| Restoration of rat calvarial defects by poly（lactide-co-glycolide）/ hydroxyapatite scaffolds loaded with bone mesenchymal stem cells and DNA complexes |
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| 引用本文： | LI Dan,WANG Wei,GUO Rui,QI YiYing,GOU ZhongRu,GAO ChangYou. Restoration of rat calvarial defects by poly（lactide-co-glycolide）/ hydroxyapatite scaffolds loaded with bone mesenchymal stem cells and DNA complexes[J]. 科学通报(英文版), 2012, 57(5): 435-444. DOI: 10.1007/s11434-011-4914-0 |
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| 作者姓名： | LI Dan WANG Wei GUO Rui QI YiYing GOU ZhongRu GAO ChangYou |
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| 作者单位： | [1]Key Laboratory of Macromolecular Synthesis and Functionalization of Ministry of Education, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027, China [2]Zhejiang-California International NanoSystems Institute, Hangzhou 310027, China [3]Department of Orthopedic Surgery, Second Affiliated Hospital, Zhejiang University, Hangzhou 310027, China [4]State Key Laboratory of Diagnosis and Treatment for Infectious Diseases, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China |
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| 基金项目： | This work was supported by the National Natural Science Foundation of China （20934003）, the Science Technology Program of Zhejiang Province （2009C14003, 2009C13020）, and the National Basic Research Program of China （2011CB606203）. |
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| 摘 要： | A composite construct comprising of a bone mesenchymal stem cell (BMSC) sheet, plasmid DNA, encoding human bone morphogenic protein-2 (hBMP-2), and poly(lactide-co-glycolide)/hydroxyapatite (PLGA/HA) sponge was designed and employed in the restoration of rat calvarial defects. To improve gene transfection efficiency, a cationic chitosan derivative, N,N,N,-trimethyl chitosan chloride (TMC), was employed as the vector. The TMC/DNA complexes had a transfection efficiency of 13% in rat BMSCs, resulting in heterogeneous hBMP-2 expression in a 10-d culture period in vitro. In vivo culture of the composite constructs was performed by implantation into rat full-thickness calvarial defects, using constructs lacking pDNA-hBMP-2 or BMSC sheets as controls. Significantly higher heterogeneous expression of hBMP-2 was detected in vivo at 2 weeks for the cell sheet/DNA complex/scaffold constructs, compared with the constructs lacking pDNA-hBMP-2 or BMSC sheets. New bone formation was evident as early as 4 weeks in the experimental constructs. At 8 weeks, partial bridging of calvarial defects was observed in the experimental constructs, which was significantly better than the constructs lacking pDNA-hBMP-2 or BMSC sheets. Therefore, the combination of the PLGA/HA scaffold with BMSC sheets and gene therapy vectors is effective at enhancing bone formation.
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| 关 键 词： | gene therapy bone regeneration bone marrow stem cells poly(lactide-co-glycolide) BMP-2 |
Restoration of rat calvarial defects by poly(lactide-co-glycolide)/hydroxyapatite scaffolds loaded with bone mesenchymal stem cells and DNA complexes |
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| Dan Li,Wei Wang,Rui Guo,YiYing Qi,ZhongRu Gou,ChangYou Gao. Restoration of rat calvarial defects by poly(lactide-co-glycolide)/hydroxyapatite scaffolds loaded with bone mesenchymal stem cells and DNA complexes[J]. Chinese science bulletin, 2012, 57(5): 435-444. DOI: 10.1007/s11434-011-4914-0 |
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| Authors: | Dan Li Wei Wang Rui Guo YiYing Qi ZhongRu Gou ChangYou Gao |
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| Affiliation: | 1,4* 1 Key Laboratory of Macromolecular Synthesis and Functionalization of Ministry of Education, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027, China; 2 Zhejiang-California International NanoSystems Institute, Hangzhou 310027, China; 3 Department of Orthopedic Surgery, Second Affiliated Hospital, Zhejiang University, Hangzhou 310027, China; 4 State Key Laboratory of Diagnosis and Treatment for Infectious Diseases, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China |
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| Abstract: | A composite construct comprising of a bone mesenchymal stem cell (BMSC) sheet, plasmid DNA, encoding human bone morphogenic protein-2 (hBMP-2), and poly(lactide-co-glycolide)/hydroxyapatite (PLGA/HA) sponge was designed and employed in the restoration of rat calvarial defects. To improve gene transfection efficiency, a cationic chitosan derivative, N,N,N,-trimethyl chitosan chloride (TMC), was employed as the vector. The TMC/DNA complexes had a transfection efficiency of 13% in rat BMSCs, resulting in heterogeneous hBMP-2 expression in a 10-d culture period in vitro. In vivo culture of the composite constructs was performed by implantation into rat full-thickness calvarial defects, using constructs lacking pDNA-hBMP-2 or BMSC sheets as controls. Significantly higher heterogeneous expression of hBMP-2 was detected in vivo at 2 weeks for the cell sheet/DNA complex/scaffold constructs, compared with the constructs lacking pDNA-hBMP-2 or BMSC sheets. New bone formation was evident as early as 4 weeks in the experimental constructs. At 8 weeks, partial bridging of calvarial defects was observed in the experimental constructs, which was significantly better than the constructs lacking pDNA-hBMP-2 or BMSC sheets. Therefore, the combination of the PLGA/HA scaffold with BMSC sheets and gene therapy vectors is effective at enhancing bone formation. |
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| Keywords: | gene therapy bone regeneration bone marrow stem cells poly（lactide-co-glycolide） BMP-2 |
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