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find Keyword "Angiopoietin 1" 2 results
  • REPAIR OF RADIAL SEGMENTAL BONE DEFECTS BY COMBINED ANGIOPOIETIN 1 GENE TRANSFECTED BONE MARROW MESENCHYMAL STEM CELLS AND PLATELET-RICH PLASMA TISSUE ENGINEERED BONE IN RABBITS

    Objective To investigate the osteogenesis effects of angiopoietin 1 (Ang-1) gene transfected bone marrow mesenchymal stem cells (BMSCs) seeded on β tricalcium phosphate (β-TCP) scaffolds (tissue engineered bone) with platelet-rich plasma (PRP). Methods  BMSCs were isolated from bone marrow tissue of rabbits. The Ang-1 gene was transfected into the BMSCs at passage 2 by lentivector, which were seeded on β-TCP scaffolds with PRP (0.5 mL) after 48 hours of transfection. Bilateral radial segmental bone defects (15 mm in length) were created in 20 3-month-old New Zealand rabbits. Then the tissue engineered bone with the Ang-1 gene transfected BMSCs (experimental group) and untransfected BMSCs (control group) were implanted into the defects in the right and left radius, respectively. X-ray, histology, immunohistochemistry, and biomechanics observations were done at 2, 4, 8, and 12 weeks after operation. Results In vitro, the transfected rate was over 90% and RT-PCR showed that the Ang-1 expression were significantly increased after transfection. The X-ray films showed that some callus formed at 4 weeks, partial bony union was observed at 8 weeks, and complete union at 12 weeks in experimental group; and bone union was not observed at 12 weeks in control group. HE staining showed that capillary appeared at 8 weeks and more capillaries were observed in new bone at 12 weeks in experimental group; only a few capillaries were observed at 12 weeks in control group. At 8 and 12 weeks, the microvascular density were (50.1 ± 7.8) /mm2 and (66.1 ± 3.5) /mm2 in experimental group and were 0 and (30.3 ± 7.2)/mm2 in control group, showing significant differences between 2 groups at 12 weeks (Z= —2.107, P=0.031). Immunohistochemistry examination showed that the positive cells can be found at 8 weeks in experimental group. And the biomechanical analysis showed that maximum loads of experimental group were significantly higher than those of control group in three-point bending test and compression test at 12 weeks (P lt; 0.05). Conclusion The tissue engineered bone with PRP and Ang-1 can increase the osteogenic properties by enhancing capillary regeneration, thus it can be used to repair radial segmental bone defects of rabbit.

    Release date:2016-08-31 05:45 Export PDF Favorites Scan
  • CONSTRUCTION OF EUKARYOTIC EXPRESSION VECTOR FOR HUMAN ANGIOPOIETIN 1 AND ITS EXPRESSION IN THE MARROW MESENCHYMAL STEM CELLS OF RABBIT

    Objective To investigate the possibility of constructing eukaryoticexpression vector for human angiopoietin 1(hAng-1),transfecting it to bonemarrow mesenchymal stem cells (MSCs) so as to repair bone defect. Methods The eukaryotic expression vector pcDNA3-hAng-1 was constructed by recombinant DNA technique, transfected into MSCs by liposome DOTAP, and selected with G418. The hAng-1 expression of mRNA and protein was detected by reverse transcript-PCR and Western Blot. Results After the recombinant eukaryotic expressionvector for hAng-1 was digested with Xho-I and BamH-I, electrophoresis revealed 1.4 kb fragment for hAng-1 gene and 5.4 kb fragment for pcDNA3 vector. In the transfected MSCs, the mRNA and protein expression of hAng-1 gene were detected with reverse transcriptPCR and Western Blot. Conclusion The constructed eukaryotic expression vector hAng-1 could be expressed in the transfected MSCs, thus to provide the basis for bone repair with tissue engineering.

    Release date:2016-09-01 09:29 Export PDF Favorites Scan
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