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10 June 2016, Volume 20 Issue 24 Previous Issue    Next Issue

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Icariin promotes osteogenic differentiation of bone marrow mesenchymal stem cells Bao Yuan, Huang Jun-ming, Jing Xing-zhi, Li Xing-yan, Dong Yong-hui, Zhang Jin-ming, Guo Feng-jin, Chen An-min 2016, 20 (24):  3501-3507.  doi: 10.3969/j.issn.2095-4344.2016.24.001 Abstract ( 368 )   PDF (2170KB) ( 215 )   Save BACKGROUND: Herba epimedii, a traditional Chinese medicine, has a long time in dealing with various orthopedic disorders. Icariin with many biological activites is one of the most important compositions of Herba epimedii. OBJECTIVE: To investigate the effects of icariin on osteogenic differentiation of mesenchymal stem cells and the underlying mechanisms. METHODS: Bone marrow mesenchymal stem cells were treated using icariin with or without osteogenic medium in vitro. Osteogenic differentiation markers, including runt-related transcription factor 2, osteocalcin and osterix, were detected by real time-qPCR. Alizarin red staining was used to measure calcium nodes generated by osteoblasts induced from bone marrow mesenchymal stem cells. The proximal tibia bone structure of rats fed with icariin (2 mg per day) for 5 weeks was detected and analyzed by MicroCT. RESULTS AND CONCLUSION: Icariin was able to promote the expression of genes related to osteogenic differentiation in the absence or presence of osteogenic induction. Icariin could obviously increase the quantity of calcium nodes when mesenchymal stem cells were cultured in the osteogenic medium. The animal experiment showed that icariin improved formation of trabecular bone. 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 rabbit anatomical three-dimensional models of large segmental tibial defects Bao Xiao-gang, Xu Guo-hua 2016, 20 (24):  3508-3515.  doi: 10.3969/j.issn.2095-4344.2016.24.002 Abstract ( 382 )   PDF (6051KB) ( 198 )   Save BACKGROUND: Tissue-engineered bone has been considered to be a promising candidate for the repair and reconstruction of load-bearing large segmental bone defects. Currently, the studies on the application of tissue-engineered bone mainly focus on cell-scaffold or cytokine-scaffold constructs, which have shed light upon the repair of large segmental bone defects. OBJECTIVE: To establish simple and convenient tissue engineering of anatomically shaped tibial bone defect models using three-dimensional rapid prototyping technology to manufacture rabbit tibia biomimetic artificial bone scaffolds. METHODS: Three-dimensional electronic models were constructed using Mimic software. Hydroxyapatite/polycaprolactone scaffolds were manufactured by fused deposition modeling equipment. Fifty rabbits aged 6 months were randomly divided into three groups: blank control (n=3), control (n=6) and experimental groups (n=6), respectively. Tibial defects ranged 1.2 cm were made in all groups. No treatment was given in blank control group. The bone defects in control and experimental groups were repaired with autogenous osteotomized bone and anatomical tissue-engineered bone, respectively, and fixed with plates and screws. RESULTS AND CONCLUSION: (1) Rabbit tibial bone measurements: tibial length was (93.77±0.59) mm, tibiofibular transverse diameter (8.36±0.13) mm, sagittal diameter (5.97±0.12) mm, average thickness of bone cortex (1.20±0.10) mm, average diameter of the medullary cavity (4.30±0.06) mm. Angle between the connection line of the midpoints of superior and inferior articular surfaces at the side of tibial bone models and the connection line of the midpoints of superior and inferior intersecting surfaces at the side of osteotomized bone models was α=(5.97±0.13)°. (2) X-ray in bone defects: at postoperative 4 and 12 weeks, no obvious displacement and angulated deformity were found in bone grafts, suggesting the good bone defect repair. (3) Histological examination: at postoperative 4 weeks, bone scaffolds were filled with new bone in the experimental group. Furthermore, considerably increased new bone formation and mineralization were observed at postoperative 12 weeks. (4) General observation: no obvious displacement and angulated deformity occurred in bone defect grafts at postoperative 4 and 12 weeks. These findings suggest that rabbit anatomical models of large segmental tibial bone defects with good stability were constructed using three-dimensional prototyping technology, which may simulate the structure and function of bone tissue and be used for guiding the new bone regeneration. 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 Morinda on receptor activator of nuclear factor-kappaB expression in osteoclasts in osteoporosis Wang Jun1, Miao De-sheng1, Lv Gang1, Mo Xin-yuan2 2016, 20 (24):  3516-3522.  doi: 10.3969/j.issn.2095-4344.2016.24.003 Abstract ( 279 )   PDF (4791KB) ( 483 )   Save BACKGROUND: Morinda has been reported to promote the proliferation, the secretion of alkaline phosphatase and osteocalcin, and mRNA expression of transforming growth factor of osteoblasts. However, little information is available addressing the effects of Morinda on receptor activator of nuclear factor-κB expression in osteoclasts in rats with osteoporosis. OBJECTIVE: To study the effects of Morinda on receptor activator of nuclear factor-κB expression in osteoclasts of osteoporosis rats. METHODS: Thirty Sprague-Dawley rats were equally and randomly divided into Morinda and 17β-estradiol groups. Rat models of osteoporosis were established by bilateral ovariectomy, and then 5 mL of Morinda decocta (1.0 mmol/L) and 17β-estradiol (1×10-6 mmol/L) were administered intragastrically to rats in Morinda and 17β-estradiol groups for 3 consecutive months, respectively. Primary osteoclasts were isolated from rats in both groups, and then cultured for 3, 6 and 9 days followed by TRAP staining and cell counting. Bone mineral density of the proximal and distal femur, urine and serum levels of Ca2+ and progesterone, and receptor activator of