Restoration of rat calvarial defects by poly(lactide-co-glycolide)/hydroxyapatite scaffolds loaded with bone mesenchymal stem cells and DNA complexes  

作　　者：LI Dan WANG Wei GUO Rui QI YiYing GOU ZhongRu GAO ChangYou 

机构地区：[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

出　　处：《Chinese Science Bulletin》2012年第5期435-444,共10页

基　　金：supported by the National Natural Science Foundation of China (20934003);the Science Technology Program of Zhejiang Province (2009C14003, 2009C13020);the National Basic Research Program of China (2011CB606203)

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.A composite construct comprising of a bone mesenchymal stem cell （BMSC） sheet, plasmid DNA, encoding human bone mor- phogenic 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 con- structs 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 for- marion was evident as early as 4 weeks in the experimental constructs. At 8 weeks, partial bridging of calvarial defects was ob- served 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.

关 键 词：骨髓基质干细胞 质粒DNA 羟基磷灰石 修复工作 骨缺损 复合物 乙醇酸 大鼠 

分 类 号：Q813.1[生物学—生物工程] Q781