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08 January 2019, Volume 23 Issue 1 Previous Issue    Next Issue

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Synergistic effect of bone morphogenetic protein 2 and transforming growth factor beta2 on osteogenic differentiation of bone marrow mesenchymal stem cells Chang Xiaopeng, Chen Tao, Zhao Yin, Liang Ming 2019, 23 (1):  1-6.  doi: 10.3969/j.issn.2095-4344.1519 Abstract ( 481 )   PDF (770KB) ( 256 )   Save BACKGROUND: It is revealed from the cell and gene level that bone morphogenetic protein 2 (BMP2) and transforming growth factor β2 (TGFβ2) synergistically promote the osteogenic differentiation of bone marrow mesenchymal stem cells, providing an experimental and theoretical basis for the bone regeneration. OBJECTIVE: To investigate the overexpression of BMP2 and TGFβ2 in vitro to synergistically promote the osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS: Primary rat bone marrow mesenchymal stem cells were obtained by enzymatic digestion. Recombinant plasmids of BMP2 and TGFβ2 were packaged by lentivirus. The lentivirus carrying the recombinant plasmids of the target genes BMP2 and TGFβ2 was transfected into the third generation of bone marrow mesenchymal stem cells. MTS (cell proliferation assay) was used to detect the proliferation of bone marrow mesenchymal stem cells in different treatment groups at different time points. Quantitative PCR was used to detect the overexpression of BMP2 and TGFβ2 at mRNA levels. Immunohistochemistry of cell slides was performed to detect the expression level of osteogenic differentiation marker RUNX2 at the protein level under two-dimensional mixed culture conditions. The cell pellet obtained by three-dimensional culture in vitro was subjected to paraffin sectioning for Vonkossa staining (silver staining) to determine the positive region and expression intensity of osteogenic differentiation, and quantitative analysis was performed. RESULTS AND CONCLUSION: (1) The proliferation of bone marrow mesenchymal stem cells transfected with BMP2 and TGFβ2 was not statistically different from that of untransfected cells (P > 0.05). (2) The BMP2 and TGFβ2 mRNA levels were significantly higher in the transfected cells than untransfected ones (P < 0.05). (3) RUNX2 immunohistochemistry under the in vitro two-dimensional culture conditions and Vonkossa staining under the in vitro three-dimensional culture showed that the BMP2 and TGFβ2 co-transfected bone marrow mesenchymal stem cells had stronger osteogenic ability than BMP2 or TGFβ2 alone transfected cells (P < 0.05). In conclusion, the overexpression of recombinant plasmids of BMP2 and TGFβ2 carried by lentivirus synergistically promotes the osteogenic differentiation of rat bone marrow mesenchymal stem cells.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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miR1-2 induces the differentiation of bone marrow mesenchymal stem cells into cardiomyocytes via Wnt/beta-catenin signal pathway Gao Wei, Li Wenwei, Ren Zengfu, Chen Shiyun 2019, 23 (1):  7-12.  doi: 10.3969/j.issn.2095-4344.1503 Abstract ( 411 )   PDF (863KB) ( 201 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) have the potential to differentiate into cardiomyocytes, but exhibit a low differentiation rate. Moreover, its differentiation mechanism is still unclear. OBJECTIVE: To investigate the role of miR1-2 in the differentiation of mouse BMSCs into cardiomyocytes and to reveal the signaling pathways involved in this process. METHODS: Mouse BMSCs were treated with miR1-2 and 5-azacytosine. The cardiomyocyte marker GATA4 in BMSCs was detected by qPCR. Apoptosis in BMSCs was detected by flow cytometry and the activity of Wnt/β-catenin signaling pathway was evaluated by measuring the upstream protein of this signal pathway. RESULTS AND CONCLUSION: After 24 hours of transfection with miR1-2 mimics, the expression of miR1-2 in BMSCs was significantly increased. After 48 hours of transfection with miR-2 mimics, there was no significant difference in the apoptotic rate of BMSCs. After 48 hours of transfection with miR1-2 mimics, the expression of GATA4 increased significantly with time. After 72 hours of transfection with miR1-2 mimics, the expression of these genes was significantly higher in the miR1-2 group than in the 5-azacytosine group and the control group. Compared with miR1-2-mimics-NC, treatment with miR1-2 mimics could significantly increase the relative expression of β-catenin and Wnt11 at mRNA and protein levels, while addition of Wnt/β-catenin pathway inhibitor LGK-974 reduced the relative expression of β-catenin and Wnt11. To conclude, miR1-2 in BMSCs has the potential to effectively induce the differentiation of BMSCs into cardiomyocytes via the Wnt/β-catenin pathway. Figures and Tables | References | Related Articles | Metrics

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Adipose tissue-derived mesenchymal stem cell conditioned medium combined with platelet-rich fibrin repairs skin injury in mice Huang Min, Yan Hong 2019, 23 (1):  18-23.  doi: 10.3969/j.issn.2095-4344.1520 Abstract ( 403 )   PDF (794KB) ( 243 )   Save BACKGROUND: Platelet-rich fibrin can promote soft tissue healing, and soluble factors secreted by mesenchymal stem cells can promote skin regeneration, but their combination effects on skin repair are little reported. OBJECTIVE: To investigate the effect of conditioned medium of adipose tissue-derived mesenchymal stem cells combined with platelet-rich fibrin on the healing of skin wound in mice. METHODS: Full-thickness skin injury models were established in C57/BL6 mice and randomly divided into four groups, 10 mice in each group. After the modeling, each group was smeared with the specific medium of adipose tissue-derived mesenchymal stem cells, the conditioned medium of adipose tissue-derived mesenchymal stem cells, the platelet-rich fibrin, the conditioned medium of adipose tissue-derived mesenchymal stem cells combined with platelet-rich fibrin, thrice daily for 7 consecutive days. The wound size was measured at 1, 3, 7 and 10 days after treatment and the wound healing rate was calculated at 3 and 7 days after treatment. Histological analysis of wound healing was performed at 14 days after treatment.  RESULTS AND CONCLUSION: Compared with the specific culture medium of adipose tissue-derived mesenchymal stem cells, the wound healing time in the other three groups was shorter. The wound healing rate of adipose-derived mesenchymal stem cell conditioned medium combined with platelet-rich fibrin group was significantly higher than that in the other three groups at 3 and 7 days after treatment (P < 0.05). The adipose-derived mesenchymal stem cell conditioned medium group and platelet-rich fibrin group showed higher healing rate than the adipose-derived mesenchymal stem cell specific medium group (P < 0.05). Histological findings showed that the aggregation of white blood cells and fibroblasts, angiogenesis, granulation tissue remodeling and epithelization in the conditioned medium of adipose mesenchymal stem cells combined with the platelet-rich fibrin group were more obvious than those in the other three groups, in which the best wound healing effect was found. These results suggest that the combination of adipose tissue-derived mesenchymal stem cell conditioned medium and platelet-rich fibrin can effectively promote the repair of skin injury in mice, and the effect is better than that of conditioned medium or platelet-rich fibrin alone.