nuclear factor-κB expression in osteoclasts of rats in both groups were determined. RESULTS AND CONCLUSION: Osteoclast fusion was reduced in Morinda group. In contrast, number of osteoclasts was increased and cells became more mature in the 17β-estradiol group. Bone mineral density of the proximal and distal femur bilaterally, urine and serum levels of Ca2+ and progesterone were significantly increased, while receptor activator of nuclear factor-κB expression was significantly decreased in osteoclasts in Morinda group compared with 17β-estradiol group (P < 0.05). These results indicate that Morinda reduces receptor activator of nuclear factor-κB expression in osteoclasts in osteoporosis rats, thereby inhibiting the development and progression 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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A modified method for in vitro isolation and cultivation of periosteal cells in rabbits Zhang Jun-wei1, Lu Hai-tao1, Yuan Feng2, Yang Yu-ming1 2016, 20 (24):  3523-3528.  doi: 10.3969/j.issn.2095-4344.2016.24.004 Abstract ( 415 )   PDF (3432KB) ( 226 )   Save BACKGROUND: Periosteum is considered as a source of seed cells for cell therapy due to its biological features. OBJECTIVE: To seek the optimal way to isolate and culture rabbit periosteal cells and identify their biological features. METHODS: Rabbit periosteum on facies medialis tibiae was taken out under aseptic conditions. Periosteal cells isolated through the digestion of type II collagenase with the explants culture method were cultured in DMEM/F12 complete medium. Cell ultrastructure was observed under an inverted microscope. Periosteal cell proliferation was determined by cell counting kit-8 assay. Cell surface antigens CD90 and CD105 were determined using flow cytometry. Osteogenic and lipogenic induction mediums were applied to induce periosteal cells to differentiate into osteocytes and adipocytes, respectively. After 2 weeks of induction, cells were harvested for alizarin red staining and oil red O staining to assay the calcium nodules and lipid droplet. RESULTS AND CONCLUSION: The digestion of type II collagenase with the explants culture method shortened the period of primary cells culture and enhanced the survival rate, which caused higher purity and stronger reproductive activity of harvested periosteal cells. Primary cultured periosteal cells grew in form of spindle spiral or parallel. Alizarin red and Oil red O staining verified the multi-directional differentiation potentiality of periosteal cells. These findings suggest that the periosteal cells with high purity, strong reproductive activity, and multi-directional differentiation potentiality can be harvested in short time using digestion of type II collagenase with the explants culture method. 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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Seven risk factors of femoral head necrosis after internal fixation in femoral neck fractures: a retrospective analysis of 199 cases Xie Kang, Gao Wei-lu, Chang Jun, Yin Zong-sheng 2016, 20 (24):  3529-3534.  doi: 10.3969/j.issn.2095-4344.2016.24.005 Abstract ( 298 )   PDF (1096KB) ( 192 )   Save BACKGROUND: The incidence of femoral head necrosis after internal fixation in femoral neck fracture is at high level, but the risk factors are still under discussion. OBJECTIVE: To retrospectively propose and analyze the relative risk factors of femoral head necrosis after internal fixation with femoral neck fracture in patients younger than 60 years old. METHODS: Data from 199 patients with femoral neck fractures, younger than 60 years, undergoing closed reduction and internal fixation were analyzed retrospectively. Seven factors including age, sex, body mass index, Garden classification, fracture seasons, reduction quality and weight-bearing time after internal fixation were statistically analyzed to evaluate their correlation with femoral head necrosis. RESULTS AND CONCLUSION: The incidence of femoral head necrosis was 13.1% among the 199 cases. Body mass index, Garden classification, fracture seasons, reduction quality were significantly associated with femoral head necrosis. The age, sex and weight-bearing time were in significantly associated with femoral head necrosis. In multivariate logistic regression analysis, Garden classification, reduction quality and fracture seasons (winter versus summer) had overt effects on development of femoral head necrosis. These results suggest that Garden classification and reduction quality make an important effect on the healing of femoral neck fracture. In addition, fracture in winter may be the risk factor of the femoral head necrosis after internal fixation in femoral neck fracture.| 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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Reliable morphological parameters for the assessment of discoid lateral meniscus in adults: an MRI study Sun Xiao-xin1, Zhou Wei2, Zuo Shu-ping3, Liu Hao1, Liu Hao1, Song Jing-feng1, Yu Xiang-qian1, Zhang Yan1, Liang Chun-yu1, Zhang Liu 1 2016, 20 (24):  3535-3540.  doi: 10.3969/j.issn.2095-4344.2016.24.006 Abstract ( 302 )   PDF (1096KB) ( 267 )   Save BACKGROUND: Currently, the morphologic changes of the discoid lateral meniscus (DLM) remain unclear, and morphological indexes used to evaluate the injuries of DLM still need to be explored. OBJECTIVE: To compare the MR imaging difference between symptomatic and asymptomatic DLM in adults, and to select reliable morphological parameters for assessing DLM. METHODS: A total of 36 patients (36 knees) with a symptomatic DLM (study group) and 34 patients (34 knees) with an asymptomatic DLM (control group) were included in this study. GE Healthcare Centricity RIS/PACKS System was used to measure the primary morphological parameters of two patients with DLM on coronal MR imaging. Intergroup comparison was performed with the Wilcoxon test. RESULTS AND CONCLUSION: On the coronal MR imaging, the width of the body portion and height of the free edge