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    08 January 2021, Volume 25 Issue 1 Previous Issue    Next Issue
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    Umbilical cord mesenchymal stem cell transplantation in the treatment of ischemic heart disease: a 3-year follow-up
    Jing Yucheng, Wang Le, Wang Xianyun, Wei Mei, Li Min, Ji Lishuang, Ma Fangfang, Liu Gang , Zheng Mingqi
    2021, 25 (1):  6-12.  doi: 10.3969/j.issn.2095-4344.2153
    Abstract ( 243 )   PDF (1368KB) ( 98 )   Save
    BACKGROUND: The traditional treatment methods for ischemic heart disease can temporarily relieve pain and improve the quality of life, but cannot restore and regenerate the myocardial tissue. Stem cells have been reported to possess the potential functions of tissue regeneration and multiplex differentiation characteristics which provide a new opportunity for the ischemic heart disease treatment. Because of the unique advantages of umbilical cord mesenchymal stem cells including easy separation, no immune rejection reaction, no ethical dilemmas and remarkable immunomodulatory effect, umbilical cord mesenchymal stem cells have been considered as one of the most ideal candidate seeds for the treatment of ischemic heart disease. 
    OBJECTIVE: To investigate the clinical efficacy of umbilical cord mesenchymal stem cells transplantation in patients with ischemic heart disease in 3-year follow-up. 
    METHODS: From January 2013 to June 2016, eight patients with coronary heart disease were admitted to the First Hospital of Hebei Medical University and randomly divided into stem cell transplantation group and control group. Four patients in the stem cell transplantation group received conventional treatment and intravenous infusion of umbilical cord mesenchymal stem cells. Four patients in the control group only received conventional treatment. At 3 years after treatment, cardiac function indexes, biochemical indexes and ST segment changes of electrocardiogram lead II were evaluated in two groups. All patients signed the informed consent. The experiment was approved by the Ethics Committee of First Hospital of Hebei Medical University. 
    RESULTS AND CONCLUSION: Patients in each group were alive after 3 years. Compared with the control group, left ventricular ejection fraction and left ventricular fraction shortening displayed an increasing trend in the stem cell transplantation group (P > 0.05). The changes of left ventricular ejection fraction and left ventricular fraction shortening indicated an important statistical improvement in the stem cell transplantation group compared with the control group (P < 0.05). There was no significant improvement in New York Heart Association classification between the two groups. The biochemical indicators revealed no significant statistical difference between the two groups (P > 0.05). ST segment changes of different electrocardiogram leads were various before and after treatment in both groups. This study can only show that human umbilical cord mesenchymal stem cells can improve the cardiac function of patients with ischemic heart disease to a certain extent, but there is no significant improvement in the classification of cardiac function, biochemical indicators, and the influence on electrocardiogram leads is not clear. Thus, the significance of this treatment method needs further study. 

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    Effects of bone marrow mesenchymal stem cell transplantation on thymus structure and function in the aging macaques
    Wang Yanying, Yang Yukun, Zhu Xiangqing, Li Ye, He Jie, Tian Chuan, He Zhixu, Pan Xinghua
    2021, 25 (1):  13-19.  doi: 10.3969/j.issn.2095-4344.2157
    Abstract ( 284 )   PDF (3268KB) ( 75 )   Save
    BACKGROUND: Thymus is an important central immune organ of human body, which is the place where T cells grow, develop and mature. Thymus is the first organ of senescence in human body, gradually atrophy and degeneration after puberty, followed by the gradual decline of immune function.
    OBJECTIVE: To investigate the effect of bone marrow mesenchymal stem cells on the structure and function of thymus in the aging macaques.
    METHODS: Bone marrow was collected from female macaques with an average age of 3 years by bone marrow aspiration. Bone marrow mesenchymal stem cells were obtained by differential adherent culture. Five young macaques with an average age of 3 years were used as the young group. Ten aging macaques with an average age of 25 years were randomly divided into elderly group (n=4) and elderly treatment group (n=6). The macaques in the elderly treatment group were infused with bone marrow mesenchymal stem cells (1 × 107 cells/kg) through the femoral vein, and were infused every other day for three consecutive times. Macaques of the young group and the elderly group were infused with the same volume of saline at the same time. The changes of output and secretion levels of the subgroup of thymocytes in the elderly treatment group after infusion, thymic index, thymic tissue structure and collagen fiber deposition in each group were analyzed.
    RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cell transplantation increased thymus index, T cell output level and reduced regulatory T cells, improved thymus secretion function, increased thymosin alpha and thymosin II secretion. The thymus parenchyma area increased; the skin and medulla junction appeared; part of the thymus tissue was regenerated and transformed to normal structure; the degree of thymus tissue fibrosis was reduced; and collagen fiber deposition was reduced. These results indicate that bone marrow mesenchymal stem cell transplantation can improve the structure and function of thymus in aging macaques. 

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    Effect of Yougui Decoction on autophagy and fate of bone marrow mesenchymal stem cells in rats with glucocorticoid-associated femoral head necrosis
    Liu Xin , Du Bin, Gao Lili, Sun Guangquan, Wang Xu, Yuan Peng, Lin Xuanye
    2021, 25 (1):  20-25.  doi: 10.3969/j.issn.2095-4344.2146
    Abstract ( 260 )   PDF (1385KB) ( 19 )   Save
    BACKGROUND: Yougui Decoction is an empirical prescription for the treatment of glucocorticoid-associated femoral head necrosis. Literature has shown that the pathogenesis of glucocorticoid-associated femoral head necrosis is associated with glucocorticoid-induced autophagy down-regulation and fate change in bone marrow mesenchymal stem cells. 
    OBJECTIVE: To investigate the effect of Yougui Decoction on autophagy and fate of bone marrow mesenchymal stem cells in model rats of glucocorticoid-associated femoral head necrosis. 
    METHODS: We used Escherichia coli endotoxin combined with high-dose dexamethasone to make the rat models of early femoral head necrosis. Forty SHR rats were randomly divided into five groups: blank control group, model group, high-dose Yougui Decoction group, medium-dose Yougui Decoction group and low-dose Yougui Decoction group. After 6 weeks of intervention, medullary cavity tissue of the rat proximal femur was taken for hematoxylin-eosin staining and immunohistochemical staining of autophagy proteins LC3 II, P53 and beclin-1. After culture and induction of bone marrow mesenchymal stem cells, alizarin red staining, bone alkaline phosphatase quantification, oil red staining and MTT determination were performed and western blot assay was used to quantitatively measure the expression of LC3 II, P53 and beclin-1 proteins. 
