Intercalary frozen autografts for reconstruction of bone defects following meta-/diaphyseal tumor resection at the extremities.

BACKGROUND: For patients with malignant limb tumors, salvage surgery can be achieved using endoprosthesis or biological reconstructions like allograft or autograft. In carefully selected patients, resected bone can be recycled after sterilization using methods like autoclaving, irradiation, pasteurization or freezing with liquid nitrogen. We evaluated the clinical outcome and complications of malignant limb tumors treated with intercalary resection and frozen autograft reconstruction. METHODS: We reviewed 33 patients whose malignant bone tumors were treated by wide resection and reconstruction with recycling liquid nitrogen-treated autografts between 2006 and 2017. Limb function, bone union at the osteotomy site and complications were evaluated. Functional outcome was assessed using the Musculoskeletal Tumor Society (MSTS) scoring system. RESULTS: The cohort comprised 16 males and 17 females, with a mean age of 35.4 years (14-76 years). The most common tumor was osteosarcoma (7 cases). Tumors were located in the humerus (5), ulna (1), femur (10) and tibia (17). The mean follow-up was 49.9 months (range 12-127 months). Of the 33 patients, 16 remained disease-free, and 3 were alive with disease. The mean size of the defect after tumor resection was 11.6 cm (range 6-25 cm). Bone union was achieved in 32 patients, with a mean union time of 8.8 months (range 4-18 months). Complications included 1 graft nonunion, 2 infections (1 superficial, 1 deep infection), 1 leg length discrepancy, 2 graft fractures and 3 local recurrences. The mean MSTS score was 87.2% (range 70-100%). CONCLUSION: Liquid nitrogen-treated tumor-bearing autograft is an effective option for biological reconstruction after meta-/diaphyseal tumor resection of long bones. This method has excellent clinical outcomes and is especially recommended for patients with no severe osteolytic bone tumors.

Inositol polyphosphate-4-phosphatase type II and rucaparib treatment inhibit the growth of osteosarcoma cells dependent on phosphoinositide 3-kinase/protein kinase B pathway.

Osteosarcoma (OS) is an aggressive malignant tumor of bone, which often occurs in children and adolescents. Currently, the effective method for the treatment of OS is still limited. The study aimed to investigate the synergistic antitumor effect of inositol polyphosphate-4-phosphatase, type-II (INPP4B) and rucaparib on OS cells. The expression levels of INPP4B in OS tissues and OS cell lines were examined by quantitative real-time polymerase chain reaction and Western blot analysis. SaOS2 and U2OS cells were then transfected with overexpression vector of INPP4B or were treated with different concentrations of rucaparib, and cell viability, cell cycle, and apoptosis were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and flow cytometry. Western blot assay uncovered the combined effects of INPP4B and rucaparib on cell cycle, apoptosis and phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) signal pathway. Further, the tumor formation was examined in vivo. Results showed that INPP4B was low expressed in OS tissues and in OS cell lines. INPP4B overexpression significantly decreased cell viability and induced apoptosis in SaOS2 and U2OS cells. Additionally, rucaparib remarkably reduced cell viability in a dose-dependent and time-dependent manner. Meanwhile, rucaparib suppressed cell cycle progression in the S phase and promoted apoptosis in a dose-dependent manner. Further, combination of INPP4B overexpression and rucaparib declined Myc, cyclin E1 and cyclin D1 expressions, enhanced Bad, Bax, and cleaved-caspase-3 expressions, and blocked PI3K/AKT signal pathway in SaOS2 and U2OS cells. Finally, combination of INPP4B overexpression and rucaparib inhibited tumor formation in vivo. The study demonstrated that INPP4B and rucaparib exhibited synergistic antitumor effect by regulating PI3K/AKT pathway in OS cells.

Diagnostic accuracy of fluorine-18 fluorodeoxyglucose positron emission tomography for suspected primary and postoperative pyogenic spondylitis.