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Contact or noncontact cocultures of articular chondrocytes with bone marrow mesenchymal stem cells: cell proliferation and differentiation Zhao Wenhui, Pi Hongtao, Feng Wanwen, Li Xiangdong, Wang Jianwei, Liu Yuepeng, Jiang Yang, Ma Jianxin, Xia Yayi, Wang Cuifang, Shao Linlin, Li Chunhui, Yu Hongyang, Liu Shanglin, Dong Yanbin, Ma Yahui 2019, 23 (1):  24-29.  doi: 10.3969/j.issn.2095-4344.1521 Abstract ( 498 )   PDF (846KB) ( 231 )   Save BACKGROUND: Coculture of articular chondrocytes with bone marrow mesenchymal stem cells (BMSCs) can contribute to proliferation and phenotype maintenance of articular chondrocytes and chondrogenic differentiation of BMSCs as a result of the microenvironment provided by the small number of articular chondrocytes. Furthermore, the coculture system decreases hypertrophic phenotype and enchondral ossification during BMSCs chondrogenic differentiation. However, the mechanisms underlying the cell-cell interactions in the coculture system of articular chondrocytes with BMSCs have not been fully clarified and necessitate further studies. OBJECTIVE: To investigate the effect of the contact coculture of articular chondrocytes with BMSCs on the cell proliferation and differentiation in vitro, and to further optimize the coculture pattern providing a basis for further exploring the interaction mechanism of articular chondrocytes and BMSCs in the coculture system.  METHODS: Articular chondrocytes and BMSCs were isolated from New Zealand white rabbits (provided by the Animal Core Facility of Nanjing Medical University, Nanjing, China) and expanded in vitro. The articular chondrocytes and BMSCs both at passage 2 were harvested and cocultured at the ratio of 1:3 for 21 days. The coculture patterns included the cell-cell contact coculture in the 24-well plates as experimental group and cell-cell noncontact coculture through a Transwell chamber as control group. Cytochemistry staining and immunocytochemistry staining were performed to observe cell morphology and to evaluate distribution of related proteins. Cell proliferation was determined using Cell Counting Kit-8. Levels of glycosaminoglycans and transforming growth factor-β3 (TGF-β3) in the medium supernatants of the experimental and control groups were detected by ELISA. Expression of type II collagen alpha 1 (COL2a1) and connexin 43 (Cx43) in the coculture system of articular chondrocytes with BMSCs were analyzed using western blot. Cell proliferation, glycosaminoglycan level and expression levels of COL2a1 and Cx43 in the two groups were evaluated following TGF-β3 supplementation in each group and articular chondrocytes were subsequently cocultured with BMSCs in the contact or noncontact manner.  RESULTS AND CONCLUSION: (1) The ability of cell proliferation gradually increased with prolonged coculture time, the mean absorbance value of the experimental group was significantly higher that that of the control group (P < 0.05). The mean levels of glycosaminoglycan and TGF-β3 and expression levels of COL2a1 and Cx43 in the experimental group were significantly higher than those in the control group respectively. (2) Following addition of 10 µg/L TGF-β3, the aforementioned indicators showed no significant changes in the experimental group (P > 0.05), whereas increased significantly in the control group, but still lower than those in the experimental group with no TGF-β3 supplementation (P < 0.05). Our results indicate that the contact coculture of articular chondrocytes with BMSCs significantly promotes cell proliferation and differentiation as compared with the noncontact coculture, and Cx43 plays an important role in cell proliferation and differentiation. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Bone morphogenetic protein 2 transfected bone marrow mesenchymal stem cells and fibrin glue to promote tendon-bone healing after anterior cruciate ligament reconstruction Zhu Lequan, Lu Jianhua, Ran Junling, Yang Weibao, Wang Hui 2019, 23 (1):  30-34.  doi: 10.3969/j.issn.2095-4344.1522 Abstract ( 396 )   PDF (699KB) ( 300 )   Save BACKGROUND: The combination of bone marrow mesenchymal stem cells and fibrin glue can improve the early rupture strength and stiffness of the tendon-bone interface after anterior cruciate ligament reconstruction, and promote the early healing of graft and bone tunnel. OBJECTIVE: To explore the effect of bone morphogenetic protein 2 transfected bone marrow mesenchymal stem cells combined with fibrin glue on the tendon-bone healing after the reconstruction of the anterior cruciate ligament. METHODS: Adenovirus vector Ad-GFP-bone morphogenetic protein 2 was transfected into bone marrow mesenchymal stem cells, and the expressions of GFP gene and bone morphogenetic protein 2 were detected after 48 hours. Fifty-four New Zealand white rabbits purchased from the Medical Animal Center of Chongqing were randomly divided into blank group, experimental group and control group, with 18 rats in each group. The anterior cruciate ligament reconstruction model was established in each rat. In the experimental group, after 48 hours of transfection, the transfected bone marrow mesenchymal stem cells were mixed with fibrin glue and injected into the tendon-bone interface. In the control group, non-transfected bone marrow mesenchymal stem cells were mixed with fibrin glue and injected into the tendon-bone interface. In the blank group, no treatment was given. After 4, 8, 12 weeks, the femur-tendon-tibia specimens were taken for histological and biomechanical examinations. RESULTS AND CONCLUSION: (1) After transfection of 48 hours, the cells expressed green fluorescence under the fluorescence microscope, and the transfection efficiency was (86.1±6.3)%. The expression of cytoplasmic bone morphogenetic protein was positive as shown by SP immunohistochemical test, and the positive band of bone morphogenetic protein 2 was detected by western blot. (2) Findings from the hematoxylin-eosin staining showed best better tendon-bone healing in the experimental and control group than the blank group, and the tendon-bone healing was further better in the experimental group than in the control group. (3) The biomechanical results showed that the maximum load and stiffness were ranked as follows: the experimental group > control group > blank group (P < 0.05). Therefore, bone morphogenetic protein 2-transfected bone marrow mesenchymal stem cells combined with fibrin glue can further enhance the tendon-bone healing after anterior cruciate ligament reconstruction.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Chondrogenic differentiation of bone marrow stem cells on a hydroxyapatite coated titanium plate Qin Liling, Yan Yan, Yu Zhuo, Qin Qiaoling, Li Wei, Liu Li 2019, 23 (1):  35-40.  