Decreased serum MMP-9 levels in patients with nontraumatic osteonecrosis of the femoral head.

BACKGROUND: Matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinases-1 (TIMP-1) are involved in the pathological mechanism of osteonecrosis of the femoral head (ONFH). This study aimed to investigate the relationship of serum MMP-9, TIMP-1, and the MMP-9/TIMP-1 ratio with disease severity in patients with nontraumatic ONFH. METHODS: Serum levels of MMP-9 and TIMP-1 among 102 nontraumatic ONFH patients and 96 healthy individuals were determined by enzyme-linked immunosorbent assay (ELISA). Imaging severity was determined using the FICAT classification system. The Harris hip score (HHS) and visual analogue scale (VAS) were used to evaluate clinical progress. The correlations of serum MMP-9 and TIMP-1 levels with imaging severity and clinical progress was evaluated statistically. The diagnostic value of MMP-9 for NONFH disease severity was evaluated by examining receiver operating characteristic (ROC) curves. RESULTS: The serum MMP-9 levels and the MMP-9/TIMP-1 ratio were significantly increased in patients with ONFH compared to normal controls, and TIMP-1 levels did not differ between the two groups. Serum MMP-9 levels and the MMP-9/TIMP-1 ratio were positively correlated with FICAT stage and VAS and were negatively correlated with the HHS score. The ROC curve results indicated that MMP-9 could be used as a potential marker of nontraumatic ONFH imaging progression. CONCLUSIONS: We hypothesize that increased MMP-9 expression and an imbalance in the MMP-9/TIMP-1 ratio play a role in the development of ONFH and are correlate with the severity of ONFH. The determination of MMP-9 can be a useful tool to assess the severity of the disease in patients with nontraumatic ONFH.

A novel serological biomarker are associated with disease severity in patients with osteoarthritis.

BACKGROUND AND AIM: Apolipoprotein D (ApoD) is a 25-30 kDa glycoprotein, as a distinct component of the human plasma lipoprotein system. Its known biological functions are mainly related to lipid metabolism. The purpose of this study was to explore the potential role of ApoD concentration in knee osteoarthritis (KOA). METHODS: This study was performed in a population of 113 KOA subjects and 97 healthy controls. Serum ApoD was detected using the commercial ELISA kit (Cusabio, Wu Han, China, Cat No. CSB-EL001935HU). Radiographic progression was evaluated using Kellgren-Lawrence (KL) classification. The clinical severity of KOA was assessed by visual analog score (VAS), Hospital for special surgery (HSS) knee score disease duration and TNF-α. Receiver-operating characteristic (ROC) curve was performed to test the potential diagnostic value of ApoD in radiographic progression. RESULTS: The serum ApoD level of patients with KOA was significantly lower than that of healthy controls. The serum ApoD level was negatively correlated with KL grades, VAS score, disease duration, TNF-α concentrations. On the contrary, it was positively correlated with HSS score. However, there was no correlation between this serological indicator and which side was affected. ROC curve analysis indicated that attenuated serum ApoD could serve as an appropriate biomarker concerning the diagnosis of KOA. CONCLUSIONS: Serum ApoD concentration was correlated with the presence and severity of KOA.

Associations of TNF-α -238 A/G and IL-10 -1082 G/A Genetic Polymorphisms With the Risk of NONFH in the Chinese Population.

The study aims to explore the roles of common polymorphisms of Tumor Necrosis Factor-α (TNF-α) (-238 A/G and -308 A/G) and IL-10 (-819 T/C and -1082 G/A) genes in the risk of non-traumatic osteonecrosis of the femoral head (NONFH). One hundred and forty-seven NONFH patients and 135 healthy individuals were selected as the case and control groups. qRT-PCR and Western blotting techniques detected mRNA as well as protein expressions of TNF-α and IL-10 of each genotype in both the case and control groups. The GA genotype and the A allele of TNF-α -238 A/G were higher in the case group than in the control group. Compared with the control group, AA, GA, and AG + AA genotypes as well as the A allele of IL-10-1082 G/A were all lower in the case group. In the case groups increased levels of TNF-α as well as decreased levels of IL-10 expression when compared with the control group. TNF-α expression of TNF-α-238 GA genotype was significantly higher than that in patients with GG genotype, while the IL-10 expression of GA and AA genotypes of IL-10-1082 was significantly lower than in that of patients with the GG genotype. TNF-α protein expression in the GA genotype was significantly higher than in the GG genotype. In relation to TNF-α -238, TNF-α protein expression of GA and AA genotypes had significantly reduced more so than the GG genotype in IL-10-1082. TNF-α-238 A/G and IL-10-1082 G/A may be involved as risk factors of NONFH. J. Cell. Biochem. 118: 4872-4880, 2017. © 2017 Wiley Periodicals, Inc.

