1.Bone formation by bone marrow stromal cell using tissue engineering methods in vitro and in vivo
Dan JIN ; Guoxian PEI ; Qian WANG
Chinese Journal of Microsurgery 2000;0(03):-
Objective To study the osteogenic capability of bone marrow stromal cell (BMSc) using tissue engineering methods Methods The osteogenic potential in vitro of cultured BMSc in a conditional medium were examined by phase contrast microscopy,histochemistry stains technique The BMSc were cultured in combination with bioactive glass ceramic (BGC) materials Then the composite were implanted into the skeletal muscle beds in rabbits All implants were exmined by gross observation and histological examination Results The BMSc showed a similar property to those of osteoblasts and could synthesized mineralized new bone tissue in vitro New bone tissue can be observed in the composites of the BMSc and BGC materials Conclusions New bone tissue can be formed by tissue engineering methods
2.New bone formation by bone marrow stromal cell combined with the bioactive glass ceramic using tissue-engineering methods
Dan JIN ; Guoxian PEI ; Qian WANG
Chinese Journal of Trauma 2001;17(3):151-154
Objective To study the biocompatibility of bioactive glass ceramic (BGC) materials with bone marrow stromal cell (BMSc) and the osteogenic capability of BMSc using tissue-engineering methods. Methods The osteogenic potential in vitro of cultured BMSc in a conditional medium was examined by histochemistry stains technique. The BMSc was cultured in combination with BGC. The attaching and extending speed of the cells to the materials, the proliferation and alkaline phosphatase activity were tested. Then the composite was implanted into the skeletal muscle beds in rabbits. All implants were examined by gross observation and histological examination. Results The BMSc showed a similar property to those of osteoblasts. BMSc can attach to and extend on BGC materials. No inhibition to celluar proliferation and ALP activity were observed by the materials. New bone can be observed in the composites of the BMSc and BGC materials. Conclusions BMSc may provide a rich cellular resource in tissue-engineered bone formation. New bone tissue can be formed by tissue engineering methods.
3.Improvement for indwelling healer tube in prevention and cure the conglutination for children patiens after the surgery operation Flexor tendon tenosynovitis
Jimin PEI ; Huisheng HE ; Dan XU
International Journal of Surgery 2009;36(8):534-536
Objective Evaluate the safety and efficacy of indwelling healer tube in prevention and cure the conglutination after the surgurey of flexor tendon tenosynovitis. Methods During the year of 2001~2006, We have 38 young patients treated with indwelling of healer tube into the local or partial of the recovering muscle and sinew, then anesthetic and sodium hyaluronate were injected in the tube in certain intervals to lubicate and prevent conglutination after the operations of joint of children flexor tendon tenosynovitis. Then let the young patients do some healing training attne earlystage after surgery. Groups of patient were set up to make comparative analysis and evaluate the effectiveness of indwelling of the healer tubes according to the recovery status of the function of arthrosis and grasp after surgery. Results The result is that the rate of choiceness of 46 sinew is 89.1% in 34 cases with indwelling healer tube after the observing period from 6 months to 2 years, whereas the other group of 44 sinew in 30 cases has the rate of choiceness of 63.6%. The comparison has the significant conclusion of statistics (P<0.05). Conclusion It is convenient and safe to use indwelling healer tube to prevent the conglutination after the operation of joint the broken finger muscle and sinew of children. Therefore it is worth popularizing and promoting.