    RESULTS AND CONCLUSION: (1) Immunohistochemistry and western blot assay results showed that Yougui Decoction significantly increased autophagy protein LC3 II, P53 and beclin 1 expression in a dose-dependent manner. (2) Alizarin red staining, oil red staining and bone alkaline phosphatase quantification suggested that Yougui Decoction could significantly interfere with the fate of bone marrow mesenchymal stem cells, up-regulate their osteogenic differentiation and down-regulate their adipogenic differentiation in a dose-dependent manner. (3) MTT results suggested that Yougui Decoction significantly improved the proliferation ability of bone marrow mesenchymal stem cells, but had no significant differences in different doses. (4) To conclude, Yougui Decoction can significantly improve the autophagy expression, change the cell fate, up-regulate osteogenic differentiation and down-regulate adipogenic differentiation of bone marrow mesenchymal stem cells in the rat models of glucocorticoid-associated femoral head necrosis, which provides certain basis for elucidating the mechanism of Yougui Decoction in treating glucocorticoid-associated femoral head necrosis.
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    Osteogenic differentiation of bone marrow mesenchymal stem cells induced by demineralized bone matrix in vitro
    Chen Junyi, Wang Ning, Chen Ximiao, Zhu Lunjing, Duan Jiangtao, Zhang Xianping, Bei Chaoyong
    2021, 25 (1):  26-31.  doi: 10.3969/j.issn.2095-4344.2128
    Abstract ( 295 )   PDF (1856KB) ( 30 )   Save

    BACKGROUND: Demineralized bone matrix contains many kinds of active factors such as bone morphogenetic protein 2, which can promote bone marrow mesenchymal stem cells to transform into chondrocytes and promote their proliferation under specific joint microenvironment.

    OBJECTIVE: To explore the osteogenic differentiation of bone marrow mesenchymal stem cells in vitro induced by demineralized bone matrix and its research value as cell carrier scaffold in the treatment of bone defect.
    METHODS: Rat femur bone marrow mesenchymal stem cells were isolated and adhered with whole bone marrow. Bone marrow mesenchymal stem cells at passage 3 were selected and cultured with complete medium and 50 mg/L demineralized bone matrix inducer. The proliferation and viability of bone marrow mesenchymal stem cells were determined by CCK-8 assay. Alkaline phosphatase activities were quantitatively measured by enzyme labeling at 7 and 14 days after culture. Calcium nodule formation was observed by alizarin red staining 21 days after culture. The expression of osteogenesis-related factors was detected by qRT-PCR 21 days after culture. Demineralized bone matrix and bone marrow mesenchymal stem cells were cultured for 21 days. The adhesion of them was observed by ordinary optical inverted phase contrast microscope and scanning electron microscope.  
    RESULTS AND CONCLUSION: (1) The number of cells on days 5 and 6 in the demineralized bone matrix group was higher than that in the complete medium group (P < 0.05). (2) Alkaline phosphatase activities were significantly higher in the demineralized bone matrix group than in the complete medium group at 7 and 14 days (P < 0.05). (3) Calcium nodules were more in the demineralized bone matrix group than in the complete medium group. (4) The expression of RUNX2, ALP, OCN and OPN was significantly higher in the demineralized bone matrix group than in the complete medium group (P < 0.05). (5) The adherence of bone marrow mesenchymal stem cells was good on the demineralized bone matrix; cells were distributed in the space between scaffolds and crawled with each other. (6) The results showed that demineralized bone matrix could induce bone marrow mesenchymal stem cells to differentiate into osteoblasts, and the adhesion was good, which provided theoretical reference for the research of tissue engineering composite materials. 

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    Cardiomyocyte-like differentiation of bone marrow mesenchymal stem cells induced by myocardial tissue lysates from different parts of the myocardium 
    Wei Yashu, Liu Hongjing, Wang Huifeng, Wang Junduo, Zhao Wenjing, Chen Weiping
    2021, 25 (1):  32-37.  doi: 10.3969/j.issn.2095-4344.2142
    Abstract ( 247 )   PDF (1616KB) ( 42 )   Save
    Abstract
    BACKGROUND: Bone marrow mesenchymal stem cells can be induced into myocardial tissue-like structure in vitro. Myocardial tissue lysates from different parts of the myocardium can be used to construct differential microenvironments. Few studies have investigated the targeted induction efficiency of bone marrow mesenchymal stem cells and the expression of related genes.
    OBJECTIVE: To investigate the effects of lysates from different parts of the myocardium on the differentiation of bone marrow mesenchymal stem cells into myocardial tissue-like structures and to assess the correlation between the expressions of related genes during induction and myocardial tissue engineering.
    METHODS:  Passage 3 bone marrow mesenchymal stem cells from Sprague-Dawley rats were cultured. Whole heart tissue lysate, atrial muscle tissue lysate, and ventricular muscle tissue lysate were used to construct different microenvironments to induce bone marrow mesenchymal stem cells to differentiate into cardiomyocyte-like cells. The morphological changes and ultrastructure of cells were observed with an inverted phase contrast microscope and transmission electron microscopy. Expression of α-actin and cTnI was detected by immunofluorescence staining. qRT-PCR was used to detect the expression of upstream molecules ANP, HCN4 and downstream molecules MLC-2v in the cAMP/PKA signaling pathway after induction.  
    RESULTS AND CONCLUSION: (1) Each induction cell group followed similar morphological changes: Cardiomyocyte-like cells were capable of autonomic beats, rhythmic contractions and relaxations; under the transmission electron microscope, there were a large number of arranged myofilaments; immunofluorescent staining showed positive expression of α-actin and cTnI. (2) The whole heart tissue lysate was able to induce the differentiation of bone marrow mesenchymal stem cells into rice grain-sized myocardial tissue-like structures with collagen fibers. (3) Atrial muscle tissue lysate could induce bone marrow mesenchymal stem cells to differentiate into atrial muscle-like cells. Autonomic beats appeared earlier, but the beat frequency and duration were shorter. ANP and HCN4 were highly expressed in atrial myocytes. (4) Ventricular muscle tissue lysate induced bone marrow mesenchymal stem cells to differentiate into ventricular muscle-like cells with no secretory granules, and MLC-2v was highly expressed in ventricular muscle-like cells. (5) To conclude, the whole heart tissue lysate can induce bone marrow mesenchymal stem cells to form myocardial tissue-like structures. Atrial muscle tissue lysate can induce bone marrow mesenchymal stem cells to differentiate into atrial muscle-like cells. Ventricular muscle tissue lysate can induce bone marrow mesenchymal stem cells to differentiate into ventricular muscle-like cells. By constructing a specific microenvironment, bone marrow mesenchymal stem cells can be differentiated to cardiomyocytes in different parts, providing laboratory data for the construction of myocardial tissue engineering. 

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    Mesenchymal stem cell calcification induced by protoscolex of two species of Echinococcus: a differential analysis
    Gui Xianwei, Jiang Huijiao, Wu Jie, Liang Xueqi, Xu Xiaodan, Wang Erqiang, Zou Hailiang, Chen Hejie, Chen Xueling, Wu Xiangwei
    2021, 25 (1):  38-43.  doi: 10.3969/j.issn.2095-4344.2119
    Abstract ( 236 )   PDF (1707KB) ( 159 )   Save
    Abstract
    BACKGROUND: The growth pattern of Echinococcus granulosus is different from that of Echinococcus alveolaris. Hepatic echinococcosis can form a complete fibrous calcified cyst wall, while hepatic alveolar echinococcosis can grow infiltratively and cannot form a complete fibrous calcified cyst wall. Bone marrow mesenchymal stem cells (BMSCs) are involved in the formation of calcified wall of hydatidosis, but the calcification characteristics of Echinococcus granulosus and Echinococcus alveolaris are different and the role of BMSCs is still unclear.
