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08 July 2016, Volume 20 Issue 29 Previous Issue    Next Issue

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Silencing of estrogen receptor beta gene influences the expressions of transforming growth factor beta1 and bone morphogenetic protein 2 in human osteoblasts Deng Ang, Zhang Hong-qi, Guo Chao-feng, Wang Yu-xiang, Gao Qi-le, Tang Ming-xing, Liu Shao-hua, Liu Jin-yang 2016, 20 (29):  4261-4268.  doi: 10.3969/j.issn.2095-4344.2016.29.001 Abstract ( 231 )   PDF (1048KB) ( 256 )   Save BACKGROUND: There are few studies concerning estrogen receptor β gene, and its mechanism of regulating the bone metabolism is still unclear now.  OBJECTIVE: To analyze the effect of estrogen receptor β (ER β) silencing on the expressions of transforming growth factor β1 (TGF-β1) and bone morphogenetic protein 2 (BMP-2) in human osteoblasts METHODS: There were three groups: blank control group (hFOB 1.19 uninfected with any retrovirus); negative control group (containing invalid interference fragment ER β-shRNA-nc); optimal RNAi group (ER β-shRNA-3). ER β-shRNA retroviral vectors in the optimal RNAi group were used to transfect human osteoblasts followed by resistance screening and cell expansion. MTT assay was used to detect the proliferative activity of ER β-silenced osteoblasts. Then under estrogen intervention, the stable inhibition rate of ER β was determined using western blot assay, and the expressions of TGF-β1 and BMP-2 in human osteoblasts after ER β silencing were detected by RT-PCR technology and western blot assay. RESULTS AND CONCLUSION: Human osteoblasts that were stably transfected by ER β-shRNA-3 retroviral vector was selected successfully, and ER β silencing had no significant influence on the cell proliferation (P > 0.05). Under the interference of estrogen, the silencing efficiency of ER β protein was (93.11±0.57)% (P < 0.05), and after ER β silencing, the expressions of TGF-β1 and BMP-2 were increased by (26.65±3.81)% and (16.62±1.71)% at mRNA level, and increased by (23.79±3.76)% and (18.08±3.20)% at protein level (both P < 0.05). In conclusion, ER β may play an important role in bone metabolism by regulating the expressions of TGF-β1 and BMP-2. 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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Insulin effects on fracture healing and cytokines in the osteotylus in experimental diabetic rats Zhou Qiang, Lu Hua, Wang Zhan-chao, Zhang Hao-jie, Jiang Lei-sheng 2016, 20 (29):  4269-4276.  doi: 10.3969/j.issn.2095-4344.2016.29.002 Abstract ( 225 )   PDF (1572KB) ( 268 )   Save BACKGROUND: Fracture healing in diabetic patients is usually unsatisfactory because of hormones and metabolic disorder, and an eventual multiple organ dysfunction resulting from high blood glucose. OBJECTIVE: To dynamically observe the changes of cytokines during the fracture healing process in diabetic rats before and after insulin treatment. METHODS: A total of 120 Sprague-Dawley rats were included in this study. Of them, 90 rats intravenously injected with 5% tetraoxypyrimidine to induce rat models of diabetes were randomized into insulin treatment and diabetes groups, respectively. The remaining 30 rats were intravenously injected with equal volume of saline and selected as control group. The next day, blood glucose was determined. Healing at 1, 4, and 8 weeks after fracture were observed by the X-ray film. Biomechanical strength of the injured right tibia was measured at 4, 6, and 8 weeks after modeling. Cytokines in the osteotylus were determined by immunohistochemical staining and in situ hybridization technique. RESULTS AND CONCLUSION: The X-ray films showed that the speed of fracture healing in the diabetes group was slower than insulin treatment and control groups. Biomechanical strength of the osteotylus in the diabetes group was significantly decreased compared with the insulin treatment and control groups. However, there were no significant differences in above-mentioned parameters between the control and insulin treatment groups. Bone morphogenetic protein 2, basic fibroblast growth factor, transforming growth factor-beta, and vascular endothelial growth factor were widely expressed in the osteotylus and their expressions in diabetes group were significantly lower and slower than those in the control and insulin treatment groups. There was no statistical difference between control and insulin treatment groups. These results indicate that osteotylus formation speed, biomechanical strength, and growth factor expressions at the fracture site in diabetes rats were decreased compared with normal rats. Insulin treatment can enhance cytokine levels at the fracture site, thereby promoting the osteoblast proliferation and fracture healing. 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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Influence of interleukin-1beta on microRNA-27b and matrix metalloproteinase-13 expression in rat chondrocytes Shi Jing, Li Tong, Zuo Can-hui, Zhi Liang, Wei Rong, Wang Wei-min, Liu Ying-jie 2016, 20 (29):  4277-4283.  