PAMAM-triamcinolone acetonide conjugate as a nucleus-targeting gene carrier for enhanced transfer activity.
The excellent transfection efficiency and viability are essential for successful gene therapy. It suggested that when bound to its glucocorticoid receptor, glucocorticoid steroid can dilate the nuclear pore complexes and facilitated the transport of pDNA into the nucleus. In this research, the two different degrees of substitution of PAMAM-triamcinolone acetonide (PAMAM-TA) conjugates were synthesised for efficient translocation of pDNA into the nucleus. The physicochemical properties of the polyplexes were investigated by agarose gel electrophoresis, Zeta-sizer and TEM. They both could form nano-size polyplexes with pDNA. The polyplexes were very stable and showed excellent buffering capacities, facilitating endosomal escape, and no obvious difference was found between them. The TA-conjugated PAMAM-mediated transfection of luciferase and EGFP genes showed better transfer activity than native PAMAM and was comparable to the PEI 25K (polyethylenimine), and lower cytotoxicity in HEK 293 and HepG 2 cells. Even with 10% serum, their transfer activity was still high relatively. In addition, confocal microscopy examination confirmed that the enhancing mechanism for enhanced gene transfer activity of PAMAM-TA conjugate may involve the nuclear translocation of the polyplex. The low substituted degree of TA to 0.22 did not interrupt its nuclear localization potency. These findings demonstrated that the TA-grafted PAMAM dendrimer is a potential candidate as a safe and efficient gene delivery carrier for gene therapy.
Ma
Kun
K
Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, PR China.
Hu
Min-Xin
MX
Qi
Yan
Y
Zou
Ji-Hong
JH
Qiu
Li-Yan
LY
Jin
Yi
Y
Ying
Xiao-Ying
XY
Sun
Hong-Ying
HY
D021581:Active Transport, Cell Nucleus; D001672:Biocompatible Materials; D001709:Biotechnology / Q000379:methods; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D050091:Dendrimers; D011992:Endosomes / Q000378:metabolism; D018390:Gene Targeting / Q000295:instrumentation* / Q000379:methods; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D049452:Green Fluorescent Proteins / Q000737:chemistry; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D046529:Microscopy, Electron, Transmission; D011073:Polyamines / Q000737:chemistry*; D014162:Transfection; D014222:Triamcinolone Acetonide / Q000737:chemistry*
D001672:Biocompatible Materials; D050091:Dendrimers; C104700:PAMAM Starburst; D011073:Polyamines; D049452:Green Fluorescent Proteins; D014222:Triamcinolone Acetonide