1.Histological changes in muscle tissues with acute injury following cryotherapy
Peng SHI ; Ruowu SHEN ; Aiyu JI
Chinese Journal of Tissue Engineering Research 2011;15(20):3793-3796
BACKGROUND: Cryotherapy of acute soft tissue injury has been widely used in clinical practice.OBJECTIVE: To observe the histological changes and treatment effect of different cryotherapies on the rats' acute damage of soft tissue. METHODS: Neonatal Wistar rats were randomized to normal, model, intermittent cryotherapy and continuous cryotherapy groups. Models of acute damage of soft tissue were established in model, intermittent cryotherapy and continuous cryotherapy groups. In intermittent cryotherapy group, the injury was treated by intermittent cryotherapy with ice bag at 4 °C; in the continuous cryotherapy group, the injury was treated by continuous cryotherapy with ice bag at 4 °C; the model group was not treated. Histological changes were observed at 48 hours. Injury degree was evaluated using injury symptom index.RESULTS AND CONCLUSION: Compared with model group, the scores of injury symptom index and histology were lower, interleukin-1β expression was reduced, and transforming growth factor-β1 (TGF-β1) expression was increased in intermittent cryotherapy and continuous cryotherapy groups (P < 0.05). Compared with intermittent cryotherapy group, the scores of injury symptom index and histology were reduced (P < 0.05), interleukin-1β expression was reduced (P < 0.05), and TGF-β1 expression was increased in continuous cryotherapy group (P < 0.05). Results demonstrated that cryotherapy can cure the acute damage of soft tissue by reducing interleukin-1β expression and raising TGF-β1 expression. Continuous cryotherapy is superior over intermittent cryotherapy.
2.Effects of p38MAPK signaling pathway on cyclic tensile stress-induced fibroblast apoptosis
Jing QIU ; Guangyun ZHANG ; Zhen TIAN ; Yue ZHANG ; Jiangbo YU ; Xiao YUAN
Chinese Journal of Tissue Engineering Research 2011;15(20):3789-3792
BACKGROUND: When the teeth affected abnormal biting force, tooth absorption and periodontium would be greatly damaged. OBJECTIVE: To study whether periodontal membrane fibroblast affected apoptosis following cyclic tensile stress stimulation and whether p38MAPK signaling pathway participated in apoptosis. METHODS: Fibroblasts at passages from 4 to 7 were randomly assigned to control, loading and SB203580 groups after synchronization. In the loading and SB203580 groups, 12% strain was applied at a loading frequency of 6 cycles per minute, i.e. 5 seconds for tension and 5 seconds for relaxation. In the SB203580 group, cells were treated with 20 mmol/L p38MAPK inhibitor SB203580 at 1 hour before loading. At 6, 12 and 24 hours after loading, cells from each group were harvested, and cell apoptosis was detected using a flow cytometry. Expression of bax mRNA was determined using reverse transcription-polymerase chain reaction. RESULTS AND CONCLUSION: Compared with the control group, apoptotic rate of fibroblasts and bax mRNA expression were increased after loading (P < 0.05), and enhanced over time, and peaked at 12 hour following loading, and then decreased gradually. Compared with the loading group, cell apoptosis was reduced at corresponding time points in the SB203580 group (P < 0.05), and bax mRNA expression was diminished. These results indicated that cells affected apoptosis after mechanics stimulation, and mitogen activated protein kinase p38MAPK signaling pathway participates in the process of apoptosis.
3.Role of Fas, FasL and Caspase-3 in artesunate-induced apoptosis of human embryonic lung fibroblasts
Changming WANG ; Hongxiu LI ; Xiaofei ZHANG
Chinese Journal of Tissue Engineering Research 2011;15(20):3785-3788
BACKGROUND: Artesunate can relieve pulmonary fibrosis, but its mechanisms are rarely reported.OBJECTIVE: To explore the effect of artesunate on apoptosis of HFL-I cells and the role of Fas, FasL and Caspase-3 in the artesunate-mediated apoptosis of HFL-I cells. METHODS: Cell counting kit-8 (CCK-8) assay was used to determine the effects of artesunate at 1, 10, 100 mg/L on the growth of HFL-I cells in vitro. Apoptosis ratio was examined by flow cytometry (FCM). The mRNA level of Fas, FasL and Caspase-3, were assessed by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS AND CONCLUSION: Artesunate had a significantly inhibitory effect on the proliferation of HFL-I cells in a dose-dependent manner in vitro. The apoptosis rate of HFL-I cells was significantly increased in the artesunate-treated group compared with the control group (P < 0.05 or P < 0.01). The mRNA levels of Fas, FasL and Caspase-3 were significantly higher in the artesunate-treated group than the control group (P < 0.05). The findings of this study demonstrate that artesunate can exert a marked anti-pulmonary fibrosis effect by up-regulating mRNA levels of Fas, FasL and Caspase-3, which can induce the growth inhibition and apoptosis in HFL-I cells.