doi: 10.3969/j.issn.2095-4344.2016.29.003 Abstract ( 234 )   PDF (1841KB) ( 306 )   Save BACKGROUND: Matrix metalloproteinase-13 is most active in the degradation of collagen type II in the extracellular matrix of cartilage. Interleukin-1 (IL-1) is thought to be the inducer of matrix metalloproteinases, and participates in the degradation and degeneration of articular cartilage. OBJECTIVE: To study the influence of IL-1β on microRNA-27b (miR-27b) and matrix metalloproteinase-13 expression of chondrocytes in rats. METHODS: Chondrocytes isolated from seven male Wistar rats were cultured and divided into IL-1β stimulation group and control group. No stimulus was given in the control group; 10 μg/L of serum free medium was used to culture rat chondrocytes in the IL-1β stimulation group. Cell growth was observed at 0, 24, and 48 hours under an inverted microscope. miR-27b and matrix metalloproteinase-13 expression in the cultured chondrocytes were detected. RESULTS AND CONCLUSION: The relative expression of matrix metalloproteinase-13 in rat chondrocytes was gradually increased when induced by IL-1β at 0, 24, and 48 hours (P < 0.05). Expression of miR-27b and miR-31 in rat chondrocytes at 24 and 48 hours induced by IL-1β gradually decreased (P < 0.05); conversely, expression of MiR-26a, miR-26b, miR-23, and miR-204 gradually increased (P < 0.05). After 48 hours of IL-1β induction, expression of miR-27b was the lowest in rat chondrocytes (P < 0.05). These findings suggest that IL-1β inhibits miR-27b expression, strengthens the expression of matrix metalloproteinase-13, and damages chondrocytes, contributing to both the onset and progression of osteoarthritis. 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 low-intensity pulsed ultrasound for articular cartilage repair Liu Yang, Liu Ning, Liu Zhao-ming, Hao Zhen-min, Wang Dong-lai 2016, 20 (29):  4284-4289.  doi: 10.3969/j.issn.2095-4344.2016.29.004 Abstract ( 362 )   PDF (1238KB) ( 219 )   Save BACKGROUND: Articular cartilage injuries can result from a variety of causes. Conventional therapy cannot obtain the optimal clinical results. Low-intensity pulsed ultrasound has been shown to promote the repair of injured articular cartilage. OBJECTIVE: To investigate the effects of low-intensity pulsed ultrasound on the repair of injured articular cartilage. METHODS: Twenty New Zealand white rabbits were used to establish knee arthritis models and equally randomized into study and control groups, respectively. Rabbits in the study group received low-intensity pulsed ultrasound treatment, and sham low-intensity pulsed ultrasound treatment was given in the control group. At 8 weeks after treatment, pathological change and histological scores in articular cartilage tissue collected from both groups were determined. Moreover, the ultrastructure and type II collagen expression of chondrocytes were determined. Matrix metalloproteinase-13 mRNA expression was detected by quantitative real-time PCR. RESULTS AND CONCLUSION: At 8 weeks after treatment, toluidine blue staining showed a disordered arrangement of cells, decreased number of cartilage cells in each layer and cluster in the control group. Light disordered arrangement of cells, decreased appearance of the superficial layer cells and the cluster phenomenon were observed in the study group. Articular cartilage tissue scores were significantly decreased in the study group compared with the control group (P < 0.05). The chondrocytes were small, enlarged intracellular mitochondria and rough endoplasmic reticulum, cytoplasmic swelling, collagen fibrils coarse, well developed Golgi apparatus, and nuclear fragmentation were observed in the control group. In addition, the normal structure of organelles disappeared and cell degeneration was observed in the control group. In the study group, the size of chondrocytes and the Golgi complex and other organelles were normal, and the protein polysaccharide granules were observed in the cytoplasm and membrane. The mRNA expression of matrix metalloproteinase-13 in the study group was significantly lower than that in the control group (P < 0.05). Type II collagen immunoreactivity in the study group was stronger than that in the control group. No incision infection, suppuration, red swelling appeared in all rabbits. Our results suggest that low-intensity pulsed ultrasound can be used for the treatment of articular cartilage injury by alleviating the degradation of collagen type II and inhibiting the expression of matrix metalloproteinase-13. 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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Association between single-nucleotide polymorphisms in interleukin-12 and susceptibility to osteoarticular tuberculosis in Guangxi Zhuang population Lu Lu, Wei Gui-jiang, Xie Ke-gong, Wang Jun-li, Lin Jia-jie, Qiao Ning-ning, Tang Yu-jin 2016, 20 (29):  4290-4296.  doi: 10.3969/j.issn.2095-4344.2016.29.005 Abstract ( 224 )   PDF (985KB) ( 285 )   Save BACKGROUND: Interleukin-12 (IL-12) may function as an immune regulator in the pathogenesis of osteoarticular tuberculosis. OBJECTIVE: To explore the association of single-nucleotide polymorphisms in IL-12A rs568408 G/A and IL-12B rs3212227 A/C with susceptibility to osteoarticular tuberculosis and serum interleukin-12 levels in Guangxi Zhuang population. METHODS: The single-nucleotide polymorphisms in IL-12A rs568408 G/A and IL-12B rs3212227 A/C polymorphisms were detected by polymerase chain reaction-single base extension technique and direct DNA sequencing in 150 patients with osteoarticular tuberculosis (disease group) and 165 healthy individuals (control group) in Guangxi Zhuang population. The genotype and allele frequencies of IL-12 and the relationship of genotypes to the susceptibility to osteoarticular tuberculosis were analyzed. In addition, the association of genotypes of single-nucleotide polymorphisms in IL-12A rs568408 G/A and IL-12B rs3212227 A/C with serum IL-12 levels were analyzed. RESULTS AND CONCLUSION: There was no significant difference in the genotype and allele frequencies of IL-12A rs568408 G/A and IL-12B rs3212227 A/C between the disease group and the control group (P > 0.05). Moreover, there was no difference in four haplotypes of IL-12 gene between the disease group and the control group (P > 0.05). Serum IL-12 levels in subjects with osteoarticular tuberculosis carrying the variant rs568408 GA/AA genotypes and wild-type rs568408 GG genotypes were similar (P > 0.05). Similarly, there was no significant difference in serum IL-12 levels between subjects with osteoarticular tuberculosis carrying the variant rs3212227 AC/CC genotypes and wild-type rs3212227 AA genotypes (P > 0.05). These findings suggest that the single-nucleotide polymorphisms in IL-12A rs568408 G/A and IL-12B rs3212227 A/C polymorphisms are not associated with susceptibility to osteoarticular tuberculosis in Guangxi Zhuang population. 