激素诱导血清病动物骨坏死模型中的脂类代谢异常

目的初步探讨血清病动物激素性骨坏死的发病机制。方法将新西兰大白兔分为三组,A组联合应用马血清和地塞米松磷酸钠诱导骨坏死模型,B组单纯应用地塞米松磷酸钠,C组应用生理盐水作为对照组,对三组实验兔股骨头进行HE、MRI观察,并对各组兔的血脂、AST和ALT进行检测,分析比较。结果A组出现了典型的股骨头缺血坏死改变,血脂、AST和ALT与另外两组相比差异有显著性。单纯应用激素组和对照组未出现股骨头缺血坏死。结论运用马血清和地塞米松磷酸钠可成功诱导出兔骨坏死模型,血管炎、血脂代谢异常和肝功能损害共同导致股骨头缺血坏死发生,短期应用大剂量激素难以诱导出骨坏死模型。     

非创伤性股骨头坏死的机制及早期治疗的研究进展

非创伤性股骨头坏死(NONFH)是临床常见的骨科难治性疾病之一,本文就其发病机制和临床治疗研究的相关进展和问题展开综述,旨在进一步提高对NONFH的临床认识和治疗水平。股骨头坏死的机制仍然没有统一的定论,但各种机制最终归于股骨头供血区域血液瘀滞,导致股骨头发生缺血坏死虽然目前还没有一种治疗方法达到理想的治疗效果,髓心减压仍然是应用最为广泛的早期治疗手段,各种改进手术如骨移植、钽棒移植、骨髓干细胞移植和生长因子移植是今后的主要趋势。     

The effect of deferoxamine on angiogenesis and bone repair in steroid-induced osteonecrosis of rabbit femoral heads

In this study, we examined whether local deferoxamine (DFO) administration can promote angiogenesis and bone repair in steroid-induced osteonecrosis of the femoral head (ONFH). Steroid-induced ONFH was induced in 65 mature male New Zealand white rabbits by methylprednisolone in combination with lipopolysaccharide. Six weeks later, the rabbits received no treatment (model group, N = 15), bilateral core decompression (CD group, N = 20) or CD in combination with local DFO administration (DFO group, N = 20). Six weeks after the surgery, vascularization in the femoral head was evaluated by ink artery infusion angiography and immunohistochemical staining for von Willebrand Factor (vWF). Bone repair was assessed by histologic analysis and micro-computed tomography (micro-CT). Immunohistochemical staining was performed to analyze the expression of vascular endothelial growth factor (VEGF), hypoxia-inducible factor-1α (HIF-1α), bone morphogenetic protein-2 (BMP-2), and osteocalcin (OCN). Ink artery infusion angiography and microvessel analysis by immuohistochemical staining for vWF showed more blood vessels in the DFO group than other groups. The expression of HIF-1α, VEGF, BMP-2, and OCN, indicated by immunohistochemical staining, was higher in the DFO group compared with other groups. Micro-CT scanning results indicated that the DFO group had larger volume of newly formed bone than the CD group. This work indicated that local DFO administration improved angiogenesis and bone repair of early stage ONFH in rabbit model, and it may offer an efficient, economic, and simple therapy for early stage ONFH.     

Lack of association of MTHFR gene polymorphisms with the risk of osteonecrosis of the femoral head in a Korean population

Some studies have suggested that coagulation disorders may be implicated in osteonecrosis of the femoral head (ONFH). The C677T polymorphism of the 5, 10-methylenetetrahydrofolate reductase (MTHFR) gene has been postulated to be a genetic risk factor for venous thromboembolism and osteonecrosis in Caucasians, but this relationship has not been established in other populations. In this study, we conducted case-control analysis of whether MTHFR polymorphisms are associated with ONFH in Korean patients. Fifteen single nucleotide polymorphisms (SNPs) were selected and genotyped in 443 ONFH patients and 273 control subjects using the TaqMan 5′ allelic discrimination assay. Comparison of ONFH and control subjects using logistic regression models revealed no statistically significant differences in the frequencies of the MTHFR polymorphisms and haplotypes. Further analysis stratified by etiology also showed no association. These results suggest that MTHFR polymorphisms play no significant role in susceptibility to ONFH in the Korean population.     

