1.Density gradient centrifugation for isolation of umbilical cord blood stem cells:Screening of separation medium
Jiqiang GUO ; Aibing LIU ; Dongping WANG ; Liming WANG
Chinese Journal of Tissue Engineering Research 2013;(23):4189-4195
10.3969/j.issn.2095-4344.2013.23.002
2.Bone marrow mesenchymal stem cells from diabetic versus normal rats for treatment of myocardial infarction
Chinese Journal of Tissue Engineering Research 2013;(27):5026-5032
BACKGROUND: Current studies have shown that bone marrow mesenchymal stem cells from normal or young people usual y serve as a source of transplanted cells in stem cel transplantation treatment of myocardial infarction. OBJECTIVE: To compare the therapeutic effects of bone marrow mesenchymal stem cells from diabetic and normal rats on myocardial infarction. METHODS: Under sterile conditions, bone marrow mesenchymal stem cells from normal and diabetic rats were harvested. Then, rat models of myocardial infarction were established and randomly divided into three groups:100 μL cellsuspension containing 105-106 bone marrow mesenchymal stem cells from F2 normal or diabetic rats was injected into myocardial infarction lesions, and 100 μL Dulbecco’s modified Eagle’s medium containing 20%fetal bovine serum was injected serving as blank control. After 1 month, hematoxylin-eosin staining for myocardial infarction lesions was performed for histomorphological observation. Bcl-2 expression was detected by immunohistochemistry method. RESULTS AND CONCLUSION: Based on cel morphology observation and flow cytometry identification, high-purity bone marrow mesenchymal stem cells could be obtained using rat femoral bone marrow adherent culture. Cel growth curve showed that normal rat bone marrow mesenchymal stem cells grew faster than those from diabetic rats. At 1 month after transplantation, histomorphological improvement was seen in the infarcted area after transplantation of normal rat bone marrow mesenchymal stem cells as compared with the other two groups. In addition, the Bcl-2 expression in the infarcted area was higher in the normal rat cel group than the the other two groups. These findings indicate that bone marrow mesenchymal stem cells from normal rats grow faster than those from diabetic rats, and the cells from normal rats have better therapeutic effects on myocardial infarction.
3.Myogenetic potential of rabbit adipose-derived stromal cells
Jing LIU ; Wanjun LIN ; Yinzhong LIU ; Xiaomian LI ; Honggang GUO
Chinese Journal of Tissue Engineering Research 2013;(27):5012-5018
BACKGROUND: Myogenetic seed cells are the fundamental for construction of tissue-engineered smooth muscle, and optimizing the amplification of seed cells is the key in further clinical application. OBJECTIVE: To analyze the myogenetic potential of rabbit adipose-derived stromal cells induced with the combination of MyoD, transforming growth factor β1 and 5-azacytidine, and to investigate a new way to induce cells. METHODS: The rabbit abdominal fats were isolated and then the adipose-derived stromal cells were separated by col agenase digestion method for myogenetic induction with different methods: 5-azacytidine induction group, MyoD+transforming growth factor β1 induction group, 5-azacytidine+MyoD+transforming growth factor β1 combination induction group. The blank control group was set. The morphological characteristics of cells were observed at 1, 4, 8, 12, 16, 20, 24 and 28 days after induction, and the col agen type Ⅲ level were detected with 4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometry and immunohistochemistry. RESULTS AND CONCLUSION: Compared with the other groups, the cel activity was higher in the combination induction group and reached peak at 16 days after induction, the number and volume of myotubes were increased at 20 days with regular arrangement, and desmin showed strong expression. Cel cycle showed the ratio of adipose-derived stromal cells in the DNA replication phase was increased in the combination induction group, the ratio of cells in the clearance period was decreased; the level of col agen type Ⅲ was increased significantly, and the difference was significant. The results indicate that 5-azacytidine combined with multiple factors can promote the differentiation of adipose-derived stromal cells into myoblasts with significant cel proliferation, which is considered as the ideal method to in vitro induction of myoblasts.
