Distribution of Small Integrin-binding Ligand, N-linked Glycoproteins (SIBLING) in the Articular Cartilage of the Rat Femoral Head

The small integrin-binding ligand, N-linked glycoprotein (SIBLING) family is closely related to osteogenesis. Until recently, little was known about their existence in articular cartilage. In this study, we systematically evaluated the presence and distribution of four SIBLING family members in rat femoral head cartilage: dentin matrix protein 1 (DMP1), bone sialoprotein (BSP), osteopontin (OPN), and dentin sialophosphoprotein (DSPP). First, non-collagenous proteins were extracted and then separated by ion-exchange chromatography. Next, the protein extracts eluted by chromatography were analyzed by Stains-all staining and Western immunoblotting. IHC was used to assess the distribution of these four SIBLING family members in the femoral head cartilage. Both approaches showed that all the four SIBLING family members are expressed in the femoral head cartilage. IHC showed that SIBLING members are distributed in various locations throughout the articular cartilage. The NH₂-terminal fragments of DMP1, BSP, and OPN are present in the cells and in the extracellular matrix, whereas the COOH-terminal fragment of DMP1 and the NH₂-terminal fragment of DSPP are primarily intracellularly localized in the chondrocytes. The presence of the SIBLING family members in the rat femoral head cartilage suggests that they may play important roles in chondrogenesis. (J Histochem Cytochem 58:1033–1043, 2010)     

人工股骨头置换与动力髋螺钉内固定用于老年股骨头转子间骨折治疗的疗效比较

目的：比较人工股骨头置换术与动力髋螺钉内固定治疗老年股骨头转子间骨折的临床疗效。方法：选择我院74例老年股骨头转子间骨折患者作为观察对象,随机分为A组（人工股骨头置换术）和B组（动力髋螺钉内固定术）,每组37例。比较两组患者的手术时间、术中出血量、术后卧床时间、术后Harris评分及术后并发症和死亡率。结果：手术时间和术后卧床时间A组明显短于B组（P〈0．05）,A组术中出血量较B组明显减少,差异显著（P〈0．05）。术后1、3个月Harris评分A组均高于B组（P〈0．05）;术后6个月Harris评分两组无明显差异（P〉0．05）。结论：与动力髋螺钉内固定术相比,人工股骨头置换术术后可更早负重、恢复好,而且并发症少,是较为理想的治疗老年股骨头转子间骨折的治疗方法。     

外后侧小切口髋关节置换术治疗股骨头缺血性坏死的临床研究

目的：观察髋关节置换术治疗股骨头缺血性坏死临床可行性和疗效。方法：后侧入路小切口行全款关节置换术,选取55例患者作为临床研究对象共62髋,进行12个月以上随访平均随访（15．5±2．5）个月。结果：术后患者功能恢复较好,假体位置及周围组织结合良好,Harris评分比术前提高45分以上,临床效果值得肯定。结论：通过外后侧入路髋关节置换治疗股骨头缺血性坏死可达到预期疗效,术前的充分准备对手术成功具有积极作用,可以作为减轻患者自身痛苦和降低医疗费用临床治疗方式。     

超高龄患者填充骨水泥型人工股骨头置换术后的凝血功能变化及意义

目的：观察骨水泥填充对人工股骨头置换术术后超高龄老年患者凝血系统的影响。方法：选择80岁以上骨水泥型人工股骨头置换术患者29例,于术前、术后当天及术后第3天空腹抽取静脉血,测定凝血功能相关指标,包括凝血酶原时间（PT）、部分凝血活酶时间（APTT）、凝血酶时间（TT）、凝血酶原活动度（PTA）、国际标准化比值（INR）、纤维蛋白原（FIB）、D二聚体（DD）、抗凝血酶Ⅲ（ATⅢ）、血小板（PLT）水平,并对结果进行比较分析。结果：患者术后当天FIB、DD、显著升高（P〈0.05）,ATⅢ降低（P〈0.05）,提示高凝状态,且纤溶亢进,此时段TT、PT延长（P〈0.05）,血小板明显降低,提示存在出血风险;术后第3天TT、PT显著延长（P〈0.05）,ATⅢ恢复到术前水平,FIB,DD水平较手术后当天下降,提示术后第3天有明显的出血倾向,凝血与纤溶系统逐渐恢复平衡。结论：骨水泥型人工股骨头置换术对80岁以上超高龄患者凝血功能有显著影响,术后当天高凝状态、纤溶亢进,存在潜在出血风险,术后第3天有明显出血倾向,提示高龄患者术后应适当补充凝血因子且须谨慎使用抗凝药物。     