    OBJECTIVE: To compare the effects of Echinococcus granulosus and Echinococcus alveolaris on the calcification of BMSCs and to preliminarily investigate the formation mechanism of echinococcosis calcifications.
    METHODS:  BMSCs of C57BL/6 mice were extracted, cultured and identified, followed by co-culture with the protoscolex of Echinococcus granulosus (BMSC+CE group) and Echinococcus alveolaris (BMSC+AE group), respectively. BMSCs cultured alone were used as control group. After 1, 4, and 7 days of co-culture, alkaline phosphatase activity was detected by a microplate reader, the expression of BMP2 and RUNX2 mRNA was detected by RT-q PCR, and the expression of BMP2, RUNX2 and phosphorylated Smad1/5/8 (P-Smad1/5/8) proteins was detected by western blot assay.  
    RESULTS AND CONCLUSION: (1) The alkaline phosphatase activity of the BMSC+CE group and BMSC+AE group was significantly higher than that of the control group at 1 and 4 days after culture (P < 0.05), and the alkaline phosphatase activity of the BMSC+CE group was significantly higher than that of BMSC+AE group (P < 0.05). (2) Western blot results showed that the expression of BMP2, RUNX2, and P-Smad1/5/8 protein in the BMSC+CE group and BMSC+AE group was significantly higher than that in the control group at 1 and 4 days after culture (P < 0.05), while the expression of BMP2, RUNX2, and P-Smad1/5/8 protein in the BMSC+CE group was significantly higher than that in the BMSC+AE group (P < 0.05). (3) RT-qPCR results showed that the expression of BMP2 and RUNX2 mRNA in the BMSC+CE group was significantly higher than that in the control group at 1, 4 and 7 days after culture (P < 0.05), and was significantly higher than that in the BMSC+AE group at 4 and 7 days after culture (P < 0.05). The expression of RUNX2 mRNA in the BMSC+AE group was significantly higher than that in the control group at 1, 4, and 7 days after culture (P < 0.05). (4) To conclude, co-culture of the protoscolex of Echinococcus alveolaris and BMSCs promotes the expression of alkaline phosphatase and RUNX2 in BMSCs by up-regulating BMP-Smad1/5/8 pathway. At the later stage of co-culture, the effect of Echinococcus alveolaris on BMSCs calcification is significantly weakened, while the effect of Echinococcus granulosus on BMSCs calcification remains unchanged, suggesting that this mechanism may be related to the different growth patterns of two kinds of hydatids.

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    Proportion and morphological characteristics of human oligodendrocyte precursor cells in different cell culture vessels
    Ye Dou, , Ma Xuexia , Guan Qian, , Luan Zuo , Yang Yinxiang , Wang Zhaoyan , Wang Qian , He Ying , Yao Ruiqin
    2021, 25 (1):  44-49.  doi: 10.3969/j.issn.2095-4344.2139
    Abstract ( 312 )   PDF (1510KB) ( 39 )   Save
    Abstract
    BACKGROUND: Oligodendrocyte precursor cell transplantation is one of the keys to the treatment of white matter damage in premature infants. At present, there is a lack of research on the comparison of oligodendrocyte precursor cells induced by neural stem cells derived from human fetal brain cultured in vitro in different vessels worldwide.   
    OBJECTIVE: To observe the morphology of human oligodendrocyte progenitor cells and pre-oligodendrocytes in different cell culture vessels (6-well plates, 24-well plates and T25 flasks). 
    METHODS:  The 6-well plates, 24-well plates and T25 flasks were used as culture vessels to culture human oligodendrocyte progenitor cells and pre-oligodendrocytes. The characteristics of human oligodendrocyte progenitor cells were identified by immunofluorescence staining and flow cytometry. The morphology of cells was observed by an ordinary light microscope. Cell counts were performed according to cell morphology and statistical analysis was performed. 
    RESULTS AND CONCLUSION: (1) The oligodendrocyte progenitor cell body was round and the bipolar protrusions were bead-like; the pre-oligodendrocyte protrusions were more than two poles, and did not bifurcate. (2) The ratios of oligodendrocyte progenitor cell morphology in the oligodontia lines were significantly higher in the 6-well plates than those in the 24-well plates and T25 flasks (P < 0.05), followed by T25 flasks and 24-well plates. Morphological ratios of pre-oligodendrocytes were significantly higher in the 24-well plates compared to the 6-well plates and T25 flasks (P < 0.01), followed by T25 flasks and 6-well plates. (3) The cells cultured in the 6-well plate had fewer dregs, and the morphology, vigor and growth were better than those of the other culture vessels. (4) According to morphological view, 6-well plates are more suitable for oligodendrocyte progenitor cell growth, and 24-well plates are more suitable for pre-oligodendrocytes growth. 
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    Supporting effect of human skeletal muscle-derived myoendothelial cells on hematopoietic stem/progenitor cells in vitro
    Gu Jingjing, Zhou Rui, Yang Tingting, Yang Xiaoping, Xu Fei, Zheng Bo
    2021, 25 (1):  50-55.  doi: 10.3969/j.issn.2095-4344.2145
    Abstract ( 223 )   PDF (1132KB) ( 24 )   Save
    Abstract
    BACKGROUND: Human skeletal muscle derived myoendothelial cells (MECs) are located in the vascular wall and co-express the markers of muscle stem cells and vascular endothelial cells (CD56+CD34+CD144+CD45-). Studies have shown that MECs are similar to mesenchymal stem cells, express the surface markers of mesenchymal stem cells and have the potential of multidirectional differentiation.  
    OBJECTIVE: To establish an in vitro culture system for human umbilical cord blood CD34+ cells with MECs as trophoblastic layer, and to evaluate the in vitro supporting effect of human skeletal muscle MECs on hematopoietic stem/progenitor cells by measuring the changes in the number, immunophenotype and colony forming ability of CD34+ cells before and after culture.
    METHODS: There were three groups in the experiment. In experimental group, human umbilical cord blood CD34+ cells were co-cultured with MECs as the nourishing layer; in control group, human umbilical cord blood CD34+ cells were co-cultured with bone marrow mesenchymal stem cells as the nourishing layer; and in blank control group, human umbilical cord blood CD34+ cells were cultured alone without the nourishing layer. The main outcome measures, including the number of human umbilical cord blood CD34+ cells, immunophenotype of blood cells and colony formation ability of hematopoietic stem/progenitor cells were analyzed and compared at 1, 2, and 4 weeks after co-culture. No detection was conducted at 5 weeks due to the lack of survived cells.
