A nonviral transfection approach in vitro: the design of a gold nanoparticle vector joint with microelectromechanical systems.
Au nanoparticles modified with 21-base thiolated-oligonucleotides have been evaluated as delivery vehicles for the development of a nonviral transfection platform. The electromigration combined with electroporation for DNA delivery in an osteoblast like cell was employed to test on microchips. Electroporation introduces foreign materials into cells by applying impulses of electric field to induce multiple transient pores on the cell membrane through dielectric breakdown of the cell membrane. On the basis of the characteristic surface plasmon of the Au particles, UV-vis absorption was utilized to qualitatively judge the efficiency of delivery. Transmission electron microscopy images and atomic absorption measurements (quantitative analysis) provided evidence of the bare Au and Au/oligonucleotide nanoparticles before and after electroporation and electromigration function. The experiments demonstrated that electrophoretic migration followed by electroporation significantly enhanced the transportation efficiency of the nanoparticle-oligonucleotide complexes as compared with electroporation alone. Most interestingly, Au capped with oligonucleotides led to optimal performance. On the other hand, the bare Au colloidal suspensions resulted in aggregation, which might be an obstacle to the internalization process. In addition, analytical results demonstrated an increase in the local particle concentrations on the cell surface that provided additional support for the mechanism underlying the improved Au nanoparticle transportation into cells in the presence of electromigration function.
Jen
Chun-Ping
CP
Department of Engineering Science, National Cheng Kung University, Tainan 701, Taiwan.
Chen
Yu-Hung
YH
Fan
Chun-sheng
CS
Yeh
Chen-Sheng
CS
Lin
Yu-Cheng
YC
Shieh
Dar-Bin
DB
Wu
Chao-Ling
CL
Chen
Dong-Hwang
DH
Chou
Chen-Hsi
CH
D016475:3T3 Cells; D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D004247:DNA / Q000737:chemistry; D004563:Electrochemistry / Q000379:methods*; D004566:Electrodes; D018274:Electroporation; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006046:Gold / Q000737:chemistry*; D051379:Mice; D046529:Microscopy, Electron, Transmission; D015233:Models, Statistical; D049329:Nanostructures / Q000737:chemistry*; D009841:Oligonucleotides / Q000737:chemistry; D010006:Osteoblasts / Q000166:cytology; D010957:Plasmids / Q000378:metabolism; D013053:Spectrophotometry; D020349:Surface Plasmon Resonance; D014162:Transfection; D014466:Ultraviolet Rays
D009841:Oligonucleotides; D006046:Gold; D004247:DNA