不同影像方法对股骨头缺血坏死的诊断价值

目的:研究早期股骨头无菌性坏死(aseptic necrosis of the femoral head,ANFH)的不同影像方法(X-ray、CT、MRI)诊断价值。方法:34例双侧股骨头行X线、CT和MRI检查,分析其对早期ANFH的显示能力。结果:34例患者均存在早期骨坏死表现。X线发现Ⅰ-Ⅱ期ANFH 14例,诊断准确率41.18%;CT发现Ⅰ-Ⅱ期ANFH 19例,诊断准确率57.35%;MRI发现Ⅰ-Ⅱ期ANFH 27例,诊断准确率为77.94%。结论:X线平片难以发现ANFH的早期表现,MR结合螺旋CT扫描既可发现早期病变,又可进行分期,是研究早期ANFH的可靠手段。     

营养动脉结扎和液氮冷冻对股骨头内局部氧分压的影响

目的：观察营养动脉结扎和液氮冷冻对股骨头内局部氧分压的影响,为后期制作极度低氧股骨头缺血坏死模型打下基础。方法：9只雄性大耳白兔随机平均分为对照组、液氮冷冻组和动脉结扎＋液氮冷冻组,对照组动物不做任何处理,液氮冷冻组动物行股骨头液氮冷冻使其发生缺血坏死,动脉结扎＋液氮冷冻组动物先结扎旋股内外侧动脉,再对股骨头进行液氮冷冻。造模后立刻活体观察股骨头内氧分压变化。结果：液氮冷冻组股骨头内氧分压较对照组下降一半左右,2组比较有显著性差异（P〈0．05）;动脉结扎＋液氮冷冻组动物股骨头内氧分压进一步下降到对照组的1／8左右,与其他2组相比有显著性差异（P〈0．05）。结论：液氮冷冻和旋股内、外侧动脉结扎均可明显降低股骨头内氧分压,两者合用具有重叠效应,可共同促使股骨头内氧分压下降。     

双极人工股骨头置换治疗老年不稳定股骨粗隆间骨折的临床研究

目的:通过观察我院老年不稳定股骨粗隆间骨折患者临床治疗资料,探讨分析采用双极人工股骨头置换治疗该疾病的临床效果.方法:将我院收治的100例该疾病患者按照手术方法平分为A组与B组两组,分别进行双极人工股骨头置换术与动力髋螺钉手术治疗,对比分析使用两种方法手术时间、出血量情况、术后并发症以及治疗效果等.结果:两组患者经各自手术治疗后,临床症状均有改善,A组患者在手术时间、出血量情况与B组患者相比,差异不显著(P＞0.05),而在术后并发症、髋关节功能对比方面,A组优于B组,差异具有显著性(P＜0.05).结论:采用双极人工股骨头置换手术治疗老年不稳定股骨粗隆间骨折患者,手术并发症少,安全可靠性高.有效改善患者髋关节功能,临床效果显著,值得临床上推广与进一步研究.     

Vitamin K2 Prevents Glucocorticoid-induced Osteonecrosis of the Femoral Head in Rats

Glucocorticoid medication is one of the most common causes of atraumatic osteonecrosis of the femoral head (ONFH), and vitamin K₂ (VK₂) has been shown to play an important and beneficial role in bone metabolism. In this study, we hypothesized that VK₂ could decrease the incidence of glucocorticoid-induced ONFH in a rat model. Using in vitro studies, we investigated how bone marrow-derived stem cells in the presence of methylprednisolone proliferate and differentiate, specifically examining osteogenic-related proteins, including Runx2, alkaline phosphatase and osteocalcin. Using in vivo studies, we established glucocorticoid-induced ONFH in rats and investigated the preventive effect of VK₂. We employed micro-CT scanning, angiography of the femoral head, and histological and immunohistochemical analyses, which demonstrated that VK₂ yielded beneficial effects for subchondral bone trabecula. In conclusion, VK₂ is an effective antagonist for glucocorticoid on osteogenic progenitors. The underlying mechanisms include acceleration of BMSC propagation and promotion of bone formation-associated protein expression, which combine and contribute to the prevention of glucocorticoid-induced ONFH in rats.     