doi: 10.3969/j.issn.2095-4344.1523 Abstract ( 309 )   PDF (796KB) ( 217 )   Save BACKGROUND: Hydroxyapatite has good biocompatibility and bioactivity and can promote adhesion, proliferation and chondrogenic differentiation of stem cells. OBJECTIVE: To investigate the effect of hydroxyapatite coated titanium plate on chondrogenic differentiation of bone marrow mesenchymal stem cells. METHODS: Hydroxyapatite coating was fabricated on the titanium surface by electrochemical deposition technique (deposition time: 3 minutes and 1.5 hours) and characterized by field emission scanning electron microscopy for coating morphology and by X-ray diffraction for the chemical composition of the coating. Bone marrow samples from four Sprague-Dawley rats of 3 weeks old (purchased from Hubei Provincial Laboratory Animal Center, Wuhan, China) were taken to culture bone marrow mesenchymal stem cells using whole bone marrow adherent method. Passage 3 cells were inoculated onto the titanium plate with or without coating followed by 4 weeks chondrogenic induction. For assessing the ability of chondrogenesis, cell counting kit-8 assay was applied to test the cell proliferation; dimethylmethylene blue colorimetric assay was used to detect the content of glycosaminoglycans; and RT-PCR was used to test the expression of chondrocyte specific genes. RESULTS AND CONCLUSION: (1) The hydroxyapatite coating was evenly distributed on the titanium plate and its structure was uniform. The X-ray diffraction peaks of hydroxyapatite powder generally matched the characteristic X-ray diffraction pattern of hydroxyapatite. (2) The hydroxyapatite-coated titanium plate improved the proliferation of bone marrow mesenchymal stem cells and showed better bioactivity compared to the non-coated titanium plate. (3) The glycosaminoglycan content was significantly higher in the hydroxyapatite coating group than the non-coated group (P < 0.01). (4) The RT-PCR results showed higher expression levels of chondrocyte specific genes (SOX9, type II collagen, type X collagen, aggrecan) in the hydroxyapatite coating group than the non-coated group (P < 0.05). The expression of type II collagen was significantly higher in the 1.5-hour group than the 0.5-hour group and non-coated group (P < 0.05). To conclude, the hydroxyapatite coating could evenly distribute on the titanium plate via electrophoretic deposition. Hydroxyapatite coated titanium surface has good biocompatibility and can promote proliferation and chondrogenic differentiation of rat bone marrow mesenchymal stem cells in vitro.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Differentiation of human umbilical cord mesenchymal stem cells into neuron-like cells: a comparison of two induction methods Wang Wu, Qi Baojun, Wu Zhongyan, Cui Yong 2019, 23 (1):  41-46.  doi: 10.3969/j.issn.2095-4344.1524 Abstract ( 357 )   PDF (774KB) ( 286 )   Save BACKGROUND: Stem cells are a promising treatment for nerve injury diseases due to its ability of differentiating into neuron-like cells. However, the differentiation efficacy is low, so we have make an advance based on the traditional induction method. OBJECTIVE: To explore the potentiality of human umbilical cord mesenchymal stem cells differentiating into neuron-like cells induced by two methods and their merits. METHODS: Human umbilical cord mesenchymal stem cells were isolated and cultured using tissue explant method. The cell morphology was observed under microscope, and the cultured cells were identified by flow cytometry. Human umbilical cord mesenchymal stem cells were differentiated into neuron-like cell under the neurotrophic factor inducing scheme. Two different methods but using the same inducers (2% B27, 20 μg/L basic fibroblast growth factor and 20 μg/L epidermal growth factor) were utilized. Traditional method: B27, basic fibroblast growth factor, epidermal growth factor and penicillin streptomycin combination were added in the DMEM/F12 medium, 3 days later all-trans retinoic acid was added, followed by 10 days of culture. Advanced method: B27, basic fibroblast growth factor, epidermal growth factor, penicillin streptomycin combination and fetal bovine serum were added in the DMEM/F12 medium, 6 days later the cells underwent trypsinization and re-seeded in the confocal capsule, B27 and fetal bovine serum were removed next day, and all-trans retinoic acid was added, cultured for 14 days. Then the differentiated nerve cells were identified by immunofluorescence detection of neurofilament protein and glial fibrillary acidic protein. Meanwhile, the positive rate of the neuromarkers in the cells was compared between two methods. RESULTS AND CONCLUSION: Primary human umbilical cord mesenchymal stem cells were isolated and cultured successfully, and highly expressed CD29 and CD105 whereas lowly expressed CD34 and CD45, which were the characteristics of the stem cell. The differentiated cells were presented as nerve cells in the morphology under microscope in both inducing methods. The results of the immunofluorescence indicated that the neurofilament protein and the glial fiber acidic protein in the differentiated cells were all expressed positive in both methods. The positive rates of neurofilament protein and glial fiber acidic protein were 18.73% and 18.01% for the traditional method, while 27.48% and 29.24% for the advanced methods, respectively (P < 0.05). In summary, human umbilical cord mesenchymal stem cells can differentiate into the neuron-like cells induced by two methods. Moreover, the advanced method is better than the traditional method in the morphology of the differentiated cells and the positive rate of neurofilament protein and glial fiber acidic protein.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Dopamine-induced biomimetic hydroxyapatite coating: preparation and biological effect on bone marrow mesenchym stem cells Xu Yanan, Yang Qiming, Li Hong, Jiang Dianming, Qiao Bo 2019, 23 (1):  47-54.  doi: 10.3969/j.issn.2095-4344.1525 Abstract ( 369 )   PDF (871KB) ( 307 )   Save BACKGROUND: Hydroxyapatite (HA) coating can be formed on different kinds of metals assisted by polydopamine (PDA). However, the preparation of HA coating with biomimetic method assisted by PDA on artificial synthetic materials is rarely reported. OBJECTIVE: To prepare the HA coating on HA/P66 surface assisted by PDA and to evaluate its effect on bone marrow mesenchymal stem cells. METHODS: First HA coating was prepared on the HA/P66 surface using the biomimetic method assisted by PDA to obtain HA-PDA-HA/P66 substrate and then verified by X-ray photoelectron spectroscopy and scanning electron microscopy. Its surface properties such as hydrophilicity and surface roughness were also characterized. Then C3H10T1/2 cells were co-cultured with HA/P66, PDA-HA/P66 and HA-PDA-HA/P66. After 6 and 12 hours of cultivation, the initial cell adhesion was observed by cell counting kit-8. After 1 day of cultivation, cell morphology was observed by scanning electron microscope. After 1 and 3 days of cultivation, cell proliferation was assessed by 4,6-diamino-2-phenyl indole staining. After osteogenic induction for 7 and 10 days, intracellular alkaline phosphatase activity was detected. After osteogenic induction for 14 days, mineralization of the extracellular matrix was detected by alizarin red staining. RESULTS AND CONCLUSION: (1) HA coating was successfully prepared on the HA/P66 substrate which was verified by X-ray photoelectron spectroscopy and scanning electron microscope. HA coating greatly increased the hydrophilicity and surface roughness of HA/P66. (2) After 6 and 12 hours of cultivation, the number of adherent cells on the surface of HA-PDA-HA/P66 was significantly higher than that in the other groups (P < 0.05). After 1 day of cultivation, the cells on the surface of HA-PDA-HA/P66 and PDA-HA/P66 showed polygonal shape with a large number of pseudopodia. (3) After 1 and 3 days of cultivation, the cell proliferation on the surface of HA-PDA-HA/P66 was significantly higher than that in the other two groups (P < 0.05). (4) After 7 and 10 days of osteogenic induction, the alkaline phosphatase activity of cells on the surface of HA-PDA-HA/P66 was significantly higher than that of the other groups. After 14 days of osteogenic induction, there were more calcium nodules on the surface of HA-PDA-HA/P66 than in the other groups. These findings indicate that HA coating can be successfully prepared on the surface of HA/P66 through the biomimetic process assisted by PDA and has good biocompatibility and bioactivity.