of the DLM was significantly increased in the study group compared with the control group (P < 0.001), while the height of the peripheral portion which connects with the capsule of the DLM was significantly decreased in the study group compared with the control group (P = 0.002). The height of the middle of the DLM and the width of the lateral femoral condyle showed no significant differences between the both groups (P > 0.05). The width ratio of the body of the DLM to the lateral femoral condyle and the height ratio of height of the free edge to the middle of the DLM were significantly increased in the study group compared with the control group (P < 0.001), while the height ratio of the peripheral portion to the middle for the DLM was significantly decreased compared with the control group (P < 0.001). These findings suggest that compared with asymptomatic discoid lateral meniscus, the width of the body portion and the height of the free edge for the DLM are increased; however, the height of the peripheral portion which connects with the capsule of the DLM is decreased. The width ratio of the body portion of the DLM to the lateral femoral condyle and the height ratio of the free edge to the middle for the DLM may be reliable morphological parameters for the assessment of the DLM in adults. 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 acellular dermal matrix and expanded flap in half auricular reconstruction with rib cartilage grafts Dong Hai-jiang, Wang Xi-mei, Wan Cheng, Li Xiang, Zhang Qiong-ge 2016, 20 (24):  3541-3548.  doi: 10.3969/j.issn.2095-4344.2016.24.007 Abstract ( 243 )   PDF (1730KB) ( 237 )   Save BACKGROUND: Traumatic auricle defects in upper 1/2 or lower 1/2, seriously involve the auricular cartilage and skin blood vessels. The autogenic rib cartilage graft and acellular dermal matrix have good histocompatibility, and expanded flap is a kind of thin and achromatic tissue for skin defect repair.  OBJECTIVE: To explore the application of acellular dermal matrix and expanded flap in half auricular reconstruction, and to find out the fine carving and anastomosis of autogenic rib cartilage graft as well as its similarities with the ear and clinical significance. METHODS: Eight cases of half auricular defects were treated with expanded flap, autogenic rib cartilage graft, fine anastomosis of autogenic rib cartilage graft and residual ear for half auricular reconstruction, during which the acellular dermal matrix was used to promote residual ear docking and skull auricle angle formation. The reconstruction was performed in three stages: first, an expander (volume, 80 mL) was subcutaneously implanted at the retro-auricular area; second, the auricular defects were reconstructed with fine rib cartilage graft, acellular dermal matrix and auriculoplasty; finally, acellular dermal matrix was used to promote residual ear docking. Then the half auricular reconstruction was evaluated by objective measurement and subjective rating. RESULTS AND CONCLUSION: Half auricular reconstruction was successful in all the eight cases without obvious complications, and the cartilage grafts were in good condition. During the follow-up, the reconstructed auricle was shaped well and formed a good involution with the residual auricle. In addition, the flange was smooth without obvious color difference and edema, and its position, size and shape were consistent with those of the contralateral one. After the follow-up of 6 months, objective indicators showed that the affected side had no significant difference from the contralateral one (P > 0.05). In conclusion, the acellular dermal matrix can obviously decrease the complications of the cartilage grafts and contribute to a good shaped auricle; and half auricular reconstruction by expanded flap, autogenic rib cartilage graft, and fine anastomosis of autogenic rib cartilage graft can achieve significant clinical effects. 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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Correlation of alveolar bone structure and incisor labial inclination in adolescents with skeletal class II malocclusion of Uygur and Han populations in Xinjiang, China Ruan Xiao-hui, Hou Peng, Wu Pei-ling 2016, 20 (24):  3549-3554.  doi: 10.3969/j.issn.2095-4344.2016.24.008 Abstract ( 395 )   PDF (982KB) ( 236 )   Save BACKGROUND: Location relationship between maxillary incisor labial inclination and the thickness of anterior alveolar bone displays ethnic and regional differences. OBJECTIVE: To study the correlation of alveolar bone structure and incisor labial inclination in adolescents with skeletal class II malocclusion in Uygur and Han populations of Xinjiang, China. METHODS: Sixty Uygur and 60 Han patients at 11-16 years of age (average 13.8 years old), 32 males and 28 females, with skeletal class II malocclusion were included. The thickness of anterior alveolar bone and incisor labial inclination were measured on lateral cephalometric films, and the results were analyzed using SPSS 17.0 statistical software. RESULTS AND CONCLUSION: In Han and Uygur populations, palatal alveolar bone thickness in maxillary incisor region was significantly negatively correlated with the inclination of maxillary incisor (r=-0.715 for Han, r=-0.651 for Uygur, P < 0.05), while labial alveolar bone thickness in mandibular incisor region was significantly positively correlated with the lower incisor inclination (r=0.902 for Han, r=0.745 for Uygur, P < 0.05). These results indicate that the palatal alveolar bone thickness in maxillary incisor region decreases with the increase of the inclination of maxillary incisor; however, the labial alveolar bone thickness in mandibular incisor region increases with the increase of the lower incisor inclination. 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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Combined use of docetaxel and bone marrow mesenchymal stem cells in human hepatoma cell line SMMC-7721 Li Jing-yun, Jiao Ting 2016, 20 (24):  3555-3561.  