A microanatomical and histological study of the fin long bones of the Devonian sarcopterygian Eusthenopteron foordi

Meunier F.J. and Laurin M. 2012. A microanatomical and histological study of the fin long bones of the Devonian sarcopterygian Eusthenopteron foordi. —Acta Zoologica (Stockholm) 93 : 88–97. A paleohistological study of the endoskeletal bones of the dorsal and pelvic fins shows that Eusthenopteron foordi had true long bones that grew in length and thickness through endochondral and periosteal ossification, respectively. Endochondral ossification required cartilaginous epiphyses with a growth plate system whose presence is confirmed by both calcified cartilage and thin endochondral bony trabeculae that overlaid the erosive bays located in hypertrophic calcified cartilage. Articulations between axial mesomeres in paired fins were diarthroses. This microanatomical organization, i.e. longitudinal growth of diaphysis and articulations between epiphyses, can be considered an exaptation for terrestrial locomotion as it can produce skeletal elements able to support strong mechanical stress.     

An Automated Technique for Double Staining Rat and Rabbit Fetal Skeletal Specimens to Differentiate Bone and Cartilage

Assessment of chemicals for their potential to cause developmental toxicity must include evaluation of the development of the fetal skeleton. The method described here is an improved and fully automated double staining method using alizarin red S to stain bone and alcian blue to stain cartilage. The method was developed on the enclosed Shandon PathcentreTM, and the quality of specimens reported here will be reproduced only if carried out on a similar processor under the same environmental conditions. The staining, maceration and clearing process takes approximately 6 days. The personnel time, however, is minimal since solutions are changed automatically and the fetuses are not examined or removed from the processor until the procedure is completed. Upon completion of processing, the bone and cartilage assessment of the specimens can be carried out immediately if required. Full evaluation of skeletal development in both the rat and the rabbit is necessary to meet the requirements of safety assessment studies. This method allows this to be accomplished on a large scale with consistently clear specimens and in a realistic time.     

关节软骨与软骨下骨改变在不同骨关节炎动物模型中的特点

目的 探讨不同骨关节炎(osteoarthritis,OA)动物模型中软骨下骨和关节软骨的病理改变特征。方法 采用三种SD大鼠骨关节炎模型,将24只6月龄雌性大鼠随机分为4组:假手术对照组(Sham,n=6),前交叉韧带切除手术组(ACLT,n=6),木瓜蛋白酶关节腔注射组(Papain,n=6),以及卵巢切除术组(OVX,n=6)。造模后8周取膝关节,胫骨平台行Micro-CT扫描分析,关节软骨行甲苯胺蓝染色、Mankin评分,比较软骨下骨和关节软骨改变情况。结果 造模操作后8周,不同OA模型的软骨破坏程度有所不同。ACLT和Papain组软骨破坏比较严重,OVX组软骨变化较轻。所有OA模型中的软骨下松质骨均发生改变,OVX组相对于Sham对照组,软骨下骨微结构显著疏松,而ACLT组与Papain组相对于Sham对照组,软骨下骨微结构没有显著改变,但相对于OVX组,有显著性差异。三种OA模型的软骨下骨板厚度都较Sham组减少。结论 三种动物模型软骨下骨和关节软骨都发生明显病理改变,并且改变有所不同。不同OA模型代表不同病理,预示着软骨下骨所发挥的作用有所不同,这为进一步研究不同类型OA发生发展的机制,以及将软骨下骨作为OA治疗的可能靶点提供了更多的依据。     