4.Alpha-lipoic acid inhibits mitochondrial oxidative stress in the rat skeletal muscle with chronic hypoxia exposure
Pin XIAO ; Yihui PANG ; Peng PENG ; Hai BO
Chinese Journal of Tissue Engineering Research 2013;(28):5216-5222
BACKGROUND: α-lipoic acid is named as “nature antioxidant” and “mitochondrial nutrition”. But it is unclear whether α-lipoic acid can be used to protect skeletal muscle with chronic hypoxia exposure, as wel as the relative mechanism. OBJECTIVE: To observe the effect of α-lipoic acid on the antioxidant enzymes and oxidative stress in rat skeletal muscle with chronic hypoxia exposure, and to investigate the relative signaling pathway of α-lipoic acid. METHODS: Thirty-six Sprague Dawley rats were randomly divided into three groups: normoxia control group, hypoxia control group, and hypoxia+α-lipoic acid group. Rats in the hypoxia control group were subjected to hypoxia exposure in normobaric hypoxic tent with 11.3% oxygen concentration. Rats in the hypoxia+α-lipoic acid group were induced by adding α-lipoic acid (0.25%) in the normal diet. Al the interventions were lasted for 4 weeks. RESULTS AND CONCLUSION: α-lipoic acid in hypoxia could markedly enhance the mitochondrial Sirtuin-3 expression, improve the mitochondrial adenosine triphosphate synthesis activity and membrane potential, up-regulate the mitochondrial state 3 respiratory rate, respiratory control ratio and ratio of phosphorus to oxygen, down-regulate the mitochondrial state 4 respiratory rate and promote and up-regulate the activity of mitochondrial antioxidant enzymes such as manganese superoxide dismutase, glutathione peroxidase and catalase, thus inhibiting mitochondrial H2O2 generation rate and reducing mitochondrial malondialdehyde level. The results indicated that α-lipoic acid could improve the efficiency of energy metabolism of chronic hypoxia skeletal muscle mitochondria and inhibit reactive oxygen generation, and it could inhibit the oxidative stress through improving antioxidant enzyme activity of mitochondria. The protection mechanism of α-lipoic acid on hypoxia skeletal muscle mitochondria may be related to the increasing of mitochondrial state 3 respiratory rate.
5.The top cited articles on bioartificial liver in Web of Science
Fan ZHANG ; Xuejun LAO ; Jianwei JIANG ; Mingrong CAO
Chinese Journal of Tissue Engineering Research 2013;(31):5723-5728
BACKGROUND:Bioartificial liver could partial y replace the major liver functions, including detoxification, synthesis, secretion and biotransformation. OBJECTIVE:To use bibliometric indexes to track study focuses on bioartificial liver, and to investigate the relationships among geographic origin, impact factors, and highly cited articles indexed in Web of Science. METHODS:A list of citation classics for bioartificial liver was generated by searching the database of Web of Science-Expanded using the terms“artificial liver support system”or artificial liver or“bioartificial liver”. The top 33 cited research articles which were cited more than 100 times were retrieved. RESULTS AND CONCLUSIONS:Of 4 144 articles published, the 33 top-cited articles were published between 1992 and 2010. The highest citations paper was published in 2002, with a total of 668 citations, mean cited 55.67 per year. The total citations of 33 articles were 6 094 times, with a mean of 12.64 citations per article. These top-cited papers came from 11 countries, of which 12 articles came from the United States. University of Rostock led the list of classics with five papers. Harvard University and Massachusetts General Hospital ranked the second with four papers each. The 33 top-cited articles were published in 18 journals, predominantly Annals of Surgery and Hepatology, fol owed by Artificial Organs and Biotechnology and Bioengineering. Our bibliometric analysis provides a historical perspective on the progress of bioartificial liver research. Articles originating from outstanding institutions of the United States and published in high-impact journals are most likely to be cited.