4.Analysis of birth defects in Xinjiang multi-ethnic region
Ruoyun QIAN ; Hong LIU ; Nan ZHONG ; Rui WANG ; Hongyun ZOU ; Jiang HE ; Wuzhong YU
Chinese Journal of Tissue Engineering Research 2011;15(20):3797-3800
BACKGROUND: Xinjiang is a multi-ethnic region with significant differences in local geographical position, economic development and climatic environment. OBJECTIVE: To analyze the occurrence and development tendency of birth defects, disease categories and disparity among different ethnic groups and regions in Xinjiang.METHODS: A stratified cluster random sampling observation was performed in 13 counties (cities) according to the status of ethnical distribution and local economics of Xinjiang. Quarter Report Sheet on Babies and The defect babies register card were filled as the scheme of Chinese birth defect monitoring, and ICD10 diagnostic code was adopted in birth defect diagnosis. The birth defects rate was calculated from January 2005 to December 2008, and the disease categories and disparity among different ethnic groups and regions in Xinjiang were analyzed. RESULTS AND CONCLUSION: The average incidence rate of birth defect was 9.74‰, which was dramatically descended in 2006 and ascended afterward yearly. The incidence rate of countryside was higher than city, and male more than female. In geography, south of Tianshan Mountain was higher than north and east in birth defect incidence. Among major ethnic groups in Xinjiang, Sibe and Uygur had the highest birth defect incidence rate, followed by Man, Hazakh, and Han. The birth defect incidence of Han, Uygur and Hazakh people showed descend tendency, Hui, Mongolia, and Man people fluctuated, yet Sibe's rate had a change of rise and fall. The first five birth defect entities were neural tube deformity, cleft lips, anencephaly, congenital hydrocephalus and cleft palate combined with cleft lips. The birth defects rates are different from ethnic groups and regions in Xinjiang.
5.Effects of combined human thymus and cord blood hematopoietic stem/progenitor cell transplantation on cellular immunity of nude mice
Qingjie ZHAO ; Qianru LI ; Ying DU ; Qingyong ZHANG ; Guoxi LI ; Xiang HU
Chinese Journal of Tissue Engineering Research 2011;15(23):4199-4202
BACKGROUND: T cells are believed to play an important role in anti-infection, anti-tumor and immune function. However, the mechanism underlying the differentiation and development remains poorly understood. OBJECTIVE: To investigate the distribution of T cells in nude mice that are jointly transplanted human thymus and cord blood and the reconstruction of the immune function. METHODS: Thirty Balb/c nu/nu nude mice were randomly divided into two groups: an experimental group and a control group. In the experimental group, human thymus tissue was transplanted into the renal capsule of nude mice. Two weeks later, freshly isolated human cord blood CD34+ cells suspension was back perfused into the nude mice via the vein. In the control group, CD34+ cells transplantation was performed directly without thymus transplantation. After 60 days of breeding, the immune function of nude mice was detected in two groups. RESULTS AND CONCLUSION: Human thymus tissue in the renal capsule of nude mice survived and expressed CD3 and HLA-DR molecule. In the experimental group, CD3+ cells which distributed in the form of dots were observed in the mouse spleen. The proportion of CD3+, CD4+, CD8+, and CD4+CD25+ cells were significantly higher in the experimental group than in the control group. The nude mice from the experimental group rejected human gastric cancer BGC823 cells, while those from the control group did not. These findings demonstrated that combined human thymus and CD34+ cell transplantation allow nude mice to acquire T cell-mediated cellular immune function and possess the ability of anti-tumor.