Efficacy of Fu-Yuan Capsule in the Healing of Fractures of the Lower End of the Radius in a Rabbit Model.

OBJECTIVE: The aim of the study was to explore the efficacy of Fu-Yuan Capsule in the healing of fractures of the lower end of the radius in a rabbit model. METHODS: After establishing a rabbit fracture model, all animals were randomly divided into the model group (n = 24), the Fu-Yuan Capsule group (n = 24), and the Shenyang Hongyao group (n = 24). The X-ray was applied to observe the course of fracture healing at 2, 4, 6, and 8 weeks after treatment. Haematoxylin-eosin staining and immunohistochemical staining were used to determine the histological change and the expression of bone morphogenetic protein-2 (BMP-2). Serum alkaline phosphatase (ALP), calcium, and phosphorus levels were detected before and after treatment. RESULTS: X-ray showed that the Fu-Yuan Capsule and Shenyang Hongyao groups exhibited abundant callus shadow areas than the model group in a time-dependent manner. In the model group, the fractures exhibited poor recovery with fibrous callus and obstructed bone marrow cavity. In the Fu-Yuan Capsule and Shenyang Hongyao groups, the fracture showed good recovery and restored normal structure with an effective remodeling of the lamellar bone. Immunohistochemical staining showed that the Fu-Yuan Capsule and Shenyang Hongyao groups had higher expressions of BMP-2 than the model group. Furthermore, serum ALP and calcium-phosphorus product in the Fu-Yuan Capsule and Shenyang Hongyao groups were higher than what they were in the model group. CONCLUSIONS: These results suggest that Fu-Yuan Capsule could promote the fracture healing through upregulating BMP-2 expression and increasing serum ALP and calcium-phosphorus product.

Astragaloside I Stimulates Osteoblast Differentiation Through the Wnt/ß-catenin Signaling Pathway.

Astragaloside I (As-I), one of the main active ingredients in Astragalus membranaceus, is believed to have osteogenic properties, but this hypothesis has not been investigated in detail. In the present work, the As-I-induced osteogenic effects and its underlying mechanism were studied in MC3T3-E1 cells. The results indicated that the cellular levels of ALP and extracellular matrix calcium increased in a dose-dependent manner by As-I. To clarify the mechanisms involved in this process, the effect of As-I on the key osteogenic-related genes was investigated. We found that As-I stimulated the expression of ß-catenin and Runx2 in MC3T3-E1 cells, which play central roles in the Wnt/ß-catenin signaling pathway, suggesting that As-I could promote osteoblastic differentiation by regulating the Wnt/ß-catenin signaling pathway. Moreover, the osteogenic effect of As-I could be inhibited by DKK-1, which is the classical inhibitor of Wnt/ß-catenin-signaling pathway. Furthermore, As-I also increased BMP-2, BGP and OPG/RANKL expression, which are also activated by Wnt/ß-catenin signaling pathway. Taken together, our findings show that As-I stimulates osteoblast differentiation through the Wnt/ß-catenin signaling pathway, which also activates the BMP pathway and RANK pathway, thus highlighting the As-I for pharmaceutical and medicinal applications such as treating bone disease. Copyright © 2016 John Wiley & Sons, Ltd.

Comparison of topical and intravenous tranexamic acid on blood loss and transfusion rates in total hip arthroplasty.