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of Buyang Huanwu Decoction combined with bone marrow mesenchymal stem cell transplantation on expression of tight junction proteins in the brain of cerebral ischemia-reperfusion rats Zhang Yunke, Che Zhiying, Li Ke 2019, 23 (1):  55-60.  doi: 10.3969/j.issn.2095-4344.1526 Abstract ( 434 )   PDF (818KB) ( 265 )   Save BACKGROUND: In acute cerebral ischemia, the tight junction between endothelial cells is damaged, and variation in the transcription or expression of tight junction proteins increases the blood-brain barrier permeability eventually triggering the formation of brain edema.  OBJECTIVE: To explore the effect mechanism by which Buyang Huanwu Decoction combined with bone marrow mesenchymal stem cell (BMSCs) transplantation treats a cerebral ischemia-reperfusion rat model via the blood brain barrier.  METHODS: Eighty rats provided by the Animal Experimental Central of Shandong Province, China, were randomized into sham operation group (n=16), model group (n=16), transplantation group (n=16), Buyang Huanwu Decoction group (n=16), and Buyang Huanwu Decoction+transplantation group (combination group, n=16). Ischemia-reperfusion rat model was established via middle cerebral artery occlusion using improved Zea Longa’s suture-occlusion method. In the Buyang Huanwu Decoction group and combination group, the rats were gavaged with 15 ml/kg Buyang Huanwu Decoction 3 days before modeling, while in the sham operation group, model group and transplantation group, the rats were gavaged with the equal volume of normal saline. The transplantation group and combination group were injected with bone marrow mesenchymal stem cell suspension (1x106 mL, 1 mL) 1 day after modeling, and the other groups were gavaged with the equal volume of normal saline. The expression of Occludin, Claudin-5, Jam-A, and ZO-1 in ischemic brain tissues was detected by western blot assay at 3 and 5 days after modeling.  RESULTS AND CONCLUSION: Compared with the sham operation group, the protein expression of Occludin, Claudin-5, Jam-A, and ZO-1 in the ischemic brain tissue of rats in the model group decreased significantly (P < 0.01). Compared with the model group, the protein expression levels of Occludin, Claudin-5, Jam-A, and ZO-1 in the rat ischemic brain tissue in the Buyang Huanwu Decoction group, transplantation group and combination group were up-regulated significantly (P < 0.05), with the most significant increase in the combination group (P < 0.01). Buyang Huanwu Decoction combined with bone marrow mesenchymal stem cell transplantation exerts a synergistic effect in blood-brain barrier repair, and keeps the tight junction with the blood-brain barrier through the upregulation of ischemic brain tissue Occludin, Claudin-5, Jam-A and ZO-1protein expression, to achieve the brain protection. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Molecular mechanism underlying the effect of adipose-derived stem cells on the proliferation of keloid fibroblasts Li Xiang, Wu Zhixian, Liu Hongwei, Liang Jie, Mo Zizeng 2019, 23 (1):  61-67.  doi: 10.3969/j.issn.2095-4344.0694 Abstract ( 486 )   PDF (927KB) ( 385 )   Save BACKGROUND: Adipose-derived stem cells have been widely used for tissue filling and repair, but the mechanism underlying skin scar inhibition and repair is still unclear.  OBJECTIVE: To explore the effect of adipose-derived stem cells on biological activity of keloid fibroblasts and its molecular mechanism. METHODS: Passage 3 human adipose-derived stem cells were seeded into the upper Transwell chamber at the density of 0, 3×104, 6×104, 1.2×105 per well (0 indicates control group). Passage 4 keloid fibroblasts at logarithmic growth phase were inoculated into the lower chamber at the density of 6×104 per well, and co-cultured with human adipose-derived stem cells at the ratio of 1:0.5, 1:1 and 1:2, respectively. After 24-hour co-culture, keloid fibroblasts from the lower chamber were used for index measurement.  RESULTS AND CONCLUSION: Human adipose-derived stem cells markedly inhibited the proliferation, migration and collagen synthesis of keloid fibroblasts, and the inhibitory effect was significantly enhanced as the proportion of adipose-derived stem cells increased. Human adipose-derived stem cells also markedly promoted cell apoptosis in keloid fibroblasts, and this effect was enhanced as the proportion of adipose-derived stem cells increased. Western blot results showed that human adipose-derived stem cells significantly suppressed keloid fibroblasts p-ERK, Bcl2 and β-catenin protein expression. Overall, adipose-derived stem cells can inhibit the proliferation and migration but promote apoptosis of keloid fibroblasts by inhibiting the ERK/β-catenin pathway. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of adipose-derived mesenchymal stem cell exosomes on the proliferation and migration of keratinocytes and the underlying mechanisms Tian Xinli, Jiang Bo, Yan Hong 2019, 23 (1):  68-73.  doi: 10.3969/j.issn.2095-4344.1527 Abstract ( 378 )   PDF (1509KB) ( 347 )   Save BACKGROUND: Skin wounds caused by burns and scalds are diseases of high morbidity and mortality rate, and it is urgent to find an effective way to facilitate wound healing. OBJECTIVE: To investigate whether adipose-derived mesenchymal stem cell exosomes can vary the proliferation and migration of keratinocytes and the potential mechanisms. METHODS: Human keratinocytes were cultured using trypsin digestion method, and identified by detection of keratinocyte specific marker protein Keratin 14. Exosomes from adipose-derived mesenchymal stem cells were obtained by ultracentrifugation and identified by the exosome specific marker proteins CD9, CD63 and CD81. The purified exosomes were added to keratinocytes at different concentrations (0, 10, 20, 50, 100 mg/L) to detect the proliferation and migration of keratinocytes at different time points (0, 12, 24, 48 hours). RESULTS AND CONCLUSION: Adipose-derived mesenchymal stem cell exosomes facilitated the proliferation and migration of keratinocytes. The greatest improvement in the proliferation of keratinocytes was detected when the mass concentration was 50 g/mL after 48 hours of treatment. Meanwhile, the phosphorylation of ERK, AKT and STAT3 was strengthened with the existence of exosomes, indicating that adipose-derived mesenchymal stem cell exosomes may promote the proliferation and migration of keratinocytes through promoting the phosphorylation of ERK, AKT and STAT3.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Artificial bone construction using bone marrow mesenchymal stem cells combined with calcium polyphosphate fiber reinforced calcium phosphate cement Cui Xintao, Yang Xiansheng, Chi Zhiyong, Guan Guofa 2019, 23 (1):  74-78.  