doi: 10.3969/j.issn.2095-4344.2016.24.009 Abstract ( 222 )   PDF (4585KB) ( 324 )   Save BACKGROUND: Docetaxel, a cell cycle specific anti-tumor drug, is a drug that is used primarily for treating breast cancer; however, its efficacy is low when used for treatment of cancer not sensitive to radiotherapy. Bone marrow mesenchymal stem cells have been shown to strengthen the effects of tumor-specific targeting chemotherapy drugs. OBJECTIVE: To investigate the effects of docetaxel combined with bone marrow mesenchymal stem cells (BMSCs) on human hepatoma cell line SMMC-7721. METHODS: BMSC cells were cultured in vitro. The logarithmic growth of SMMC-7721 hepatoma cells were randomly divided into blank control group, BMSCs group and combined treatment group (combined treatment of BMSCs and docetaxel). SMMC-7721 cell cycle was detected using flow cytometry. Cell growth rate of SMMC-7721 was determined by MTT assay. mRNA and protein expressions of tumor suppressor genes PTEN and p53 were detected by reverse transcription polymerase chain reaction and western blot assay. RESULTS AND CONCLUSION: Combined treatment of docetaxel and BMSCs inhibited SMMC-7721 cell proliferation. Compared with the blank control group, the number of cells at the G0/G1 phase was significantly increased in BMSCs group and combined treatment group. The cell growth rate of SMMC-7721 was significantly inhibited in BMSCs group compared with the blank control group, and that was further inhibited in combined treatment group (P < 0.05). mRNA and protein expression of PTEN and p53 were significantly increased in combined treatment group compared with BMSCs group and blank control group (P < 0.05). Our results suggest that BMSCs inhibit the growth of SMMC-7721 cells, and combined use of docetaxel and BMSCs strengthens the antitumor effect of BMSCs. 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 pre-vascularization of tissue-engineered constructs on the survival of transplanted pancreatic islets Zhao Gao-ping1, 2, Deng Shao-ping1, Huang Xiao-lun2, Yang Mao-zhu2, Wei Ling-ling2, Li Shu-rong2, Wei liang2 2016, 20 (24):  3562-3567.  doi: 10.3969/j.issn.2095-4344.2016.24.010 Abstract ( 300 )   PDF (6393KB) ( 256 )   Save BACKGROUND: Pancreatic islet transplantation via portal vein system leads to the apoptsis of a number of islet cells due to local hypoxia, thereby affecting transplant outcomes. OBJECTIVE: To explore the effect of pre-micrvascularization network of tissue-engineered constructs on the survival of transplanted islets and the feasibility of xenogenic islet transplantation. METHODS: A 5-mm-long cylindrical silicone tube filled with Matrigel TM matrix surrounding the superficial epigastric vessel was placed in the groin of diabetic mice. After the syngeneic islets with 300 islet equivalents (IEQ) were transplanted into the silicone chamber on days 0, 14 and 28 post-chamber implantation, respectively, the recovery time of blood glucose was observed. The islets with the quantity of 100 IEQ, 200 IEQ and 300 IEQ, respectively, were transplanted on day 28 post-implantation and then the blood glucose level was determined. Moreover, the survival of human pancreatic islets with 1 000 IEQ transplanted into the pre-vascularizated chamber or under the renal capsule of diabetic mice, followed by the treatment of anti-CD45RB and/or anti-CD40L (MR-1) was analyzed. RESULTS AND CONCLUSION: An abundant micro-vascularized network was established in the silicone chamber on day 28 post-implantation. The time of the blood glucose returning to normal level in diabetic mice was negatively correlated with the time required for pre-vascularization and the number of implanted islets. No islet grafts implanted in the silicone chamber and treated by anti-CD45RB survived for long term. However, one of seven (14.3%) grafts survived for long term, which was not significantly different from the transplantation under the renal capsule group (n=8, MST > 71 days, P > 0.05). The tissue-engineered pre-vascularization network markedly extends the survival time of the islet grafts before transplantation. The transplantation of the xenogenic pancreatic islets into the vascularized silicone chamber might be a promising method in the future clinical application. 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 Tougu Xiaotong Capsule and its disassembled recipes on chondrocyte degeneration of rats via Wnt/beta-catenin signaling pathway Wu Zhui-le, Chen Xing-qiang, Chen Chun-rong, Yu Chao, Zhu Xiao-qin, Zhuang Zhi-qiang 2016, 20 (24):  3568-3573.  doi: 10.3969/j.issn.2095-4344.2016.24.011 Abstract ( 283 )   PDF (5320KB) ( 204 )   Save BACKGROUND: Previous studies have showed that Tougu Xiaotong Capsule (TGXTC) exerts better effects on osteoarthritis, by regulating Rho/Rock signaling pathway, inhibiting signal transduction of chondrocyte mitochondrial apoptosis pathway, varying the rate and pattern of subchondral bone remodeling and improving the arrangement of subchondral bone collagen fibers and calcium-phosphate crystallization. OBJECTIVE: To observe the effects of the serum containing TGXTC and its disassembled recipes on chondrocyte degeneration of rats via Wnt/β-catenin signal pathway, and to explore the main therapeutic method for osteoarthritis in the TGXTC. METHODS: Forty Sprague-Dawley rats were randomly assigned to receive the treatment of TGXTC, Bushen Rougan (BSRG), Huoxue Qufeng (HXQF) and normal saline, respectively, according to the dose conversion methods of animal to animal and animal to human. Then various drug-containing serums were prepared for the following cellular experiment. After culture and passage, chondrocytes from Sprague-Dawley rats at passage 3 were divided into five groups: blank control, model, TGXTC, BSRG, HXQF groups. Cells in the latter four groups were cultured in appropriate drug-containing serums (normal saline serum for the model group) for 72 hours, following intervention with interleukin-1β for 24 hours. Cells in the blank control group were cultured in normal saline serum. Afterwards, cells in all the five groups