The objective of this study was to determine whether topical tranexamic acid (TXA) carried similar hemostatic effect compared with intravenous TXA in total hip arthroplasty (THA). Three hundred and three THA patients were enrolled and randomized into 3 groups: no TXA group, topical and intravenous TXA group. The results showed that the topical and intravenous TXA group had reduced but similar blood transfusion rates (5.88% v. s. 5.94%, P = 0.816). No significant difference was detected in total blood loss between the two TXA groups [(963.4 ± 421.3) ml vs. (958.5 ± 422) ml P = 0.733]. We conclude that topical use of TXA was equally effective and safe compared with intravenous TXA in reducing blood loss and transfusion rate following THA without substantial complications.

Correlation of serum and local CXCL13 levels with disease severity in patients with non-traumatic osteonecrosis of femoral head.

OBJECTIVE: The primary aim of the present study was to explore the potential correlation of serum / local CXCL13 expressions and disease severity in non-traumatic osteonecrosis of the femoral head (NT-ONFH). METHODS: In total, NT-ONFH patients (n = 130) together with healthy controls (HCs, n = 130) were included in this investigation. Radiographic progression was evaluated based on the imaging criteria outlined in the ARCO classification system. To assess the diagnostic value of serum CXCL13 in relation to radiographic progression, Receiver operating characteristic (ROC) curve analysis was conducted. Serum CXCL13 levels were quantified utilizing ELISA in all participants. Furthermore, local protein/mRNA expressions of CXCL13 were examined employing immunohistochemistry, western blot, as well as RT-PCR techniques. Clinical severity was appraised using the visual analogue scale (VAS), Harris Hip Score (HHS), and Western Ontario as well as McMaster Universities Osteoarthritis Index (WOMAC). RESULTS: The findings revealed a significant reduction in serum CXCL13 levels among NT-ONFH patients in contrast with HCs. Moreover, both mRNA and protein expressions of CXCL13 were markedly decreased in the necrotic area (NA) than the non-necrotic area (NNA) as well as the healthy femoral head tissues. Additionally, serum CXCL13 levels were substantially lower among patients classified as ARCO stage 4 than those at ARCO stage 3. The concentrations of CXCL13 in stage 3 patients were notably diminished relative to those at ARCO stage 2. Notably, serum CXCL13 levels demonstrated a negative association with ARCO grade. Furthermore, these levels were also inversely linked to VAS scores as well as WOMAC scores while displaying a positive association with HHS scores. The findings of ROC curve suggested that reduced serum CXCL13 levels could be an underlying indicator for ARCO stage. CONCLUSIONS: The reduced levels of either serum CXCL13 or local CXCL13 were intricately linked to disease severity for patients with NT-ONFH.

Serum glutathione peroxidase 4 as a novel biomarker for nontraumatic osteonecrosis of the femoral head: A retrospective case-control study.

There are no serum biomarkers available in nontraumatic osteonecrosis of the femoral head in clinical practice. This study aimed to evaluate the clinical value of serum glutathione peroxidase 4 in nontraumatic osteonecrosis of the femoral head. This retrospective study analyzed serum glutathione peroxidase 4 levels and clinical data of 80 patients with nontraumatic osteonecrosis of the femoral head and 80 healthy controls between August 2021 and May 2022. Serum glutathione peroxidase 4 levels were analyzed using an enzyme-linked immunosorbent assay. The Association Research Circulation Osseous classification system determined disease progression. Clinical severity was assessed by Harris hip score and visual analogue scale. Correlations between serum glutathione peroxidase 4 and disease progression as well as clinical severity were evaluated statistically. The diagnostic accuracy of serum glutathione peroxidase 4 in nontraumatic osteonecrosis of the femoral head was determined using receiver operating characteristic analysis. The baseline characteristics of participants between 2 groups were comparable. Patients with nontraumatic osteonecrosis of the femoral head displayed a decreased glutathione peroxidase 4 level compared with healthy controls (11.87 ±â€…2.76 µU/mL vs 16.54 ±â€…4.89 µU/mL, P < .01). The levels of glutathione peroxidase 4 were inversely correlated with Association Research Circulation Osseous stage (P < .01) and visual analogue scale scores (P < .01), and positively correlated with Harris score (P < .01). Receiver operating characteristic analyses showed that area under curves of glutathione peroxidase 4 was 0.808 (95% CI 0.721-0.858) and 0.847 (95% CI 0.743-0.951) with regard to diagnosis and collapse prediction in nontraumatic osteonecrosis of the femoral head, respectively. Serum glutathione peroxidase 4 could serve as a novel biomarker for diagnosing nontraumatic osteonecrosis of the femoral head and predicting collapse of the femoral head.

