Objective: To detect the effect of hBMP-2 transfected rMSCs on bone repair and the capability of the new biomaterial in enhancing bone repair. Methods: Auto-rMSCs were cultured and transfected with hBMP-2 by liposome....Objective: To detect the effect of hBMP-2 transfected rMSCs on bone repair and the capability of the new biomaterial in enhancing bone repair. Methods: Auto-rMSCs were cultured and transfected with hBMP-2 by liposome. All the transfected and un-transfected riSCs were attached to Allo-DBMs. These new biomaterials were implanted in muscle bags and segmental radius defects of the New Zealand white rabbits, and some controlled material groups were established for comparison. All the biomaterials and the controlled materials were assessed by gross observation, radiographical and histological methods. Results: The osteoblasts could be seen in the biomaterials with transfected rMSCs, which have been implanted in muscle bags. There was no sign of bone formation in the biomaterials with untransfected rMSCs and the single DBM groups. With the segmental bone defects, all the transfected, untransfected and single DBM biomaterials could work, but the healing of the biomaterial with transfected hBMP-2 was the fastest and most effective. Conclusion: Delivery of rMSCs with transfected hBMP-2 genemay generate osteo.morphogenesis and promote skeletal repair in vivo.展开更多
To promote efficient screening of active angiogenic drugs from traditional medicines, we constructed a humanembryonic kidney-293 cell model using vascular endothelial growth factor （VEGF） gene promoter as the drug t...To promote efficient screening of active angiogenic drugs from traditional medicines, we constructed a humanembryonic kidney-293 cell model using vascular endothelial growth factor （VEGF） gene promoter as the drug target. Inthis model, VEGF gene promoter may regulate the expression of the luciferase reporter gene by responding to thestimulation of drug molecules. This cell model allows rapid and efficient screening of vascular-inducing activecomponents from several drug monomer molecules. Furthermore, we used rat bone marrow mesenchymal stem cells（rMSCs） to conduct a preliminary study on the activity of alantolactone. Using simvastatin as a positive control, weinvestigated the effects of alantolactone on the expression of vascular-related cell marker molecules such as VEGF andα-smooth muscle actin （α-SMA） in rMSCs. According to our results, 0.1, 1, 3 and 5 μM of alantolactone upregulated thetranscriptional luciferase gene activity of VEGF promoter, and a significant difference from that in the control group wasobserved. Among them, 3μM of alantolactone showed the better effect than that of 3 μM of simvastatin （P = 0.036） andother concentrations of alantolactone and simvastatin showed similar effects. Compared with that in the control group,rMSCs induced with 1μM alantolactone for 3 days showed a significant increase in the relative mRNA expressions ofVEGF and α-SMA genes. However, these effect of 5 μM alantolactone were weaker than those of 5 μM simvastatin （P 〈0.05）; rMSCs treated with 1 μM alantolactone for 3 days showed brighter green fluorescence （FITC marker） of α-SMAand VEGF in situ expression than that observed in the control group and similar fluorescence intensity than that ofsimvastatin group in an immunoradiometric assay. The above results demonstrate the reliability of the highly efficientsystem for screening of active drug molecules and confirmed the vascular induction function of alantolactone at the geneand protein levels.展开更多
To develop a novel degradable poly (L-lactic acid)/β-tricalcium phosphate (PLLA/β-TCP) bioactive materials for bone tissueengineering, β-TCP powder was produced by a new wet process. Porous scaffolds were prepared ...To develop a novel degradable poly (L-lactic acid)/β-tricalcium phosphate (PLLA/β-TCP) bioactive materials for bone tissueengineering, β-TCP powder was produced by a new wet process. Porous scaffolds were prepared by three steps, i.e. solventcasting, compression molding and leaching stage. Factors influencing the compressive strength and the degradation behaviorof the porous scaffold, e.g. weight fraction of pore forming agent-sodium chloride (NaCl), weight ratio of PLLA: β-TCP,the particle size of β-TCP and the porosity, were discussed in details. Rat marrow stromal cells (RMSC) were incorporatedinto the composite by tissue engineering approach. Biological and osteogenesis potential of the composite scaffold weredetermined with MTT assay, alkaline phosphatase (ALP) activity and bone osteocalcin (OCN) content evaluation. Resultsshow that PLLA/β-TCP bioactive porous scaffold has good mechanical and pore structure with adjustable compressive strengthneeded for surgery. RMSCs seeding on porous PLLA/β-TCP composite behaves good seeding efficacy, biocompatibility andosteoinductive potential. Osteoprogenitor cells could well penetrate into the material matrix and begin cell proliferation andosteogenic differentiation. Osseous matrix could be formed on the surface of the composite after culturing in vitro. It isexpected that the PLLA/β-TCP porous composites are promising scaffolds for bone tissue engineering in prosthesis surgery.展开更多
文摘Objective: To detect the effect of hBMP-2 transfected rMSCs on bone repair and the capability of the new biomaterial in enhancing bone repair. Methods: Auto-rMSCs were cultured and transfected with hBMP-2 by liposome. All the transfected and un-transfected riSCs were attached to Allo-DBMs. These new biomaterials were implanted in muscle bags and segmental radius defects of the New Zealand white rabbits, and some controlled material groups were established for comparison. All the biomaterials and the controlled materials were assessed by gross observation, radiographical and histological methods. Results: The osteoblasts could be seen in the biomaterials with transfected rMSCs, which have been implanted in muscle bags. There was no sign of bone formation in the biomaterials with untransfected rMSCs and the single DBM groups. With the segmental bone defects, all the transfected, untransfected and single DBM biomaterials could work, but the healing of the biomaterial with transfected hBMP-2 was the fastest and most effective. Conclusion: Delivery of rMSCs with transfected hBMP-2 genemay generate osteo.morphogenesis and promote skeletal repair in vivo.