    RESULTS AND CONCLUSION: (1) The number of human umbilical cord blood CD34+ cells increased by MECs as the nourishing layer compared with bone marrow mesenchymal stem cells as the nourishing layer at 1, 2, and 4 weeks; however, there was no significant difference between the two groups (P > 0.05). (2) The cell immunophenotype by flow cytometry analysis indicated that only in the 2nd week, the expression of CD34+CD33- in human umbilical cord blood CD34+ cells in the control group was significantly higher in the experimental group (P < 0.05). Other subtypes of immunophenotypes CD45+, CD14+, CD10+/CD19+ showed no significant difference between the two groups (P > 0.05). (3) The colony formation capacity of hematopoietic stem/progenitor cells showed no significant difference between the experimental and control groups at 1, 2, and 4 weeks (P > 0.05). (4) Due to the non-nourishing layer culture system, the number of human umbilical cord blood CD34+ cells decreased significantly in the 1st week, and no cells survived in the 2nd week. Therefore, blood cell immunophenotype and colony analysis could not be performed. (5) To conclude, human skeletal muscle MECs as trophoblasts are the same as human bone marrow mesenchymal stem cells, which have a hematopoietic support in vitro.

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    Human umbilical cord mesenchymal stem cells overexpressing interleukin 8 receptor inhibit inflammation and promote vascular repair
    Zhu Bingbing, He Haibin, Deng Jianghua, Wang Wenqiang, Mu Xiaoling
    2021, 25 (1):  61-66.  doi: 10.3969/j.issn.2095-4344.2124
    Abstract ( 272 )   PDF (2083KB) ( 21 )   Save
    Abstract
    BACKGROUND: Vascular injury is a common complication after balloon dilatation. The development of umbilical cord mesenchymal stem cells (UC-MSCs) provides a new method for treating vascular injury.
    OBJECTIVE: To investigate the mechanism underlying the repair of damaged blood vessels by human UC-MSCs (hUC-MSCs) transfected with interleukin-8RA/B (IL-8RA/B) adenovirus. 
    METHODS: hUC-MSCs and human umbilical vein endothelial cells (hUVECs) were collected and transfected with adenovirus vectors containing human IL-8RA and/or IL-8RB cDNAs and green fluorescent protein. A rat model of carotid artery injury was established. Sprague-Dawley rats were randomly divided into four groups: IL-8RA/B-hUCMSCs group, Il-8ra/B-hUVECs group, Null-hUCMSCs group, and control group, followed by injection of 0.5×106 corresponding cells 
    (500 μL) and same volume of normal saline via the tail vein respectively at 1, 3, and 5 hours post-surgery. After 30 minutes of injection, the carotid artery was taken and the expression of green fluorescent protein was observed. After 24 hours, the serum levels of inflammatory and anti-inflammatory factors were measured by ELISA; and the infiltration of neutrophil cells and mononuclear macrophages was observed by immunohistochemistry. After 14 days, Evans blue staining was used to observe vascular endothelialization and fibrosis. After 28 days, the neointimal hyperplasia was observed by hematoxylin-eosin staining. 
    RESULTS AND CONCLUSION: (1) After 30 minutes of IL-8RA/B-hUC-MSCs infusion, the expression of green fluorescent protein was observed in the injured vascular intima, and the fluorescence expression was higher than that of the other three groups. (2) After 24 hours of IL-8RA/B-hUC-MSCs infusion, the expression of inflammatory factors in the serum was significantly lower than that of the other three groups, while the expression of anti-inflammatory factor interleukin-10 was higher than that of the other three groups (P < 0.05). In addition, inflammatory cell infiltration in the IL-8RA/B-hUC-MSCs group decreased significantly. (3) hUC-MSCs overexpressing interleukin-8 receptor promoted re-endothelialization of injured vessels and reduced vascular fibrosis after 14 days of infusion. (4) IL-8RA/B-hUC-MSCs reduced vascular neointimal hyperplasia after 28 days of infusion. (5) Interleukin-8 receptor enhances the targeted homing ability of hUC-MSCs, allowing MSCs to migrate to the site of vascular injury, inhibit inflammation, reduce neointimal hyperplasia, and promote vascular repair.
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    Isolation, culture and biological characteristics of high-purity orofacial-bone-derived mesenchymal stem cells of the rats
    Cheng Bingkun, Liang Jianfei, Qin Dongze, Cheng Zixu, Zhao Yunzhuan
    2021, 25 (1):  67-72.  doi: 10.3969/j.issn.2095-4344.2151
    Abstract ( 226 )   PDF (1932KB) ( 41 )   Save
    BACKGROUND: It is widely accepted to recruit mesenchymal stem cells from the jaws to repair the defects of the jaws. However, there are relatively few researches on orofacial-bone-derived mesenchymal stem cells, mainly due to the difficulty in separating mesenchymal stem cells from the jaws. 
    OBJECTIVE: To establish the methods of in vitro isolation and culture of rat orofacial-bone-derived mesenchymal stem cells and observe and study its related biological characteristics.  
    METHODS: The mandibles of rats were dissected. The attached muscles were stripped and cut into pieces. Cortical bone was loosened by digesting with collagenase II. The migration and adherent growth ability of mesenchymal stem cells was used to isolate orofacial-bone-derived mesenchymal stem cells. Cell morphology was observed by inverted microscope. Surface markers of the cell were detected by flow cytometry. Cell proliferation was detected by MTT assay and cell growth curve was drawn. Fibroblast colony forming rate was calculated by colony formation. Osteogenic and lipogenic induction experiments were conducted to study the multi-directional differentiation potential of cells. 
    RESULTS AND CONCLUSION: Cells isolated by collagenase digestion and bone slice culture were positive for CD29, CD44 and Sca-1, and negative for CD31, CD34 and CD45. Cell proliferation test showed that the growth curves of orofacial-bone-derived mesenchymal stem cells exhibited incubation period, logarithmic phase and platform period. In addition, the cells had a strong ability of proliferation, and the cell clone formation rate was 20% and the cells in DNA synthesis stage accounted for 52.5%. Alizarin red and oil red O staining showed positive reaction after osteogenic and lipogenic induction, indicating that the cells have the potential of multi-directional differentiation. It is concluded that the method of bone fragment culture after digestion with collagenase II could separate orofacial-bone-derived mesenchymal stem cells sufficiently and purely. Besides, the orofacial-bone-derived mesenchymal stem cells show strong proliferative and osteogenic differentiation capacities. Thus, it provides abundant source of seed cells for bone tissue engineering of maxillofacial represented by bone defects repairing of implants.
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    Immunomodulatory effect of human adipose derived mesenchymal stem cells on skin transplantation between different mouse strains
    Hua Hong, Xie Tongling, Hao Guiliang
    2021, 25 (1):  73-77.  doi: 10.3969/j.issn.2095-4344.2143
    Abstract ( 247 )   PDF (1568KB) ( 17 )   Save

    BACKGROUND: Skin transplantation is one of the most effective methods for treating large-area burns. How to effectively suppress the immune rejection after allogeneic skin transplantation is a problem that needs to be solved urgently.

    OBJECTIVE: To investigate the effect of human adipose derived mesenchymal stem cells (hADSCs) on the immunoregulation of skin grafts in different strains of mice.  
