Adsorption of plasmid DNA onto N,N'- (dimethylamino)ethyl-methacrylate graft-polymerized poly-L-lactic acid film surface for promotion of in-situ gene delivery.
The surface of biodegradable poly-L-lactic acid (PLLA) film was modified with N,N'-(dimethylamino)ethyl-methacrylate (DMAEMA) via UV-induced graft copolymerization, and plasmid DNA molecules were adsorbed onto the surface of modified PLLA film by electrostatic interactions with cationic DMAEMA polymer. We characterized the structure of the modified PLLA film surface by Fourier transform infrared attenuated total reflection (FTIR-ATR) spectroscopy and X-ray photoelectron spectroscopy (XPS). The weight-average molecular weight (Mw) of grafted DMAEMA polymer chains was estimated from the elution time of gel filtration chromatography. C.I. Acid Orange 7 dyeing results indicated that graft density of DMAEMA on PLLA film increased with the UV irradiation time and then reached a saturated value. DNA adsorption density was proportioned to graft density of DMAEMA. Mouse fibroblast L929 cell line was cultured on modified PLLA films, and cell viability and gene transfection efficiency were monitored after 2 days culture. It was found that the DMAEMA grafted PLLA film had obvious cytotoxicity to the cells. On the contrary, cytotoxicity of the surface was highly decreased after adsorption with plasmid DNA. This DNA adsorbed DMAEMA modified PLLA showed the ability to deliver DNA into mammalian cells cultured on the surface with high-transfection efficiency at a low DNA amount. The present results suggest that the DMAEMA grafted PLLA has potentiality to be used as a safe and effective gene delivery system in gene-activated materials.
Jiang
Tingting
T
State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, Nanjing 210093, China.
Chang
Jianbin
J
Wang
Chunming
C
Ding
Zhi
Z
Chen
Jiangning
J
Zhang
Junfeng
J
Kang
En-Tang
ET
D000327:Adsorption; D000818:Animals; D002460:Cell Line; D002621:Chemistry / Q000379:methods*; D004247:DNA / Q000737:chemistry*; D018014:Gene Transfer Techniques; D019344:Lactic Acid / Q000737:chemistry*; D046911:Macromolecular Substances / Q000737:chemistry; D008689:Methacrylates / Q000737:chemistry*; D051379:Mice; D010957:Plasmids / Q000378:metabolism*; D011091:Polyesters; D011108:Polymers / Q000737:chemistry*; D013052:Spectrometry, X-Ray Emission; D017550:Spectroscopy, Fourier Transform Infrared; D055672:Static Electricity; D013499:Surface Properties
D046911:Macromolecular Substances; D008689:Methacrylates; D011091:Polyesters; D011108:Polymers; D019344:Lactic Acid; C033616:poly(lactide); D004247:DNA; C049840:2-(dimethylamino)ethyl methacrylate