doi: 10.3969/j.issn.2095-4344.1502 Abstract ( 308 )   PDF (678KB) ( 249 )   Save BACKGROUND: Calcium phosphate cement is a research focus of biomedical materials and has been widely used in clinical practice. However, the low compressive strength and bending strength of calcium phosphate cement limit its applications, and as the artificial bone, the lack of osteoinductive activity is also a major defect. Therefore, the enhancement of calcium phosphate cement has become a hot spot in this field. OBJECTIVE: To explore the feasibility of constructing artificial bone by bone marrow mesenchymal stem cells combined with calcium phosphate fiber/bone cement. METHODS: Calcium phosphate cement/calcium polyphosphate fiber composite scaffold materials at a reasonable proportion were prepared, in which calcium polyphosphate fibers accounted for 10%, 20%, 30%, 40%, 50% and 60% of the total mass respectively. Then, compression test, three-point bending test and torsion test were performed. The maximum compressive load and bending load were recorded, so as to determine the proportion of calcium phosphate/calcium polyphosphate fibers under the best strength. Bone marrow mesenchymal stem cells were isolated and cultured by density gradient centrifugation and induced by osteogenesis. The cultured cells were inoculated onto the calcium polyphosphate fiber/calcium phosphate cement composite scaffold at the most suitable proportion. The growth of bone marrow mesenchymal stem cells on the composite scaffold was observed by scanning electron microscope. RESULTS AND CONCLUSION: The hardness and toughness of the composite scaffold material reached the best when the calcium phosphate fibers accounted for 20% of the total mass of the scaffold. Bone marrow mesenchymal stem cells grew well on the composite scaffold under the scanning electron microscope. Approximately 7 days after culture, the bone marrow mesenchymal stem cells were completely filled in the space of the composite scaffold. To conclude, the calcium polyphosphate fiber plays the role of “steel bar” in the composite scaffold, enhancing the strength and toughness of the scaffold material. The addition of bone marrow mesenchymal stem cells changes the osteogenesis from a single bone conduction mode to an osteogenic mode with bone conductivity, bone induction and bone generation, which enhances the biocompatibility of the scaffold material.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Immune properties of human CD200+ sub-population from human placenta-derived mesenchymal stem cells Liu Ting, Ma Xiaona, Ma Haibin, Yang Tingting, Jin Yiran, Liang Xueyun 2019, 23 (1):  79-84.  doi: 10.3969/j.issn.2095-4344.1528 Abstract ( 367 )   PDF (811KB) ( 322 )   Save BACKGROUND: In different studies, mesenchymal stem cells exhibit contradictory immune characteristics. This may be because mesenchymal stem cells are a group of complex cells and different subgroups exhibit different immune characteristics.  OBJECTIVE: To isolate and identify a sub-population of placenta-derived mesenchymal stem cells with high positive surface antigen of CD200, and to investigate the immune characteristics of this sub-population.  METHODS: Flow cytometry was utilized to sort CD200+ placenta-derived mesenchymal stem cells. Phenotypic characterization and multilineage differentiation of the sorted cells were determined. Cytokine secretion, involving interleukin-6 and interleukin-8, was measured by ELISA. Mixed lymphocyte proliferation assay was used to determine the immune characteristics of CD200+ placenta-derived mesenchymal stem cells.  RESULTS AND CONCLUSION: The CD200+ placenta-derived mesenchymal stem cells exhibited homogeneous spindle shape, and highly expressed CD44, CD73, CD90 and CD105, negatively expressed CD34, CD45, CD80 and CD86. After induced differentiation, the cells were positively stained by oil red O staining and alizarlin red staining, which were the markers of adipocytes and osteoblasts. The results of ELISA indicated that the expression level of interleukin-6 and interleukin-8 in the CD200+ placenta-derived mesenchymal stem cells was strikingly higher than placenta-derived mesenchymal stem cells and CD200- placenta-derived mesenchymal stem cells. Compared with normal and CD200- placenta-derived mesenchymal stem cells, CD200+ placenta-derived mesenchymal stem cells inhibited T cell proliferation significantly, lowered the levels of interferon-γ and interleukin-2, and upregulated the level of interleukin-10. We found that CD200+ placenta-derived mesenchymal stem cells have better immunoregulatory capability compared with normal and CD200- placenta-derived mesenchymal stem cells, which may provide an ideal theoretical basis for clinical use. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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CD4+T and Treg immunomodulatory functions after culture with the supernatant of human placenta mesenchymal stem cells Li Weiwei, Li Xiaofeng, Hou Ping, Li Jianping 2019, 23 (1):  85-89.  doi: 10.3969/j.issn.2095-4344.0696 Abstract ( 357 )   PDF (673KB) ( 166 )   Save BACKGROUND: Treg cells belong to inhibitory CD4+T cells. Human placenta mesenchymal stem cells (hPAMSCs) have T cell immunomodulatory function. OBJECTIVE: To investigate the CD4+T and Treg cells immunomodulatory ability after culture with the supernatant of hPAMSCs. METHODS: hPAMSCs were isolated and cultured, and the stable cell lines were then established. Passage 4 hPAMSCs and its supernatant were collected, and the expression of cell surface markers CD90, CD73, CD45 and HLA-DR were detected by flow cytometry. Cell activation cocktail stimulated peripheral blood mononuclear cells from healthy donor were co-cultured with hPAMSCs supernatant for 6 hours. The percentage of CD4+T cells and Treg cells was detected by flow cytometry. ELISA was used to test the level of transforming growth factor β in the cell supernatant. RESULTS AND CONCLUSION: Successfully established primary hPAMSCs cell lines had stable passage. Supernatant of hPAMSCs significantly inhibited the percentage of CD4+T cells (P < 0.05), while significantly enhanced the percentage of Treg cells (P < 0.05). Meanwhile, the level of transforming growth factor β was significantly up-regulated (P < 0.01). In conclusion, the supernatant of hPAMSCs has the immunomodulatory function via inducing the conversion from CD4+T to Treg cells, which is expected to be a new therapeutic treatment for immune disorders. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Transplantation of endothelial progenitor cells with RNA interference targeting transforming growth factor beta1 inhibits pulmonary fibrosis in rats Peng Qiufeng, Gao Jingzhen, Ye Kui, Xing Aimin 2019, 23 (1):  90-95.  doi: 10.3969/j.issn.2095-4344.1529 Abstract ( 405 )   PDF (873KB) ( 386 )   Save BACKGROUND: Abundant animal experiments have shown that transforming growth factor β1 (TGF- β1) gene is a hotspot of pulmonary fibrosis and an important target for drugs.  OBJECTIVE: To explore the therapeutic effect of the transplantation of endothelial progenitor cells with RNA interference targeting TGF-β1 in the rats with pulmonary fibrosis.  