Peroxisome proliferator-activated receptor-gamma gene polymorphisms are not associated with osteonecrosis of the femoral head in the Korean population

Osteonecrosis of the femoral head (ONFH) is a multifactorial disease to which certain individuals are more at risk. Altered lipid metabolism is one of the major risk factors for osteonecrosis, especially corticosteroid therapy and alcoholism. Peroxisome Proliferator-Activated Receptor-gamma (PPARgamma) plays a crucial role in differentiation of mesenchymal cells to adipocytes, lipid homeostasis, and bone metabolism. To investigate the possible association between PPARgamma gene variants and susceptibility to ONFH, we genotyped three common polymorphisms (-796A > G, +34C > G[Pro12Ala], and +82466C > T[His477His]) in 448 ONFH patients and 336 control subjects. Genotypes, allele frequencies, and haplotypes of the polymorphisms in the complete set of patients as well as in subgroups by sex or etiology were not significantly different from those in the control group. This suggests that the examined polymorphisms and haplotypes of the PPARgamma gene are unlikely to be associated with susceptibility to ONFH.     

Hypoxia-Inducible Factor Prolyl Hydroxylase Inhibitor Prevents Steroid-Associated Osteonecrosis of the Femoral Head in Rabbits by Promoting Angiogenesis and Inhibiting Apoptosis

The purpose of this study was to investigate the preventive effect of ethyl 3,4-dihydroxybenzoate(EDHB) on steroid-associated femoral head osteonecrosis(ONFH) in a rabbit model. New Zealand white rabbits were randomly divided into two groups (prevention group and model group), each containing 24 rabbits. Osteonecrosis was induced by lipopolysaccharide(LPS) combined with methylprednisolone(MPS). The prevention group received an intraperitoneal injection of EDHB at 50 mg/kg body weight every other day starting three days before establishing rabbit models of osteonecrosis, for a total of nine doses. Osteonecrosis was verified by haematoxylin-eosin (HE) staining. The expression of HIF-1α and VEGF was analyzed by immunohistochemistry. Angiogenesis, apoptosis and microstructural parameters were also analyzed. The rabbit models of osteonecrosis were successfully established and observed by HE staining. Histopathological observations indicated that EDHB reduced the rate of empty lacunae and the incidence of osteonecrosis. Immunohistochemical staining for HIF-1α and VEGF suggested that EDHB therapy inhibited degradation of HIF-1α and promoted expression of VEGF. Ink artery infusion angiography and microvessel density analysis revealed that there were more microvessels in the prevention group than in the model group. The TUNEL apoptosis assay suggested that EDHB intervention could reduce the number of apoptotic cells in avascular osteonecrosis of the femoral head. Micro-CT scanning indicated that the treatment group had better microstructural parameters than the model group. EDHB prevents steroid-associated osteonecrosis of the femoral head in rabbits by promoting angiogenesis and inhibiting apoptosis of bone cells and hematopoietic tissue.     

Icariin promotes angiogenesis in glucocorticoid‐induced osteonecrosis of femoral heads: In vitro and in vivo studies

The injury and dysfunction of the femoral head microvascular endothelial cells are associated with the pathogenesis of glucocorticoid‐induced osteonecrosis of the femoral head (ONFH). Reports indicate that icariin (ICA) can enhance vascular roles and also inhibit endothelial cell dysfunction. However, it still remains unclear whether ICA can promote angiogenesis in glucocorticoid‐induced ONFH. In this study, we investigate this hypothesis through in vitro and in vivo experiments. Results showed that 0.1 mg/mL hydrocortisone significantly suppressed bone microvascular endothelial cells (BMECs) proliferation while ICA at 10^?5 mol/L reversed this inhibition. ICA significantly promoted BMECs migration, tube formation, the angiogenesis‐related cytokines expression and the activation of Akt. Furthermore, ICA enhanced Bcl‐2 expression but diminished Bax expression. According to in vivo results, rats with ICA treatment exhibited a lower ratio of empty lacunae, higher volume of blood vessels and more CD31‐positive cells. This study revealed that ICA promotes angiogenesis of BMECs in vitro and improves femoral head blood vessel volume of rats treated with glucocorticoid, suggesting the efficacy of ICA in the prevention of glucocorticoid‐induced ONFH.     

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.     

Circulating microRNA signature of steroid‐induced osteonecrosis of the femoral head

Objectives

Steroid‐induced osteonecrosis of the femoral head (ONFH) is a common orthopaedic disease of which early detection remains clinically challenging. Accumulating evidences indicated that circulating microRNAs (miRNAs) plays vital roles in the development of several bone diseases. However, the association between circulating miRNAs and steroid‐induced ONFH remains elusive.

