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14 October 2016, Volume 20 Issue 42 Previous Issue    Next Issue

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Comparison of physicochemical properties of five kinds of cancellous bones Gao Huan-yu, Li Yan-lin, Xiao Yu, Li Long-teng, Wang Guo-liang 2016, 20 (42):  6237-6243.  doi: 10.3969/j.issn.2095-4344.2016.42.001 Abstract ( 290 )   PDF (1130KB) ( 903 )   Save BACKGROUND: Existing evidence has shown that xenogeneic bone derived graft materials have the ability to repair bone defects, but little report is on the comparison of composition of xenogeneic bone and human bone. OBJECTIVE: To analyze the chemical composition, mineral phase, calcium and phosphorus contents, total protein contents, and appearance of the cancellous bones of the tibia from pig, bovine and human. METHODS: Cancellous bones of the tibia from pig, Banna miniature pig, bovine, young and old people were cut into bone particles of 0.5 cm×0.5 cm×0.5 cm. After degreasing or drying treatment, chemical composition, mineral phase, calcium and phosphorus contents, total protein contents of bone particles were detected using Fourier transform infrared spectroscopy, X-ray diffraction, plasma emission spectroscopy, scanning electron microscopy, and Kjeldahl method, respectively. Nano Measurer 1.2 and Photoshop cs6 were used to measure pore diameter and porosity rate based on the results of scanning electron microscope. RESULTS AND CONCLUSION: The main inorganic phase of five kinds of cancellous bones was hydroxyapatite and the main chemical composition was phosphate and carbonate. The calcium content of the bovine cancellous bonewas lower than that of the other cancellous bones (P < 0.05) and the phosphorus content of the cacellous bone of young man was higher than that of the pig and bovine cancellous bones (P < 0.01). The total protein content in the cancellous bones of normal pig and Banna miniature pig was higher than that in the other three kinds of cancellous bones (P < 0.05). The pore diameter of five kinds of cancellous bones was 400 to 600 μm and the porosity rate was 60% to 70%. Moreover, the porosity rate of the bovine cancellous bone was the highest. Therefore, we conclude that there are some certain differences in the organic and inorganic components of the pig or bovine cancellous bones as compared with humans. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of osteopontin on the expression of hyaluronic acid in human knee osteoarthritic chondrocytes Luo Wei1, Zhang Fang-jie1, Li Yu-sheng1, Lei Guang-hua1, 2 2016, 20 (42):  6244-6251.  doi: 10.3969/j.issn.2095-4344.2016.42.002 Abstract ( 193 )   PDF (2112KB) ( 361 )   Save BACKGROUND: Both osteopontin and hyaluronic acid involve in the pathological process of osteoarthritis, resulting in the abnormal expression levels of various cytokines and enzymes. However, the relationship between the high expression of osteopontin and hyaluronic acid in chondrocytes remains unclear. OBJECTIVE: To investigate the effect of osteopontin on the expression of hyaluronic acid in human knee osteoarthritic chondrocytes in vitro by modulating the level of osteopontin. METHODS: Chondrocytes from human knee osteoarthritic cartilage were cultured in vitro, and were then divided into three groups: blank control group without any treatment; osteopontin group and and pontin siRNA group were treated with 1 mg/L recombinant human osteopontin and osteopontin siRNA, respectively. Expression levels of osteopontin, hyaluronic acid synthase 1, 2 and 3 mRNA were detected by real-time PCR, and the levels of hyaluronic acid were measured using ELISA. RESULTS AND CONCLUSION: Compared with the blank control group, the mRNA expressions of hyaluronic acid synthase 1, 2 and 3 were remarkably increased in the osteopontin group, while siRNA made the significantly inhibitory effects on the hyaluronic acid synthase 1, 2 and 3 mRNA expressions (P < 0.05). The level of hyaluronic acid in chondrocytes in the osteopontin group was significantly higher than that in the other two groups (P < 0.05). Our results suggest that osteopontin induces the synthesis of hyaluronic acid in osteoarthritic chondrocytes through upregulating the hyaluronic acid synthases expression levels. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of cytochrome c on early steroid-induced femoral head necrosis Ji Hao-yu1, Wang Xiao-long2 2016, 20 (42):  6252-6258.  doi: 10.3969/j.issn.2095-4344.2016.42.003 Abstract ( 250 )   PDF (2206KB) ( 264 )   Save BACKGROUND: Studies have shown that apoptosis is closely related to the pathogenesis of steroid-induced femoral head necrosis. The release of cytochrome c plays a very important role in the process of apoptosis. OBJECTIVE: To observe the effects of cytochrome c on early steroid-induced femoral head necrosis. METHODS: Twenty-four healthy adult male New Zealand rabbits at 5 months old were randomly divided into model group and control group (n=12 per group). Models of early steroid-induced femoral head necrosis were established by intragluteal injection of hormone combined with ear vein injection of horse serum. In the control group, rabbits were given ear vein injection of the same amount of physiological saline. At 2, 4 and 6 weeks after model establishment, histopathological changes of bilateral femoral head were observed by optical microscope, and the ratio of empty lacuna was calculated. Apoptosis of osteocytes was determined by TUNEL assay, and apoptotic index was calculated. Immunohistochemical method was used to determine cytochrome c and to calculate cytochrome c-positive expression rate. RESULTS AND CONCLUSION: (1) The ratio of empty lacuna and apoptotic index: The model of early steroid-induced femoral head necrosis was successfully established in the experiment. Compared with the control group, ratio of empty lacuna, apoptotic index and expression rate of cytochrome c in osteocytes were significantly higher in the model group (P < 0.05 or P < 0.01). (2) Correlation analysis: Ratio of empty lacuna was significantly positively associated with apoptotic index at various time points in the model group (r=0.856, P < 0.01). Expression rate of cytochrome c was significantly positively associated with apoptotic index at various time points in the model group (r=0.824, P < 0.01). (3) These findings confirm that cytochrome c-involved apoptosis of osteocytes may play an important role in steroid-induced femoral head necrosis. Expression rate of cytochrome c in osteocytes is remarkably positively associated with the occurrence of steroid-induced femoral head necrosis in rabbits. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Platelet-rich plasma for the repair of damaged articular cartilage in a rabbit model of knee osteoarthritis Wu Xiao, Tan Ya-jun, Xian Jie, He Ben-xiang 2016, 20 (42):  6259-6264.  doi: 10.3969/j.issn.2095-4344.2016.42.004 Abstract ( 280 )   PDF (1393KB) ( 409 )   Save BACKGROUND: Platelet-rich plasma (PRP) containing high levels of platelet-derived growth factor for knee osteoarthritis has achieved good clinical results; however, the effects of RPR on the repair of damaged articular cartilage are still controversial. OBJECTIVE: To study the effects of RPR on the repair of damaged articular cartilage in a rabbit model of knee osteoarthritis. METHODS: The model of osteoarthritis in the rabbit right knee was established by Hulth’s method. Autologous PRP (0.5 mL) (PRP group), sodium hyaluronate (0.5 mL) (sodium hyaluronate group), and normal saline (model group) were injected into the right knee joint cavity, respectively. The morphology of articular surface and nitric oxide contents in knee joint fluid were observed and determined at 8 weeks after surgery. RESULTS AND CONCLUSION: The morphology of articular cartilage in the PRP group was better than that in the other three groups. Mankin scores of articular cartilage and nitric oxide contents of knee joint fluid in the PRP group were significantly decreased compared with the model and sodium hyaluronate groups that in (P < 0.05), while increased compared with the sham-operation group (P < 0.05). Our results suggest that repair effects of PRP on the damaged articular cartilage are superior to sodium hyaluronate treatment. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Erk activation and proliferation in osteoarthritc chondrocytes after continuous passive motion Muyesaier•Maimaitiniyazi, Hu Yi, Ren Yun-ping, Zhang Yong, Xiong Dao-hai 2016, 20 (42):  6265-6270.  doi: 10.3969/j.issn.2095-4344.2016.42.005 Abstract ( 263 )   PDF (850KB) ( 219 )   Save BACKGROUND: Whether continuous passive motion improves osteoarthritis by enhancing the proliferation ability of chondrocytes is rarely reported. OBJECTIVE: To analyze the therapeutic outcomes of continuous passive motion in rabbits with osteoarthritis and the underlying mechanism. METHODS: Thirty-six New Zealand white rabbits were randomly allotted into three groups (n=12 per group). Rabbits in control group only underwent capsulotomy with no harm to the cartilage; osteoarthritis models were established in the rabbits of model and treatment groups using Hulth method. At 1 day after modeling, the treatment group rabbits were treated with continuous passive motion, 8 hours daily for consecutive 8 weeks. Interleukin-1 and tumor necrosis factor α levels in the synovial fluid were detected by ELISA; collagen type II expression and the proliferation ability of chondrocytes were detected by MTT assay; Erk signaling pathway activation was determined using western blot assay. RESULTS AND CONCLUSION: In the model group, interleukin-1 and tumor necrosis factor α levels in the synovial fluid were significantly increased, and the expression level of collagen type II mRNA was remarkablely down-regulated. Continuous passive motion significantly downregulated interleukin-1 and tumor necrosis factor α levels and up-regulated the collagen type II mRNA level (P < 0.01). The model group showed significantly decreased proliferation ability of chondrocytes and down-regulated Erk signaling pathway activation, while after continuous passive motion, all above indicators were significantly improved (P < 0.01). These results indicate that the continuous passive motion can alleviate osteoarthritis probably by influencing interleukin-1 and tumor necrosis factor α levels, proliferation ability of chondrocytes, and collagen type II expression, as well as regulating Erk signaling pathway activation. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Changes in proteoglycan components in degenerative intervertebral disc after intragastric administration of Liuwei Dihuang Wan Wang Le, Xu Wu-ji 2016, 20 (42):  6271-6277.  doi: 10.3969/j.issn.2095-4344.2016.42.006 Abstract ( 194 )   PDF (938KB) ( 391 )   Save BACKGROUND: Proteoglycan, chondroitin sulfate and keratan sulfate, and hyaluronic acid are important substances in the maintenance of intervertebral discs of normal structure. They impact the physiological function of intervertebral disc. OBJECTIVE: To observe the influence of Liuwei Dihuang Wan on proteoglycan in rabbit models of intervertebral disc degeneration, and to study the efficacy of Liuwei Dihuang Wan on prevention and treatment of intervertebral disc degeneration. METHODS: 80 New Zealand rabbits were randomly divided into Liuwei Dihuang Wan group, model group, sham surgery group and blank control group (n=20 per group). Animal models of intervertebral disc degeneration were established in the model and Liuwei Dihuang Wan groups. In the sham surgery group, surgery via the same approach to expose the lumbar spine was conducted followed by layer-by-layer suturing. In the Liuwei Dihuang Wan group, rabbits were given 10 mg/kg Liuwei Dihuang Wan by lavage, once a day. In the model group, sham surgery group and blank control group, rabbits were given an equal volume of physiological saline, once a day, by natural feeding. At 2, 4, 6 and 8 weeks, five rabbits from each group were sacrificed, and intervertebral disc specimens were taken to measure proteoglycan components in the intervertebral disc.  