    METHODS:  Isolated hADSCs were cultured to the 3rd generation. Sixty ICR neonatal mice, 2-4 days of age, were randomly divided into four groups (n=15). The skin tissues of ICR neonatal mice were transplanted into adult C57BL/6 mice to establish a different strain of mouse skin graft immune rejection model. PBS and low dose (5×104), medium dose (10×104), high dose (20×104) hADSCs were injected into the model mice through tail vein, and the survival time of transplanted skin in each group was recorded. On the 7th day after operation, five mice from each group were randomly selected to remove their spleen and serum, and the expression of immune factors interleukin-10, tumor necrosis factor-α and interferon-γ were detected by RT-PCR and ELISA respectively. The transplanted part of the skin was taken to make pathological sections for observing the infiltration of lymphocytes.   
    RESULTS AND CONCLUSION: Compared with the PBS group, the survival time of the skin was prolonged in the low dose hADSCs group; however, there was no significant difference between the two groups (P > 0.05). Compared with the PBS and low dose hADSCs groups, the survival time of the skin was significantly increased in the medium and high dose groups (P < 0.01), and no significant difference was found between the medium and high dose groups (P > 0.05). Compared with the PBS group, the relative expression of tumor necrosis factor-α and interferon-γ in the spleen and serum was significantly decreased in the low, medium and high dose hADSCs groups (P < 0.05), whereas the level of interleukin-10 was significantly elevated in the medium and high dose hADSCs groups (P < 0.05). To conclude, the appropriate dose of hADSCs can significantly prolong the survival time of transplanted skin between different strains of mice, by regulating the expression of related immune factors in the recipient mice.

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    Transplantation of islet-like cells induced by human umbilical cord mesenchymal stem cells via different ways for the treatment of type 1 diabetic mice
    Guo Xuan, Xie Jun, Suo Jinrong, Li Yingrui, Huang Lei, Ma Munan, Li Jingjing, Fu Songtao
    2021, 25 (1):  78-83.  doi: 10.3969/j.issn.2095-4344.2144
    Abstract ( 326 )   PDF (1809KB) ( 33 )   Save
    BACKGROUND: Transplanting islet-like cells induced by human umbilical cord mesenchymal stem cells (hUC-MSCs) into type 1 diabetic mice can reduce blood glucose level and improve the symptoms of diabetes mellitus. However, there are few reports on intraperitoneal transplantation.
    OBJECTIVE: To study the therapeutic effect of transplantation of islet-like cells induced by hUC-MSCs in different ways for the treatment of type 1 diabetic mice.
    METHODS: The hUC-MSCs were isolated and cultured by tissue explants adherent method and differentiated into islet-like cells. The 3 of 15 male C57BL/6J mice were used as normal group, and the remaining mice were taken to prepare a mouse model of type 1 diabetes using intraperitoneal injection of streptozotocin. After successful modeling, nine model mice were randomly divided into diabetes group, tail vein-islet-like cells group, and abdomen-islet-like cells group, with three mice in each group. After 10 days of modeling, the normal group and diabetic group were not treated. The tail vein-islet-like cells group was injected with 5×105 cells/0.4 mL islet-like cells via the tail vein and the abdomen-islet-like cells group was intraperitoneally injected with 5×105 cells/0.4 mL islet-like cells. During the treatment, the blood glucose and insulin levels were measured twice a week; glucose tolerance test was performed at 28 days after cell transplantation; and fasting insulin level was detected at 42 days after cell transplantation. 
    RESULTS AND CONCLUSION: (1) Compared with the diabetic group, in the tail vein-islet-like cells group, the blood glucose level began to decrease on the 10th day after transplantation and maintained until the 31st day, and the insulin level and glucose tolerance significantly improved (P < 0.05). However, there was no significant improvement in blood glucose level, insulin level and glucose tolerance in the abdomen-islet-like cells group. (2) To conclude, transplantation of hUC-MSCs induced islet-like cells for the treatment of type 1 diabetic mice via tail vein is an ideal transplantation method, and the effect of intraperitoneal injection is unsatisfactory.
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    Construction of a stable Atg5 gene knockdown cell line in RAW 264.7 cells by lentivirus infection
    Xian Guoyan, Chen Weishen, Zhang Ziji, Ye Yongyu, Pan Baiqi, Zheng Linli, Sheng Puyi
    2021, 25 (1):  84-89.  doi: 10.3969/j.issn.2095-4344.2149
    Abstract ( 378 )   PDF (1449KB) ( 33 )   Save
    BACKGROUND: The autophagy of macrophage cells plays a key role in regulating the immune system and inflammatory response, while knockdown Atg5 gene can specifically inhibit the autophagy. There are some shortcomings of traditional siRNA transfection methods such as transience and incompleteness. It is of great value and significance to construct a stable Atg5 gene knockdown cell line in RAW 264.7 cells for studying the macrophage autophagy and immune inflammation-related diseases and their pathogenesis. 
    OBJECTIVE: To construct RAW 264.7 macrophage cell line stably knocking down the Atg5 gene by lentivirus infection, and to provide chassis cells for studying macrophage autophagy and immune inflammation-related diseases and their pathogenesis.
    METHODS: Recombinant expression vector (HBLV-m-Atg5-shRNA-GFP-Puro) with green fluorescence signal, Puro resistance gene, and knockdown Atg5 gene was constructed and transfected into 293T cells to obtain lentivirus plasmid system. The green fluorescence signal of the acquired lentivirus infected RAW 264.7 cells was observed under an inverted fluorescence microscope. Purinomycin resistance screening and flow cytometry were used to obtain high-purity infected cells. The RAW 264.7 cell line stably knocking down the Atg5 gene was identified by real-time quantitative polymerase chain reaction and western blot assay. 
    RESULTS AND CONCLUSION: DNA sequencing results showed that Atg5 gene interfering sequences were correctly inserted into the expression vector, and the HBLV-m-Atg5-shRNA-GFP-Puro expression vector was successfully constructed. After infecting RAW 264.7 cell line with lentivirus plasmid, green fluorescence was observed under an inverted fluorescence microscope. Green fluorescent protein positive cell groups were observed by flow cytometry. The results of real-time quantitative polymerase chain reaction and western blot assay showed that the expression level of the Atg5 gene in RAW 264.7 cell line was significantly decreased (P < 0.05), indicating that the RAW 264.7 cell line with stable knockdown of Atg5 gene was successfully constructed.
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    Effect of umbilical cord mesenchymal stem cell transplantation for treating systemic lupus erythematosus in a tree shrew model
    Ruan Guangping, Yao Xiang, Cai Xuemin, Li Zian, Pang Rongqing, Pan Xinghua
    2021, 25 (1):  90-95.  doi: 10.3969/j.issn.2095-4344.2140
    Abstract ( 227 )   PDF (2498KB) ( 25 )   Save
    BACKGROUND: Systemic lupus erythematosus is an autoimmune disease with unknown causes. To establish a tree shrew model of systemic lupus erythematosus is helpful to understand its pathogenesis and provide evidence for stem cell transplantation in the treatment of autoimmune diseases.  
    OBJECTIVE: To establish a tree shrew model of systemic lupus erythematosus and to assess the therapeutic effect of umbilical cord mesenchymal stem cell transplantation.  