Materials and methods

miRNA microarray was performed to identify the differentially abundant miRNAs in the serums of systemic lupus erythematosus (SLE) patients with steroid‐induced ONFH as compared with SLE control and healthy control group. We predicted the potential functions of these differentially abundant miRNAs using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses and reconstructed the regulatory networks of miRNA‐mRNA interactions.

Results

Our data indicated that there were 11 differentially abundant miRNAs (2 upregulated and 9 downregulated) between SLE‐ONFH group and healthy control group and 42 differentially abundant miRNAs (14 upregulated and 28 downregulated) between SLE‐ONFH group and SLE control group. We also predicted the potential functions of these differentially abundant miRNAs using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses and reconstructed the regulatory networks of miRNA‐mRNA interactions.

Conclusions

These findings corroborated the idea that circulating miRNAs play significant roles in the development of ONFH and may serve as diagnostic markers and therapeutic targets.     

Decreased Serum Pituitary Adenylate Cyclase Activating Polypeptide (PACAP) Levels May Reflect Disease Severity in Patients with Non-traumatic Osteonecrosis of Femoral Head

To investigate serum pituitary adenylate cyclase activating polypeptide (PACAP) levels in relation to disease progression in patients diagnosed with non-traumatic osteonecrosis of femoral head (ONFH). A total of 102 non-traumatic ONFH patients and 95 healthy controls were enrolled in this study. Serum PACAP levels were examined using enzyme-linked immunosorbent assay (ELISA). Radiographic progression of ONFH was detected using the Association Research Circulation Osseous (ARCO) classification system. Harris hip score (HSS) and visual analogue scale (VAS) were analyzed to understand clinical severity. Serum levels of the ONFH marker IL-33 and β‐CTX were also detected. ROC curve analysis was performed to investigate the potential diagnostic value of serum PACAP in the radiographic progression of ONFH. Serum PACAP levels were significantly decreased in non-traumatic ONFH patients when compared to healthy controls. ARCO stage 4 showed markedly lower serum PACAP levels than stage 3. ARCO stage 3 had significantly decreased serum PACAP levels compared to stage 1/2. Serum PACAP levels were negatively correlated with the ARCO classification. In addition, serum PACAP levels were positively associated with HSS score and negatively related to VAS score. Last, PACAP levels were negatively correlated with serum IL-33 and positively associated with β-CTX. ROC curve analysis indicated that PACAP may serve as an early indicator for the radiographic progression of ONFH. Reduced serum PACAP levels may reflect disease severity in non-traumatic ONFH patients.

     

Metabolomic study of the bone trabecula of osteonecrosis femoral head patients based on UPLC–MS/MS

The severity and/or progression of osteonecrosis of the femoral head (ONFH) are commonly assessed by radiography, nuclear magnetic resonance image which aren’t invariably correlated to severity of disease and may be disturbed by other factors. Consequently, exploring the novel biochemical signatures of ONFH may be beneficial for diagnosing and understanding this disease. In this work, a bone trabecula metabolomics was undertaken to determine the expression pattern of low molecular mass metabolites in patients of femoral head necrosis based on the ultra-performance liquid chromatography/time-of-flight tandem mass spectrometry (UPLC/TOF MS/MS). Histological study showed that necrotic bone was characterized by necrosis, fibrosis and lacuna, but adjacent “normal” bone was pathologically normal. Principal component analysis in combination with orthogonal partial least-squares discrimination analysis was used to find out changed metabolites. MS/MS was used to speculate the corresponding molecule. Both osteonecrotic bone trabecula (ONBT) and adjacent “normal” bone trabecula (ANBT) showed higher levels of amino acids, such as proline, arginine, glutamine, dipeptides and lower levels of antioxidants. Most disrupted lipids, such as fatty acid esters, glycerophospholipids, sphingolipids, were found in osteonecrotic zone. The area under the receiver operating characteristic curve of combinational biomarkers (d-arginine, l-proline, l-carnitine, inosine) in ONBT and ANBT was 0.996 and 0.950, respectively. Our findings might provide a significant insight to understand the metabolic mechanism and diagnosis biomarkers of ONFH in the future.     

Prevalence of Clinical Anxiety,Clinical Depression and Associated Risk Factors in Chinese Young and Middle-Aged Patients with Osteonecrosis of the Femoral Head

Objective

To investigate the prevalence of clinical anxiety and clinical depression in Chinese young and mid-aged patients with osteonecrosis of the femoral head (ONFH) and to analyze their potential risk factors.