Reduced serum and local LncRNA MALAT1 expressions are linked with disease severity in patients with non-traumatic osteonecrosis of the femoral head.

OBJECTIVE: This study was performed to illustrate the potential relationship between reduced serum and local LncRNA MALAT1 expressions with disease severity in patients with non-traumatic osteonecrosis of the femoral head (ONFH). METHODS: A total of 104 patients with non-traumatic ONFH and 100 healthy controls were consecutively recruited from our hospital. Serum and local LncRNA MALAT1 expressions were detected using real-time polymerase chain reaction (RT-PCR). Radiographic progression was defined by Ficat classification. Clinical severity was evaluated by Visual Analog Scale (VAS) and Harris Hip Score (HHS). Receiver operating characteristic (ROC) curve was carried out to determine the diagnostic value of MALAT1 in the radiographic progression. RESULTS: Serum LncRNA MALAT1 expressions were significantly lower in non-traumatic ONFH patients than in healthy controls. In addition, local MALAT1 expressions in non-traumatic ONFH tissue were significantly lower in the affected area than in the non-affected area. Ficat grade 4 has significantly lower serum and local LncRNA MALAT1 expressions in comparison with grade 3, and Ficat grade 3 showed markedly decreased serum and local LncRNA MALAT1 expressions compared with grade 2. Serum and local LncRNA MALAT1 expressions were significantly and negatively associated with VAS and positively related to the HHS. Further ROC curve analysis indicated that serum MALAT1 may act as a decent indicator in the diagnosis of non-traumatic ONFH. CONCLUSIONS: Decreased serum and local MALAT1 expressions may reflect disease severity in non-traumatic ONFH patients.

Attenuated serum vasoactive intestinal peptide concentrations are correlated with disease severity of non-traumatic osteonecrosis of femoral head.

BACKGROUND AND OBJECTIVE: The neuropeptide vasoactive intestinal peptide is a 28-amino acid neuropeptide that has been shown to stimulate bone repair and angiogenesis. The purpose of this study was to explore the potential role of serum VIP concentration in osteonecrosis of femoral trauma (ONFH). METHODS: One hundred five patients diagnosed with non-traumatic ONFH and 103 healthy individuals were enrolled in our study. Serum VIP, tumor necrosis factor-α (TNF-α), interluekin-1 beta (IL-1ß), and macrophage colony-stimulating factor (M-CSF) levels also were detected using the commercial ELISA kit. Radiographic progression was evaluated using FICAT classification. The clinical severity of ONFH was assessed by visual analog score (VAS) and Harris Hip Score (HHS). Receiver-operating characteristic (ROC) curve was performed to test the potential diagnostic value of VIP in radiographic progression. RESULTS: The serum VIP level of patients with non-traumatic ONFH was significantly lower than that of healthy controls. There was no significant difference between the alcohol group, the steroid-induction group, and the idiopathic group. Serum VIP levels were significantly higher in ONFH patients with femoral head pre-collapse stage than collapse stage. Serum VIP levels were significantly lower. FICAT 4 non-traumatic ONFH patients had significantly lower serum concentrations of VIP when compared with FICAT 3 and FICAT 2. Moreover, serum VIP levels were significantly lower in ONFH patients with FICAT 3 than FICAT 2. Serum VIP levels were negatively related to FICAT stage. In addition, serum VIP levels were negatively associated with VAS score and positively associated with HHS score. Last, we found serum VIP levels were negatively associated with serum TNF-α and IL-1ß levels. ROC curve analysis indicated that decreased serum VIP could serve as a decent biomarker with regard to the diagnosis of radiographic progression. CONCLUSION: Attenuated serum VIP concentrations are correlated with disease severity of non-traumatic ONFH. Decreased serum VIP may serve as a potential indicator of non-traumatic ONFH.

Clinical observation and finite element analysis of cannulated screw internal fixation in the treatment of femoral neck fracture based on different reduction quality.