6.Effects of different concentrations of recombinant human erythropoietin on proliferation of neural stem cells cultured in vitro
Zhengmin XUE ; Meng HU ; Changhai ZHANG ; Xiancheng ZHANG ; Xiaopeng ZHOU
Chinese Journal of Tissue Engineering Research 2011;15(23):4194-4198
BACKGROUND: Recombinant human erythropoietin (rhEPO) is a glycoprotein. Recent studies have demonstrated that rhEPO regulates many functional activities of neural cells. OBJECTIVE: To investigate the effects of different concentrations of rhEPO on proliferation of neural stem cells (NSCs) cultured in vitro. METHODS: Newborn Sprague-Dawley rat NSCs were harvested and cultured with serum-free culture medium containing different concentrations (5, 50, 500 U/mL) of rhEPO and 20 μg/L basic fibroblast growth factors (5, 50, and 500 U/mL rhEPO groups) and serum-free culture medium only containing 20 μg/L basic fibroblast growth factors (control group). After 7 days of culture, the cloning efficiency of NSCs was calculated. After 10 days of culture, neuron specific enolase (NSE)-and glial fibrillary acidic protein (GFAP)-immunoreactive cells were quantified. RESULTS AND CONCLUSION: In the rhEPO groups, cells proliferated rapidly, and the number of NSC microspheres was greater, in particular in the 50 U/mL rhEPO group, compared with the control group. NSCs grew faster in the 50 U/mL rhEPO group than in the control group. The number of NSE-and GFAP-immunoreactive cells was greater in the 50 U/mL rhEPO group than in the control group (P<0.01). These findings suggest that rhEPO promotes the in vitro culture and proliferation of NSCs, in particular 50 U/mL rhEPO.
7.Expression of vascular endothelial growth factor and its receptor in the epididymal sperm of adolescent male rats
Qingyan AI ; Yufeng ZHAO ; Yanmei WANG ; Naizhou MIAO ; Li MA ; Jiazhou YANG
Chinese Journal of Tissue Engineering Research 2011;15(28):5221-5224
BACKGROUND: Vascular endothelial growth factor (VEGF) and its receptor fms-like tyrosine kinase-1 (Flt-1) have a very important position in the field of male reproduction. However, it is still unclear about their expression meaning and regulatory mechanism in the reproductive system. OBJECTIVE: To study the expression and location of VEGF and Flt-1 in the epididymal sperm of adolescent male rats. METHODS: The expression of VEGF and Flt-1 was detected in 10 adolescent SD rats. The concentration of the sperm was (30-40)×109/L. Immunofluorescent staining was used for VEGF and Flt-1 expression and location in the sperm. RESULTS AND CONCLUSION: Immunofluorescent staining showed that VEGF and Flt-1 were both localized in the acrosome of sperm head, as well as in the neck, middle and principal segment of sperm tail. Specific expression patterns of VEGF and Flt-1 in the sperm of rats display that they may participate in spermiotelcosis, relevant to movement, capacitation and acrosome reaction of the sperm.
8.Vein transplantation of olfactory ensheathing cells for spinal cord injury
Kaibin WEI ; Yuliang LIU ; Hong LIU ; Xingang Lü ; Wenzheng ZHANG ; Jun LI
Chinese Journal of Tissue Engineering Research 2011;15(27):5021-5025
BACKGROUND: Olfactory ensheathing cell transplantation is a better theray for spinal cord injury, and it becomes one of the most promising treatment methods. Local transplantation is applied currently, with the disadvantages of complex operation, large trauma, repeated transplantation. Looking for a simple and effective way for cell transplantation becomes a hotspot for scholars from various countries.OBJECTIVE: To investigate the effect and possibility of transplantation of olfactory ensheathing cells for treatment of spinal cord injury.METHODS: Wistar rats with T 10 spinal cord hemisecti on were divided into 4 groups: intramedullary local transplantation group (A),vein transplantation group (B), D/F12 transplantation group (C) and control group (D). The functional recovery of rats with spinal cord injury was observed with combined behavioral score at different phases. The tissue sections of each group were made at 5 and 10 weeks postoperatively to observe the axon regeneration and the survival of olfactory ensheathing cells.RESULTS AND CONCLUSION: The experiment showed that the rats transplanted with OECs at injured site and through the vein had more improvement in functional recovery and histological changes than the other two groups. The effect between A group and B group had not significant difference. The method of treating spinal cord injury by transplanting OECs via the vein not only simplifies the operation and avoids many complications but also has good curative effect similar to local transplantation.