4.Changes of CGRP and NPY expression in tissue-engineered bone with fascia flap
Jiande CUI ; Guoxian PEI ; Dan JIN
Medical Journal of Chinese People's Liberation Army 1981;0(06):-
Objective To evaluate the expressive levels of calcitonin gene related peptide (CGRP) and neuropeptide Y (NPY) in tissue-engineered bone with fascia flap in vivo. Method A segmental bone defect 1.5cm in length was made at the both radius of 12 healthy New Zealand rabbits. The defects were repaired by implantation in two ways: for left radius, engineered bone with fascia flap was implanted (served as experiment group), and for right radius only engineered bone was implanted (served as control group). 3, 6 and 12 months after implantation, the expressions of CGRP and NPY in the new bone were determined with immunohistochemistry and semi-quantified using image analysis software. Results CGRP and NPY expressions in the both groups were significantly increased in a time-dependent manner (P
5.HUMAN BONE MORPHOGENETIC PROTEIN 7 EXPRESSION IN RABBIT BONE MARROW STEM CELL TRANSFERRED BY RETROVIRAL VECTOR MEDIATED hBMP_7 GENE
Dan JIN ; Weisen ZENG ; Guoxia PEI
Medical Journal of Chinese People's Liberation Army 1981;0(06):-
The human bone morphogenetic protein 7 (hBMP 7) gene was reconstructed in retroviral vector and transferred into incasing cells PT67 by liposome mediated method.The clones of the cells transferred with BMP 7 were selected by G418, and targeted rabbit bone marrow stem cells were infected with the virus granules which secreted from PT67 cells and also selected by G418. The mRNA and protein of BMP 7 gene in transferred cells were analyzed with hybridization in situ and immunohistochemistry. BMP 7 retrovirus vetor was successfully reconstructed. Cells transferred by PLNCX 2 hBMP 7 expressed abundant human BMP 7 mRNA and protein in the cytoplasm. However positive findings were not found in those cells that were not transferred. It may be used to increase the osteogenic capability of BMSc in the study of bone tissue engineering.
6.Feasibility of chitosan-beta-tricalcium phosphate as an injectable bone tissue engineering scaffold material
Wenjun CHENG ; Dan JIN ; Guoxian PEI
Medical Journal of Chinese People's Liberation Army 1983;0(02):-
0.05).Under SEM,the BMSCs showed good adhesion to beta-TCP with obvious proliferation.Conclusion BMSCs can grow and proliferate well on the compound BMSCs/beta-TCP and beta-TCP has good biocompatibility with BMSCs in vitro,which may be used as a good scaffold material for injectable tissue engineering bone.
7.Association of the degradation of the hybrid layer and the durability of dentin adhesion.
Dan-Dan PEI ; Xue-Qing HUANG ; Cui HUANG
Chinese Journal of Stomatology 2011;46(1):58-60
Acid Etching, Dental
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adverse effects
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Chondroitin ABC Lyase
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chemistry
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Dental Bonding
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Dentin
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chemistry
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ultrastructure
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Dentin-Bonding Agents
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chemistry
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Humans
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Hydrolysis
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Matrix Metalloproteinase Inhibitors
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pharmacology
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Matrix Metalloproteinases
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metabolism
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Microscopy, Electron, Transmission
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Surface Properties
8.Designation and silencing function of the small interfering RNA of HLA-A2
Jiansheng LIU ; Gang ZHAO ; Dan PEI ; Deyu ZHENG
Chinese Journal of Tissue Engineering Research 2014;(29):4605-4610
BACKGROUND:Human leukocyte antigen (HLA), the major histocompatibility complex of human, plays an important function in the transplant rejection. Decreasing the expression of HLA wil prolong the survival time of transplants.
OBJECTIVE:To design smal interfering RNA (siRNA) of HLA-A2 and to detect the effect of siRNA-HLA-A2 on the expression of HLA-A2.
METHODS:Four kinds of siRNA-HLA-A2 domains were designed, and recombinant lentivirus expression vector were formed. The 293T cells, highly expressing HLA-A2, were infected in vitro. Then the knockout efficacy of four domains was detected to select the highly efficient siRNA-HLA-A2 target sequences. The human embryo lung fibroblasts were cultured in vitro and infected with the lentivirus carrying the target sequence. The infecting efficiency of LV-siRNA-HLA-A2 was observed under the fluorescence microscope and the silence function of this siRNA in human embryo lung fibroblasts was detected by western blot analysis.
RESULTS AND CONCLUSION:According to the mRNA sequence of HLA-A2 in Genbank, three siRNAs were designed and synthesized. In vitro, the over expression of HLA-A2 in 293K cells was successful y silenced. The HLA-A2 expression in human embryo lung fibroblasts was also efficiently silenced after the human embryo lung fibroblasts were infected by the highly efficient siRNA of HLA-A2. The efficacy was up to 80%.