METHODS: (1) The siRNA plasmid directed to TGF-β1 was constructed and transfected into endothelial progenitor cells. It was divided into blank group, control group and TGF-β1-siRNA transfection group. The expression of TGF-β1 at 3 and 28 days after transfection was detected by western blot assay. (2) Eighty Wistar rats provided by Beijing Vital River Laboratory Animal Technology Co. Ltd. were divided into control, model, endothelial progenitor cell, and TGF-β1 silent groups. The rat trachea in the latter three groups was injected with 0.3 mL of bleomycin (5 mg/kg) to establish the rat model of pulmonary fibrosis. At 24 hours after modeling, the control and model groups received the injection of 20 μL of normal saline via caudal vein, and the endothelial progenitor cell, and TGF-β1 silent groups subjected to the injection of 20 μL 3×106/L endothelial progenitor cell suspension and TGF-β1 silent endothelial progenitor cell suspension, respectively. (3) After 28 days of transplantation, the levels of interleukin-10, interleukin-6, and tumor necrosis factor-α in the rat bronchoalveolar lavage fluid were detected by ELISA. The pathological changes of lung tissue were observed by hematoxylin-eosin staining. Expression levels of TGF-β1, Smad-2, and Smad-7 were detected by RT-PCR, and western blot assay. RESULTS AND CONCLUSION: (1) The protein expression of TGF-β1 in the TGF-β1-siRNA transfection group was significantly lower than that in the blank and control groups at 3 and 28 days after transfection (P < 0.05). (2) Compared with the model group, the pulmonary fibrosis was relieved in the endothelial progenitor cell group with reduced alveolar interval thickness and further attenuated in the TGF-β1 silent group. (3) The levels of interleukin-10, interleukin-6, and tumor necrosis factor-α in the endothelial progenitor cell group were significantly lower than those in the model group (P < 0.05). These factor levels in the TGF-β1 silent group were lower than those in the endothelial progenitor cell group (P > 0.05). (4) Compared with the model group, the mRNA and protein expression levels of TGF-β1 and Smad-2 in the endothelial progenitor cell and TGF-β1 silent groups were significantly decreased, the levels of Smad-7 were significantly increased (both P < 0.05). All above levels in the TGF-β1 silent group were significantly superior to those in the endothelial progenitor cell group (P < 0.05). (5) To conclude, TGF-β1 silent endothelial progenitor cell transplantation has a protective effect against pulmonary fibrosis in rats and probably inhibits pulmonary fibrosis by reducing Smad-2 and enhancing Smad-7 expression. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Expression of the clock gene in embryonic and maternal tissues of mice Yan Yindi, Luo Xuguang, Yang Yanping, Li Hairong, Cui Huilin, Cao Ximei 2019, 23 (1):  96-102.  doi: 10.3969/j.issn.2095-4344.0607 Abstract ( 573 )   PDF (987KB) ( 349 )   Save BACKGROUND: Expression and mechanisms of periodic rhythm emergence during mammalian development are not fully understood. OBJECTIVE: To study the spatio-temporal expression characteristics of clock gene in mouse embryos at the gestational age, maternal livers, and maternal skeletal muscles during the development of the mouse.  METHODS: The whole embryos from pregnant 10-17-day ICR mice were collected. The mouse heart, lung, liver and skeletal muscle of body underwent immunohistochemical staining by anti-CLOCK and -Per1 antibodies. Total RNA was extracted using TRIZol from embryos, quadriceps and liver. BMAL1, CLOCK, Per1, myogenin, Tcap and MAZ mRNA were examined by RT-qPCR. RESULTS AND CONCLUSION: The levels of BMAL1, CLOCK and Per1 expression in mouse embryo in the early developmental stages were lower. Immunohistochemical analysis showed that the heart of the early embryos did not express CLOCK at gestational days 10 and 11. However, apparent signals against Per1 and weak expression of CLOCK were observed in the maternal uterus tissue (decidua) of embryos at gestational days 10 and 11. At gestational day 13, CLOCK positive cells showed C-shaped patterns in the wall of the left and right bronchi. At the same time, CLOCK positive cells were detected in the bottom of left atrium and Per1 positive cells were detected in the left ventricular myocardium. At gestational day 14, CLOCK and Per1 positive cells were easily observed in skeletal muscle of body wall. At gestational day 15, immunohistochemical analysis showed a small number of Per1 positive cells were observed in the bottom of right atrium. The cross section of bronchial branches in the lungs increased obviously. At the same time, the expression levels of BMAL1, CLOCK and Per1 mRNA in mouse embryo were increased, especially Per1. However, circadian molecular rhythms could not be found. We found the level of Tcap expression at gestational day 15 significantly increased. The data suggest that sarcomeres in muscle develop rapidly. During development from gestational day 16 to gestational day 17, CLOCK and Per1 positive cells were also observed in the airway smooth muscle, the wall of left atrium and skeletal muscle of body. During the development, CLOCK and Per1 positive cells could not be seen in the liver. The level of myogenin expression in mouse embryo in the early developmental stages was lower. The level of MAZ expression is the highest than myogenin and Tcap in embryos. MAZ may concern with the development and differentiation of skeletal muscle. It is indicated that circadian clock development in embryos is closely correlated with the cellar differentiation process. Mouse embryos begin to express key circadian genes and have the capacity to express active circadian regulatory cycles during development. The clock gene is a positive regulator of myogenesis and the development of heart and liver. The right ventricle shows a relatively slow pace of maturation. However, the present results indicate that synchrony does not occur during prenatal development despite exposure to maternal rhythms. It may be affected by maternal skeletal muscles of mice. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Immunomodulatory properties of mesenchymal stem cells and their application in organ transplantation Jiang Shanshan, Wang Feng, Yu Limei 2019, 23 (1):  103-109.  doi: 10.3969/j.issn.2095-4344.1530 Abstract ( 575 )   PDF (1608KB) ( 274 )   Save BACKGROUND: Allogeneic organ transplantation is one of the most effective therapies to treat end-stage organ failure. However, it is an important problem to effectively overcome the transplantation reaction and induce the immune tolerance of transplantation. The current use of immunosuppressant drugs in clinical practice still has some problems such as limited effect on reducing chronic rejection and high toxicity, and causing chronic infection and tumor. In recent years, the immunomodulatory properties of mesenchymal stem cells have attracted attention in immune tolerance after organ transplantation. OBJECTIVE: To review the immunomodulatory properties of mesenchymal stem cells and their research progress in organ transplantation. METHODS: CNKI and PubMed databases were retrieved by using computer to search relevant articles about immunomodulatory properties of mesenchymal stem cells and their application in organ transplantation. The key words were “mesenchymal stem cells; immune regulation; organ transplantation; rejection reaction; immune tolerance” in Chinese and English, respectively. RESULTS AND CONCLUSION: Mesenchymal stem cells isolated from bone marrow, adipose tissue, umbilical cord, amniotic membrane and other tissues not only have self-replication and multidirectional differentiation potentials, but also have low immunogenicity and conspicuous immunomodulatory properties. Mesenchymal stem cells can inhibit the proliferation and activation of T cells and B cells, and inhibit the proliferation, cytotoxic effect and cytokine secretion of NK cells. Immune tolerance is induced by mesenchymal stem cells through regulating the function and activity of dendritic cells and macrophages. The basic and clinical researches have shown good long-term therapeutic effects and promising application prospect on the anti-rejection response to kidney, liver, heart and skin transplantation.