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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Expression of AMP-activated protein kinase in subcultured rat endplate chondrocytes Zhao Quan-lai, Zheng Quan, Xu Hong-guang, Shen Xiang, Wang Hong, Liu Ping, Wang Ling-ting, 2016, 20 (29):  4297-4302.  doi: 10.3969/j.issn.2095-4344.2016.29.006 Abstract ( 267 )   PDF (906KB) ( 206 )   Save BACKGROUND: Endplate cartilage degeneration initiates intervertebral disc degeneration. AMP-activated protein kinase (AMPK) regulates the formation and degradation of cartilage. OBJECTIVE: To explore the role of AMPK in an in vitro natural degeneration model of chondrocytes derived from endplate of rat intervertebral discs. METHODS: Morphology of in vitro subcultured endplate chondrocytes of rat intervertebral discs at passages 0, 2, and 5 were observed under an inverted microscope following cytoskeleton staining. Chondrocyte phenotype, proliferation, and the cartilage marker genes (type II collagen, proteoglycan, SOX-9, matrix metalloproteinase-3 and -13), and AMPK phosphorylation were determined by toluidine blue staining, MTT assay, real-time PCR analysis, and western blot assay, respectively. RESULTS AND CONCLUSION: The altered morphology, decreased proliferation ability, and phenotype loss were observed in chondrocytes with increased passage number. Gene expression of type II collagen, proteoglycan, SOX-9 was significantly decreased; while gene expression of matrix metalloproteinase-3 and -13 was significantly increased in endplate chondrocytes at passage 5 compared with those at passages 0 and 2. AMPK phosphorylation in endplate chondrocytes at passage 5 was significantly decreased. These findings indicate that AMPK phosphorylation is involved in in vitro natural degeneration of chondrocytes derived from the endplate of rat intervertebral discs, and the degeneration of endplate chondrocytes and intervertebral discs can be inhibited through the regulation of AMPK activity. 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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Serum and tissue levels of matrix metalloproteinase-1 and its tissue inhibitor in patients with intervertebral disc degeneration Deng Bin, Wang Ye-xin, Meng Chun-yang 2016, 20 (29):  4303-4310.  doi: 10.3969/j.issn.2095-4344.2016.29.007 Abstract ( 278 )   PDF (2176KB) ( 271 )   Save BACKGROUND: Matrix metalloproteinases are now generally considered to be able to degrade all extracellular matrices. Hypersecretion of matrix metalloproteinases or reduction in tissue inhibitors of matrix metalloproteinases leads to destruction of the dynamic balance of extracellular matrix. OBJECTIVE: To elucidate the role of matrix metalloproteinase-1 and tissue inhibitor of matrix metalloproteinase-1 in the pathogenesis and progression of intervertebral disc degeneration. METHODS: A total of 60 patients with intervertebral disc degeneration were included. Mild, moderate, and severe degeneration signals appeared on MRI imaging of the patients. Meanwhile, 20 patients with vertebral fracture, mainly cervical spine fracture, were selected as the control group. Venous blood samples were collected before the surgery; the intervertebral disc specimens were sequentially collected. RESULTS AND CONCLUSION: Serum and tissue levels of matrix metalloproteinase-1 in patients with intervertebral disc degeneration were significantly increased compared with the control group (P < 0.05), and furthermore those were significantly increased in patients with severe disc degeneration compared with patients with mild and moderate disc degeneration (P < 0.05). However, serum and tissue levels of tissue inhibitors of matrix metalloproteinases did not differ significantly between the disc degeneration and control groups (P > 0.05). These results indicate that hypersecretion of matrix metalloproteinase-1 occurs in patients with intervertebral disc degeneration; however, the expression of tissue inhibitor of matrix metalloproteinase-1 is not correlated with 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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Clinical effects of renal transplantation with kidneys from donors dying of organophosphate poisoning Dong Jian-hui, Li Hai-bin, Sun Xu-yong, Qin Ke, Liao Ji-xiang, Li Mei-si, Huang Xiao-dan, Huang Chen, Huang Ying, Cao Song, Gao Zhao, Li Zhuang-jiang, Nie Feng, Yang Jian-jun 2016, 20 (29):  4311-4318.  