9.Effects of ?-lipoic acid on bone metabolism in lead-poisoned juvenile rats
Dan PEI ; Deyu ZHENG ; Wei SHAN ; Shujian QIN
Journal of Xi'an Jiaotong University(Medical Sciences) 2004;0(05):-
Objective To determine the effects of ?-lipoic acid on bone metabolism in lead-poisoned rats.Methods Totally 31 SD rats were randomly divided into two groups,8 rats in blank control group and 23 in lead group that received gastric lavage with lead acetate(230 mg/L).After 40 days,3 rats from each group were taken for the measurement of lead in the blood and bone.The rats in lead group were then randomly divided into four groups: lead control group and three lead groups with different concentrations of ?-lipoic acid [30,60 and 100 mg/(kg?d)],with 5 rats in each.At the end of the experiment(80 d),the rats were killed and the samples of whole blood,serum and bone were collected to detect osteocalcin content with radioimmunoassay and alkaline phosphatase expression with immunohistochemistry staining.Results ① Blood and bone lead contents in each ?-lipoic treatment group were significantly lower than those in lead control group(P
10.Experimental study on the repair of goat tibia defects with revascularized tissue engineered bone
Bin CHEN ; Guoxian PEI ; Ke WANG ; Dan JIN ; Kuanhai WEI
Chinese Journal of Tissue Engineering Research 2006;10(5):164-169
BACKGROUND: It is a key point to revascularize the tissue-engineered bone during the repairing of large bone defect. Fascia flap is commonly used in clinic to accelerate the blood supply of implant.OBJECTIVE: To observe the feasibility of repairing goat tibia defects with tissue-engineered bone and accelerating revascularization with fascia flaps.DESIGN: Randomized and controlled animal experiment SETTING: Department of Traumatic Orthopaedics, Nanfang Hospital,Southern Medical University.MATERIALS: Totally 36 goats with the body mass of 14.5-15.5 kg of either gender were enrolled.METHODS: This experiment was conducted at the Department of Traumatic Orthopaedics, Nanfang Hospital, formerly the First Military Medical University of Chinese PLA from December 1999 and December 2003.Bone and periosteum defects 20 mm long were made and fixed with plate of left tibia in 36 goats. They were randomly divided into four groups: Group A in which the defects were filled with coral hydroxyapatite (CHAP), Group B I CHAP+ bone marrow stroma cells (BMSc); Group C with fascia flaps;Group D with nothing. Next, the bone regeneration and the revasculariza tion were evaluated. Radionuclide bone imaging was done 2, 4, 8 weeks after operation. After X-ray examination, the index of optical density of Xray films and histology of the implants were analyzed at 4, 8, 12 weeks after operation, and the biomechanical characters were studied 12 weeks postoperatively.MAIN OUTCOME MEASURES: Gross observation and X-ray, radionuclide bone imaging, biomechanical and histological observation RESULTS: Totally 36 goats entered the stage of result analysis. ① Gross observation of the repair sample of bone defects of the animals in each group: there was no osteogenesis postoperatively at each time point in the blank control group . In Group B, at week 8 to 12, there was no obvious osteogenesis and callus formation on the surface of the materials. In Group C,At weeks 8 to 12, bone defects were filled gradually, many bone callus processes were seen on the surface of the materials , centralizing and enwrapping the materials. The osteogenetic process in the Group C were superior to that of theGroup B. ②Examination result with -901/SA PET-CT scanners: It was seen by naked eyes that at weeks 2 to 8 in the Group A,the radioactivity concentration at region of interesting (ROI) of the operation side had obvious increasing trend, and similar trend of changing appeared in the Group B and Group C, but the ROI counts and T/NT value in the Group B were both lower than those in the Group C. The decreasing trend in the Group A was lower than that in the Group B. ③) Radiological results: the osteogenesis volume through measuring absorbance in the order from large to small was Group C, Group B, and Group A[At week 12, they were (4.180±0.192), (3.480±0.453), (2.959±0.682)respectively ].④Biomechanical results: there were significant difference of loading and bending stress in the Group C, Group B and Group A [ The loading was (758.333±88.754), (530.214±65.297), (359.667±60.715)N , respectively; and the bending stress was (13.937±2.199), (10.123±1.243),(6.223±0.945)N/mm2, respectively ].⑤)Histological results: Slices at various time points in the blank control group showed no bone tissue. In the other three 3 groups, with the prolongation of time, the osteagenetic range and quality were in the order of Group C, Group B and Group A.CONCLUSION: The fascia flaps can accelerate the revascularization process in the formation of tissue-engineered bone so that the capability of tissue engineered bone to repair the large bone defects may be enhanced.