), cortical bone density (CoD), cortical thickness (Cort.Th), trabecular bone volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), trabecular spacing standard deviation (Tb.Sp SD), and bone strength index (BSI, mg²/mm⁴) at the distal tibia in 133 females and 146 males (15 to 20 years of age). We used a general linear model to determine differences by age‐ and sex‐group and age × sex interactions (p < 0.05). Across age categories, ToD, CoD, Cort.Th, and BSI were significantly lower at 15 and 16 years compared with 17 to 18 and 19 to 20 years in males and females. There were no differences in ToA, TrD, and BV/TV across age for either sex. Between sexes, males had significantly greater ToA, TrD, Cort.Th, BV/TV, Tb.N, and BSI compared with females; CoD and Tb.Sp SD were significantly greater for females in every age category. Males' larger and denser bones confer a bone‐strength advantage from a young age compared with females. These structural differences could represent bones that are less able to withstand loads in compression in females. © 2010 American Society for Bone and Mineral Research     

Building better bone: The weaving of biologic and engineering strategies for managing bone loss

Segmental bone loss remains a challenging clinical problem for orthopaedic trauma surgeons. In addition to the missing bone itself, the local tissues (soft tissue, vascular) are often highly traumatized as well, resulting in a less than ideal environment for bone regeneration. As a result, attempts at limb salvage become a highly expensive endeavor, often requiring multiple operations and necessitating the use of every available strategy (autograft, allograft, bone graft substitution, Masquelet, bone transport, etc.) to achieve bony union. A cost‐sensitive, functionally appropriate, and volumetrically adequate engineered substitute would be practice‐changing for orthopaedic trauma surgeons and these patients with difficult clinical problems. In tissue engineering and bone regeneration fields, numerous research efforts continue to make progress toward new therapeutic interventions for segmental bone loss, including novel biomaterial development as well as cell‐based strategies. Despite an ever‐evolving literature base of these new therapeutic and engineered options, there remains a disconnect with the clinical practice, with very few translating into clinical use. A symposium entitled “Building better bone: The weaving of biologic and engineering strategies for managing bone loss,” was presented at the 2016 Orthopaedic Research Society Conference to further explore this engineering‐clinical disconnect, by surveying basic, translational, and clinical researchers along with orthopaedic surgeons and proposing ideas for pushing the bar forward in the field of segmental bone loss. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:1855–1864, 2017.

     

重组合异种骨植骨修复骨囊肿所致骨缺损

2001年10月～2003年9月,笔者共收治28例骨囊肿患者,均采用病灶刮除,瘤腔灭活和重组合异种骨植骨治疗,获得满意疗效,体会如下。     

感染性骨缺损的治疗及研究进展

感染性骨缺损由于存在感染及骨缺损双重病变,治疗棘手,疗程长,且易出现肌肉萎缩、局部瘢痕而致肢体功能受到严重影响.近年来随着外固定技术、显微外科技术、生物材料技术及骨组织工程技术等的发展,感染性骨缺损的治疗取得明显进步,短缩了治疗时间,且效果显著,笔者对其研究进展综述如下.     

The benefit of bone marrow concentrate in addition to a glass‐reinforced hydroxyapatite for bone regeneration: An in vivo ovine study

This study evaluates the ability of a Glass Reinforced Hydroxyapatite Composite (GRHC), in a new microporous pellet formulation with autologous bone marrow concentrate (BMC), to enhance bone regeneration and new bone formation. Ninety non‐critical sized bone defects were created in the femurs of nine Merino breed sheep and randomly left unfilled (group A), filled with GRHC pellets alone (group B) or filled with GRHC pellets combined with BMC (group C). The sheep were sacrificed at 3 weeks (three sheep), 6 weeks (three sheep) and 12 weeks (three sheep) and histological analysis (Light Microscopy‐LM), scanning electron microscopy (SEM) and histomorphometric analysis (HM) were performed. At 3, 6, and 12 weeks, HM revealed an average percentage of new bone of 48, 72, 83%; 25, 73, 80%, and 16, 38, 78% for Groups C, B and A respectively (significantly different only at 3 weeks p < 0.05). LM and SEM evaluation revealed earlier formation of well‐organized mature lamellar bone in Group C. This study demonstrates that the addition of a bone marrow concentrate to a glass reinforced hydroxyapatite composite in a pellet formulation promotes early bone healing. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:1176–1182, 2017.

     

Massive bone reconstruction with heat‐treated bone graft loaded autologous bone marrow‐derived stromal cells and β‐tricalcium phosphate composites in canine models

Bone marrow‐derived stromal cells (BMSCs) contain mesenchymal stem cells that are capable of forming various mesenchymal tissues. We hypothesized that BMSCs and β‐tricalcium phosphate (β‐TCP) composites would promote the remodeling of large‐sized autologous devitalized bone grafts; therefore, the aim of this study was to evaluate the effects of the composites on the remodeling of autologous devitalized bone grafts. Autologous BMSCs cultured in culture medium containing dexamethasone (10^?7 M) were loaded into porous β‐TCP granules under low‐pressure. Theses BMSC/TCP composites were put into the bone marrow cavity of autologous heat‐treated bone (femoral diaphysis, 65‐mm long, 100°C, 30 min) and put back to the harvest site. In the contralateral side, β‐TCP without BMSC were used in the same manner as the opposite side as the control. Treatment with the BMSC/TCP composites resulted in a significant increase in thickness, bone mineral density, and matured bone volume of the cortical bone at the center of the graft compared to the control. Histological analysis showed matured regenerated bone in the BMSC loaded group. These results indicate that BMSC/TCP composites facilitated bone regeneration and maturation at the graft site of large‐sized devitalized bone. This method could potentially be applied for clinical use in the reconstruction of large bone defects such as those associated with bone tumors. © 2013 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 31:1308–1316, 2013

     

Oxyphenbutazone (Tanderil®) in Surgery for Herniated Discs: A Double Blind Trial

Background and purpose In detection of glenoid labrum pathology, MR arthrography (MRA) has shown sensitivities of 88-100% and specificities of 89-93%. However, our practice suggested that there may be a higher frequency of falsely negative reports. We assessed the accuracy of this costly modality in practice.

Patients and methods We retrospectively reviewed MRA reports of 90 consecutive patients with clinical shoulder instability who had undergone shoulder arthroscopy. All had a history of traumatic anterior shoulder dislocation and had positive anterior apprehension tests. All underwent arthroscopy and stabilization during the same procedure. We compared the findings, using arthroscopic findings as the gold standard in the identification of glenoid labrum pathology.

Results 83 of the 90 patients had glenoid labrum tears at arthroscopy. Only 54 were correctly identified at MRA. All normal glenoid labra were identified at MRA. This gave a sensitivity of 65% and a specificity of 100% in identification of all types of glenoid labrum tear. 74 patients had anterior glenoid labral tears that were detected at an even lower rate of sensitivity (58%).

Interpretation The sensitivity of MRA in this series was substantially lower than previously published, suggesting that MRA may not be as reliable a diagnostic imaging modality in glenohumeral instability as previously thought. Our findings highlight the importance of an accurate history and clinical examination in the management of glenohumeral instability. The need for MRA may not be as high as is currently believed.