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Arginine-chitosan nanoparticles for siRNA transport

Small interfering RNAs (siRNAs) can be used as in gene therapies for e.g. cancers such as leukemia. However, the application of these is limited by a lack of efficient drug delivery. Therefore, in the presented study, chitosan was functionalized with arginine to form nanoparticles, loaded with siRNA, and their properties as siRNA vectors were investigated.

A NOVEL FORM OF ARGININE-CHITOSAN AS NANOPARTICLES EFFICIENT FOR SIRNA DELIVERY INTO MOUSE LEUKEMIA CELLS

Luo, J.; Chen, J.; Liu, Y.; He, Y.; Dong, W. A Novel Form of Arginine-Chitosan as Nanoparticles Efficient for siRNA Delivery into Mouse Leukemia Cells. Int. J. Mol. Sci. 2023, 24, 1040. https://doi.org/10.3390/ijms24021040

Small interfering RNAs (siRNA) are double-stranded RNAs with a length of 21-25 bp that are able to bind mRNA of specific genes and thus inactivate them. This makes it particularly interesting for gene therapeutics. However, the application of siRNA is limited by a lack of efficient transport to target cells especially in so-called solid tumor cells such as leukemia cells. Transport of siRNA can occur through viral and non-viral vectors. Viral vectors show good transport efficiency, but the risk of mutations makes them medically too unsafe. An alternative are non-viral vectors, e.g. liposomes, polymers or polypeptides. Their transport efficiencies have been significantly improved by the advent of nanoparticles.

Chitosan in particular is of interest as a non-viral vector. Due to its cationic properties, it is able to bind negatively charged oligonucleotides. It also shows good biocompatibility, biodegradability and low toxicity. In previous studies, it was also observed that chitosan in drug transport had improved drug stability. Therefore, in the presented study, chitosan (480-776 Da) was modified with arginine to form nanoparticles (NPs) and investigated for their ability to transport siRNA to mouse L1210 leukemia cells. For this purpose, chitosan and arginine were physically mixed, then freeze-dried, and the resulting NPs were characterized. To obtain the ideal siRNA-NP ratio, different ratios of siRNA and arginine-chitosan NPs (Arg-CS NPs) were investigated.

RESULTS

  • Arginine was successfully covalently bound to chitosan via EDC/NHS method, detection via FTIR
  • Arg-CS was identified as NP by AFM and TEM, they showed a size of 75.76±12.07 nm
  • Based on the change of zeta potential from 0.57±0.14 mV to 16.63±1.83 mV, it was demonstrated that Arg-CS NPs are able to encapsulate siRNA in vitro via electrostatic interaction
  • size Arg-CS NPs siRNA complexes distributed between 124.83±11.5 nm and 1261.90±438.88 nm
  • good biocompatibility of Arg-CS NPs siRNA complexes in a ratio between 20:1 and 50:1 against L1210 leukemia cells
  • Successful transfection of L1210 cells by knockout of Rhoa genes in L1210 cells→Transport of siRNA via Arg-CS NPs possible

Conclusions: In the presented study, arginine and chitosan were successfully synthesized into Arg-CS nanoparticles. These were shown to be able to successfully encapsulate siRNA and safely transport Rhoa siRNA into L1210 leukemia cells.

Link to article: https://www.mdpi.com/1422-0067/24/2/1040

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drug delivery, transfection, chitosan, nanoparticles, cancer

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