doi: 10.3969/j.issn.2095-4344.2016.29.008 Abstract ( 366 )   PDF (1988KB) ( 204 )   Save BACKGROUND: Delayed graft function (DGF) occurs frequently in kidney transplants from donation after cardiac death if creatinine level is high in kidney recipients. OBJECTIVE: To analyze the clinical effects of renal transplantation with kidneys from donors dying of cardiac death in organophosphate poisoning. METHODS: Data were collected from kidney transplants from two donors dying of cardiac death in organophosphate poisoning. After some donor maintenance, donor organ were obtained and perfused with impulse type machine. Recipients were treated with intervention of immunity induction, anti-rejection drugs and infection prevention drugs during and after renal transplantation. Pathological data of donor kidney zero needle biopsy, DGF after kidney transplantation, complication rate (such as acute rejection), renal allograft recovery situation, the survival rate of recipients and kidney transplants were collected and analyzed. RESULTS AND CONCLUSION: Needle biopsy results from four donor kidneys showed that glomerular morphology was normal, but there were edema and degeneration in kidney tubules in some degree. Donor DGF rate was 75% (3/4), acute rejection rate was 0% (0/4), perioperative period donor kidney and recipient survival rate were 100% (4/4). All recipients showed a good result of transplanted kidney, their creatinine and urea nitrogen were at low level, and had no proteinuria. One recipient died of severe pulmonary infection 4 months after surgery. For some organophosphate poisoning donors dying of cardiac death, donor kidney quality can be improved by suitable donor maintenance and high-quality donor kidney preservation using machine perfusion. Kidney transplants from donors dying of cardiac death in organophosphate poisoning who receive the maintenance of organ function may be a promising candidate for renal transplantation due to a severe lack of kidney donor sources. 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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Construction of recombinant lentiviral vector and interfering carrier for tumor necrosis factor alpha stimulated gene 6 and its effect on proliferation and apoptosis of human keloid fibroblasts Chen Zhao, Li Xiao-jing, Wang Hui 2016, 20 (29):  4319-4327.  doi: 10.3969/j.issn.2095-4344.2016.29.009 Abstract ( 292 )   PDF (2707KB) ( 226 )   Save BACKGROUND: Current research has shown that tumor necrosis factor α stimulated gene 6 (TSG-6) has anti-inflammatory effect, and the scar formation can be inhibited by local injection of TSG-6 protein at the early stage of trauma. However, the mechanism of this effect is still unclear. OBJECTIVE: To construct the lentiviral expression vector and shRNA vector for human TSG-6, with stable overexpression, transfection and interference, and to explore the effect of TSG-6 on proliferation and apoptosis of keloid fibroblast cell lines.  METHODS: Human keloid fibroblast cells were isolated from the keloid’s tissue by enzyme digestion and identified by immunocytochemistry assay. Lentiviral vectors pLVX-puro-TSG-6 and pLVX-shRNA1-TSG-6 were constructed and transfected into human keloid fibroblast, exclusively. Expression levels of TSG-6 mRNA and protein were detected by RT-PCR and western blot assay. MTT assay and flow cytometry were used to estimate the cell proliferation and apoptosis in each group after transfection. In addition, expression of Bcl-2, p53 and active-caspase-3 were detected by western blot assay in each group. RESULTS AND CONCLUSION: (1) Human keloid fibroblasts were separated successfully. Under the light microscope, cells were spindle. Immunohistochemical staining for vimentin was performed in the fifth passage of cells, with the positive rate of 100%. Cells were negative for cytokeratin. (2) Recombinant lentiviral vectors and stably transfected cell lines were successfully established. TSG-6 gene expression was altered apparently. Compared with the control group, cell proliferation was delayed and apoptotic rate was noticeably increased in TSG-6 gene overexpression group. Cell proliferation increased and apoptotic rate decreased in the TSG-6 gene intervention group (P < 0.05). (3) Western blot assay results demonstrated that Bcl-2 expression reduced, P53 and Active-caspase-3 expression significantly increased in the TSG-6 gene overexpression group (P < 0.05). (4) These finding showed that TSG-6 could inhibit proliferation and induce apoptosis in keloid fibroblasts. Its mechanism may be associated with the down-regulation of Bcl-2 protein expression, up-regulation of P53 protein expression and increased Caspase-3 activity. 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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Optimization of culture conditions for oligodendrocytes of the rat cerebral cortex Yang Kai, Li Yi-peng, Liu Ying-fu, Cheng Yuan-chi, Tang Feng-wu, Liang Bing, Xu Zhong-wei, Chen Xu-yi 2016, 20 (29):  4328-4333.  doi: 10.3969/j.issn.2095-4344.2016.29.010 Abstract ( 395 )   PDF (1597KB) ( 246 )   Save BACKGROUND: Oligodendrocytes are mostly differentiated from oligodendrocyte precursor cells. A suitable medium and cell seeding density have a significant impact on the process of the isolation of oligodendrocyte precursor cells to obtain oligodendrocytes. OBJECTIVE: To explore the optimization of oligodendrocyte culture conditions. METHODS: Oligodendrocyte precursor cells isolated from the newborn rats 48 hours after birth were cultured in DMEM/high glucose medium or DMEM/F12 medium using seeding densities of 2×104 cells/cm2, 4×104 cells/cm2, 8×104 cells/cm2, 16×104 cells/cm2, 32×104 cells/cm2, and 64×104 cells/cm2, respectively. Oligodendrocyte precursor cells were induced to differentiate into oligodendrocytes at 72 hours after cell adhesion. Morphology of differentiated oligodendrocyte precursor cells were observed under a light microscope, and the differentiation results were identified