    METHODS:  Tree shrews were grouped and intraperitoneally injected pristane, lipopolysaccharide, and their combination. At 3 weeks after injection, 12 tree shrew models were divided into treatment group and model control group (n=6 per group). An additional 6 models were selected as a normal control group. In the treatment group, each tree shrew was injected with 1×106 DiR-labeled umbilical cord mesenchymal stem cells through caudal vein. Two weeks later, the heart, liver, spleen, lung and kidney of tree shrews were taken for pathological sections. The sections received hematoxylin-eosin staining, kidney Masson staining and immune complex detection. Simultaneously, the heart, liver, spleen, lung and kidney of the three groups of tree shrews were taken for in vitro imaging.   
    RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining showed pathological changes of the heart, liver, spleen, lung and kidney in the model control group; and there were a lot of immune complex deposits in renal tissue in the model control group. The pathological changes in the treatment group improved, and the structure recovered to close to the normal control group. (2) In vitro imaging showed that DiR-labeled cells were mainly distributed in the lung, liver and spleen of the tree shrews in the treatment group. The fluorescence intensity of tree shrews in the treatment group was significantly higher than that in the normal control group and model control group (P < 0.05). (3) Results demonstrated that intraperitoneal injection of pristane and lipopolysaccharide is the best method to induce pathological changes of systemic lupus erythematosus in tree shrews. The pathological changes after treatment with umbilical cord mesenchymal stem cells have improved, indicating that umbilical cord mesenchymal stem cells have certain treatment effects on tree shrew models of systemic lupus erythematosus. 
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    Regulatory effects of ADT-OH on the proliferation of neural precursor cells
    Zhou Liyu, Ma Yanxia, Qi Shibin, Saijilafu, Wei Shanwen, Ni Li
    2021, 25 (1):  96-100.  doi: 10.3969/j.issn.2095-4344.2156
    Abstract ( 249 )   PDF (1592KB) ( 24 )   Save
    BACKGROUND: Researchers believe that hydrogen sulfide (H2S), as an important cell protective molecule, may become a new treatment method to restore the physiological function of diseased cells or organ systems through the artificial regulation of endogenous H2S biosynthesis or in vitro administration of H2S donor. ADT-OH is a slow-release donor of H2S that can improve the survival rate of hippocampal nerve cells with glutamate-induced injury, but studies on the proliferation of cerebral cortical neural precursor cells are rare.
    OBJECTIVE: To investigate the effect of ADT-OH on the proliferation of neural precursor cells in embryonic cerebral cortex.
    METHODS:  Neural precursor cells from cerebral cortical ventricular zone and subventricular zone of embryonic mice at embryonic 14.5 days were isolated. Neural precursor cells from one fetal mouse were inoculated into one well (24-well plate), and cultured with the medium containing 100 μmol/L ADT-OH. The size and number of neural spheres per well were measured at 3 days after culture. The proliferation rate of cultured neural precursor cells was detected by BrdU labeling. The proliferation of the cells was further verified by immunofluorescence staining with the specific antibody Ki67. The expression of cyclin D1 was finally detected by western blot assay.
    RESULTS AND CONCLUSION: Our experimental results showed that ADT-OH could promote the formation of neural spheres, and further detection by BrdU and Ki67 antibody showed that ADT-OH could promote the proliferation rate of neural precursor cells. Meanwhile, the expression of cyclin D1, a proliferation-related gene, was up-regulated in neural precursor cells after ADT-OH treatment. Overall, ADT-OH may promote the proliferation of neural precursor cells by regulating the expression of cyclin D1.
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    Two kinds of stem cell nasal transplantation for treating white matter injury in premature rat infants
    Zang Jing, Luan Zuo, Wang Qian, Yang Yinxiang, Wang Zhaoyan, Wu Youjia, Guo Aisong
    2021, 25 (1):  101-107.  doi: 10.3969/j.issn.2095-4344.2150
    Abstract ( 202 )   PDF (1850KB) ( 19 )   Save
    BACKGROUND: Stem cell transplantation has a significant neuroprotective effect on neurological diseases. Current transplantation methods such as arteriovenous transplantation and brain stereotactic transplantation are not suitable for clinical application in preterm infants.
    OBJECTIVE: To explore the feasibility of nasal transplantation of human umbilical cord mesenchymal stem cells and human neural stem cells for the treatment of white matter injury in premature rat infants.
    METHODS: Human umbilical cord mesenchymal stem cells were prepared from human umbilical cord tissue, and human neural stem cells were prepared from human embryonic brain tissue. In vitro migration of two kinds of cells was assessed by Transwell method. Forty 3-day-old Sprague-Dawley rats were randomly divided into sham operation group, model control group, human umbilical cord mesenchymal stem cell transplantation group and human neural stem cell transplantation group, with 10 rats in each group. Rats in all groups except the sham operation group were treated with right common carotid artery ligation and hypoxia for 90 minutes to establish a rat model of white matter injury in the preterm infant. Totally 1×106 cells were delivered intranasally in the transplantation group at 3 days after injury. Each nostril was infused with 5×105, and each nostril was infused once. On day 7 after injury, MBP immunofluorescence staining was used to detect the expression of myelin basic protein in the white matter of the brain to identify the damage of the white matter injury model. At 24 hours after transplantation, human umbilical cord mesenchymal stem cell migration was detected by anti-HuNu immunohistochemical method and human neural stem cell migration was detected by CM-Dil fluorescent labeling method. 
    RESULTS AND CONCLUSION: (1) On day 7 after modeling, compared with the normal side, the positive area of MBP decreased in cingulate band, corpus callosum and external capsule of the affected side in the model of brain white matter injury in preterm infants (P < 0.05), indicating a successful modeling. (2) In vitro experiments showed that the migration rate of human neural stem cells was the same as that of human umbilical cord mesenchymal stem cells. (3) At 24 hours after the nasal transplantation, human umbilical cord mesenchymal stem cells migrated to the cortex, corpus callosum and hippocampus on the normal side and the damaged side, and human neural stem cells migrated to the damaged cortex, corpus callosum and hippocampus, and human umbilical cord mesenchymal stem cells migrated more than human neural stem cells. (4) Overall, these findings indicate that 24 hours after the nasal transplantation, human umbilical cord mesenchymal stem cells could survive and migrate to the normal side and the injury side, and human neural stem cells could survive and migrate to the injury side; and the migration of human umbilical cord mesenchymal stem cells was more extensive than that of human neural stem cells.
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    Transcription factor NKX2-5: molecular structure and biological function in regulating cardiovascular precursor cells
    Wang Hongshu, Han Shen, Liu Zu, Li Xiaofang, Zhong Chongbin, Li Lifeng, Li Yaxiong, Jiang Lihong
    2021, 25 (1):  108-115.  doi: 10.3969/j.issn.2095-4344.2135
    Abstract ( 325 )   PDF (706KB) ( 44 )   Save
    BACKGROUND: NKX2-5 is an important transcriptional regulator during mammalian heart development. Studies from model animals such as rats, mice, and zebrafish have shown that the absence or abnormal function of NKX2-5 affects heart development, leading to pathological manifestations similar to human congenital heart disease. However, the regulatory role and specific mechanisms of NKX2-5 in cardiac development are still unclear.