OBJECTIVE: Femoral neck fracture is one of the most common bone types. The effect of reduction quality on hip joint function and complications after screw internal fixation is not fully understood. To investigate the clinical efficacy and mechanical mechanism of positive buttress, anatomical reduction, and negative buttress in the treatment of femoral neck fracture after cannulated screw fixation. METHODS: Retrospective analysis of patients with femoral neck fracture treated with three cannulated screws internal fixation in our hospital from January 2013 to December 2018. According to the quality of fracture reduction, the patients were divided into positive buttress group, anatomical reduction group, and negative buttress group. Basic information such as injury mechanism, time from injury to surgery, Garden classification and Pauwels classification was collected, Harris scores were performed at 3 months, 6 months, and 12 months after surgery, and postoperative complications (femoral head necrosis, femoral neck shortening, and femoral neck nonunion) were collected. At the same time, three groups of finite element models with different reduction quality were established for stress analysis, their stress clouds were observed and the average displacement and stress of the three groups of models were compared. P < 0.05 was used to represent a statistically significant difference. RESULTS: A total of 225 cases of unilateral femoral neck fractures were included and followed up for an average of 4.12 ± 0.69 years. There was no significant difference in age, gender, side, injury mechanism, time from injury to surgery, BMI, Garden classification, Pauwels classification, and follow-up time among the three groups (P > 0.05). However, there was significant difference in Harris score at 6 and 12 months after operation among the three groups (P < 0.05), which was higher in the positive buttress group and anatomical reduction group than in the negative buttress group. In addition, the incidence of osteonecrosis of the femoral head in the negative buttress group (32.2%) was greater than that in the anatomical reduction group (13.4%) and the positive buttress group (5.4%) (P < 0.05). In addition, the incidence of femoral neck nonunion and femoral neck shortening in the negative buttress group was also higher than that in the anatomical reduction positive buttress group (P < 0.05). The finite element results showed that the stress and fracture end displacement in the negative buttress group were greater than those in the positive buttress group (P < 0.05). CONCLUSION: Both positive buttress and anatomical reduction in the treatment of femoral neck fracture with cannulated screw internal fixation can obtain better clinical effect and lower postoperative complications. Positive brace support and anatomic reduction can limit the restoration of femoral stress conduction. Therefore, it is not necessary to pursue anatomical reduction too deliberately during surgery, while negative buttress reduction should be avoided.

Bioinformatics analysis of microRNA linked to ubiquitin proteasome system in traumatic osteonecrosis of the femoral head.

MicroRNAs (miRNAs) have been suggested to act critical roles in the pathophysiology of traumatic osteonecrosis of the femoral head (TONFH). Unfortunately, their roles in the development of TONFH are still ambiguous. The purpose of this study is to identify promising miRNA biomarkers in traumatic osteonecrosis development.We conducted a comprehensive bioinformatics analysis using microarray datasets downloaded from the Gene Expression Omnibus database, and compared the expression of miRNAs in the serum of TONFH patients with controls. Next, we performed target prediction, function enrichment analysis, and protein-protein interaction network analysis based on differentially expressed (DE) miRNAs.We identified 26 DE miRNAs that may contribute to the pathophysiology of TONFH. The miRNAs were linked to ubiquitin proteasome system including conjugating protein ligase activity, ubiquitin-protein ligase activity and ubiquitin mediated proteolysis 5 pathway, and we exposed miR-181a-5p and miR-140-5p as promising biomarkers in TONFH.A predicting model consisting of 5 miRNAs may help discriminating high-risk patients who might develop TONFH after femur neck fracture. Among DE miRNAs, MiR-181a-5p and miR-140-5p may contribute to the development femoral head osteonecrosis after femur neck fracture via ubiquitin proteasome system.

[Impacting bone graft via surgical hip dislocation approach versus core decompression and bone graft for avascular necrosis of femoral head at ARCO stage â ¢].