OBJECTIVE: Fluorine-18 fluorodeoxyglucose positron emission tomography (18F-FDG PET) and PET/CT have been suggested for confirming or excluding musculoskeletal infection but the diagnostic value of this tool for pyogenic spondylitis remains to be confirmed. This meta-analysis was performed to verify the accuracy of 18F-FDG PET and PET/CT in diagnosing suspected pyogenic spondylitis by performing a systematic review and meta-analysis. METHODS: We conducted a comprehensive literature search of PubMed, Embase and Cochrane Library to retrieve diagnostic accuracy studies in which suspected pyogenic spondylitis was assessed with 18F-FDG PET or PET/CT. The pooled sensitivity, specificity, likelihood ratios, diagnostic odds ratio (DOR), summarized receiver operating characteristic curve (sROC) and the area under the sROC (AUC) were calculated by using Stata software. RESULTS: A total of 18 eligible studies (660 patients) with suspected pyogenic spondylitis were included in the quantitative analysis. 18F-FDG PET and PET/CT illustrated relatively high sensitivity (0.91, 95% CI: 0.84-0.95) and specificity (0.90, 95% CI: 0.79-0.95) for the diagnosis of pyogenic spondylitis. The pooled DOR and AUC were 86.00 (95% CI, 31.00-240.00) and 0.96 (95% CI, 0.94-0.97), respectively. For diagnosing pyogenic spondylitis without previous spine surgery, the pooled sensitivity, specificity, DOR and AUC were 0.93 (95% CI, 0.85-0.97), 0.91 (95% CI, 0.77-0.97), 136 (95% CI, 35-530) and 0.97 (95% CI, 0.95-0.98), respectively. For diagnosing postoperative pyogenic spondylitis, the pooled sensitivity, specificity, DOR and AUC were 0.85 (95% CI, 0.71 to 0.93), 0.87 (95% CI, 0.66 to 0.96), 38 (95% CI, 9 to 167) and 0.92 (95% CI, 0.89 to 0.94), respectively. CONCLUSION: 18F-FDG PET and PET/CT presented satisfactory accuracy for diagnosing pyogenic spondylitis. The diagnostic effect of this nuclear imaging method for pyogenic spondylitis without previous spine surgery seems to be better than that for the postoperative ones. However, whether 18F-FDG PET and PET/CT could become a routine in patients with suspected pyogenic spondylitis remains to be confirmed. LEVEL OF EVIDENCE: Level I evidence, a summary of meta-analysis.

Mechanism of immune infiltration in synovial tissue of osteoarthritis: a gene expression-based study.

BACKGROUND: Osteoarthritis is a chronic degenerative joint disease, and increasing evidences suggest that the pathogenic mechanism involves immune system and inflammation. AIMS: The aim of current study was to uncover hub genes linked to immune infiltration in osteoarthritis synovial tissue using comprehensive bioinformatics analysis and experimental confirmation. METHODS: Multiple microarray datasets (GSE55457, GSE55235, GSE12021 and GSE1919) for osteoarthritis in Gene Expression Omnibus database were downloaded for analysis. Differentially expressed genes (DEGs) were identified using Limma package in R software, and immune infiltration was evaluated by CIBERSORT algorithm. Then weighted gene co-expression network analysis (WGCNA) was performed to uncover immune infiltration-associated gene modules. Protein-protein interaction (PPI) network was constructed to select the hub genes, and the tissue distribution of these genes was analyzed using BioGPS database. Finally, the expression pattern of these genes was confirmed by RT-qPCR using clinical samples. RESULTS: Totally 181 DEGs between osteoarthritis and normal control were screened. Macrophages, mast cells, memory CD4 T cells and B cells accounted for the majority of immune cell composition in synovial tissue. Osteoarthritis synovial showed high abundance of infiltrating resting mast cells, B cells memory and plasma cells. WGCNA screened 93 DEGs related to osteoarthritis immune infiltration. These genes were involved in TNF signaling pathway, IL-17 signaling pathway, response to steroid hormone, glucocorticoid and corticosteroid. Ten hub genes including MYC, JUN, DUSP1, NFKBIA, VEGFA, ATF3, IL-6, PTGS2, IL1B and SOCS3 were selected by using PPI network. Among them, four genes (MYC, JUN, DUSP1 and NFKBIA) specifically expressed in immune system were identified and clinical samples revealed consistent change of these four genes in synovial tissue retrieved from patients with osteoarthritis. CONCLUSION: A 4-gene-based diagnostic model was developed, which had well predictive performance in osteoarthritis. MYC, JUN, DUSP1 and NFKBIA might be biomarkers and potential therapeutic targets in osteoarthritis.

Therapeutic Effects of Mechanical Stress-Induced C2C12-Derived Exosomes on Glucocorticoid-Induced Osteoporosis Through miR-92a-3p/PTEN/AKT Signaling Pathway.