were collected for detecting expression of Wnt 4, β-catenin and matrix metalloproteinase 13 at mRNA and protein levels using real-time PCR and western blot assay, respectively. RESULTS AND CONCLUSION: Compared with the blank control group, the expression of Wnt 4, β-catenin, matrix metalloproteinase 13 was significantly increased in the model group. Compared with the model group, the expression of Wnt 4, β-catenin, matrix metalloproteinase 13 in the TGXTC, BSRG and HXQF groups were decreased significantly, sequenced as TGXTC group < BSRG group < HXQF group. These results indicate that TGXTC plays a synergistic protection against interleukin-1β induced degeneration of chondrocytes. In addition, BSRG as a disassembled recipe of TGXTC is the main therapeutic method for 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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Local injection of ligustrazine influences the expression of osteoprotegerin in the maintenance phase in rats with orthodontic tooth movement Guo Rui, Ruan Qiang, Lu Wei, Wu Xiao-dan, Zhao Gang 2016, 20 (24):  3574-3580.  doi: 10.3969/j.issn.2095-4344.2016.24.012 Abstract ( 248 )   PDF (5057KB) ( 500 )   Save BACKGROUND: Ligustrazine has been shown to restore the function of the femoral head via the revascularization, increased blood flow, the absorption of necrotic bone, and bone regeneration. OBJECTIVE: To study the effects of ligustrazine on remodeling of periodontal tissues and the expression of osteoprotegerin in the maintenance phase in rats with orthodontic tooth movement. METHODS: Thirty-two healthy male Wistar rats were included and equally randomized into four groups. Maxillary left first molar mesialization was performed through traction of 50 g force for 21 days to establish the rat model of tooth movement. 5, 10, 15 mg/L ligustrazine (50 µL) were locally injected into the first molar periosteum in model rats on the day before removing the orthodontic forcing device. Same volume of saline was injected in the control group. The injection was administered every other day. At 1 and 4 weeks after injection, the distance of tooth movement, the recurrence distances and percentage were determined and calculated. The pathological changes in periodontal tissues were observed by immunohistochemistry and hematoxylin-eosin staining. The width of the parodontium and number of osteoblasts were observed under an optical microscope. RESULTS AND CONCLUSION: The recurrence distance in the control group was increased compared with the experimental group, while the number of osteoblasts and osteoprotegerin immunoreactivity were decreased. Good width of the parodontium and small recurrence trend were found in 10 mg/L ligustrazine group. These findings indicate that ligustrazine promotes the proliferation of osteoblasts and enhances the expression of osteoprotegerin, which is beneficial to the retention of teeth after orthodontic surgery. 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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Bone morphogenetic proteins-2/Osterix signaling pathway regulates the differentiation of preosteoblasts Li Cheng-lin, Chen Shu-lan, Ren Wei-wei 2016, 20 (24):  3581-3587.  doi: 10.3969/j.issn.2095-4344.2016.24.013 Abstract ( 365 )   PDF (1076KB) ( 279 )   Save BACKGROUND: Bone formation and development are reported to be regulated by bone morphogenetic protein 2 (BMP2)-induced Osterix expression. OBJECTIVE: To investigate the regulatory effect of BMP2/Osterix signaling pathway on differentiation of preosteoblasts in mice. METHODS: mRNA and protein expression of Osterix was determined by real-time RT-PCR and western blot assay, respectively at various time points after mouse preosteoblasts were treated with BMP2. pcDNA3.1/myc-Osterix eukaryotic expression vector was constructed and transducted into preosteoblasts, and then mRNA expression of alkaline phosphatase, bone sialoprotein, and matrix extracellular phosphoglycoprotein was detected by real-time RT-PCR after transduction and BMP2 treatment. RESULTS AND CONCLUSION: Osterix mRNA expression was up-regulated when treated with BMP2 in mouse preosteoblasts, and reached the peak at 24 hours. In addition, the protein expression of Osterix was increased after BMP2 treatment. Alkaline phosphatase, bone sialoprotein, and matrix extracellular phosphoglycoprotein mRNA expression was up-regulated after transfection of mouse preosteoblasts with pcDNA3.1/myc-Osterix eukaryotic expression vector and BMP2 treatment. Our results indicate that BMP2 regulates the synthesis of genetic markers of osteogenesis, such as alkaline phosphatase, matrix extracellular phosphoglycoprotein via up-regulating Osterix expression in mouse preosteoblasts, suggesting BMP2/Osterix signaling pathway plays a critical role in bone development. 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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Preload-induced stress relaxation and membrane current changes in the denervated intact smooth muscle Ni Zhi-zhan, Li Jie-yao, Zhang Pei, Xia Li, Ye Si-qi, Annie Christel Bell, Fan Shou-yan, Piao Ling-hua, Chen Zhi-bin, Gao Ling-feng, Wang Yang 2016, 20 (24):  3588-3594.  doi: 10.3969/j.issn.2095-4344.2016.24.014 Abstract ( 212 )   PDF (1754KB) ( 234 )   Save BACKGROUND: Muscle stretch test is a method for testing mechanical properties of denervated muscle under diverse loads. OBJECTIVE: To observe changes of spontaneous tension wave in myography and membrane electric current in aorta smooth muscle samples under several preload conditions. METHODS: Denervated intact smooth muscle samples were taken from the Kunming mouse aorta and urinary bladder wall. Smooth muscle samples were fixed on a micro positioning device and the first stretch was induced for a passive tension up to 1 g, this position was determined as the initial length (L0). Intermittently, with increasing sample length from L0 every 5 minutes, the 1st and 10th stretch were recorded as low- (L0+1) and high-preload (L0+10). 