RESULTS AND CONCLUSION: With the extension of feeding time, Liuwei Dihuang Wan could increase sugar amino polysaccharide content, chondroitin sulfate/acid ratio of keratin, and hyaluronic acid content in the intervertebral disc degeneration model, and stabilized the proteoglycan content. Thus, it delayed the degeneration of the intervertebral disc to a certain extent. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of nitric oxide and nitric oxide synthase inhibitors on mitochondrial function of nucleus pulposus cells Zhou Jian-guo1, Yang Cao2, Xiong Li-ming2 2016, 20 (42):  6278-6283.  doi: 10.3969/j.issn.2095-4344.2016.42.007 Abstract ( 253 )   PDF (838KB) ( 213 )   Save BACKGROUND: Nitric oxide can interfere with the function of mitochondria, and accelerate the intervertebral disc damage and degeneration by interfering with the release of inflammatory cytokines. Nitric oxide is an important inflammatory cell medium leading to degeneration of intervertebral disc induced by pressure and other external factors. OBJECTIVE: To investigate the regulatory effect of nitric oxide and nitric oxide synthase inhibitor niacinamide on mitochondrial function and its association with biological behavior of rabbit nucleus pulposus. METHODS: Cultured nucleus pulposus cells of rabbit lumbar intervertebral disc were randomly divided into six groups: normal blank control group, 10 μmol/L sodium nitroprusside group, 100 μmol/L sodium nitroprusside group, 200 μmol/L sodium nitroprusside group, 0.05 g/L nicotinamide group (100 μmol/L sodium nitroprusside+0.05g/L nicotinamide), and 0.5 g/L nicotinamide group (100 μmol/L sodium nitroprusside and 0.5 g/L nicotinamide). Different doses of nitric oxide donor sodium nitroprusside and nicotinamide were added in the medium of each group. Three days after intervention, cell proliferation activity, intracellular ATP concentration, cell nitric oxide synthase activity, cellular reactive oxygen species level, and mitochondrial membrane potential were detected respectively. RESULTS AND CONCLUSION: (1) After 3 days of rabbit nucleus pulposus cells intervened by different concentrations of sodium nitroprusside, intracellular nitric oxide synthase content increased with sodium nitroprusside volume increase, and ATP concentration decreased along with sodium nitroprusside volume increase; there were significantly differences between the normal control group and sodium nitroprusside groups (P < 0.01). (2) Reactive oxygen species could be increased in the sodium nitroprusside group. Niacinamide groups indicated a dose-dependent manner to improve the increase of cellular reactive oxygen species levels with sodium nitroprusside intervention (P < 0.01). (3) In the sodium nitroprusside groups, nucleus pulposus cell membrane potential decreased. In the niacinamide groups, sodium nitroprusside- induced decline in mitochondrial membrane potential was reduced (P < 0.01). (4) Niacinamide groups also indicated a dose-dependent manner to improve the proliferative activity of nucleus pulposus cells as compared with sodium nitroprusside groups (P < 0.01). Significant differences were determined between the two groups (P < 0.01). (5) Results suggest that the excess nitric oxide can damage mitochondrial metabolic function of rabbit nucleus pulposus cells and cause cell energy metabolism. Niacinamide can reverse these damages by inhibiting nitric oxide synthesis, thereby contributing to the prevention against intervertebral disc degeneration. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Excellence of differential velocity adherent procedure in primary culture of nucleus pulposus cells of intervertebral disc Hu Yong-kai, Sun Hao-lin, Qi Long-tao, Li Chun-de 2016, 20 (42):  6284-6289.  doi: 10.3969/j.issn.2095-4344.2016.42.008 Abstract ( 404 )   PDF (2268KB) ( 332 )   Save BACKGROUND: As the main function cell of intervertebral disc, nucleus pulposus cells are the focus of studying the degenerative mechanism; thereby, it is crucial to maintaining the physiological function of nucleus pulposus cells in vitro and the stability of the cell phenotype. OBJECTIVE: To study the excellence of differential velocity adherent procedure in primary culture of nucleus pulposus cells of rat intervertebral disc through comparison. METHODS: Twenty male Wistar rats aged 4 weeks were enrolled, and then nucleus pulposus cells of intervertebral disc were isolated and cultured in vitro; cell passage culture was performed in different groups when the primary cells were merged to 90%. Differential velocity adherent group cells adhered for 30 minutes, and non-adherent cells were aspirated and transferred to new culture dish after readjusting the concentration; the controls received no intervention. Passages 1 and 2 cells in the differential velocity adherent group were isolated and purified by the same procedure. The morphology of three generations of cells in the two groups was compared, the purity of the identification was detected by immunohistochemistry, the cell viability was detected by cell counting kit-8 and the cell growth curve was drawn. RESULTS AND CONCLUSION: Inverted phase contrast microscope and hematoxylin-eosin staining revealed that the cell homogeneity of the differential velocity adherent group was significantly higher than that of the control group, and there were more kinds of fibroblast-like cells in nucleus pulposus cells in the control group. Identification and purity analysis of collagen type II showed that the cytoplasm of two groups were both stained brown, indicating