by immunofluorescence staining after 7-day induced differentiation. RESULTS AND CONCLUSION: Morphology of oligodendrocyte precursor cells were recognized when cultured in DMEM/high glucose medium or DMEM/F12 medium using seeding densities of 2×104 cells/cm2, 4×104 cells/cm2, and 8×104 cells/cm2, respectively. Immunofluorescence staining showed that myelin basic protein-positive cells were found after 7-day induced differentiation, and the positive cell number were 16.40±3.30, 49.95±2.33, and 76.95±4.86 in DMEM/F12 medium, and 12.65±2.53, 32.10±1.17, and 54.05±1.56 in DMEM/high glucose medium (P < 0.05). These findings indicate that DMEM/F12 medium is more suitable for culturing oligodendrocyte precursor cells compared with DMEM/high glucose medium to some extent. The number of differentiated oligodendrocytes was gradually increased with the enhanced seeding density of oligodendrocyte precursor cells, and the seeding densities from 4×104 to 8×104 cells/cm2 were appropriate for the observation of cell morphology. 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 cyclic tensile stress on caspase-12 expression in chondrocytes Chen Zhu-ke, Li Xiao-fei, Zhang Hai-ning 2016, 20 (29):  4334-4340.  doi: 10.3969/j.issn.2095-4344.2016.29.011 Abstract ( 293 )   PDF (2551KB) ( 205 )   Save BACKGROUND: Excessive mechanical stimulation can lead to endoplasmic reticulum stress. Endoplasmic reticulum stress plays an important role in the occurrence and development of degenerative diseases. Caspase-12 is a specific molecule of endoplasmic reticulum stress, and the intensity of endoplasmic reticulum stress can be reflected by caspase-12 expression. OBJECTIVE: To observe the expression rule of Caspase-12 in chondrocytes under the cyclic tensile stress. METHODS: Human chondrocytes were used as test subjects. After group assignment, mechanical loading system was used. The loading group and corresponding Z-ATAD-FMK inhibitor group received 2, 12, 24, 36, and 48 hours of mechanical stimulation. The blank group (0 hour) and the paired Z-ATAD-FMK inhibitor group received the same process as the loading group except the loading. After loading, cell morphology and growth were observed under the microscope. RT-PCR and western blot assay were used to detect the expression changes of caspase-12 gene and protein in each group. RESULTS AND CONCLUSION: (1) Morphological observation results demonstrated that apoptosis appeared at 2 hours after loading, peaked at 24 hours. With prolonged time, cell growth showed the trend along stress, but apoptosis weakened. It is indicated that cyclic tensile stress could make chondrocyte apoptosis. Endoplasmic reticulum stress might be activated. The model of cells in vitro was established successfully. (2) Caspase-12 gene and protein expression showed consistent trend, and peaked at 24 hours, which showed significant differences as compared with the blank and inhibitor groups (P < 0.05). (3) Cyclic tensile stress can induce chondrocyte endoplasmic reticulum stress, and affect caspase-12 expression. 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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Application of mandibular kinesiography in dental occlusion Liu Yu, Zhang Qi, Li Tian-shu, Fu Gui-yuan, Xu Lin, Yi Long, Wang Jia-ying, Güzelay, He Yuan, Gao Lu 2016, 20 (29):  4341-4348.  doi: 10.3969/j.issn.2095-4344.2016.29.012 Abstract ( 354 )   PDF (2007KB) ( 299 )   Save BACKGROUND: Maxillofacial malformation limits the function of the mandible, produces the overload of the temporalmandibular joint leading to oral diseases. OBJECTIVE: To measure the characteristics of mandibular movement in populations with various types of dental occlusion. METHODS: Thirty-three volunteers (1 male and 32 females, mean age 21.71 years) meeting the inclusion criteria were enrolled from Dalian Medical University, China and divided into four groups: normal control (n=10), Angle’s class I, II, III malocclusion (n=10, 8, 5). The protrusion, lateral, opening motion trace of mandibular central incisor and the right mandibular first molar, and the trace from mandibular postural position to intercuspal position were measured using the mandibular kinesiograph. The features of different occlusal patterns were analyzed using SPSS 17.0 software. RESULTS AND CONCLUSION: In the trace of protrusion, lateral, opening and closing motion, there were significant differences in the trace of the right mandibular first molar’s Max.Slant between Angle’s class II malocclusion group and the other three groups (P < 0.05). In the trace of open wide and close fast, there were significant differences in the trace of mandibular central incisor in vertical distances between normal control group and Angle’s class I, III malocclusion groups (P < 0.05); there were significant differences in the trace of mandibular central incisor in Max.Slant between normal control group and Angle’s class I malocclusion group (P < 0.05); there were significant differences in the trace of the right mandibular first molar in vertical distances between Angle’s class II malocclusion group and the other three groups (P < 0.05); there were significant differences in the trace of the right mandibular first molar in Max.Opening velocity between normal control group and Angle’s class II, III malocclusion groups (P < 0.05). There were no significant differences in free space between the four groups (P > 0.05). These results indicate that there are differences in the trace of mandibular central incisor and the trace of the right mandibular first molar in range and speed between different occlusal patterns. We can draw a conclusion that malocclusion has effects on the direction, range and speed of mandibular movement. 