6.Hypoglycemic effect of bone marrow mesenchymal stem cells in vitro differentiated into islet-like cells
Liming ZHANG ; Renhao WANG ; Meirong WAN ; Quan WEN ; Xiaoyun LIU ; Yong SHAO
Chinese Journal of Tissue Engineering Research 2013;(31):5699-5704
BACKGROUND:Islet and islet cel transplantation for the treatment of diabetes has achieved effect, but the research is limited dut to the shortage of islet and immune rejection. OBJECTIVE:To observe the effect of transplantation of islet-like cells that in vitro differentiated from bone marrow mesenchymal stem cells on the treatment of diabetes in rats. METHODS:The rat bone marrow mesenchymal stem cells were induced with basic fibroblast growth factors and hepatocyte growth factors, and then received immunocytochemistry staining to detect the induction. The Sprague Dawley rats received intraperitoneal injection of streptozotocin to establish the diabetes models. After modeling, the rats were randomly divided into control group and experimental group (transplanted with induced islet-like cells). The experimental group was transplanted with the induced islet-like cells through renal capsule, and the control group was transplanted with normal saline in the same dose. The blood glucose and body mass of the diabetes rats were observed after transplantation. RESULTS AND CONCLUSION:The bone marrow mesenchymal stem cells could differentiate into islet-like cells after in vitro induced with basic fibroblast growth factors and hepatocyte growth factors. There was no significant change in blood glucose of the control group after transplantation (P>0.05), and the blood glucose of the rats in the experimental group was significantly decreased compared with the control group (P<0.05). The bone marrow mesenchymal stem cells can differentiate into islet-like cells after in vitro induced with the induction system containing basic fibroblast growth factors and hepatocyte growth factors, and the islet-like cells have a certain ability of insulin secretion. The transplantation of induced islet-like cells after transplanted into the diabetes rats through renal capsule can decrease the blood glucose level of the rats.
7.Platelet-rich plasma effects on grainy fat transplantation
Feng LI ; Kun LI ; Jie LI ; Weidong TIAN
Chinese Journal of Tissue Engineering Research 2013;(31):5645-5651
BACKGROUND:The higher long-term absorption rate greatly influence the widely application of fat transplantation. Platelet-rich plasma contains a high concentration of growth factors, which benefits to the tissue healing and regeneration. OBJECTIVE:To observe the effects of platelet-rich plasma on grainy fat transplantation and to investigate the mechanisms preliminarily. METHODS:Ten 6-week-old nude mice were prepared. The right or left dorsal subcutaneous tissues were randomly selected as the platelet-rich plasma group (0.5 mL fat granule+0.1 mL platelet-rich plasma), and the contralateral side was regarded as the control group (0.5 mL fat granule+0.1 mL phosphate-buffered saline). At 10 and 90 days after implantation, five nude mice were selected from each group, and then the mice were sacrificed to obtain the grafts in each group for general appearance observation, volume determination and histological detection. Furthermore, we isolated adipose-derived stem cells from human subcutaneous fat tissue during the in vitro experiment. Cel counting kit-8 and real-time PCR were used to evaluate the influence of platelet-rich plasma on adipose-derived stem cel proliferation and adipogenic differentiation in vitro, respectively. RESULTS AND CONCLUSION:Comparison of the grafts obtained at 10 and 90 days after implantation, the residual volume in the platelet-rich plasma group was significantly larger than that in the control group (P<0.05), Moreover, more normal adipocytes and capil ary formation were observed in the platelet-rich plasma group (P<0.05). For in vitro experiment, platelet-rich plasma could significantly improve adipose-derived stem cel proliferation, and the expressions of adipogenic-related genes were up-regulated in platelet-rich plasma-induced adipose-derived stem cells. Al results demonstrate that platelet-rich plasma can improve the survival of fat grafts,which might be closely related to that platelet-rich plasma can promote the proliferation and adipogenic differentiation of adipose-derived stem cells and the revascularization in grafted fat tissue.
