Fluorescent Nanodiamond-Assisted Delivery of siRNAs
Date Issued
2010
Date
2010
Author(s)
Lee, Chia-Ying
Abstract
Delivery technique holds the key to successful treatment of cancer and many diseases. For example, gene therapy is a promising method which manipulates the defective genes by delivery nucleotides into cells to treat inherited and acquired diseases that are currently considered incurable. It has been exercising in many clinical trials. RNA interference has been considered one of the most promising therapeutic platforms by introducing siRNA into the cell and switch off certain disease-relating genes. There are many vectors reported to be useful for genetic delivery, however, most of them have the concerns of cytotoxicities due to the component molecule and the induction of long-term damage. For these reasons, an efficient delivery system for siRNA remains to be developed.Type Ib fluorescent nanodiamonds (FNDs) can emit no photobleaching and no photoblinking fluorescence from their nitrogen-vacancy point defects, at a spectral range well suited for long term observation in living cells. They have good biocompatibility and can be easily functionalized for specific or nonspecific binding with biomolecules. In this study, based on the experiences of oral cancer, we propose to use cell and animal based oral cancer models to address the biological responses elicited by FNDs, and to simultaneously assess the best combinations of functionalization and cargo loading for future applications. We combined the FND with siRNA and common transfection reagents, applied with mild centrifugation to explore the gene-inhibition efficiency. In the same time, we studied the uptake mechanism and cellular response to the internalization of bare and cargo loaded FNDs. Besides, we tested the genetic delivery efficiency of FND-optimized siRNA complexes into xenografe bioluminescence-marked oral cancer animal models. From our results, FND showed dramatic improvement in silencing effect with good biocompatibility. And FND may be internalized into cells by macropinocytosis besides normal endocytosis pathways. We found that centrifugation facilitated the entrance of FND by increasing the contact between the particles and cellular surface, thus triggered more macropinocytosis responses to include more siRNA-combined FNDs. We provide the first and fundamental knowledge for a well-grounded development in biomedical applications of the FNDs. This is a powerful and promising achievement for subsequent improvements towards the final achievement of revolutionary new capabilities in the diagnosis, tracing and curing of cancer, as well as the treatment of other difficult diseases.
Subjects
gene therapy
fluorescent nano-diamond
siRNA
SDGs
Type
thesis
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