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 mesenchymal stem cells conditioned media on the transforming growth factor-beta signaling pathway in normal and hypertrophic scar fibroblasts Wu Yan, Yuan Xiao-huan, Yang Lan, Zhao Xiao-jin, Li Hou-zhong, Wang Ying, Jin Hong 2016, 20 (29):  4349-4354.  doi: 10.3969/j.issn.2095-4344.2016.29.013 Abstract ( 330 )   PDF (966KB) ( 251 )   Save BACKGROUND: Numerous studies have shown that mesenchymal stem cells (MSCs) can effectively attenuate the fibrosis of damaged heart, lung and kidney by secreting various bioactive factors. OBJECTIVE: To evaluate the anti-fibrotic therapeutic potential of bone marrow MSCs conditioned media in vitro. METHODS: Normal fibroblasts and hypertrophic scar fibroblasts were treated with bone marrow MSCs conditioned media, then transforming growth factor-β and collagen production were analyzed by ELISA, and mRNA expression level of Smad7 and hydroxyproline content were detected by RT-PCR and colorimetry, respectively. RESULTS AND CONCLUSION: Bone marrow MSCs conditioned media significantly inhibited the production of both transforming growth factor-β and collagen in hypertrophic scar fibroblasts (P < 0. 01), and up-regulated the mRNA expression level of Smad7 (P < 0. 01), a major inhibitory regulator in the SMAD family. However, the normal fibroblasts were scarcely influenced by bone marrow MSCs conditioned media. These findings indicate that bone marrow MSCs conditioned media is considered a promising candidate for the treatment of hypertrophic scars, which may provide new theoretical supports to reduce cutaneous scarring. 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 low power microwave irradiation (20 W) on the proliferation and differentiation of skeletal muscle satellite cells in a rabbit model of femoral fracture with internal fixation Zou Yu-zhen, Wang Gang, Xu Yi-ming, Bai Yue-hong 2016, 20 (29):  4355-4361.  doi: 10.3969/j.issn.2095-4344.2016.29.014 Abstract ( 322 )   PDF (1051KB) ( 279 )   Save BACKGROUND: Low power microwave irradiation has been shown to promote the healing of fractures with internal fixation; however, its action mechanisms on the skeletal muscle around the fracture site are unclear. OBJECTIVE: To study the effects of low power microwave irradiation (20 W) on the proliferation ability of skeletal muscle satellite cells in a rabbit model of femoral fracture with internal fixation. METHODS: Forty male New Zealand rabbits were used to establish femoral fracture followed by internal fixation models, and then were equally randomized into spontaneous recovery and microwave treatment groups. Low power microwave irradiation (20 W) was given for 30 consecutive days in the microwave treatment group on day 4 after modeling, while no microwave irradiation was given in the spontaneous recovery group. Rabbit thigh muscles adjacent to the implant were obtained to isolate skeletal muscle satellite cells. Immunohistochemical staining, hematoxylin-eosin staining and quantitative RT-PCR were used to evaluate the ability of the proliferation and differentiation of skeletal muscle satellite cells. RESULTS AND CONCLUSON: Hematoxylin-eosin staining showed that there was no significant difference in the morphology and histology of skeletal muscle tissues between the spontaneous recovery and microwave treatment groups. However, the relative mRNA expression of MyoG in the cultured skeletal muscle satellite cells in vitro and the number of α-sarcometric actin-postive cells in the microwave treatment group were significantly increased compared with the spontaneous recovery group (P < 0.05). The proliferative ability of skeletal muscle satellite cells was inhibited at the early stage, but not at the later stage. Our results suggest that low power microwave irradiation (20 W) can promote the proliferation and differentiation of skeletal muscle satellite cells around the implant in a rabbit model of femoral fracture with internal fixation, and thereby confirm the efficacy and safety of low power microwave irradiation for the internal fixation of fractures. 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 20-minute warm ischemia on coronary endothelial function of pig donor hearts after cardiac death Sun Xiao-ke, Zhang Wei-hua, Qin Guang-qi, Liao Qiu-ming, Stig Steen, Qiao Chen-hui 2016, 20 (29):  4362-4367.  doi: 10.3969/j.issn.2095-4344.2016.29.015 Abstract ( 333 )   PDF (843KB) ( 252 )   Save BACKGROUND: Myocardial and coronary endothelial injury occurs in donor hearts due to warm ischemia during cardiac transplantation. Coronary endothelial structure and function play a critical role in long-term outcomes for patients after cardiac transplantation.  OBJECTIVE: To study the effect of hypoxia-induced warm ischemia (20 minutes) on coronary endothelial function of porcine donor hearts after cardiac death (DCD). METHODS: Sixteen healthy Swedish domestic pigs were randomized into control (n=6), DCD (n=5), and DCD plus cold storage (n=5) groups, respectively. A DCD model in pigs was established using the method of hypoxia-induced 20-minute warm ischemia in the DCD and DCD plus cold storage groups. Isolation of the heart left anterior descending coronary artery or combined with heart preservation pretreatment for 4 hours was performed in the DCD and DCD plus cold storage groups. The maximum coronary endothelium-dependent relaxation was determined in the three groups. RESULTS AND CONCLUSION: There were no significant differences in the maximum coronary endothelium-dependent relaxation and the minus logarithmic of substance concentration induced 50%maximal relaxation among three groups (P > 0.05). These results indicate that 20-minute warm ischemia cannot lead to obvious coronary endothelial dysfunction. In addition, DCD combined with 4-hour cold storage does not affect coronary endothelial function. 