    OBJECTIVE: To review the molecular structure, functional effects, and upstream and downstream regulatory molecules of NKX2-5.
    METHODS: The search terms “NKX2-5, congenital heart disease, CHD, heart development” were used for literature retrieval in the PubMed database (website https://www.ncbi.nlm.nih.gov/pubmed/). The deadline for publication was April 1, 2019. By downloading and reading the retrieved literature, articles irrelevant to NKX2-5 and with duplicate opinions and similar conclusions were excluded. Finally 97 eligible articles were taken for review. 
    RESULTS AND CONCLUSION: (1) NKX2-5 plays a key role in the regulation of cardiac development. Abnormal functions or gene mutations of NKX2-5 can lead to abnormal cardiac development and dysfunction, which are closely related to the occurrence of human congenital heart disease. (2) The function of NKX2-5 depends on its functional domains. NKX2-5 enters the nucleus after transcription and translation, and activates or inhibits downstream regulatory molecules through binding with CO-factors or alone to specific DNA sequences, thereby regulating the proliferation, migration, differentiation and function of cardiac precursor cells and regulating the correct cardiac development process and the normal development and function of the cardiac conduction system. (3) The transcription, translation, nucleation process, transcription activation or inhibition of NKX2-5 is also regulated by various methods. These regulatory factors regulate NKX2-5 from the aspects of chromatin conformation, promoter and enhancer functions, RNA uncoiling, post-transcriptional modification, and nuclear localization.
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    Related factors regulating osteogenic differentiation of bone marrow mesenchymal stem cells through Wnt/β-catenin signaling pathway
    Wu Ming, Zhang Yan
    2021, 25 (1):  116-122.  doi: 10.3969/j.issn.2095-4344.2158
    Abstract ( 259 )   PDF (769KB) ( 51 )   Save
    BACKGROUND: Bone marrow mesenchymal stem cells induce osteogenesis and inhibit adipocyte differentiation, which is the key to the prevention and treatment of osteoporosis and the source of seed cells for bone tissue repair engineering. Wnt signaling pathway plays an important role in bone formation.
    OBJECTIVE: To review the related factors and molecular mechanisms that regulate osteogenic differentiation of bone marrow mesenchymal stem cells through Wnt/β-catenin signaling pathway.
    METHODS: CNKI, PubMed and Wanfang Medical Database were retrieved by computer for related literatures published from inception to February 2020. The Chinese key words were “bone marrow mesenchymal stem cells, osteoporosis, osteogenic differentiation, osteocytes, extracellular matrix, osteoarthritis, Wnt, β-catenin”. The English key words were “BMSCs, osteoarthritis, Wnt/β-catenin, Wnt”. Finally, 62 articles were included in the review.
    RESULTS AND CONCLUSION: The relevant factors can directly or indirectly regulate the osteogenic differentiation of bone marrow mesenchymal stem cells through the Wnt/β-catenin signaling pathway. There are positive promotion, negative inhibition and even loop regulation due to the interaction between various factors. The Wnt/β-catenin signaling pathway is a key signal for osteogenic differentiation of bone marrow mesenchymal stem cells. The systematic analysis of the molecular mechanism of osteogenic differentiation of bone marrow mesenchymal stem cells provides a theoretical basis for bone tissue engineering, and can realize the application of bone tissue engineering faster.
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    Role and hotspots of stem cell-derived exosome in the repair of traumatic brain injury
    Sun Tianjing , Liu Sijia , Xie Fangke , Huang Xiaofei , Zhang Ji , Jiang Xuheng , Feng Hua, Yu Anyong
    2021, 25 (1):  123-127.  doi: 10.3969/j.issn.2095-4344.2127
    Abstract ( 236 )   PDF (611KB) ( 44 )   Save
    BACKGROUND: The treatment and prognosis of traumatic brain injury are difficult points in clinical work at present. More and more studies have shown that exosomes derived from stem cells show unique advantages in traumatic brain injury, which may have greater neurotherapeutic potential in the treatment of traumatic brain injury, and may essentially restore neurovascular function and promote neuronal regeneration, thereby providing new ideas for the treatment of traumatic brain injury.
    OBJECTIVE: To review the application and advantages of exosomes derived from stem cells in traumatic brain injury.
    METHODS: A computer-based search was performed in the PubMed and CNKI databases for articles addressing exosomes. The keywords were “Traumatic brain injury, Exosomes, Stem cells” in English and Chinese, respectively. Finally, 53 articles were included for review.
    RESULTS AND CONCLUSION: (1) A large number of traumatic brain injury animal experiments proved that nerve cells and stem cells can secrete exosomes. (2) Exosomes derived from nerve cells and stem cells play a positive role in the treatment of traumatic brain injury, but the exosomes derived from stem cells are more effective in nerve, vascular regeneration and neurofunctional recovery. (3) The exosomes derived from stem cells may open up a new approach to traumatic brain injury.
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    Bone marrow mesenchymal stem cells and tongue squamous carcinoma cells: promoting growth or targeting biotherapy
    Xu Zhuqing, Wang Zhuhui, Zhang Xinyu, Zheng Jianjin
    2021, 25 (1):  128-132.  doi: 10.3969/j.issn.2095-4344.2152
    Abstract ( 239 )   PDF (701KB) ( 22 )   Save
    BACKGROUND: Currently, the treatment methods for tongue squamous cell carcinoma mainly adopt comprehensive sequence therapy mainly based on surgery, but the therapeutic effect is still unsatisfactory. In recent years, tumor targeted biotherapy has attracted increasing attention. Stem cell technology enables therapeutic factors to be selectively delivered to tumor sites to play corresponding roles. Bone marrow mesenchymal stem cells show great advantages in this respect. However, the effect and action mechanism of bone marrow mesenchymal stem cells on tongue squamous cell carcinoma are still unclear.
    OBJECTIVE: To review the research progress on the effect of bone marrow mesenchymal stem cells on tongue squamous carcinoma cells, and clarify its function and discuss its mechanism.
    METHODS: The computer was used to search literatures about bone marrow mesenchymal stem cells in PubMed database, Wanfang database and CNKI database. The key words were “bone marrow mesenchymal stem cells, tumor cells”, “bone marrow mesenchymal stem cells, tongue squamous cell carcinoma”, “bone marrow mesenchymal stem cells, proliferation”, “bone marrow mesenchymal stem cells, migration, invasion” in Chinese, and “bone marrow mesenchymal stem cells, cancer cells” and “bone marrow mesenchymal stem cells, tongue squamous cell carcinoma”, “bone marrow mesenchymal stem cells, proliferation”, “bone marrow mesenchymal stem cells, migration, invasion” in English. A total of 43 articles were included for analysis. 
    RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells have strong proliferation characteristics and can migrate directionally and promote invasion of tongue squamous carcinoma cells, suggesting that even if bone marrow mesenchymal stem cells have the function of promoting defect repair, and can also express therapeutic factors by gene modification and migrate to tumor growth site to play a therapeutic role, they cannot be used for repair treatment of postoperative defects of tongue squamous cell carcinoma. Targeted biological therapy of mesenchymal stem cells is a brand-new and promising field, but its safety still needs to be taken into account, and its mechanism of action and potential risks still need our continuous exploration.
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    Effect of cytokines and platelet-rich plasma on tendon derived stem cells
    Yu Chenghao, Zhang Yi, Qi Chao, Chen Jinli, Gao Jiake, Yu Tengbo
    2021, 25 (1):  133-140.  doi: 10.3969/j.issn.2095-4344.2148
    Abstract ( 238 )   PDF (688KB) ( 33 )   Save
    BACKGROUND: Tendon derived stem cells exist in the tendon and have unique functions for tendon repair. Different cytokines have different effects on the proliferation and differentiation of tendon derived stem cells. Platelet-rich plasma refers to the blood product obtained from the whole blood through gradient centrifugation and stratification, which contains a variety of cytokines that could help to promote the regeneration of ligaments and tendons.
    OBJECTIVE: To investigate the latest progress of effects of cytokines and platelet-rich plasma on proliferation and differentiation of tendon derived stem cells.
    METHODS: Using “tendon derived stem cells, tendon stem/progenitor cells, tendon stem cell, platelet-rich plasma, ligament injury” as keywords in English and “tendon derived stem cells, platelet-rich plasma, ligament injury” in Chinese, the first author searched PubMed, CNKI, and Wanfang for relevant articles published from 2007 to 2019. Literature unrelated to the purpose of the study and repetitive literature were excluded, and 83 articles that meet the criteria were included for review. 
    RESULTS AND CONCLUSION: Tendon derived stem cells are ideal cells for the treatment of tendon injury in cell transplantation. Its proliferation and differentiation are influenced by cytokines. Platelet-rich plasma contains a large number of cytokines, which can stimulate the proliferation and differentiation of tendon derived stem cells and have the potential to become a carrier of cell transplantation. Exploring the relationship between cytokines and proliferation and differentiation of tendon stem cells will provide a new approach for the clinical application of tendon derived stem cells.
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    Future hot spots for cartilage repair: laminin promotes stem cell proliferation
    Zhang Xiaobo, Zhang Jie, Sun Yu
    2021, 25 (1):  141-145.  doi: 10.3969/j.issn.2095-4344.2133
    Abstract ( 239 )   PDF (764KB) ( 20 )   Save
    BACKGROUND: The characteristics of laminin that can promote the proliferation of stem cells have been widely concerned.
    OBJECTIVE: To review the interactions between laminin and many different stem cells, and provide reliable theoretical basis for chondrogenic research and application of stem cells.
    METHODS: Wanfang, CNKI, PubMed and Web of Science databases were searched for articles related to mechanism of laminin, changes in stem cell behaviors, and cartilage regeneration published from January 2010 to October 2019. The retrieval terms were “laminin” and “steam cells” in Chinese and English. Duplicated and poorly related articles were excluded, and finally 57 articles were included for review.
    RESULTS AND CONCLUSION: (1) The structural characteristics of laminin were summarized. The spatiotemporal changes of laminin during cartilage development and degradation were analyzed. At the same time, the distribution of laminin expression in natural cartilage tissue and tissue engineered cartilage tissue was compared. (2) The effects of laminin on the proliferation of various stem cells, including embryonic stem cells, induced pluripotent stem cells and adult stem cells, were described. (3) The possible hotspots on the combination of laminin and stem cells for cartilage regeneration were proposed, with the attempt of providing theoretical basis for cartilage repair and regeneration in the future.
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    Application and function of autologous blood concentrate in tissue regeneration
    Wang Ning, Zhong Weijian
    2021, 25 (1):  146-151.  doi: 10.3969/j.issn.2095-4344.2154
    Abstract ( 264 )   PDF (740KB) ( 59 )   Save
    BACKGROUND: Autologous blood concentrates are widely applied in clinical practice. They are easy to prepare, and can promote the regeneration of soft and hard tissues. Their application prospects can be broad in periodontal treatment, oral implant therapy and many other fields.
    OBJECTIVE: To review the development, biological characteristics and clinical application of autohemoconcentrate.
    METHODS: The first author searched the related literature published on PubMed, CNKI and Wanfang databases from 2004 to 2019. The key words were “autologous blood concentration, platelet-rich plasma, platelet-rich fibrin” in English, and “blood concentration, platelet-rich plasma, platelet-rich fibrin” in Chinese. The titles and abstracts were selected, and the full text was consulted. Finally, 59 articles were retained for careful reading and analysis.
    RESULTS AND CONCLUSION: Containing huge amount of platelets and leukocytes, autologous blood concentrate could produce a highly positive effect on the regeneration of soft and hard tissues and the reduction of postoperative inflammatory response. When it is in use alone or in combination with other bone transplantation materials, the effect of bone augmentation is positive. However, the degradation rate is high. How to control its absorption rate requires further study. 
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    Therapeutic effect of stem cells in chronic temporal lobe epilepsy: a systematic review of animal studies
    Liu Yali, Wang Huan, Yan Qiong, Wang Gang, Hou Boru, Wang Dengfeng, Ma Bin, Ren Haijun
    2021, 25 (1):  152-158.  doi: 10.3969/j.issn.2095-4344.2134
    Abstract ( 208 )   PDF (680KB) ( 24 )   Save
    BACKGROUND: Stem cell therapy has emerged as a novel treatment for chronic temporal lobe epilepsy that has been widely used in various animal models of epilepsy. However, there is yet no consensus in the therapeutic effect in existing experimental studies.  
    OBJECTIVE: To systemically evaluate the therapeutic efficacy of stem cell therapy in animal models of chronic temporal lobe epilepsy, providing experimental evidence and new ideas for further clinical trials. 
    METHODS:  A systematic search of CNKI, WanFang, PubMed, Embase and Web of Science was performed for animal studies related to stem cell therapy for chronic temporal lobe epilepsy published from inception to May 2018. Two reviewers independently extracted data, assessed methodological quality and evaluated evidence quality by using CERQual tool.   
    RESULTS AND CONCLUSION: Eight animal studies were eligible for inclusion, including 284 animals suffering chronic temporal lobe epilepsy. Between-study heterogeneity was substantial, so a qualitative description was performed. Stem cell therapy has some improvement in the seizure frequency and duration of chronic temporal lobe epilepsy in animals, which can promote the recovery from epilepsy in most animals. Other outcomes include memory and learning, stem cell migration, differentiation and integration. The exact efficacy of stem cells cannot be determined due to the lack of rigor, characterization, and evidence quality. We therefore do not have absolute confidence to recommend the clinical use of stem cell therapy for chronic temporal lobe epilepsy. Thus, it is necessary to conduct high-quality preclinical studies to further evaluate the efficacy of stem cell transplantation for chronic temporal lobe epilepsy and its clinical conversion risk before implementation of clinical trials.
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