Introduction: Osteoporosis is a common bone disease in which the bone loses density and strength and is prone to fracture. Bone marrow mesenchymal stem cells (BMSCs) are important in bone-related diseases. Exosomes, as mediators of cell communication, have potential in cell processes. Previous studies have focused on muscle factors' regulation of bone remodeling, but research on exosomes is lacking. Methods:  In order to confirm the therapeutic effect of mechanically stimulated myocytes (C2C12) derived exosomes (Exosome-MS) on the Glucocorticoid-induced osteoporosis(GIOP) compared with unmechanically stimulated myocytes (C2C12) derived exosomes (Exosomes), we established a dexamethasone-induced osteoporosis model in vivo and in vitro. Cell viability and proliferation were assessed using CCK8 and EDU assays. Osteogenic potential was evaluated through Western blotting, real-time PCR, alkaline phosphatase activity assay, and alizarin red staining. Differential expression of miRNAs was determined by high-throughput sequencing. The regulatory mechanism of miR-92a-3p on cell proliferation and osteogenic differentiation via the PTEN/AKT pathway was investigated using real-time PCR, luciferase reporter gene assay, Western blotting, and immunofluorescence. The therapeutic effects of exosomes were evaluated in vivo using microCT, HE staining, Masson staining, and immunohistochemistry. Results:  In this study, we found that exosomes derived from mechanical stress had a positive impact on the proliferation and differentiation of bone marrow mesenchymal stem cells (BMSCs). Importantly, we demonstrated that miR-92a-3p mimics could reverse dexamethasone-induced osteoporosis in vitro and in vivo, indicating that mechanical stress-induced mouse myoblast-derived exosomes could promote osteogenesis and prevent the occurrence and progression of osteoporosis in mice through miR-92a-3p/PTEN/AKT signaling pathway. Conclusion:  Exosomes derived from mechanical stress-induced myoblasts can promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells through miR-92a-3p/PTEN/AKT signaling pathway, and can have a therapeutic effect on glucocorticoid-induced osteoporosis in mice in vivo.

Study on biomechanical analysis of two-level cervical Mobi-C and arthrodesis.

OBJECTIVE: To investigate the range of motion (ROM) index of a two-level cervical arthroplasty. METHODS: Seven human cadaveric spines were biomechanically examined from C2 level to T1 level under intact status and the following conditions: 2-level arthroplasty (C4-C6) employing Mobi-C devices (MM group), 2-level anterior cervical discectomy and fusions (2-ACDFs) (FF group), and both as a hybrid surgery (HS) (MF group and FM group). Multidirectional flexibility examination was conducted according to the Panjabi hybrid testing protocol. Unconstrained intact moments of ±1.5 NM were performed for axial rotation (AR) flexion/extension (FE), and lateral bending (LB). RESULTS: No statistical differences were found between the intact spine and MM group at the operative- and adjacent-level kinematics in the three loading conditions, except that C4-C5 ROM significantly increased in the axial rotation loading (P<0.05). Compared with the intact spine, MF group led to a significant decrease at the arthrodesis segment ROM C5-C6 in the three loading (P<0.05), with corresponding significantly increased at C4-C5 in FE and AR (P<0.05). FM group resulted in a significant decrease in ROM C4-C5 (P<0.05) with corresponding significantly increased at C5-C6 in FE, AR and LB (P<0.05). There was not any difference for non-operative level kinematics between MF group and FM group and intact spine. Compared with the intact spine, FF group led to a significant decrease at the arthrodesis-levels (P<0.05) and marked increase at the non-operative level kinematics. CONCLUSION: A two-level Mobi-C and Hybrid construct generated better biomechanical conditions. This study suggested that two-level cervical total disc replacement or HS could become an alternative approach for therapy of two-level consecutive cervical spondylosis.

[Application of the proximal tibiofibular joint resection on the limb salvage for osteosarcoma of the proximal tibia].

OBJECTIVE: To evaluate the proximal tibiofibular joint resection on the limb salvage for osteosarcoma of the proximal tibia. METHODS: Between August 1995 and January 2004, 11 patients with osteosarcoma of proximal tibia and tibiofibular joint involved underwent resection and endoprosthetic proximal tibial replacement. Seven patients were male and 4 were female. The patients ranged in age from 14 to 23 years (mean 18 years). The surgical stage were IIB. RESULTS: The follow-up period varied from 2 to 9 years (mean 59 months). Three patients died of pulmonary metastases, 1 patient was still alive with pulmonary metastasis, 1 patient underwent amputation for local recurrence. There was 1 early skin necrosis, 2 transient palsy of the common peroneal nerve, and 2 thrombus of lower limb vein. The mean postoperative Musculoskeletal Tumor Society score of the knee joint was 70% (range 55% - 86%), the mean postoperative range of motion was 85 degrees (range 0 degrees - 120 degrees ), and the extensor lag varied from 0 degrees to 20 degrees . CONCLUSIONS: Limb salvage for osteosarcoma of the proximal tibia with proximal tibiofibular joint resection and custom-made prosthesis reconstruction has had excellent results, however, the prevention and treatment of relevant complications should be given more attention.

Comparative Study of Modified Posterior Operation to Treat Kümmell's Disease.