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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Construction strategies for tissue-engineered ligaments Di Ya-long, Han Chang-xu, Zhao Liang, Ren Yi-zhong 2016, 20 (29):  4368-4375.  doi: 10.3969/j.issn.2095-4344.2016.29.016 Abstract ( 231 )   PDF (675KB) ( 285 )   Save BACKGROUND: Until now, musculoskeletal bioengineering used in the orthopedic surgery and regenerative medicine are developing. OBJECTIVE: To summarize the construction strategies for tissue-engineered knee ligaments. METHODS: A computer-based online search was conducted in PubMed to screen the relevant literatures using the terms “knee ligament” AND “tissue engineering” OR “regenerative medicine”. The literature retrieval, data assessment and extraction were performed by both authors independently. The relevant literatures were included through reviewing preclinical and clinical studies to display the preclinical and clinical medicine progresses in the construction strategies for tissue-engineered ligaments. RESULTS AND CONCLUSION: Included 224 literatures focus on the construction strategies for tissue-engineered ligaments, including stem cell therapy, growth factors, biomaterials and scaffolds, and mechanical stimulation pattern. Construction techniques for tissue-engineered ligaments have some potential advantages, such as rapid function recovery, good tenacity, and reduced relapse. However, rigorous and high-level randomized controlled clinical trials are needed for the translation of preclinical results into successful clinical trials. 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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Establishment and application of a finite element model of the foot structure Song Ya-wei, You Chuan-bao, Yu Wen-bing 2016, 20 (29):  4376-4382.  doi: 10.3969/j.issn.2095-4344.2016.29.017 Abstract ( 310 )   PDF (753KB) ( 266 )   Save BACKGROUND: The finite element analysis used to study the biomechanical properties of foot structure contributes to overcome the disadvantages of traditional mechanical analysis of specimens. OBJECTIVE: To summarize some representative finite element models of the foot and review the establishment methods of finite element models of the foot with biomechanical properties. METHODS: A computer-based online search was conducted in CNKI, Wanfang and PubMed databases by using the key words of “finite element model, biomechanic, foot structure” from 1999 to 2015. The language was limited to both Chinese and English. Establishment methods of finite element models of the foot structure containing obtaining of model establishment data, establishment of solid model, setting methods of properties of model materials. The application of finite element analysis of the foot structure in sports biomechanics and medical orthotics was summarized to point the characteristics and disadvantages of finite element analysis application in biomechanics. RESULTS AND CONCLUSION: An increasing number of studies on foot biomechanics provide the platform for finite element analysis of the foot applied in biomechanical analysis. Many key technologies, including self-adaption image segmentation method, hexahedron grid product method and establishment of finite element models with various materials have to be utilized to converse the medical images into digital models of finite element analysis during establishment of finite element models. Results of model establishment may be affected by limitation conditions, accuracy of image collection, calculation results of finite element analysis. The reliability of finite element models can be increased by modify technologies and verification methods. 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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Current status of the relationship of digit and phalange length ratios to athletic ability Liang Zhi-dong 2016, 20 (29):  4383-4388.  doi: 10.3969/j.issn.2095-4344.2016.29.018 Abstract ( 283 )   PDF (883KB) ( 274 )   Save BACKGROUND: At present, it is a hot spot to study the relationship between digit or phalange length ratios and athletic ability. However, the results from various studies cannot be verified each other because there are controversies about the research objects and the research technique in the area. Therefore, it is necessary to point a right direction for the research in this field. OBJECTIVE: To explore the relationships of digit and phalange length ratios to athletic ability and the existing problems by literature search strategies and logic thinking methods. METHODS: A computer-based online search was conducted in Elsevier Science-direct, PubMed, EM, CNKI, and CJFD databases by using the key words of “digit length ratio, 2D:4D, phalange length ratio, athletic ability” from 1990 to 2015. The language was limited to both Chinese and English. 