3% CaCl2 and 0.05% nitrendipine were dropped on the samples before L0+1 and L0+10 stretch were recorded. Membrane current changes were evaluated by glass microelectrodes and MultiClamp 700B Amplifier and pClamp 10 analyzer software. The membrane current changes after L0+1 and L0+10 were analyzed. RESULTS AND CONCLUSION: Prolonged stress relaxation phase was shortened with increasing preload in smooth muscle preparations. The stress relaxation phase in smooth muscle samples from urinary bladder wall was shorter than aorta samples. It was revealed that there were compliance differences between two kinds of smooth muscle samples. In aorta samples, myogenic spontaneous contraction amplitude during stress relaxation and membrane current were significantly increased with increasing preload. Increased amplitude and frequency of membrane current in aorta smooth muscle samples under high calcium concentration (3% CaCl2) were inhibited by L-type calcium channel blockade (0.05% nitrendipine). Collectively, increased preload in smooth muscle samples induced a directly decrease of compliance with an enlargement of myogenic spontaneous contraction amplitude. It appeared vital in particular in aorta samples. Membrane current significantly enhanced in spontaneous contraction period suggested the mechanical stretching ionic inward flow involved in this period. Membrane current amplitude was increased by high-preload under elevated calcium conditions; however, this change was suppressed by L-type calcium channel blockade. This study indicates quick stretching influences not only mechanically-gated channels in smooth muscle, but also the activation of L-type calcium channel. 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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Isokinetic muscle strength of the shoulder flexor and extensor muscles in adolescent softball players Ma Tao, Gao Bing-hong 2016, 20 (24):  3595-3601.  doi: 10.3969/j.issn.2095-4344.2016.24.015 Abstract ( 351 )   PDF (1071KB) ( 214 )   Save BACKGROUND: The isokinetic muscle strength test system can quantitatively test the shoulder flexor and extensor muscles and objectively analyze the relationship between size and balance of flexor and extensor. Therefore, it could be used for strength training monitoring as well as prediction and rehabilitation of sports injuries. OBJECTIVE: To understand shoulder flexor and extensor strength characteristics of adolescent softball players, and find out shortcomings in the shoulder flexor and extensor strength to provide guidance for adolescent athletes’ strength training and point out possible sports injury risks caused by imbalanced strength. METHODS: ISOMED2000 isokinetic testing system (D&R, Germany) was employed to test the isokinetic strength of shoulder flexor and extensor of 12 adolescent female softball players from Shanghai Sports University, China. RESULTS AND CONCLUSION: With the increase of test speed, the bilateral shoulder flexor and extensor peak torque, the flexor and extensor average power ratio, and the flexor and extensor powers were reduced, but the flexor and extensor average power was on the rise. Under the same test speed, flexor and extensor peak torque, the flexor and extensor peak torque ratio, the flexor and extensor average power, the flexor and extensor average power ratio and the flexor powers of the throwing shoulder were significantly larger than those of the non-throwing shoulder (P < 0.05), while the extensor peak torque and the extensor average power of the throwing and non-throwing shoulders showed no significant differences (P > 0.05). Under the same test speed, bilateral shoulder homonymous flexor strength ratio was larger than that of the extensor, and the extensor power of the throwing shoulder was less than that of the non-throwing shoulder. The ratio of homolateral shoulder flexor and extensor ranged from 70% to 80%. These results indicate that there is a significant difference between the flexor strength of the bilateral shoulders in adolescent softball players, but the bilateral extensor strength is similar. Flexor fast strength and extensor maximum strength of the throwing shoulder are lower. Besides, the isokinetic muscle strength test system can be used to evaluate the flexor and extensor strength in adolescent softball athletes, based on which coaches can give targeted strength training in adolescent softball 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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Clinical epidemiological study on anterior cruciate ligament rupture and reconstruction Chen Lian-xu, Fu Li-gong 2016, 20 (24):  3605-3608.  doi: 10.3969/j.issn.2095-4344.2016.24.016 Abstract ( 337 )   PDF (1067KB) ( 213 )   Save BACKGROUND: Anterior cruciate ligament (ACL) reconstruction is the primary treatment method for ACL rupture. Currently, studies on ACL reconstruction involve histology and embryology, anatomical structure, biomechanics, reconstruction materials, operating technology, and rehabilitation after reconstruction. However, clinical epidemiological studies describing ACL rupture and reconstruction remain scarce. OBJECTIVE: To analyze the clinical epidemiological characteristics of ACL rupture and reconstruction to provide guidance for prevention and treatment of ACL rupture. METHODS: Data of 352 patients for age, gender, cause and mechanism of injury, treatment time, and the impact of ACL rupture on menisci and articular cartilage were gathered. Meanwhile, the events during surgery, operation methods and reconstruction materials were analyzed. RESULTS AND CONCLUSION: ACL rupture mostly occurred in young men, and happened more often to the left knee; male patients got hurt in basketball, football and accidental injuries, while female patients got hurt in the accidental injuries, badminton and skiing injuries, internal rotation with valgus stress accounted for the predominant injury mechanism. The ACL reconstruction was mostly performed within 1-3 months after ACL rupture, often accompanied by meniscal and articular cartilage damage. Lateral meniscus injury incidence was relatively stable, medial meniscus injury incidence increased significantly over the half year after ACL rupture. Most articular cartilage injury occurred to patellar cartilage. A significant increase in medial condylar cartilage damage over 1 year after ACL rupture was often observed. Anatomic single-bundle ACL reconstruction was the primary surgical approach, the resident ridge and the lateral bifurcate ridge could be used to position bone tunnel and autogenous semitendinosus and gracilis tendon were the most commonly used reconstruction materials. Our results indicate that anatomic ACL reconstruction should be performed as early as possible in restore knee joint stability and prevent secondary injury of the medial meniscus and cartilage of medial femoral condyle. 