9.Preparation and mechanism of anticoagulatent biomedical polymer materials with blood compatibility
Chinese Journal of Tissue Engineering Research 2011;15(29):5507-5510
BACKGROUND: Biomedical materials contact internal environment of human body, and sometimes are implanted into organism. Therefore, they should have biocompatibility, chemical stability, suitable physical mechanical function and simple processing and molding, but no toxicity.OBJECTIVE: To investigate the preparation of biomedical polymer anticoagulant materials in the aspects of bioinert material, biological active surface, albumin structure and application in anticoagulation.METHODS: A computer-based online search of PubMed and Wanfang database was performed for articles related to preparation of biomedical polymer anticoagulant materials published between 1969 and 2010.RESULTS AND CONCLUSION: Currently preparation of anticoagulant materials commonly utilizes bioinert surface or bioactive surface alone, which has obtained good effects, but the biocompatibility, such as blood compatibility, cannot be retained for a long period of time. The combination of bioinert surface and bioactive surface plus albumin, natural constitutions in human blood may be the trend of anticoagulant materials development. Polyethylene glycol with high bioinert property in combination with albumin recognition factor cibacron blue with high bioactivity can be used to prepare active modifier, which is used to modify polyurethane.
10.Drug release, morphology and cytotoxicity of enantiomeric poly(L-Lactic acid)-poly(ethylene glycol)-poly(L-Lactic acid)/poly(D-Lactic acid)-poly(ethylene glycol)- poly(D-Lactic acid) stereocomplex hydrogel
Zhili TIAN ; Yao WU ; Rong LIU ; Yanfeng XIAO ; Bin HE ; Zhongwei GU ; Fang WU
Chinese Journal of Tissue Engineering Research 2011;15(29):5501-5506
BACKGROUND: Recently biodegradable hydrogel has been extensively used to delivery anticancer drug and bioactive macromolecule. However, to protect the activity of the bioactive macromolecule, we need to obtain series of hydrogel which have milder hydrogelation conditions and shorter hydroglation time.OBJECTIVE: To prepare enantiomeric poly(L-Lactic acid) (PLLA)-poly(ethylene glycol (PEG)-PLLA/ poly(D-Lactic acid) (PDLA)-PEG-PDLA stereocomplex hydrogel which has shorter hydroglation time, to physically encapsulate a model drug-lysozyme and sustained release it from the hydrogel. METHODS: Triblock copolymers of PLLA-PEG-PLLA and PDLA-PEG-PDLA were synthesized by ring-opening polymerization of L(D)-lactide using PEG as the initiator and Sn(Oct)2 as the catalyst. The triblock copolymers were characterized by 1H nuclear magnetic resonance, FT-IR and X-Ray diffractometry. A hydrogel was prepared from an aqueous mixture of PLLA20-PEG227-PLLA20 and PDLA21-PEG227-PDLA21 (10 wt% concentration) at room temperature for 12 hours. X-Ray diffractometry test was used to research the gelation mechanism. The release profile of the lysozyme as a model drug from the hydrogel was tested. The morphology of the freeze-dried hydrogel was investigated by scanning electron microscope. The cytotoxicity of the hydrogel was evaluated by 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide) assay.RESULTS AND CONCLUSION: Triblock copolymers of PLLA-PEG-PLLA and PDLA-PEG-PDLA were obtained. Both the PEG and PLA blocks in the copolymers could crystallize, but the crystallization of the PEG block was predominant. The stereocomplex formation between the PLLA and PDLA blocks within the hydrogel was confirmed by the X-Ray diffractometry analysis. The release profile of the lysozyme from the hydrogel exhibited a sustained-release pattern with a duration period of 7 days. The hydrogel exhibited a 3D interconnected porous structure with 50-100 μm pore size after being freeze-dried. The mouse fibroblast cell viability percentage was 99.3% after the cells contacted with the 100% extracted liquid for 72 hours.