The present study aimed at examining the curative effect of modified posterior operation on treatment of Kümmell's disease. About 30 patients of Kümmell's disease with complete image and clinical data treated during June 2004 to July 2013 were conducted with anterior and posterior approaches, respectively. Kyphotic Cobb angle, fractured vertebra wedge angle, and the anterior and posterior heights of fractured vertebra were all measured through x-ray before and after operation, and the pain visual analog scale (VAS) was determined for evaluating the effect of operations. The injury and restoration of neurological function were assessed using Frankel classification. Patients in group A were treated with anterior operation, whereas group B was posterior operation. Postoperatively, VAS score, kyphotic Cobb angle, anterior vertebra height, and pathologic vertebra wedge angle were all significantly improved in patients with Kümmell's disease receiving modified posterior operation (group B). Similar results were also observed in patients with anterior operation. The improvement of VAS and correction rate of kyphotic Cobb angle indicated equivalent effects of posterior and anterior operations. Meanwhile, alleviated neurological function damage was observed in 2 groups. Relevant factor analysis illustrated that there was no significant correlation of the severity and improvement rate of pain symptoms with age, medical history, anterior and posterior vertebra heights, kyphotic Cobb angle, and pathological vertebra wedge angle. Compared with traditional anterior approach, modified posterior operation, adopting transpedicular vertebral body grafting combined with vertebral pedicle screw fixation, could produce equivalent effects on kyphosis correction, pain relief, and improvement of neurological function in patients with Kümmell's disease.

Development of Alendronate-conjugated Poly (lactic-co-glycolic acid)-Dextran Nanoparticles for Active Targeting of Cisplatin in Osteosarcoma.

In this study, we developed a novel poly (lactic-co-glycolic acid)-dextran (PLD)-based nanodelivery system to enhance the anticancer potential of cisplatin (CDDP) in osteosarcoma cells. A nanosized CDDP-loaded PLGA-DX nanoparticle (PLD/CDDP) controlled the release rate of CDDP up to 48 h. In vitro cytotoxicity assay showed a superior anticancer effect for PLD/CDDP and with an appreciable cellular uptake via endocytosis-mediated pathways. PLD/CDDP exhibited significant apoptosis of MG63 cancer cells compared to that of free CDDP. Approximately ~25% of cells were in early apoptosis phase after PLD/CDDP treatment comparing to ~15% for free CDDP after 48h incubation. Similarly, PLD/CDDP exhibited ~30% of late apoptosis cells comparing to only ~8% for free drug treatment. PLD/CDDP exhibited significantly higher G2/M phase arrest in MG63 cells than compared to free CDDP with a nearly 2-fold higher arrest in case of PLD/CDDP treated group (~60%). Importantly, PLD/CDDP exhibited a most significant anti-tumor activity with maximum tumor growth inhibition. The superior inhibitory effect was further confirmed by a marked reduction in the number of CD31 stained tumor blood vessels and decrease in the Ki67 staining intensity for PLD/CDDP treated animal group. Overall, CDDP formulations could provide a promising and most effective platform in the treatment of osteosarcoma.

Icariin protects against glucocorticoid-induced osteoporosis in vitro and prevents glucocorticoid-induced osteocyte apoptosis in vivo.

Icariin is the major active ingredient in Herba epimedii which is a commonly used Chinese herbal medicine for the treatment of osteoporosis. The present study aims to evaluate the osteoprotective effect of Icariin in glucocorticoid-induced osteoporosis in vivo and investigate the effect of Icariin on glucocorticoid-induced osteocyte apoptosis in vitro. A total of 48 female Sprague-Dawley rats were used. Glucocorticoid-induced osteoporosis was induced by daily injections of dexamethasone (0.1 mg/kg, daily, s.c.) for 60 days, whereas sham animals were injected daily with vehicle. At the end of the osteoporosis development period, osteoporotic rats were randomized to receive: vehicle (n = 8), Icariin (5,125 mg/kg, i.g.; n = 8), or alendronate (0.03 mg/kg, s.c.; n = 8) for 12 weeks. Sham animals were treated with vehicle for 12 weeks. At the beginning and at the end of treatments, animals were examined for bone mineral density. Serum bone-alkaline phosphatase and carboxy-terminal collagen cross links were measured. Primary osteocytes were isolated, and apoptosis was determined by trypan-blue assay. Interaction between Icariin and estrogen receptor and prosurvival signaling pathways activated by Icariin were also investigated. Icariin showed a comparable efficacy with alendronate in increasing bone mass. Icariin significantly increased bone-alkaline phosphatase (bone formation marker) and reduced carboxy-terminal collagen cross links (bone resorption marker). In vitro studies demonstrated that Icariin significantly prevented GC-induced apoptosis in osteocytes by activating ERK signaling via estrogen receptor. Our results suggest that Icariin might exert osteoprotective effect by maintaining osteocyte viability, thereby, regulating bone remodeling. Furthermore, our study provides preclinical evidence for the efficacy of Icariin for management of Glucocorticoid-induced osteoporosis.