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Physical stimulation methods promote myocardial differentiation and maturation of stem cells Zhang Zeqian, Wu Jiaqi, Fan Yubo, Zheng Lisha 2019, 23 (1):  110-117.  doi: 10.3969/j.issn.2095-4344.1531 Abstract ( 410 )   PDF (979KB) ( 309 )   Save BACKGROUND: Methods for improving the myocardial differentiation of stem cells have become a hot topic in the treatment of heart diseases. OBJECTIVE: To summarize and analyze the physical stimulation methods and research advances in improving the myocardial differentiation of stem cells. METHODS: A online computer search of PubMed, Chinese Journals Full-text Database (CNKI) and other literature databases was conducted using the keywords of “stem cell, cardiomyocyte, mechanical stimulation, electrical stimulation, substrate, three-dimensional culture, maturity” in English and Chinese, respectively. Research literature related to normative, logical and rigorous methods as well as physical factors for stimulating myocardium maturation differentiated from stem cells. Case studies, studies with no rational standard design program, review with no rigorous inclusion criteria, and studies having nothing to do with the topic were excluded. RESULTS AND CONCLUSION: Through the literature search, a variety of physical stimuli, mechanisms of action and research progress have been summarized to improve myocardial differentiation and maturity of stem cells. The use of stem cell transplantation for the treatment of heart disease has brought new methods for clinical use, but there are still some problems to be solved, such as transplanted cell death, immune rejection, arrhythmia and tumorigenesis, which have limited the clinical application of stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Differentiation of human pluripotent stem cells into dopaminergic neurons: security risk for heterogeneity Chen Li, Hu Lan, Peng Yanan, Yang Liu, Shen Hui, Wang Tan, Zhao Zhenqiang 2019, 23 (1):  118-124.  doi: 10.3969/j.issn.2095-4344.1532 Abstract ( 429 )   PDF (781KB) ( 805 )   Save BACKGROUND: The transplantation of human pluripotent stem cell-derived nerve cells for the treatment of Parkinson’s disease has made great progress and the relevant research is at the subclinical stage. However, there are still many problems to be solved. As one of the existing problems, heterogeneity of differentiation can cause tumor formation and dyskinesia in the host after transplantation, which brings potential and unpredictable risks to patients. OBJECTIVE: To summarize the current status of human pluripotent stem cells differentiated into dopaminergic neurons, the principle and process of differentiation design, the induction process and the research status of heterogeneous differentiation, laying a theoretical foundation for the clinical transformation of transplantation therapy. METHODS: The keywords were “iPSC AND Parkinson’s disease, induced pluripotent stem cells AND Parkinson’s disease, ES cells AND Parkinson’s disease, embryonic stem cells AND Parkinson’s disease, pluripotent stem cells AND Parkinson’s Disease” in English and Chinese, respectively. Relevant articles published from 1980 to 2018 were retrieved by the first author in the PubMed, CNKI, and WanFang. Literature addressing the treatment of Parkinson’s disease with induced pluripotent stem cell-derived neurons in recent years was reviewed, and finally 46 articles were retained. RESULTS AND CONCLUSION: A variety of induction differentiation protocols can be used to induce the differentiation of pluripotent stem cells into A9 dopaminergic neurons. Cell transplantation can promote the behavioral and limb function recovery of different Parkinson's disease models. However, in addition to the differentiation of A9 dopaminergic neurons, the current differentiation protocol products include different neurons such as A10 dopaminergic neurons and serotonergic neurons. There is no differentiation scheme that can achieve uniform differentiation. Further optimizing the differentiation conditions of human pluripotent stem cells in vitro and homogenizing differentiation into A9 dopaminergic neurons may further improve the behavioral performance of Parkinson’s disease model animals and promote the clinical transformation of the cell therapy.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Mesenchymal stem cells for acute liver injury: homing to the liver and directed differentiation Sun Ting, Li Fan, Du Lianfang 2019, 23 (1):  125-131.  doi: 10.3969/j.issn.2095-4344.1501 Abstract ( 408 )   PDF (794KB) ( 199 )   Save BACKGROUND: Liver disease is the main cause of mortality and morbidity that are rising globally. Mesenchymal stem cells are characterized with self-renewal, high proliferation, multi-directional differentiation and no immunogenicity. Mesenchymal stem cells exhibit great advantages in the treatment of acute liver injury. OBJECTIVE: To briefly conclude the transplanted routes, safety, efficacy and potential risks of mesenchymal stem cells in the treatment of acute liver injury. METHODS: We retrieved relevant articles published from 2010 to 2017 in the WanFang, CNKI and PubMed databases. The key words were “mesenchymal stem cells, acute liver injury, cell transplantation” in Chinese and English, respectively. After removal of the repetitive and old views, 61 eligible articles were further analyzed. RESULTS AND CONCLUSION: Researches on mesenchymal stem cell transplantation for the treatment of acute liver injury mainly concentrate on fundamental studies and animal experiments. The transplanted routes of mesenchymal stem cells may influence the therapeutic effect of transplanted cells. The mechanisms by which mesenchymal stem cells are used to treat acute liver injury focus on paracrine effect, cell transformation and immunoregulatory effect. It is a developmental tendency to improve the homing rate of mesenchymal stem cells to the liver and hepatic differentiation ability of mesenchymal stem cells, which maybe provides a new strategy for treatment of acute liver injury.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Interactions between bone marrow stromal stem cells and nano-hydroxyapatite Li Lu, Shu Jingyuan, Zheng Lixia, Cui Yingying, Niu Yueyue, Wang Qian, Wang Qingshan 2019, 23 (1):  132-138.  