Transcutaneous iontophoretic delivery of STAT3 siRNA using layer-by-layer chitosan coated gold nanoparticles to treat melanoma.
Overexpression of signal transducer and activator of transcription 3 (STAT3) protein prevents apoptosis and enhances proliferation of melanocytes. The aim of this study was to investigate the feasibility of using layer-by-layer assembled gold nanoparticles (LbL-AuNP) as a carrier for iontophoretic delivery of STAT3 siRNA to treat melanoma. Chitosan coated AuNP (AuNP-CS) were prepared by direct reduction of HAuCl4 in the presence of chitosan. The AuNP-CS were then sequentially layered with siRNA and chitosan to form AuNP-CS/siRNA/CS. STAT3 siRNA replaced with scrambled siRNA or sodium alginate were used as controls. The average particle size and zeta-potential of the prepared LbL-AuNP were 150±10nm (PDI: 0.41±0.06) and 35±6mV, respectively. In vitro studies in B16F10 murine melanoma cells showed that AuNP-CS/siRNA/CS inhibited the cell growth by 49.0±0.6% and 66.0±0.2% at 0.25nM and 0.5nM STAT3 siRNA concentration, respectively. Fluorescence microscopy and flow cytometry studies showed a time dependent cell uptake of the LbL-AuNP up to 120min. Clathrin mediated endocytosis was found to be the predominant cell uptake mechanism for LbL-AuNP. STAT3 siRNA loaded LbL-AuNP reduced the STAT3 protein expression by 47.3% in B16F10 cells. Similarly, apoptosis assay showed 29% and 44% of early and late apoptotic events, respectively after treatment with STAT3 siRNA loaded LbL-AuNP. Confocal microscope and skin cryosections showed that application of 0.47mA/cm(2) of anodal iontophoresis enhanced the skin penetration of LbL-AuNP to reach viable epidermis. In conclusion, layer-by-layer chitosan coated AuNP can be developed as a carrier for iontophoretic delivery of STAT3 siRNA to treat melanoma.
Department of Pharmacy, Birla Institute of Technology and Science (BITS) Pilani, Hyderabad Campus, Shameerpet, Hyderabad 500078, India.
Suman
S
Labala
Department of Pharmacy, Birla Institute of Technology and Science (BITS) Pilani, Hyderabad Campus, Shameerpet, Hyderabad 500078, India.
Anup
A
Jose
Department of Pharmacy, Birla Institute of Technology and Science (BITS) Pilani, Hyderabad Campus, Shameerpet, Hyderabad 500078, India. Electronic address: vamsi@hyderabad.bits-pilani.ac.in.
Venkata Vamsi Krishna
VV
Venuganti
D000279:Administration, Cutaneous; D000818:Animals; D048271:Chitosan / Q000737:chemistry*; D004337:Drug Carriers / Q000737:chemistry*; D016503:Drug Delivery Systems; D020868:Gene Silencing; D006046:Gold / Q000737:chemistry*; D007478:Iontophoresis; D008546:Melanoma, Experimental / Q000188:drug therapy* / Q000473:pathology; D053768:Metal Nanoparticles / Q000737:chemistry*; D051379:Mice; D008856:Microscopy, Fluorescence; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics; D050796:STAT3 Transcription Factor / Q000037:antagonists & inhibitors*; D013552:Swine; D014407:Tumor Cells, Cultured
D004337:Drug Carriers; D034741:RNA, Small Interfering; D050796:STAT3 Transcription Factor; D006046:Gold; D048271:Chitosan
B16F10 cells; Chitosan; Gold nanoparticles; Layer-by-layer assembly; Melanoma; STAT3 siRNA; Skin penetration