that they were the nucleus pulposus chrondrocytes. The positive rate of differential velocity adherent group was significantly higher than that of the control group (P < 0.05). The cell growth curve of cell counting kit-8 showed cells in the two groups all passed by the latency phase within 2 days, then to the logarithmic phase of 3 days, and entered the lag phase, while the growth rate of the control group was more rapid during the latency and the early logarithmic phases. These findings suggest that differential velocity adherence method is a practical and effective procedure for the isolation and purification of primary cultured rat nucleus pulposus cells. Through the primary culture, twice differential velocity adherence procedure, the passage 3 rat nucleus pulposus cells are metabolic exuberant, consistent with the phenotype, the cell purity is higher, and the logarithmic growth phase can be used as the optimal time for studying the mechanism of intervertebral disc cells. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Hydroxyapatite-bone morphogenetic protein ossicular prosthesis versus allogeneic cartilage ossicular prosthesis for ossicular chain damage Zhang Xue-hong, Liang Xiao-wei, Lu Hong-lue, Yu Shu-jie, Wang Hong-mei, Zhang Yu-li 2016, 20 (42):  6290-6295.  doi: 10.3969/j.issn.2095-4344.2016.42.009 Abstract ( 275 )   PDF (1847KB) ( 259 )   Save BACKGROUND: There is a great dispute on the choice of repair materials for ossicular chain damage. OBJECTIVE: To explore the effects of new hydroxyapatite-bone morphogenetic protein ossicular prosthesis versus allogeneic cartilage ossicular prosthesis in patients with ossicular chain damage. METHODS: Sixty patients with chronic otitis media were equally assigned into a control group and a test group. Two groups of patients underwent tympanoplasty and ossicular chain reconstruction with allogeneic cartilage ossicular prosthesis or hydroxyapatite-bone morphogenetic protein ossicular prosthesis, respectively. Twelve months after surgery, follow-up results were compared between two groups. RESULTS AND CONCLUSION: Air conduction value and air-bone gap value were both improved significantly in the two groups after surgery (P < 0.05), and especially, the postoperative air conduction value was better in the test group than the control group (P < 0.05). Implant detachment was found in three cases of the control group, whereas did not occur in the test group. Improved hearing was found in 28 cases in the test group with an improvement rate of 93% and 25 cases in the control group with an improvement rate of 83%. There was a significant difference in the hearing improvement between the two groups (P < 0.05). Overall, two kinds of prostheses can share similar effects on the ossicular chain repair. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Regulatory function of microRNA-182 in apoptosis of nucleus pulposus cells Qiu Xiao-wen, He Xi-jing, Wang Dong, Li Hao-peng, Zhao Bo 2016, 20 (42):  6296-6301.  doi: 10.3969/j.issn.2095-4344.2016.42.010 Abstract ( 279 )   PDF (789KB) ( 293 )   Save BACKGROUND: Previous study has found that hsa-miR-182 is probably related to the apoptosis-related genes such as cytochrome C (Cycs C) and calcineurin subunit CnB (PPP3R1) in nucleus pulposus cells. OBJECTIVE: To determine whether miR-182 plays a regulatory role in nucleus pulposus cell apoptosis by detecting the relative gene expression levels after transfecting miR-182 with Cycs C and PPP3R1 into nucleus pulposus cells via plasmid delivery. METHODS: After a bioinformatics prediction about miR-182, miR-182 and target genes were transfected into the nucleus pulposus cells, and at the same time, blank control group was established. Then the expression levels of the target genes were detected through cell lysis. RESULTS AND CONCLUSION: miR-182 significantly inhibited the expression of Cycs C in nucleus pulposus cells compared with the blank control group (P < 0.05). Compared with the blank control group, miR-182 made no inhibitory effect on the expression of PPP3R1. These findings suggest that miR-182 may play a regulatory part in nucleus pulposus cell apoptosis by inhibiting the expression of Cycs C. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Thyroid hormones affect alveolar bone density Zhang Xiang1, Qiu Ze-wen2, Xu Jing1, Qu Zhe1, Ma Lan1, Pan Xiao-qing1, Zhang Dan-di1 2016, 20 (42):  6302-6307.  doi: 10.3969/j.issn.2095-4344.2016.42.011 Abstract ( 319 )   PDF (884KB) ( 331 )   Save BACKGROUND: Overproduction of thyroid hormones is shown to increase the activities of osteoblasts and osteoclasts, stimulating bone resorption and transformation. Inadequate compensation of increased bone resorption by bone transformation results in an increased loss of bone mass. OBJECTIVE: To investigate the effects of hyperthyroidism on the density of the alveolar bone. METHODS: Twenty-four New Zealand rabbits were equally randomized into hyperthyroidism group and control group. Rabbits in the hyperthyroidism group or control group were daily injected intraperitoneally with 50 μg/kg levothyrocine diluted in physiological saline solution or equal volume of physiological saline. At 8 weeks after treatment, serum levels of thyroid hormones (FT3 and FT4), alkaline phosphatase, magnesium and calcium, phosphorus were determined; meanwhile, the bone densities of the lumbar vertebra, mandible, bilateral distal femur were determined by dual energy X-ray absorptiometry and the correlation analysis was performed. RESULTS AND CONCLUSION: At 8 weeks after treatment, serum levels of FT3, FT4, alkaline phosphatase calcium, phosphorus, and magnesium in the hyperthyroidism group were significantly increased (P < 0.01), while the bone densities of the lumbar vertebra, mandible, bilateral distal femur were significantly decreased (P < 0.05), compared with the control group. Bone density of the mandible was positively correlated with the bone density the lumbar vertebra and bilateral distal femur. These findings suggest that the changes in FT3 and FT4 are sufficient for the diagnosis of hyperthyroidism. Hyperthyroidism results in the decreased density of the alveolar bone, indicating the occurrence of osteoporosis. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Toll-like receptor 4 antagonist protects against Wallerian degeneration after peripheral nerve injury Xiong Le1, Zhang Bei1, Shen Ruo-wu2, Ji Ai-yu3, Sun Guang-qiang1, Bian Hong-lin3, Zhang Feng-yu1, Wang Yi1, Huang Heng4, Li Hua-qiao1, Zhou Shan-yu5, Shen Zhao-kang6, Wang Zhong1 2016, 20 (42):  6308-6316.  doi: 10.3969/j.issn.2095-4344.2016.42.012 Abstract ( 262 )   PDF (1115KB) ( 261 )   Save BACKGROUND: The mechanism underlying Wallerian degeneration following peripheral nerve injury is complex. Immune regulation on Wallerian degeneration is beneficial for early repair of perpheral nerve injury. OBJECTIVE: To investigate the effects of Toll-like receptor 4 (TLR4) antagonist on Wallerian degeneration and axonal regeneration after early peripheral nerve injury in rats. METHODS: Fifty male Wistar rats were recruited and randomly divided into treatment group (n=20), model group (n=20) and sham group (n=10). The right sciatic nerves of rats in treatment and model groups were cut and sutured end-to-end, while the sciatic nerves of rats in sham group were only exposed. In the treatment group rats were intravenously injected with 0.15 mg/kg TAK-242 via tail vein 1 hour preoperatively and 7 days postoperatively, and the rats in the other two groups were given intravenous injection of the same volume of normal saline. The sciatic nerves were removed at 24 hours, 3, 4 and 7 days after surgery. RESULTS AND CONCLUSION: Real-time PCR indicated that the mRNA expressions of interleukin-1β and monocyte chemoattractant-1 were significantly increased in the model group compared with the sham group at 24 hours after surgery (both P < 0.001), while the expressions were significantly decreased after TAK-242 injection (both P < 0.001). Immunofluorescence showed that compared with the model group, down-regulated expression of CD68+ and iba1+ cells appeared in the treatment group at 3 days after surgery (P < 0.01, P < 0.05). Luxol fast blue staining revealed that demyelination at the sciatic nerve stump appeared in both model and treatment groups at postoperative 7 days, but myelin debris clearance in the treatment group was significantly reduced compared with the model group (P < 0.05). Hematoxylin-eosin staining showed that a lot of inflammatory cells, Schwann cells and regenerated nerve fibers at the sciatic nerve stump were found in the model group, while there were few inflammatory cells, Schwann cells and regenerated nerve fibers in the treatment group at 7 days after surgery. Immunohistochemistry found that the expression of growth-associated protein-43 in the treatment group was significantly lower than that in the model group at 4 days postoperatively (P < 0.05). Besides, compared with the model group, a significantly decreased sciatic functional index was found in the treatment group at 20, 30 and 40 days after surgery (P < 0.05). These results show that TLR4 antagonists delay early nerve regeneration in rats after sciatic nerve injury probably by inhibiting the TLR4 signaling pathway. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of hyaluronic acid on scar formation in the acellular nerve allograft Liu Ying-wei, Zhang Wan-li, Chi Cheng-tao, Xu Qing-yu, Lu De-zhi 2016, 20 (42):  6317-6323.  doi: 10.3969/j.issn.2095-4344.2016.42.013 Abstract ( 357 )   PDF (977KB) ( 499 )   Save BACKGROUND: In terms of the histocompatibility, immune rejection and scar formation after repair, acellular nerve allograft is closer to autologous nerve cells. At present, hyaluronic acid has been applied for autologous peripheral nerve repair; however, research on the nerve allograft is rarely reported. OBJECTIVE: To explore the effect of hyaluronic acid on the anastomotic scar in acellular nerve allograft repair of rat sciatic nerve defect. METHODS: Thirty-six Sprague-Dawley rats were randomly divided into three groups (n=12 per group). The rat model of nerve defect of 10 mm was established by cutting the sciatic nerve of the left hind leg and then given nerve allograft combined with the injection of hyaluronic acid at anastomosis (experimental group), only nerve allograft (control group) and autologous nerve graft (nerve autograft group), respectively. Afterwards, the healing of the proximal anastomosis was observed and scar components were assessed. RESULTS AND CONCLUSION: Gross observations showed that the rat skin and muscle fascia had no significant differences in healing among groups, while the surrounding tissue adhesion in the experimental group was milder than that in the control group (P < 0.05). Masson staining found that collagen deposition in the epinerium could be observed in each group. In the experimental group, a small amount of collagen fibers arranged orderly in the epineurium; in the control group numerous collagen fibers accumulated and arranged irregularly; in the nerve autograft group, sparse epineurial collagen fibers appeared in an order arrangement. The gray value of collagen type I in the experimental group was higher than that in the control group (P < 0.05), while the gray value of collagen type III was lower than that in the control group (P < 0.05). No significant differences were found in the sum gray values of collagen type I and III among groups (P > 0.05). These findings indicate that in the peripheral nerve repair, hyaluronic acid abrogates the scar formation by increasing the deposition of collagen type III and reducing the deposition of collagen type I. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Neuron-secreted factors promote the growth of skeletal muscle cells Cai Ming-xuan1, Wang Mei-chen1, Xu Zhi-cheng1, Yan Nan2, Meng Xin-yu1, Wang Zheng-dong3 2016, 20 (42):  6324-6329.  doi: 10.3969/j.issn.2095-4344.2016.42.014 Abstract ( 324 )   PDF (790KB) ( 342 )   Save BACKGROUND: There is no effective treatment for muscle atrophy caused by peripheral nerve damage. Skeletal muscle cells, a structural unit of muscle contraction, can be used for studies on muscle atrophy when cultured in vitro. OBJECTIVE: To investigate the promotion effect of neuron-secreted factors on the growth of skeletal muscle cells in vitro. METHODS: Skeletal muscle cells primary cultured in vitro were divided into two groups: experimental group with neuron-secreted factors, and control group with common culture medium, respectively. Afterwards, the number of skeletal muscle cells and expression level of alpha actin were detected by hematoxylin-eosin staining and immunohistochemical staining, respectively. RESULTS AND CONCLUSION: The number of skeletal muscle cells and expression level of alpha actin in the experimental group were significantly higher than those in the control group (P < 0.05). In conclusion, neuron-secreted factors have the ability of promoting the growth of skeletal muscle cells and may be helpful for denervated muscle atrophy. Subject headings: Tissue Engineering; Satellite Cells, Skeletal Muscle; Actinin; Neurons; Cells, Cultured 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | Related Articles | Metrics

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Simulation of acoustic response of microvessel containing microbubble in ultrasound field based on finite element analysis and lumped parameter model Niu Chuan-xiao, Guo Sheng-wen, Qiu Lin, Lao Yong-hua, Jiang Xing-jun 2016, 20 (42):  6330-6336.  doi: 10.3969/j.issn.2095-4344.2016.42.015 Abstract ( 288 )   PDF (1020KB) ( 577 )   Save BACKGROUND: Exploration on nonlinear acoustic response of the contrast agent microbubble contained in microvessel under ultrasound excitation is of great significance to maximizing ultrasonic energy deposition, promoting the development of quantitative imaging algorithm, revealing the damage mechanism or evaluating the targeted therapy, and overcoming the limitations of the traditional methods that are mainly used in large-size vessels, and measuring microvessel elasticity. OBJECTIVE: To build a microvessel containing an ultrasound microbubble, revealing the internal mechanism among ultrasound, microbubble, blood flow and microvessel. METHODS: Based on the finite element analysis and the lumped parameter model, three-dimensional microvessel containing microbubble model was built and simulated on Comsol Multiphysics 4.4 platform. RESULTS AND CONCLUSION: Microbubble exhibited slower radial motion compared with axial motion due to vascular wall limitation, but maximum displacement and stress were found near the microbubble center because of the oscillation coupling of the microbubble with the vascular wall. Under the same ultrasound pressure, the excitation frequency increased, accompanied by decreased and stabilized microvessl constriction and dilation; under the same frequency, with the enhancement of ultrasound pressure, the local microbubble oscillation lasted longer. With the increase of Young’s modulus of the microvessel wall, the frequency of microbubble oscillation was reduced, while the amplitude increased. All these findings indicate that the frequency of microbubble oscillation increased with the reduction of microvessel size, while its amplitude decreased. The frequency of microbubble oscillation increased with the enhancement of ultrasound excitation, while the amplitude decreased. On the contrary, ultrasound pressure affected the dynamic characteristics of microbubble and microvessel. In particular, it was the first to demonstrate that the elasticity of microvessel has approximate linear positive correlation with the amplitude of microbubble oscillation, which reveals the relationship between microvessel elasticity and microbubble response so as to provide theoretical basis for indirect measurement of microvessel elasticity. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Preparation of an electronic moxibustion apparatus which simulates the mechanism of traditional moxibustion: application security and environmental effects Yang Xu-ming, Tang Jia-ming 2016, 20 (42):  6337-6342.  doi: 10.3969/j.issn.2095-4344.2016.42.016 Abstract ( 342 )   PDF (1055KB) ( 962 )   Save BACKGROUND: An electronic moxibustion apparatus that simulates the mechanism of traditional moxibustion therapy is designed and developed to overcome shortcomings of traditional moxibustion therapy, such as non-standardized treatment, limited body position, and production of "smoke" and "ash" during the treatment. OBJECTIVE: To develop a kind of electronic moxibustion apparatus which simulates the traditional moxibustion therapy and has obtained the patent so as to industrialize and modernize moxibustion therapy. METHODS: Bio-waves launched from far infrared materials are beneficial to the human body. An electronic moxibustion apparatus including time temperature controller and moxibustion head was developed, and the closed loop control technique was used to precisely control the temperature and time. Thermal radiation infrared spectrum that moxibustion head generated could simulate the emission spectrum of moxibustion in combination with wormwood oil coating or drug interval, thereby achieving the therapeutic effect similar to traditional moxibustion. RESULTS AND CONCLUSION: The prototype of DZWJY-1 electronic moxibustion instrument has been developed and its safety and electromagnetic compatibility have been detected in Shanghai Testing & Inspection Institute for Medical Devices, China. Relevant clinical trials are ongoing to verify the efficacy. Preliminary clinical evidence shows that the electronic moxibustion apparatus has the same therapeutic effect to the traditional moxibustion therapy, which realizes the safety and environmental protection of moxibustion therapy and provides objective, standardized and modernized evidence for moxibustion therapy 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 References | Related Articles | Metrics