358 relevant articles were screened, and 42 eligible articles were included finally. RESULTS AND CONCLUSION: Currently, studies show that there are some relationships between digit phalange length ratio or phalange length ratio and athletic ability. There are controversies about the research results due to the diverse study methods and techniques. It is necessary to define the digit phalange length ratio or phalange length ratio in the human study. The most advanced techniques should be considered to reduce the measurement bias. Moreover, multi-angle, multi-level studies should be performed to reveal the relationship of digit phalange length ratio or phalange length ratio to athletic ability. Human digit phalange length ratio or phalange length ratio, particularly 2D:4D can reflect the aerobic exercise capacity of athletes. 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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Skeletal muscle sarcopenia: molecular mechanisms, prevention and treatment Wang Jin-yue 2016, 20 (29):  4389-4394.  doi: 10.3969/j.issn.2095-4344.2016.29.019 Abstract ( 323 )   PDF (838KB) ( 280 )   Save BACKGROUND: There is an urgent need to explore the loss of skeletal muscle mass during aging (sarcopenia), and its molecular mechanisms of action, and prevention and treatment strategies. OBJECTIVE: To summarize the latest progress in molecular mechanisms of sarcopenia, and to provide a fundamental for promoting functional recovery and regeneration of skeletal muscle. METHODS: A computer-based online search was conducted in PubMed and Wanfang databases from 2005 to 2015 to screen the relevant literatures using Chinese and English key words “sarcopenia, skeletal muscle, mechanism, therapy”, respectively. Consequently, 31 eligible literatures were collected, summarized, and analyzed. RESULTS AND CONCLUSION: Sarcopenia is closely correlated with oxidative stress caused by mitochondrial respiratory failure, inhibition of activating factors for regulating satellite cells, reduction in insulin secretion, decreased sensitivity, protein synthesis, and low protein diet. There are common features and molecular mechanisms in sarcopenia and disuse muscle atrophy. Further exploration of exercise and diet strategies for the treatment of sarcopenia is required. 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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Lisfranc injury in an athletic population: diagnostic and therapeutic strategies Ru Jiang-ying, Niu Yun-fei 2016, 20 (29):  4395-4401.  doi: 10.3969/j.issn.2095-4344.2016.29.020 Abstract ( 277 )   PDF (977KB) ( 202 )   Save BACKGROUND: Lisfranc injury is a concealed or low-energy damage in the athletic population. The optimal treatment strategies for Lisfranc injury in the athletes, especially for high-level or professional athletes, remain controversial. Improvement and development in treatment for Lisfranc injury are ongoing. OBJECTIVE: To summarize the diagnostic and therapeutic strategies and problems in surgery in Lisfranc injuries in the athletic population. METHODS: A computer-based online search was conducted in PubMed and Web of science databases from June 1909 to June 2014 to screen the relevant articles regarding the diagnostic and therapeutic strategies for Lisfranc injury using the key words “Lisfranc, injury, athletes”. The irrelevant and duplicate articles were excluded, and finally 43 articles were reviewed. RESULTS AND CONCLUSION: With the improvement and development in the therapeutic methods for Lisfranc injury, suture button fixation and bioabsorbable screw technology, as novel treatment strategies, have the potential to help restore and/or preserve stability at the tarsometatarsal joints, to avoid the potential risk for internal fixation irritation or the need for removal of hardware after fixation. However, more multi-center, prospective, randomized controlled clinical trials are required for seeking the optimal treatment for Lisfranc injury. For the athletes with Lisfranc injury, the best treatment option, removal timing of internal fixation devices, and the proper postoperative function exercise performed according the conditions of patients are vital for restoring the professional sports level. 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 role of growth factors in the repair of skeletal muscle injury Huang He-ping 2016, 20 (29):  4402-4408.  doi: 10.3969/j.issn.2095-4344.2016.29.021 Abstract ( 304 )   PDF (710KB) ( 213 )   Save BACKGROUND: Growth factors involved in the regulation of cellular processes play an important role in the wound healing and tissue regeneration. OBJECTIVE: To elaborate the role of a variety of cellular processes involving growth factors in the repair of skeletal muscle injury, and to provide the references for the treatment and rehabilitation strategies, and the synthesis of biomaterials with growth factor for the skeletal muscle after injury. METHODS: A computer-based online search was conducted in PubMed, Mendeley, Google Scholar, and CNKI databases from 1995 to November 2015 to screen the relevant literatures using the keywords “skeletal muscle, damage repair, insulin-like growth factor, epidermal growth factor, growth factor”. Data screening, processing, and summary were performed. RESULTS AND CONCLUSION: Fifty-one eligible literatures were included. Exercise training promotes the repair and regeneration of the injured skeletal muscle cells and the recovery of the function by activating satellite cells in the sarcolemma and basement membrane to produce the numerous myoblasts. The repair involves the complex biological process regulated by growth factors. Exogenous growth factors up-regulate the mRNA expression of endogenous growth factors, stimulate the proliferation of the myoblasts, accelerate the fusion between myotubes and muscle fibers, promote the repair of skeletal muscle injury, inhibit the formation of scars, thereby enhancing the healing quality. 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