OBJECTIVE: To compare the effectiveness of sequestrum clearance and impacting bone graft via surgical hip dislocation approach and core decompression and bone graft for avascular necrosis of the femoral head (ANFH) at Association Research Circulation Osseous (ARCO) stage Ⅲ. METHODS: A clinical data of 60 patients (69 hips) of non-traumatic ANFH at ARCO stage Ⅲ, which met the inclusion criteria between October 2013 and April 2016, was retrospectively analyzed. Among them, 24 patients (28 hips) were treated with sequestrum clearance and impacting bone graft via surgical hip dislocation approach (group A); and 36 patients (41 hips) were treated with core decompression, sequestrum clearance, impacting bone graft, and nonvascular fibular allograft supporting (group B). There was no significant difference in gender, age, disease duration, affected side, type and stage of the ANFH, and preoperative Harris hip score and visual analogue scale (VAS) score between the two groups ( P>0.05). After operation, the function of the hip was evaluated by Harris hip score, imaging examination was performed to observe the femoral head shape and evaluate whether the hip preserving success. RESULTS: The incisions of two groups healed by first intention. All patients were followed up. The follow-up time was 12-48 month (mean, 25.8 months) in group A and 12-54 months (mean, 26.4 months) in group B. At last follow-up, 5 hips in group A were classified as clinical failure, femoral head survival rate was 82.1%, the median survival time was 43 months. While 19 hips in group B were classified as clinical failure, femoral head survival rate was 53.7%, the median survival time was 42 months. There was significant difference in survival curve distribution between the two groups ( χ 2= 4.123, P=0.042), and the surgical procedures of group A was superior to group B. In the two groups, the Harris hip scores at last follow-up were significantly higher than preoperative ones ( P<0.05), and VAS scores were significantly lower than preoperative ones ( P<0.05). There was no significant difference in Harris hip score and VAS score at last follow-up between the two groups ( P>0.05). All grafted bones got fusion according to the X-ray films, and there was no significant difference in the fusion time between the two groups ( t=0.752, P=0.456). In group A, greater trochanter bone cutting were healed well; and the heterotopic ossification around the hip joint occurred in 1 case. CONCLUSION: The surgery of impacting bone graft via surgical hip dislocation approach and core decompression and bone graft can be applied to treat ANFH at ARCO stage ⅢA which was mild collapse and satisfactory effectiveness can be obtained. While for the patients of ANFH at ARCO stage Ⅲ B with severe collapse, the hip survival rate of the former is better than that of the latter.

Identification of Key Genes and Pathways Associated with Sex Differences in Osteoarthritis Based on Bioinformatics Analysis.

Sex differences have been suggested to play critical roles in the pathophysiology of osteoarthritis (OA), resulting in sex-specific prevalence and incidence. However, their roles in the development of OA remain largely unknown. The aim of this study was to screen out key genes and pathways mediating biological differences between OA females after menopause and OA males. First, the gene expression data of GSE36700 and GSE55457 were downloaded from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) between sexes were identified using R software, respectively. The overlapping DEGs were obtained. Then, protein-protein interactive (PPI) network was constructed to further analyze interactions between the overlapping DEGs. Finally, enrichment analyses were separately performed using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes tools. In our results, a total of 278 overlapping DEGs were identified between OA females after menopause and OA males, including 219 upregulated and 59 downregulated genes. In the PPI network, seven hub genes were identified, including EGF, ERBB2, CDC42, PIK3R2, LCK, CBL, and STAT1. Functional enrichment analysis revealed that these genes were mainly enriched in PI3K-Akt signaling pathway, osteoclast differentiation, and focal adhesion. In conclusion, the results in the current study suggest that pathways of PI3K-Akt, osteoclast differentiation, and focal adhesion may play important roles in the development of OA females after menopause. EGFR, ERBB2, CDC42, and STAT1 may be key genes related to OA progression in postmenopausal women and may be promising therapeutic targets for OA.

LY404039 Inhibits Proliferation and Metastasis of Osteosarcoma Cells via PI3K Signaling Pathway.