Study on the Microstructure of Human Articular Cartilage/Bone Interface

For improving the theory of gradient microstructure of cartilage/bone interface, human distal femurs were studied. Scanning Electron Microscope (SEM), histological sections and MicroCT were used to observe, measure and model the microstructure of cartilage/bone interface. The results showed that the cartilage/bone interface is in a hierarchical structure which is composed of four different tissue layers. The interlocking of hyaline cartilage and calcified cartilage and that of calcified cartilage and subchondral bone are in the manner of “protrusion-pore” with average diameter of 17.0 μm and 34.1 μm respectively. In addition, the cancellous bone under the cartilage is also formed by four layer hierarchical structure, and the adjacent layers are connected by bone trabecula in the shape of H, I and Y, forming a complex interwoven network structure. Finally, the simplified structure model of the cartilage/bone interface was proposed according to the natural articular cartilage/bone interface. The simplified model is a 4-layer gradient biomimetic structure, which corresponds to four different tissues of natural cartilage/bone interface. The results of this work would be beneficial to the design of bionic scaffold for the tissue engineering of articular cartilage/bone.     

两种不同术式对老年股骨颈骨折治疗对比

目的：探讨全髋关节置换术与股骨头置换术治疗对比。方法：回顾性的分析对2007年1月-2008年12月年我院接收的70例60岁以上的老年股骨颈骨折的进行股骨头置换和全髋关节置换两组不同手术方式的临床资料。结果：两组股骨颈骨折治疗的手术时间、住院时间及术后并发症及治疗效果无明显统计学差异,两组术后半年到一年半的Harris评分较术前评分有显著的统计学意义。结论：人工股骨头置换术和髋关节置换是治疗老年股骨颈骨折的有效方法,可根据患者的自身情况、疾病特点等选择不同术式。     

两种不同术式对老年股骨颈骨折治疗对比

目的:探讨全髋关节置换术与股骨头置换术治疗对比。方法:回顾性的分析对2007年1月-2008年12月年我院接收的70例60岁以上的老年股骨颈骨折的进行股骨头置换和全髋关节置换两组不同手术方式的临床资料。结果:两组股骨颈骨折治疗的手术时间、住院时间及术后并发症及治疗效果无明显统计学差异,两组术后半年到一年半的Harris评分较术前评分有显著的统计学意义。结论:人工股骨头置换术和髋关节置换是治疗老年股骨颈骨折的有效方法,可根据患者的自身情况、疾病特点等选择不同术式。     

糖蛋白PAGE分离后的糖基显色法

鸡卵清蛋白通过SDS-PAGE分离后,用过碘酸-希夫(PAS)显色法可以使卵清蛋白显红色,而用糖苷酶去除卵清蛋白的糖基可以去除相应的红色。考马斯亮兰染色结果显示,去糖基后的卵清蛋白分子量略有减小。可见,PAS可方便地使糖蛋白呈现红色。     

Processing of Whole Femoral Head Allografts: A Method for Improving Clinical Efficacy and Safety

Femoral heads removed during primary hip replacement surgery are widely utilised as a source of allograft bone. Despite evidence that processing these grafts to remove blood and marrow elements improves both the clinical performance and safety of these allografts, many are transplanted without any processing being applied at all. The goal of this study was to investigate the efficiency of an allograft processing protocol which incorporates pasteurisation, (3h, 56–60°C) centrifugation, (1850g, 2 × 15min, 40°C) sonication, and repeated washing in warm (56–60°C, 19h) distilled, sterile water to remove blood and marrow elements from the graft. The protocol also involves applying heat treatment to the grafts which has been demonstrated to inactivate many pathogenic viruses. Following the processing procedure, the grafts are lyophilised and sterilised with ethylene oxide gas. The amount and rate of removal of 4 different components of blood and marrow from 6 whole femoral head allografts were measured. These were lipid, soluble protein, elastase and chloride ions. Lipid removal was assessed gravimetrically by solvent extraction of dried samples, soluble protein by the Bradford assay, elastase by radioimmunoassay and choride ion content by a modified commercially available colorimetric assay. Removing lipid from grafts has been shown to increase the rate of incorporation when the graft is used clinically. Elastase was studied as a marker of leukocyte removal, as evidence suggests the majority of potentially infective transmissible spongiform encephalopathy (TSE) activity resides in a sub-population of leukocytes. Soluble protein was studied as a marker of plasma removal, as a smaller amount of TSE infectivity resides here. Chloride removal was measured as this is a necessary pre-requisite to terminal sterilisation with ethylene oxide. The results showed that the protocol removed 74.5% (range: 68.0–90.8) of the lipid content, 96.4% (range: 94.8–98.4) of the soluble protein content, 97.7% (range: 97.1–100) of the elastase content and 98.8% (range: 98.0–99.2) of the chloride ion content. We have shown that processing designed to improve the clinical efficiency and safety of bone allografts can be accomplished without compromising the structural and biological properties of the graft.     