doi: 10.3969/j.issn.2095-4344.0693 Abstract ( 479 )   PDF (957KB) ( 174 )   Save BACKGROUND: As a kind of bioactive material, nano-hydroxyapatite has received extensive attention for its excellent biological performance. Its effect on the proliferation, differentiation, mineralization and cytotoxicity of bone marrow stromal stem cells is an important factor affecting its osteogenesis performance, and it is also a current hot topic in the field of bone tissue engineering. OBJECTIVE: To explore and analyze the effect of nano-hydroxyapatite on bone marrow stromal stem cells and to provide guidance for further study of tissue engineered bone. METHODS: Relevant literature published from January 1, 2006 to July 26, 2018 was retrieved by the first author in PubMed, CNKI, VIP, WanFang and Google Scholar. The keywords were “hydroxyapatite, nano-hydroxyapatite, biological matrix material, bone marrow stromal stem cells, osteogenic activity, biocompatibility, cytotoxicity, bone defect, bone tissue engineering, tissue engineered bone” in English and Chinese, respectively. Sixty-six eligible articles were finally reviewed. RESULTS AND CONCLUSION: A large number of studies have shown that the nano-hydroxyapatite and its composite materials with bone marrow stromal stem cells have good affinity, adhesion and biocompatibility, and can induce the proliferation and differentiation of bone marrow stromal stem cells. Nano-hydroxyapatite is an ideal material for bone defect repair, which can be widely used in bone defect repair and surface modification of implants, thereby promoting early new bone formation and strengthening the binding between implants and bone tissues. Nano-hydroxyapatite composite materials are also used to develop biological artificial ligaments and guided bone tissue regeneration membrane materials as well as to treat femoral head necrosis. Further investigations on the interaction between nano-hydroxyapatite and bone marrow stromal stem cells will certainly promote the research progress and clinical application of tissue-engineered bone.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Paracrine actions of bone marrow mesenchymal stem cells: angiogenesis, immunoregulation, and inflammatory regulation Zhang Xin, Xie Jiabing 2019, 23 (1):  139-143.  doi: 10.3969/j.issn.2095-4344.0527 Abstract ( 563 )   PDF (686KB) ( 203 )   Save BACKGROUND: Although bone marrow mesenchymal stem cells have the inherent ability to differentiate into different types of tissue cells, there is increasing evidence that bone marrow mesenchymal stem cells may regulate the repair process of damaged tissues through a variety of paracrine factors. OBJECTIVE: To review the progress in the paracrine effect of bone marrow mesenchymal stem cells in orthopedics. METHODS: A search of Web of Science and PubMed was performed for articles regarding the paracrine effect of bone marrow mesenchymal stem cells in orthopedics. Then, 49 eligible articles were included after systematical summarization and analysis. RESULTS AND CONCLUSION: Studies have shown that bone marrow mesenchymal stem cells play a vital role in the angiogenesis, immunoregulation, and inflammatory regulation, which is expected to be a new therapy for bone diseases. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Cytokines and cartilage tissue engineered scaffold for chondrogenic differentiation of adipose-derived stem cells: roles, difficulties and intrinsic mechanisms Liu Xiaonan, Qi Yiying, Xu Tengjing, Yang Quanming, Yu Xinning, Dai Xuesong 2019, 23 (1):  144-150.  doi: 10.3969/j.issn.2095-4344.0677 Abstract ( 321 )   PDF (825KB) ( 181 )   Save BACKGROUND: Cartilage tissue engineering is currently a research hotspot, which can overcome the problems of autologous cartilage injury and insufficient material sources encountered in autologous cartilage transplantation. It is of great significance to seek sufficient sources of stem cells with strong in vitro proliferation ability and multi-directional differentiation potential as cartilage seed cells. OBJECTIVE: To review the progress of chondrogenic differentiation of adipose-derived stem cells as well as the main existing problems. METHODS: We extensively reviewed experimental research and clinical trial regarding the chondrogenic differentiation of adipose-derived stem cells in the PubMed, WanFang, and CNKI from 2000 to present. The keywords were “adipose-derived stem cells/ASCs, chondrocyte, cartilage, cytokine” in English and Chinese, respectively. All the retrieved literatures were synthesized and analyzed in detail or concisely according to the literature reference value.  RESULTS AND CONCLUSION: Existing studies regarding the chondrogenic differentiation of adipose-derived stem cells mainly focus on exogenous cytokines, including bone morphogenetic protein, transforming growth factor β, insulin-like growth factor 1, growth differentiation factor 5, basic fibroblast growth factor, and dexamethasone, as well as their interactions. Cartilage tissue-engineered scaffolds cannot only promote the differentiation and proliferation of adipose-derived stem cells, but also compensate for the lack of extracellular matrix and accelerate the production of tissue blocks for transplantation. At present, to induce the chondrogenic differentiation of adipose-derived stem cells is mainly in exploratory stage, and further explorations on relevant mechanism are needed to introduce the chondrogenic differentiation of adipose-derived stem cells into the clinical use. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | Related Articles | Metrics

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Induced pluripotent stem cells in dental tissue regeneration: effect and application Shao Miaomiao, Liu Zhongxi, Xu Nuo, Liu Qinghua, Wang Dong, He Jianya, Li Xiaojie 2019, 23 (1):  151-157.  doi: 10.3969/j.issn.2095-4344.1533 Abstract ( 353 )   PDF (812KB) ( 194 )   Save BACKGROUND: The research of regenerative medicine is based on stem cells. The emergence of induced pluripotent stem cells blazes a trail for stem cell research, which is considered to be a major breakthrough in stem cell research in the last decade. Reprogramming adult somatic cells into a pluripotent stem cell-like state avoids dilemmas regarding ethics and immune rejection, which helps achieve personalized cell-based therapy and revolutionize disease research methods and treatment strategies. Recent studies have found that induced pluripotent stem cells have great application values in the field of dental regenerative medicine. Although this research is still at its infancy, induced pluripotent stem cells have shown huge potential in tooth tissue regeneration. OBJECTIVE: To review the main progress about induced pluripotent stem cell sources and induction conditions as well as its related application in oral tissue medicine. METHODS: A computer-based online retrieval of PubMed and CNKI was performed to search relevant articles about induced pluripotent stem cells and oral tissue engineering published from 2010 to 2018, with the keywords of “induced pluripotent stem cell (iPSCs), dental tissue regeneration, adult somatic cells” in English and Chinese, respectively. After removal of repetitive or irrelevant articles, 62 articles finally met the criteria for review. RESULTS AND CONCLUSION: Studies on induced pluripotent stem cells open a new direction for stem cell research. Although the source of induced pluripotent stem cells is extensive, the origins of oral tissue appear to have unique advantages over other tissues. In addition, induced pluripotent stem cells bypass numerous ethical and legal concerns that have hindered embryonic stem cell research. Existing studies have shown that induced pluripotent stem cells can differentiate into dental tissues, periodontal tissue and alveolar bone tissue, which have potential utility in oral tissue regeneration. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Instructions for Authors (2019)-Chinese Journal of Tissue Engineering Research (CJTER) 2019, 23 (1):  158-164.  doi: 10.3969/j.issn.2095-4344.1534 Abstract ( 361 )   PDF (1435KB) ( 210 )   Save Related Articles | Metrics