Osteosarcoma is one of the most aggressive primary malignant bone tumors, and the effect of first-line therapeutic schedule had no improvement in past few years. Therefore, it is extremely urgent to develop new drugs against osteosarcoma. In this study, human osteosarcoma U2 OS cells as a model in vitro were treated with LY404039 and dimethyl sulfoxide. CCK-8 proliferation test, transwell assay, and flow cytometry were used to assess cell proliferation, invasion/migration, and apoptosis, respectively. Moreover, western blotting was performed to determine the relative expression of protein related to apoptosis and PI3K signaling pathway. The proliferation of U2 OS cells was significantly inhibited 72 h after LY404039-treatment by CCK-8 proliferation test. Matrigel invasion analysis showed the numbers of invasive cells were significantly decreased in LY404039-treated group compared with the NC group. Furthermore, LY404039 led to the remarkable reduction of the number of U2 OS cells passing through the microwells of the transwell chamber. The expression levels of anti-apoptotic protein BCL-2 decreased, and the expression of pro-apoptotic proteins Active Caspase3, Bax increased concurrently after LY404039 stimulation using western blot. The phosphorylation of AKT and mTOR were significantly inhibited in U2 OS cells treated with LY404039. Also, the activity of the downstream proteins such as p70S6K and Cyclin D1 were also significantly decreased. LY404039 could inhibit proliferation and metastasis of osteosarcoma cells, which might be attributed to the enhancement of apoptosis via PI3K pathway.

Association of MMP-8 rs2012390 and rs11225394 polymorphisms with osteonecrosis of the femoral head risks: Evidence from a meta-analysis.

BACKGROUND: The association of MMP-8 rs2012390 and rs11225394 polymorphisms with osteonecrosis of the femoral head (ONFH) risks was investigated in several studies with conflicting results. We performed the meta-analysis to evaluate the association between them. METHODS: Potentially relevant literatures were searched from the electronic databases of PubMed, Web of Science, and Cochrane Library. All databases were searched up to May 6, 2018. The strength of associations of the MMP-8 rs2012390 and rs11225394 polymorphisms with ONFH risk was assessed by crude odds ratios (ORs) with their 95% confidence intervals (CIs) under different genetic models. RESULTS: A total of 1469 cases diagnosed with ONFH and 1211 healthy controls were included in the current meta-analysis. A remarkable association between rs11225394 in the MMP-8 gene and an increased risk of ONFH was found (allele model: OR = 1.33, 95% CI = 1.09-1.61, P = .005; heterozygote model: OR = 1.39, 95% CI = 1.13-1.71, P = .002; dominant model: OR = 1.40, 95% CI = 1.14-1.73, P = .002, respectively). Meanwhile, a significant association between MMP-8 rs2012390 and the decreased risk of ONFH was found in heterozygote model (OR = 0.63, 95% CI = 0.51-0.77, P < .00001). CONCLUSION: The meta-analysis results showed a remarkable association between rs11225394 in MMP-8 gene and an increased risk of ONFH and a significant association between MMP-8 rs2012390 and the decreased risk of ONFH.

Differentially Expressed MicroRNAs in Conservatively Treated Nontraumatic Osteonecrosis Compared with Healthy Controls.

Deregulation of microRNAs (miRNAs) contributes to nontraumatic osteonecrosis of the femoral head (ONFH-N), but the differentially expressed circulating miRNAs in patients with ONFH-N receiving nonsurgical therapy are unknown. This study aimed to determine the miRNAs expression profile of patients with ONFH-N receiving conservative treatments. This was a case-control prospective study of 43 patients with ONFH-N and 43 participants without ONFH-N, enrolled from 10/2014 to 10/2016 at the Department of Orthopedics of the Linyi People's Hospital (China). The two groups were matched for age, gender, and living area. Microarray analysis and quantitative RT-PCR were used to examine the differentially expressed miRNAs. Bioinformatics was used to predict miRNA target genes and signaling pathways. Microarray and quantitative RT-PCR revealed that nine miRNAs were downregulated and five miRNAs were upregulated in ONFH-N (n = 3) compared with controls (n = 3). Bioinformatics showed that calcium-mediated signaling pathway, regulation of calcium ion transmembrane transporter activity, cytoskeletal protein binding, and caveolae macromolecular signaling complex were probably regulated by the identified differentially expressed miRNAs. In the remaining 80 subjects (n = 40/group), miR-335-5p was downregulated (P = 0.01) and miR-100-5p was upregulated (P = 0.02) in ONFH-N compared with controls. In conclusion, some miRNAs are differentially expressed in conservatively treated ONFH-N compared with controls. Those miRNAs could contribute to the pathogenesis of ONFH-N.

Attenuated serum adiponectin levels are associated with disease severity in patients with non-traumatic osteonecrosis of the femoral head.