8.Allogeneic bone combined with autologous bone marrow stem cell transplantation for the treatment of benign bone tumors and tumor-like lesions
Yingfei LIU ; Tao WANG ; Pingde ZHANG
Chinese Journal of Tissue Engineering Research 2013;(31):5652-5658
BACKGROUND:Al ogenic bone is a clinical commonly used bone graft material, but the osteoinductive capacity is the biggest problem. OBJECTIVE:To evaluate the effect of al ogeneic bone combined with autologous bone marrow stem cells on the repair of bone defects after scraping or resection of benign bone tumors and tumor-like lesions. METHODS:Sixty-five cases of benign bone tumors (including patients with tumor-like lesions) were divided into two groups according to bone graft. There were 35 cases in the composite bone marrow stem cells for bone graft group, and 20-40 mL red bone marrow were extracted from anterosuperior iliac spine or iliac spine on both sides according to the expected amount of bone graft, then the bone marrow stem cells were isolated, purified, cultured and amplified for standby, and the bone marrow stromal stem cells and al ogeneic bone particles were ful y blended before bong graft. After tumor scraping or resection, the blended bone marrow stromal stem cells and al ogeneic bone particles were implanted into the bone defect region. In the bone graft group, the bone defect was implanted with al ogeneic bone soaked with saline for half an hour. X-ray examination was performed at 1, 3, 6 and 12 months after treatment to compare the fuzzy boundary and the time for disappear, and the postoperative complications were observed. RESULTS AND CONCLUSION:Al the 62 patients were fol owed-up for more than 12 months. The fuzzy boundary time and disappear time in the composite bone marrow stem cells for bone graft group were shorter than those in the bone graft group (P<0.05). In the composite bone marrow stem cells for bone graft group, one case appeared rejection and healed after treated with immunosuppressive agents for 2 weeks, and no complication observed in two groups. The results indicate that al ogeneic bone composite autologous bone marrow stem cells for bone graft can promote bone fusion and bone defect healing.
9.Three-dimensional visualizational reconstruction of pedicled island skin flap with arterial arch at the superior border of abductor hallucis muscle
Wei TAN ; Dongsheng LI ; Xu LI ; Wenhua HUANG
Chinese Journal of Tissue Engineering Research 2013;(31):5639-5644
BACKGROUND:At present, design of pedicled island skin flap with arterial arch at the superior border of abductor hal ucis muscle mainly depends on clinician’s abstract thinking, which makes it difficult for quantitative analysis and exchange learning, and makes it limited in clinical application. OBJECTIVE:To perform the three-dimensional reconstruction of pedicled island skin flap with arterial arch at the superior border of abductor hal ucis muscle, and to establish visible models of the flap. METHODS:One fresh adult cadaver specimen was col ected and perfused with the mixture of lead oxide-gelatine through anterior tibial artery and posterior tibial artery, and then received spiral CT scanning. The branch of anterior medial mal eolus artery, medial tarsal artery and the superficial branch of medial plantar artery as wel as its anastomotic branch were reconstructed with mimics10.0 software, in order to observe the constitute, track-way and distribution of arterial arch at the superior border of abductor hal ucis muscle. According to the feature of anatomical structures, reconstruction was conducted in skin and vessel to obtain three-dimensional arterial skin flap structure. RESULTS AND CONCLUSION:The layers of three-dimensional reconstruction were distinctively displayed, and the arterial arch at the superior border of abductor hal ucis muscle was displayed wel with strong physical sense and clear relationship between skin and arteries. The morphology of various anatomical structures could be observed clearly on the three-dimensional reconstruction image, especial y the formation of arterial arch at the superior border of abductor hal ucis muscle and its surface projection. Three-dimensional reconstruction of pedicled island skin flap with arterial arch at the superior border of abductor hal ucis muscle may provide great value for basic research, clinical experiments and surgical planning, and provide visual reference for the individual operation design and can be able to enhance the successful rate of surgery.
10.Application of continuous thermodilution method in beagle models with pulmonary arterial hypertension
Dandan CHEN ; Daxin ZHOU ; Lihua GUAN ; Fadong CHEN ; Lili DONG ; Juying QIAN ; Junbo GE
Chinese Journal of Tissue Engineering Research 2013;(24):4509-4514
10.3969/j.issn.2095-4344.2013.24.020