Methods

Two hundred and sixteen Chinese patients with ONFH were consecutively enrolled in this cross-sectional study from January 2010 to December 2010. The Zung self-rating anxiety scale (SAS) and the Zung self-rating depression scale (SDS) were used to assess the prevalence of clinical anxiety and clinical depression. An additional questionnaire containing seventeen items of potential risk factors was completed by all patients. Binary logistic regression analysis was employed to reveal potential risk factors of anxiety and depression.

Results

The prevalence of clinical anxiety and clinical depression was 20.4% and 21.8% in Chinese young and middle-aged patients with ONFH, respectively. Binary regression analysis showed that independent risk factors correlated with high incidence of clinical anxiety included involved femoral head (OR = 3.168, 95% CI: 1.496 - 6.708) and stages of ONFH (OR_{IV-V / II} = 5.383, 95% CI: 1.664-17.416). Independent risk factors correlated with high incidence of depression included gender (OR = 2.853, 95% CI: 1.467-5.778), comorbid diseases (OR = 4.243, 95% CI: 1.940-9.278) and stages of the disease (OR _IV-V/II = 16.963, 95% CI: 4.404-65.331).

Conclusions

Patients with bilateral ONFH are inclined to have clinical anxiety, while female patients and patients with comorbid diseases might tend to get clinical depression. Advanced stages of ONFH are independent risk factors for both clinical anxiety and clinical depression.     

骨源性细胞因子在激素性股骨头坏死发生发展中的研究进展

股骨头坏死是骨外科常见的难治性疾病,其机制仍有待研究.目前为止,医源性糖皮质激素是非创伤性股骨头坏死的主要原因.激素的长期使用可导致股骨头骨细胞凋亡、血液循环障碍所致缺血缺氧,最终导致股骨头塌陷.激素性股骨头坏死的发生发展与骨组织中细胞直接接触和其间接分泌的细胞因子调控相关.本文综述了骨组织中成骨细胞分泌的核因子κB受...     

Activation of cannabinoid receptor 2 alleviates glucocorticoid-induced osteonecrosis of femoral head with osteogenesis and maintenance of blood supply

In glucocorticoid (GC)-induced osteonecrosis of the femoral head (ONFH), downregulated osteogenic ability and damaged blood supply are two key pathogenic mechanisms. Studies suggested that cannabinoid receptor 2 (CB2) is expressed in bone tissue and it plays a positive role in osteogenesis. However, whether CB2 could enhance bone formation and blood supply in GC-induced ONFH remains unknown. In this study, we focused on the effect of CB2 in GC-induced ONFH and possible mechanisms in vitro and in vivo. By using GC-induced ONFH rat model, rat-bone mesenchymal stem cells (BMSCs) and human umbilical vein endothelial cells (HUVECs) to address the interaction of CB2 in vitro and in vivo, we evaluate the osteogenic and angiogenic effect variation and possible mechanisms. Micro-CT, histological staining, angiography, calcein labeling, Alizarin red staining (ARS), alkaline phosphatase (ALP), tartrate-resistant acid phosphatase (TRAP) staining, TUNEL staining, migration assay, scratch assay, and tube formation were applied in this study. Our results showed that selective activation of CB2 alleviates GC-induced ONFH. The activation of CB2 strengthened the osteogenic activity of BMSCs under the influence of GCs by promotion of GSK-3β/β-catenin signaling pathway. Furthermore, CB2 promoted HUVECs migration and tube-forming capacities. Our findings indicated that CB2 may serve as a rational new treatment strategy against GC-induced ONFH by osteogenesis activation and maintenance of blood supply. Subject terms: Prognostic markers, Calcium and phosphate metabolic disorders     

早期激素性股骨头坏死模型的建立

目的:建立早期股骨头坏死模型,为研究其发病机制及合理治疗方法的提供可靠的模型基础。方法:用单一剂量脂多糖LPS(10μg/kg)联合三次甲强龙MPS(10 mg/kg)注射,每次间隔24h,诱导建立兔早期股骨头坏死模型,通过影像学及组织病理性学方法评估模型建立情况。结果:4周后,模型组死亡1例,模型组16例X线检查未见异常表现,2例X线提示股骨头密度不均,未出现股骨头塌陷,18例HE染色示骨细胞空骨陷窝增多,脂肪细胞体积变大,数量增多。结论:用脂多糖(LPS)联合甲强龙可成功诱导建立兔早期股骨头坏死模型,模型成功率高、死亡率低。     