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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High frequency ultrasonic imaging of the dorsal Lisfranc ligament in the adults Chen Pei-jin, Mayinuer Maimaitiming, Hou Xia, Zhang Ai-hong, Liu Chun-hong 2016, 20 (24):  3609-3614.  doi: 10.3969/j.issn.2095-4344.2016.24.017 Abstract ( 339 )   PDF (1269KB) ( 223 )   Save BACKGROUND: As a novel noninvasive method, high frequency ultrasound has been used for the detection of skeletal muscle widely. OBJECTIVE:To detect the thickness of the dorsal Lisfranc ligament between the first (medial) cuneiform (C1) and second metatarsal (M2) using high frequency ultrasound to evaluate the feasibility and clinical application value of high frequency ultrasound for the detection of the Lisfranc joint. METHODS: Both feet of 100 healthy volunteers were included and the thickness of dorsal Lisfranc ligament and the distance between C1 and M2 were examined using high frequency ultrasound. RESULTS AND CONCLUSION:The display rate of high frequency ultrasound for the dorsal Lisfranc ligament in the 100 healthy volunteers was 100%. There was no significant difference in the thickness of the dorsal Lisfranc ligament between both feet or sexes in adults (P > 0.05); as did the distance between C1 and M2 (P > 0.05). These findings suggest that visual dorsal Lisfranc ligament is detected by high frequency ultrasound. The measurement of the distance between C1 and M2 by high frequency ultrasound may be a feasible, noninvasive, low-cost, and early diagnostic method for Lisfranc injuries. 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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Methotrexate prevents knee intraarticular adhesion via endoplasmic reticulum stress signaling pathway Chen Hui, Li Xiao-lei, Yan Lian-qi, Sun Yu, Liang Yuan, Zhao Shuai, Zhu Geng-yao, Dai Ji-hang, He Jun, Wang Jing-cheng 2016, 20 (24):  3615-3620.  doi: 10.3969/j.issn.2095-4344.2016.24.018 Abstract ( 283 )   PDF (1194KB) ( 253 )   Save BACKGROUND: The pathogenesis of knee intraarticular adhesion is yet unknown. Excessive proliferation of fibroblasts is considered to cause knee intraarticular adhesion. OBJECTIVE: To study the preventive effects of methotrexate on knee intraarticular adhesion through fibroblast apoptosis induced by endoplasmic reticulum stress. METHODS: The viability of the cultured fibroblasts treated with methotrexate (10-5-10-9mol/L) or PBS was determined after 24 hours. Fibroblast apoptosis was detected by Hoechst33342 staining. Endoplasmic reticulum stress- and apoptosis-related proteins, including cleaved-PARP, CHOP, Bax and Bcl-2, were determined by western blot assay. Eighteen healthy male New Zealand white rabbits were used to establish the knee intraarticular adhesion models, and equally randomized into three groups, and received topical application of 2 or 1 g/L methotrexate, or normal saline (control). The preventive effects of methotrexate on knee intraarticular adhesion and CHOP expression in scar tissue were observed. RESULTS AND CONCLUSION: Methotrexate inhibited the proliferation and viability of fibroblasts in a dose-dependent manner. The number of apoptotic fibroblasts was significantly increased compared with control group. Protein expression of cleaved-PARP, CHOP, and bax was increased, while protein expression of bcl-2 was decreased with time. The animal experiment showed that preventive effects of   2 g/L methotrexate on knee intraarticular adhesion were superior to 1 g/L methotrexate treatment. CHOP expression in the scar tissue in the methotrexate groups was higher than the control group and that was higher in high-dose methotrexate group. Our results suggest that methotrexate prevents knee intraarticular adhesion via endoplasmic reticulum stress-induced fibroblast apoptosis. 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 molecular regulatory networks involved in the neural crest cell development: a bioinformatic analysis Wang Jun-fang, Wang Jia-qi, Bi Qing-wei, Yin Han-wen, Zhang Guo-liang 2016, 20 (24):  3621-3627.  doi: 10.3969/j.issn.2095-4344.2016.24.019 Abstract ( 325 )   PDF (1303KB) ( 210 )   Save BACKGROUND: Development of neural crest cells, which is regulated by various genes, plays an important role in the formation of central nervous system, heart, craniofacial organs and other tissues. However, the relationship among these genes is unclear. OBJECTIVE: To investigate the molecular regulatory networks involved in the process of neural crest cell development based on a bioinformatic analysis. METHODS: Totally 500 differentially expressed genes during the process of neural crest cell development were obtained from the GEO on-line database. Then the DAVID and STRING on-line databases were used to evaluate the relationship among these genes. RESULTS AND CONCLUSION: Totally 500 differentially expressed genes during the process of neural crest cell development could be enriched into different subgroups based on the analysis of DAVID database, including “transforming growth factor β signal pathway”, “WNT signal pathway”, “homeobox gene” , “neural crest cell differentiation” and “neural tube development”. Additionally, 12 genes molecular networks were built based on the analysis of STRING database, such as DLX5, MSX2, SNAIL2, PAX7, SHH, SOX9, NOG, GSC, KAL1, bone morphogenetic protein 5, fibroblast growth factor 8 and WNT3a .These genes exhibited interactions by co-expression, activation and antagonism. Therefore, many genes involved in the process of neural crest cell development were interacted and formed the networks. These findings imply that we should understand these neural crest-related diseases from a holistic view of the signaling pathway and molecular regulatory networks. 