OBJECTIVE: Decreased adiponectin (APN) levels have been detected in patients with osteonecrosis of the femoral head (ONFH). The scope of this study was aimed to explore the relationship between serum APN levels and disease severity in nontraumatic ONFH patients. PATIENTS AND METHODS: Ninety two nontraumatic ONFH patients and 92 healthy controls were enrolled for this study following the estimation of sample size. Serum APN concentrations were examined by enzyme-linked immunosorbent assay (ELISA). The radiographic progression was determined by Ficat grading system. The clinical severity was evaluated by visual analog scale (VAS), Harris hip scores (HSSs) and Western Ontario and McMaster University Osteoarthritis Index (WOMAC) scores. RESULTS: Serum APN levels were significantly lower in ONFH patients than in healthy controls. Serum APN levels were significantly lower in patients with Ficat stage 4 ONFH than in patients with stage 3 ONFH, and APN levels in patients with stage 3 were lower compared with stage 2. Serum APN levels were negatively correlated with the Ficat grading system. In addition, serum APN levels were also negatively related to VAS and WOMAC scores and positively associated with HSSs. CONCLUSION: Decreased serum APN levels may reflect disease severity of nontraumatic ONFH.

Long non-coding RNA HOTAIR inhibits miR-17-5p to regulate osteogenic differentiation and proliferation in non-traumatic osteonecrosis of femoral head.

BACKGROUND AND AIM: The biological functions of non-coding RNAs (ncRNAs) have been widely identified in many human diseases. In the present study, the relationship between long non-coding RNA HOTAIR and microRNA-17-5p (miR-17-5p) and their roles in osteogenic differentiation and proliferation in non-traumatic osteonecrosis of femoral head (ONFH) were investigated. METHODS: The expression levels of HOTAIR and miR-17-5p in the mesenchymal stem cells (MSCs) derived from patients with non-traumatic ONFH and osteoarthritis (OA) were examined by real-time PCR. BMP-2 induced human MSCs from bone marrow (hMSC-BM) were used for osteogenic differentiation. RESULTS: It was observed that the expression level of miR-17-5p was lower and the level of HOTAIR was higher in samples of non-traumatic ONFH compared with OA. HOTAIR downregulation induced by si-HOTAIR led to the increase of miR-17-5p expression and the decrease of miR-17-5p target gene SMAD7 expression. The values of osteogenic differentiation markers, including RUNX2 and COL1A1 mRNA expression and ALP activity, were also elevated by si-HOTAIR. However, the increase of these values was canceled by miR-17-5p inhibitor or SMAD7 upregulation. CONCLUSION: HOTAIR played a role in regulating osteogenic differentiation and proliferation through modulating miR-17-5p and its target gene SMAD7 in non-traumatic ONFH.

Identification of aberrantly expressed of serum microRNAs in patients with hormone-induced non-traumatic osteonecrosis of the femoral head.

OBJECTIVE: The non-translation RNA-microRNA (miRNA) has been demonstrated to correlate to various disease occurrence in body. Serum miRNA was gradually considered as molecular markers for disease diagnosis. This study was designed to analyze differential serum miRNAs level in hormone-induced non-traumatic osteonecrosis of the femoral head (hormone-NOFH) patients. METHODS: We selected 30 patients with hormone-NOFH as case group, and 30 healthy volunteers were recruited as control group. miRCURYTM LNA miRNA chip and quantitative RT-PCR were used to examine differential miRNAs expression. Correlation assay was performed between miRNAs and NOFH trait. RESULTS: We found that 9 miRNAs were upregulated while 3 miRNAs were downregulated in hormone-TOFH patient serum by result of miRNA chip. QRT-PCR assay revealed that the level of miR-423-5p was significantly increased and miR-10a-5p was significantly decreased. Using Spearman correlation analysis, we observed that miR-423-5p serum level is positive association to FHC levels whereas miR-10a-5p has no association with FHC levels. Furthermore, miR-423-5p is negatively correlated to its downstream molecule-adiponectin. CONCLUSION: We report a miRNA profile of hormone-NOFH and provide a new perspective to understand this intricate disease. This novel information suggests the potential roles of miR-423-5p in the diagnosis, prognosis biomarkers, or therapy targets of hormone-NOFH.