Biomaterial-loaded allograft threaded cage for the treatment of femoral head osteonecrosis in a goat model

The purpose of this study was to evaluate the efficacy of core decompression with a biomaterial-loaded allograft threaded cage (ATC) for the treatment of femoral head osteonecrosis in an established goat model. First, bilateral early-stage osteonecrosis was induced. After core decompression, the remaining goats were randomly divided into three groups: Group A, the goats were left without any treatment; Group B, the goats were treated with implanting a composite of autologous bone and decalcified bone matrix (DBM); Group C, the goats were treated using insertion of ATC loaded with DBM and autogenous bone graft. Then radiographic, histological and biomechanical analysis were taken in each group at 5, 10, and 20 weeks postoperation. In Group A, the classical signs of osteonecrosis of the femoral head were identified 10 weeks after the induction. Twenty weeks later, the density, surface and biomechanical stability of the femoral head were normal in Group C, while an irregular surface and an inhomogeneous microstructure or variation of density/hardness were identified in Group B. The specimens revealed a continuous trabaecular bone structure throughout the cage and extensive bone ingrowth and remodeling in Group C, while fibrous tissue was evident in Group B. Core decompression with a biomaterial loaded ATC almost uniformly delays or arrests the progression of the disease before articular collapse, and it could help to get the balance between bone resorption and new bone formation, strengthen structural mechanics of the femoral head, provide structure support of articular cartilage.     

Association analysis of tissue factor pathway inhibitor polymorphisms and haplotypes with osteonecrosis of the femoral head in the Korean population

Thrombophilia and hypofibrinolysis have been implicated in the pathogenesis of osteonecrosis of the femoral head (ONFH). Tissue factor pathway inhibitor (TFPI), a multivalent protease inhibitor, is an important regulator of the tissue factor-mediated blood coagulation pathway. Mutations of the TFPI gene can increase the risk of thrombin generation and venous thrombosis. The aim of this study was to evaluate the association of TFPI gene polymorphisms with ONFH. All exons and their boundaries of the TFPI gene, including the -1500 bp promoter region, were directly sequenced in 24 Korean individuals and four sequence variants were identified. These four polymorphisms [-51096 G > A (C-399T), -50984A > G (T-287C), + 24999A > G (Int7 -33T > C), + 37339T > A] were genotyped in 474 ONFH patients and 349 control subjects. The association of genotyped SNPs with ONFH was not found in the present study. The haplotype AAAT of TFPI was significantly associated with total, alcohol-induced, and idiopathic ONFH (p = 0.003, 0.021, and 0.007, respectively), and the haplotype GAAT was significantly associated with total and alcohol ONFH (p = 0.022 and 0.009, respectively). In addition, a new SNP + 37339 T > A in the 3'-UTR of the TFPI gene, was found in the Korean population. To date, this study is the first to show that haplotypes of the TFPI gene are associated with an increased susceptibility for ONFH. The results suggest that genetic variations in TFPI may play an important role in the pathogenesis and risk factors of ONFH.     

抑制JAK/STAT信号通路可减轻激素诱导的MRL/lpr狼疮鼠股骨头坏死

股骨头坏死(osteonecrosis of femoral head, ONFH)是系统性红斑狼疮(systematic lupus erythematosus, SLE)并发症之一,其发病机制复杂,治疗棘手,是SLE致残的主要原因。已有研究证实,JAK/STAT信号通路参与SLE的病理过程,而JAK/STAT激活也被发现与ONFH的发生有关。我们推测并证实,JAK/STAT信号通路在SLE-ONFH发生发展中发挥了重要作用。30只雌性MRL/lpr小鼠随机分为3组：模型组(脂多糖/24 h, 2次+甲基强的松龙/24 h, 3次)、对照组(加等量PBS)和治疗组(模型组+JAK1/2抑制剂巴瑞替尼/d, 6周),每组各10只。比较各组小鼠抓力的结果表明,模型组小鼠在第4周与第6周时,抓力值较对照组明显减少(P<0.05);治疗组小鼠在第6周时,抓力值优于模型组(P<0.05)。造模第6周处死小鼠取双侧股骨头,观察股骨头形态及HE染色病理改变。结果表明,对照组小鼠股骨头呈球型,透亮,骨质坚硬,无软骨缺损;模型组小鼠股骨头呈不规则型,粗糙,色泽灰暗,股骨头有部分缺损;治疗组...