Efficient gene delivery to mesenchymal stem cells by an ethylenediamine-modified polysaccharide from mulberry leaves.
This study investigates the use of a natural polysaccharide isolated from mulberry leaves as a nonviral gene vector. Ethylenediamine is chemically grafted to the backbone of a polysaccharide from mulberry leaves (MPS) to acquire nucleic acid binding affinity. A particle-size observation indicates that the cationic mulberry leaf polysaccharide (CMPS) can efficiently combine with plasmid transforming growth factor β1 (TGF-β1) to form nanoscaled particles. In addition, the electrophoresis assay indicates a retarded plasmid migration when the CMPS/pTGF-β1 weight ratio is increased to 30:1. The in vitro cell transfection experiment is performed based on bone marrow mesenchymal stem cells (MSCs) derived from rat femurs and tibias, and the findings reveal that the complex with a CMPS/pTGF-β1 weight ratio of 50:1 exhibits the highest cell transfection effect, which is significantly higher than that of branched poly(ethyleneimine) (PEI) (25 kDa; p = 0.001, Student's t-test) and slightly higher than Lipofectamine 2000. Moreover, the cytotoxicity assay also demonstrates that all of these tested complexes and the plasmid TGF-β1 are nontoxic to mesenchymal stem cells (MSCs). The results of the living cell imaging confirm that more of the CMPS/plasmid TGF-β1 nanoparticles can be taken up and at a faster rate by the MSCs than by the positive control Lipofectamine 2000; these data are consistent with the transfection efficiency data. Together, these results suggest that the CMPS/pTGF-β1 nanoparticle can potentially be developed into a promising alternative for the transfer of therapeutic genes into cells.
Department of Pharmaceutics and Tissue Engineering, School of Pharmacy, Jiangsu University, Zhenjiang 212001, PR China.
Wen-Wen
WW
Deng
Xia
X
Cao
Miao
M
Wang
Yan
Y
Yang
Wei-Yan
WY
Su
Ya-Wei
YW
Wei
Zhen
Z
Ou-Yang
Jiang-Nan
JN
Yu
Xi-Ming
XM
Xu
D000818:Animals; D002412:Cations; D016923:Cell Death / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003432:Cross-Linking Reagents / Q000494:pharmacology; D024202:Electrophoretic Mobility Shift Assay; D005029:Ethylenediamines / Q000737:chemistry*; D018014:Gene Transfer Techniques; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D031625:Morus / Q000187:drug effects / Q000378:metabolism*; D053758:Nanoparticles / Q000648:ultrastructure; D010316:Particle Size; D018515:Plant Leaves / Q000187:drug effects / Q000378:metabolism*; D011134:Polysaccharides / Q000737:chemistry*; D051381:Rats; D017550:Spectroscopy, Fourier Transform Infrared; D055672:Static Electricity; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology
D002412:Cations; D003432:Cross-Linking Reagents; D005029:Ethylenediamines; D011134:Polysaccharides; D053773:Transforming Growth Factor beta1