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Correlation between the MRI-based grading system and F wave as well as H-reflex in patients with lumbar disc herniation Li Xiao, Lin Cai-na, Luo Hai-jie, Wan Qing, Ruan Yu-ting, Zhang Xin-sheng, Wu Shao-ling, Ma Chao 2016, 20 (42):  6343-6350.  doi: 10.3969/j.issn.2095-4344.2016.42.017 Abstract ( 482 )   PDF (1000KB) ( 326 )   Save BACKGROUND: Lumbar spine MRI and electrophysiological test are reliable methods for evaluating nerve root injury caused by lumbar disc herniation. OBJECTIVE: To analyze the correlation between the MRI-based grading system and the latency and frequency of F wave as well as latency and amplitude of H-reflex in patients with lumbar disc herniation. METHODS: MRI imaging of the lumbar spine was performed with a 3.0-T imager and a dedicated TCL coil to classify lumbar disc herniation and nerve root compression. F wave and H reflex were detected on the patient bilateral tibial nerves using Oxford myoelectricity evoked potential instrument. RESULTS AND CONCLUSION: Spearman correlation analysis showed that the MRI-based grading of patients with lumbar disc herniation had a negative correlation with F wave frequency (r=-0.594 0, P < 0.000 1), and a positive correlation with F wave latency (r=0.825 6, P < 0.000 1) and H-reflex latency (r=0.875 0, P < 0.000 1), but no correlation with H-reflex amplitude (R=0.117 4, P=0.257 3). With MRI grading increased, F wave frequency was decreased, and F wave and H-reflex latency were prolonged gradually, indicating aggravating nerve root compression. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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The risk factors and plantar pressure characteristics of hallux valgus Huang Ping, Qian Nian-dong, Qi Jin, Chen Bo, Deng Lian-fu 2016, 20 (42):  6351-6356.  doi: 10.3969/j.issn.2095-4344.2016.42.018 Abstract ( 435 )   PDF (942KB) ( 430 )   Save BACKGROUND: Complex anatomic deformity of the foot due to hallux valgus results in the changes of plantar biomechanics. OBJECTIVE: To analyze the risk factors of hallux valgus and to detect the plantar pressure of hallux valgus in natural walking. METHODS: The dynamic plantar pressures of 50 patients and 30 age-, height- and weight-matched normal persons were respectively tested by the Footscan USB2 system (RSscan International, Belgium). All subjects were detected during barefoot walking over three times, to obtain the dynamic plantar pressure curves and characteristic parameters for further analysis. RESULTS AND CONCLUSION: Among patients with hallux valgus, women accounted for 94% (all wearing pointed high-heeled shoes), men accounted for 6% (none wearing pointed high-heeled shoes), and 96% had a genetic history. During the natural walking, the first metatarsal pressure of hallux valgus feet was increased significantly, which markedly differed from the normal people. The average peak pressure, average load, and average impulse of the first metatarsal in hallux valgus patients were significantly increased compared with the normal controls (P < 0.05 or P < 0.01). These results show that wearing high-heeled pointed shoes is another important factor of the hallux valgus in addition to genetic factors. Pressure, load, impulse of the first metatarsal in hallux valgus patients are markedly increased contributing to the occurrence and development of the hallux valgus. All above findings provide an important basis for the prevention, treatment and functional rehabilitation of hallux valgus. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Velocity and target location influence the upper limb reaching movements: three-dimensional motion analysis combined with surface electromyograph Li Li-fang, Mao Yu-rong, Huang Dong-feng, Li Le 2016, 20 (42):  6357-6362.  doi: 10.3969/j.issn.2095-4344.2016.42.019 Abstract ( 305 )   PDF (757KB) ( 306 )   Save BACKGROUND: Three-dimensional (3D) motion analysis system combined with electromyograph cannot only objectively analyze movement function of the upper limb, but also exert an important role in exploring the exercise-related factors and the underlying mechanism. OBJECTIVE: To explore the influence of movement velocity and target location on kinematics and muscle activity in upper limb reaching. METHODS: Twelve healthy young adults reached for three different location targets (frontal, ipsilateral and contralateral) at self-selected and fast speeds to with the dominant right hand, respectively. Kinematic parameters and muscle activity were recorded by Vicon 3D motion analysis system and Noraxon wireless electromyograph, synchronously. RESULTS AND CONCLUSION: Reach path ratio became smaller when moving faster (P < 0.001), while mean velocity, peak velocity and the time percentage of peak velocity increased (P < 0.001). Posterior deltoid, biceps brachii and triceps brachii activation was higher during fast speed movement (P < 0.001). Muscle activation of anterior deltoid was lower in ipsitralateral reaching than contralateral (P=0.001) and frontal reaching (P < 0.001), and posterior deltoid was higher than contralateral reaching (P=0.019). Biceps brachii (P=0.039) and triceps brachii (P < 0.001) activation was also higher in ipsitralateral reaching than frontal reaching. These results suggest that moving fast can contribute to more muscle activation, high velocity, smooth trajectory and increased movement quality. Posterior deltoid, elbow extension and flexion muscle activation are high in ipsitralateral reaching, while high anterior deltoid activation in contralateral reaching; therefore, target location result in muscle synergies. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Research progress of metabolism and physiological functions of the lacunar-canalicular system Shi Li-jun1, Gao Fu-qiang2, Sun Wei2 2016, 20 (42):  6363-6370.  doi: 10.3969/j.issn.2095-4344.2016.42.020 Abstract ( 416 )   PDF (845KB) ( 415 )   Save BACKGROUND: Interstitial fluid flow around the osteocytes caused by a series of physiological activities plays an important role in the osteocyte metabolism and signal transduction. OBJECTIVE: To review and summarize the research progress of bone structures and physiological functions, then further illustrate how the mechanical loads make an impact on bone tissue. METHODS: A computer-based search was conducted in PubMed database for articles related to the bone microstructure, metabolism and osteocyte signal transduction published from January 2009 to December 2015. The keywords were “osteocyte, mechanical load, permeability, lacunar-canalicular system, interstitial fluid flow, mechanotransduction, signaling pathways” in English. Data were screened firstly, dated articles or literatures with wrong research methods were excluded, and totally 40 eligible articles were enrolled. RESULTS AND CONCLUSION: Bone tissue can quickly adapt to the change of mechanical environment to guarantee enough osteocytes in the functional bone area indicating that osteocytes regulate the bone absorption and formation by responding to stress, which is closely related to osteocyte physiological characters. Osteocytes exist in mineralized matrix, and the special microstructures make it possible to receive mechanical loads and transform the mechanical signals into chemical signals aimed at regulating the bone absorption or formation.So the interstitial fluid flow in the lacuna-canalicular system occures, namely load-induced fluid flow, when the mechanical load is distributed on the bone. This fluid flow affects the bone tissue through two mechanisms: regulating osteocyte metabolism and participating in the mechanotransduction. In conclusion, the mechanical load plays a vital role in maintaining health bone and regulating bone adaptation. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Three-dimensional scaffold materials for cell culture and their application in tumor tissue engineering Han Xiao-dong1, Wang Lei2, Zheng Jun2, He Xiao-long2, Ji Jin-shan1, Fu Zhao-ying1 2016, 20 (42):  6371-6377.  doi: 10.3969/j.issn.2095-4344.2016.42.021 Abstract ( 291 )   PDF (959KB) ( 873 )   Save BACKGROUND: The use of three-dimensional cell culture techniques can better simulate the cellular microenvironment, providing new tools for tissue engineering research. OBJECTIVE: To analyze the biomaterial selection and application characteristics in three-dimensional cell culture as well as applications in tumor tissue engineering. METHODS: We searched Wanfang database and PubMed database 1998-2015 years for relevant literature using keywords of “three-dimensional cultures; scaffold; cell growth; cell differentiation; tumor tissue engineering” in Chinese and English, respectively. RESULTS AND CONCLUSION: The selection and application of three-dimensional scaffold materials is one of the keys. So far, scaffold materials, such as collagen gels, gelatin sponge, agarose, chitosan, demineralized bone matrix, cannot provide the extracellular matrix similar to the micro-environment in which seed cell growth and proliferation are not affected, and the ability to secrete type II collagen and glycosaminoglycan is decreased, although they can provide three-dimensional space for seed cells. Biomimetic scaffold characterized as little trauma and strong plasticity gradually shows its unique advantages. Three-dimensional culture conditions raise pro-angiogenic growth factor secretion from tumor cells, and this feature is positively correlated with the occurrence of in vivo tumor angiogenesis. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics

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Vibration training improves senile osteoporosis: the assessment of safety and effectiveness Lou Yan-tao 2016, 20 (42):  6378-6384.  doi: 10.3969/j.issn.2095-4344.2016.42.022 Abstract ( 281 )   PDF (877KB) ( 433 )   Save BACKGROUND: Osteoporosis is increasingly threatening the elderly survival quality and life. Numerous studies have shown that the vibration training is a new method to improve osteoporosis. OBJECTIVE: To overview the effect of vibration training on the skeletal system, and provide a new idea for osteoporosis prevention. METHODS: The first author searched the PubMed and CNKI databases using the following format: (Vibration training [Title]) OR (Vibration mode [Title]) OR (Vibration pattern [Title]) and (“Vibration training” [Title]) or (“Vibration mode”[Title]) in English and Chinese, respectively, in April 2015. Finally, 38 eligible articles were included according to their titles and abstracts. RESULTS AND CONCLUSION: Bases on overseas and domestic researches about the effect of whole body vibration on the skeletal system, we summarize the mechanical load, vibration stress principles and outcomes of whole body vibration training as well as factors involvied in osteoporosis such as the stimulation site, stimulation type, exercise load, exercise duration and vibration frequency. In addition, the safety of vibration training and its preceding studies are also generalized. These results suggest that elderly osteoporosis can be improved through the vibratory equipment. We can improve bone loss and living quality of the elderly by vibration at different frequencies and duration thresholds vibration at using the vibratory equipment. 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程 Figures and Tables | References | Related Articles | Metrics