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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Relationship of neutrophils lymphocyte ratio and red blood cell distribution width with idiopathic facial palsy Shu Xiang-ning, Ma Yue-wen 2016, 20 (24):  3628-3600.  doi: 10.3969/j.issn.2095-4344.2016.24.020 Abstract ( 280 )   PDF (953KB) ( 212 )   Save BACKGROUND: Similarly with C-reactive protein, red blood cell distribution width can reflect the inflammatory process. Red blood cells involved in inflammatory process leads to the increase in red blood cell distribution width due to various inflammation factor receptors existing on the surface of red blood cells. OBJECTIVE: To analyze the relationship between the neutrophils lymphocyte ratio, red blood cell distribution width and the pathogenesis and severity of idiopathic facial paralysis. METHODS: The severity of idiopathic facial paralysis was assessed using the House and Brackmann Facial Nerve Grading System (HB) and Sunnybrook System (SB) in 30 included patients with idiopathic facial paralysis (disease group). The patients were graded as general (score 30-100) and severe paralysis (score 0-29) according to SB scores, or as mild (grade II/III), moderate (grade IV), severe paralysis (grade V/VI) according to HB grades. In addition, 30 healthy volunteers were selected as control group. Neutrophils lymphocyte ratio (NLR) and red blood cell distribution width (RDW) were compared between groups. The correlation of SB and HB scores with neutrophils lymphocyte ratio (NLR) and red blood cell distribution width was analyzed. RESULTS AND CONCLUSION: Neutrophils lymphocyte ratio was higher in the disease group than the control group (P < 0.01). A significant difference in red blood cell distribution width was found between different stages of disease severity in patients. Red blood cell distribution width was negatively correlated with SB scores (P < 0.01), while was positively correlated with HB grades (P < 0.01). Our results suggest that (1) neutrophils lymphocyte ratio is increased when idiopathic facial paralysis occurs; (2) red blood cell distribution width is positively correlated with the severity of idiopathic facial paralysis, which indicates that inflammation is involved in the pathogenesis of idiopathic facial paralysis, and red blood cell distribution may be a marker for assessing the severity of idiopathic facial paralysis. 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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Regulation of osteoarthritis hypoxia-inducible factor-2 alpha regulatory mechanism and application prospect Yang Chao, Zhang Lei, Zhou Li-wu, Zhao Jian-ning 2016, 20 (24):  3634-3641.  doi: 10.3969/j.issn.2095-4344.2016.24.021 Abstract ( 288 )   PDF (964KB) ( 359 )   Save BACKGROUND: Hypoxia inducible factor-2 α (HIF-2α) is one of the key regulators in articular cartilage degeneration and endochondral osteogenesis. OBJECTIVE: To review the regulatory mechanism and application development of HIF-2α in articular cartilage degeneration. METHODS: A computer-based retrieval of PubMed and CNKI databases was performed for relevant literatures published from January 1990 to October 2015 ,using the key words of “hypoxia inducible factor-2α; osteoarthritis; cartilage” in Chinese and English, respectively. RESULTS AND CONCLUSION: HIF-2α is a transcription factor encoded by the Epas1 gene, which is one of the key regulators in articular cartilage degeneration and endochondral osteogenesis by directly regulating the expressions of various catabolic genes to strengthen cartilage destruction in osteoarthritis. It has been reported that HIF-2α acts on the chondrocytes by regulating the expressions of Wnt/β-Catenin, nuclear factor-κ B, and interleukin-6. 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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Ciliopathies: cellular mechanism and translational application Na Yu-yan, Liu Wan-lin 2016, 20 (24):  3642-3648.  doi: 10.3969/j.issn.2095-4344.2016.24.022 Abstract ( 350 )   PDF (1199KB) ( 273 )   Save BACKGROUND: Cilium is an antenna-like organelle that plays a vital role in all organs generation and maintenance. Ciliopathies refer to multisystem diseases caused by disorders of the cilia structure and function. OBJECTIVE: To introduce current understanding of the structure and function of the cilium, with a focus on the study progress of ciliopathies. METHODS: PubMed database was searched by computer using the keywords of “primary cilium, motion cilium, ciliopathy”. Totally 371 relevant articles published from 1959 to 2015 were retrieved. After excluding repetitive and irrelevant articles, we enrolled 32 English literatures regarding cilia structure, function and ciliopathies in accordance with the inclusion criteria. RESULTS AND CONCLUSION: Ciliopathies almost involve every system of the human body, such as the brain, eyes, liver, kidney, bone and reproductive system. Most ciliopathies are related to human developmental or degenerative diseases, which characterized by wide-ranging clinical and genetic heterogeneity, such as primary ciliary dyskinesia, ploycystic kidney disease, infertility, blindness and tumor. In view of the medical importance of the cilium, scholars have devoted to studying the underlying cellular mechanisms. These in turn have revealed that ciliopathies are not only caused by defective assembly of the cilium, but also by impaired ciliary signaling. 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