文摘To promote efficient screening of active angiogenic drugs from traditional medicines, we constructed a humanembryonic kidney-293 cell model using vascular endothelial growth factor （VEGF） gene promoter as the drug target. Inthis model, VEGF gene promoter may regulate the expression of the luciferase reporter gene by responding to thestimulation of drug molecules. This cell model allows rapid and efficient screening of vascular-inducing activecomponents from several drug monomer molecules. Furthermore, we used rat bone marrow mesenchymal stem cells（rMSCs） to conduct a preliminary study on the activity of alantolactone. Using simvastatin as a positive control, weinvestigated the effects of alantolactone on the expression of vascular-related cell marker molecules such as VEGF andα-smooth muscle actin （α-SMA） in rMSCs. According to our results, 0.1, 1, 3 and 5 μM of alantolactone upregulated thetranscriptional luciferase gene activity of VEGF promoter, and a significant difference from that in the control group wasobserved. Among them, 3μM of alantolactone showed the better effect than that of 3 μM of simvastatin （P = 0.036） andother concentrations of alantolactone and simvastatin showed similar effects. Compared with that in the control group,rMSCs induced with 1μM alantolactone for 3 days showed a significant increase in the relative mRNA expressions ofVEGF and α-SMA genes. However, these effect of 5 μM alantolactone were weaker than those of 5 μM simvastatin （P 〈0.05）; rMSCs treated with 1 μM alantolactone for 3 days showed brighter green fluorescence （FITC marker） of α-SMAand VEGF in situ expression than that observed in the control group and similar fluorescence intensity than that ofsimvastatin group in an immunoradiometric assay. The above results demonstrate the reliability of the highly efficientsystem for screening of active drug molecules and confirmed the vascular induction function of alantolactone at the geneand protein levels.
基金This study was financially supported by 863 Hj-Tech ResearchDevelopment Program of China(2002AA326080)The Fund for Youth Teacher of Education Mlinistry of China(2002123).
文摘To develop a novel degradable poly (L-lactic acid)/β-tricalcium phosphate (PLLA/β-TCP) bioactive materials for bone tissueengineering, β-TCP powder was produced by a new wet process. Porous scaffolds were prepared by three steps, i.e. solventcasting, compression molding and leaching stage. Factors influencing the compressive strength and the degradation behaviorof the porous scaffold, e.g. weight fraction of pore forming agent-sodium chloride (NaCl), weight ratio of PLLA: β-TCP,the particle size of β-TCP and the porosity, were discussed in details. Rat marrow stromal cells (RMSC) were incorporatedinto the composite by tissue engineering approach. Biological and osteogenesis potential of the composite scaffold weredetermined with MTT assay, alkaline phosphatase (ALP) activity and bone osteocalcin (OCN) content evaluation. Resultsshow that PLLA/β-TCP bioactive porous scaffold has good mechanical and pore structure with adjustable compressive strengthneeded for surgery. RMSCs seeding on porous PLLA/β-TCP composite behaves good seeding efficacy, biocompatibility andosteoinductive potential. Osteoprogenitor cells could well penetrate into the material matrix and begin cell proliferation andosteogenic differentiation. Osseous matrix could be formed on the surface of the composite after culturing in vitro. It isexpected that the PLLA/β-TCP porous composites are promising scaffolds for bone tissue engineering in prosthesis surgery.