Mallory氏胰岛细胞染色法的改良

目的:为了更清晰地显示胰岛的A细胞、B细胞和D细胞。方法:zenker液固定,分别用苏木精-伊红和改良的Mallory氏法染色。结果:苏木精-伊红染色法,不易区分胰岛内三种细胞,而采用改良的Mallory氏染色方法,能够清晰地显示A细胞、B细胞和D细胞。结论:改良的Mallory氏染色法能够清楚的显示胰岛内三种细胞结构,为实验教学提供了优良的切片标本。     

Sensitivity Analysis and Optimal Shape Design for Bone-Prosthesis Interfaces in a Femoral Head Surface Replacement

Abstract

A numerical optimization procedure has been applied for the shape optimal design of a femoral head surface replacement. The failure modes of the prosthesis that were considered in the formulation of the objective functions concerned the interface stress magnitude and the bone remodelling activity beneath the implant. In order to find a compromising solution between different requirements demanded by the two objective functions, a two step optimization procedure has been developed. Through step I the minimization of interface stress was achieved, through step 2 the minimization of bone remodelling was achieved with constraints on interface stresses.

The results obtained provided an optimal design that generates limited bone remodelling activity with controlled interface stress distribution.

The computational procedure was based on the application of the finite element method, linked to a mathematical programming package and a design sensitivity analysis package.     

p63 IHC染色2+截断值可作为骨巨细胞瘤的诊断标记

为评估骨巨细胞瘤(giant cell tumor of bone, GCTOB)中p63表达在鉴别骨巨细胞瘤(GCTOB)中的作用,本研究采用横断面研究,检测50个巨细胞丰富的骨病变中p63的免疫组织化学表达,并评价p63在各肿瘤中的染色强度和程度。免疫组织化学分析显示,94.12%的骨巨细胞瘤(GCTOB)、14.29%的骨肉瘤(osteosarcoma, OSA)、60.00%的非骨化性纤维瘤(non-ossifyingfibroma, NOF)、25.00%的动脉瘤样骨囊肿(aneurysmal bone cyst, ABC)、71.43%的软骨母细胞瘤(chondroblastoma, CB)、66.67%的棕色瘤(Brown tumor)为阳性,而腱鞘巨细胞瘤(giant cell tumor of the tendon sheath, GCTTS)中未检测出p63阳性。对于p63的染色强度,可以观察到58.82%的骨巨细胞瘤(GCTOB)、28.57%的软骨母细胞瘤(CB)和20.00%的非骨化性纤维瘤(NOF)呈现强染色。对于p63的染色程度:观察到58.82%的骨巨细胞瘤(GCTOB)、28.57%的软骨母细胞瘤(CB)和14.29%的骨肉瘤(OSA)尾弥漫性染色,而中等染色在骨巨细胞瘤(GCTOB)中为17.65%,非骨化性纤维瘤(NOF)为20.00%,棕色瘤(Brown tumor)为33.33%。大量骨巨细胞瘤(94.12%)为p63阳性,这比任何其他巨细胞丰富的病变多得多。然而,p63的阳性染色并不是骨巨细胞瘤(GCTOB)特异性的。本研究结论初步表明,p63是鉴别骨巨细胞瘤(GCTOB)与其他骨巨细胞瘤的一种敏感的(97.6%)和相对特异的标志物。考虑到反应的强度和程度,本研究建议采用综合评分方法对p63 IHC染色判读富含巨细胞的病灶,并将2+截断值作为骨巨细胞瘤(GCTOB)的诊断标记。