A fast and sensitive method for measuring the integrity of siRNA-carrier complexes in full human serum

Buyens, Kevin; Lucas, Bart; Raemdonck, Koen; Braeckmans, Kevin; Vercammen, Jo; Hendrix, Jelle; Engelborghs, Yves; Smedt, Stefaan C. De; Sanders, Niek N.

doi:10.1016/j.jconrel.2007.10.024

Cited by 119 publications

(120 citation statements)

References 21 publications

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“…The release of siRNA from liposomal formulations in the peritoneal fluids was evaluated using Fluorescence Correlation Spectroscopy (FCS). As we previously demonstrated, FCS is an elegant technique to follow the complexation behavior of small nucleic acids to nanoparticles, both in buffer and in living cells [38,51]. The data in Fig 7. clearly suggest better complexation and slower release of siRNA for the non-PEGylated lipoplexes over the PEGylated ones.…”

Section: Release Of Sirna From Liposomes In the Peritoneal Fluidsmentioning

confidence: 79%

“…For this purpose, we utilize state of the art fluorescence techniques that were previously developed in our laboratory, namely Single Particle Tracking (SPT) and Fluorescence Correlation Spectroscopy (FCS) to respectively gain information on the aggregation of nanoparticles and the release of siRNA in undiluted biofluids [36,38]. The results are compared to measurements of the same nanoparticles dispersed in human serum.…”

Section: Introductionmentioning

confidence: 99%

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Colloidal stability of nano-sized particles in the peritoneal fluid: Towards optimizing drug delivery systems for intraperitoneal therapy

et al. 2014

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Intraperitoneal (IP) administration of nano-sized delivery vehicles containing small interfering RNA (siRNA) is recently gaining attention as an alternative route for the efficient treatment of peritoneal carcinomatosis. The colloidal stability of nanomatter following IP administration has, however, not been thoroughly investigated yet. Here, enabled by advanced microscopy methods such as Single Particle Tracking (SPT) and Fluorescence Correlation Spectroscopy (FCS), we follow the aggregation and cargo release of nano-scaled systems directly in peritoneal fluids from healthy mice and ascites fluid from a patient diagnosed with peritoneal carcinomatosis. The colloidal stability in the peritoneal fluids was systematically studied in function of the charge (positive or negative) and Poly-Ethylene Glycol (PEG) degree of liposomes and polystyrene nanoparticles, and compared to human serum. Our data demonstrate strong aggregation of cationic and anionic nanoparticles in the peritoneal fluids, while only slight aggregation was observed for the PEGylated ones. PEGylated liposomes, however, lead to a fast and premature release of siRNA cargo in the peritoneal fluids. Based on our observations, we reflect on how to tailor improved delivery systems for IP therapy.

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Section: Release Of Sirna From Liposomes In the Peritoneal Fluidsmentioning

confidence: 79%

Section: Introductionmentioning

confidence: 99%

Colloidal stability of nano-sized particles in the peritoneal fluid: Towards optimizing drug delivery systems for intraperitoneal therapy

et al. 2014

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“…Furthermore, the PEG-siPlex loaded microbubbles are expected to be suitable for systematic applications as ultrasound in combination with microbubbles is considered as a safe and already used in the clinic for diagnostic purposes. Additionally, PEGsiPlexes are known not to aggregate in serum which is important to avoid blockage of small blood capillaries by aggregates [49]. The developed siRNA delivery system may also allow the treatment of patients with metastasized tumours.…”

Section: Resultsmentioning

confidence: 99%

Ultrasound assisted siRNA delivery using PEG-siPlex loaded microbubbles

Vandenbroucke

Lentacker

Demeester

et al. 2008

Journal of Controlled Release

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109

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Short interfering RNA (siRNA) attracts much attention for the treatment of various diseases. However, its delivery, especially via systemic routes, remains a challenge. Indeed, naked siRNAs are rapidly degraded, while complexed siRNAs massively aggregate in the blood or are captured by macrophages. Although this can be circumvented by PEGylation, we found that PEGylation had a strong negative effect on the gene silencing efficiency of siRNA-liposome complexes (siPlexes). Recently, ultrasound combined with microbubbles has been used to deliver naked siRNA but the gene silencing efficiency is rather low and very high amounts of siRNA are required. To overcome the negative effects of PEGylation and to enhance the efficiency of ultrasound assisted siRNA delivery, we coupled PEGylated siPlexes (PEG-siPlexes) to microbubbles. Ultrasound radiation of these microbubbles resulted in massive release of unaltered PEG-siPlexes. Interestingly, PEG-siPlexes loaded on microbubbles were able to enter cells after exposure to ultrasound, in contrast to free PEG-siPlexes, which were not able to enter cells rapidly. Furthermore, these PEG-siPlex loaded microbubbles induced, in the presence of ultrasound, much higher gene silencing than free PEG-siPlexes. Additionally, the PEG-siPlex loaded microbubbles only silenced the expression of genes in the presence of ultrasound, which allows space and time controlled gene silencing.

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“…Challenges to efficient delivery include nanoparticle dissociation via serum proteins (10,11), cellular uptake (12), endosomal escape (13), and appropriate intracellular disassembly (14,15). To address some of these challenges, single-parameter studies that evaluate the effect of chemical structure on a single biological property or on delivery performance have been carried out (16)(17)(18)(19)(20)(21).…”

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confidence: 99%

Multiparametric approach for the evaluation of lipid nanoparticles for siRNA delivery

Alabi

Love

Sahay

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

136

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Nanoparticle-mediated siRNA delivery is a complex process that requires transport across numerous extracellular and intracellular barriers. As such, the development of nanoparticles for efficient delivery would benefit from an understanding of how parameters associated with these barriers relate to the physicochemical properties of nanoparticles. Here, we use a multiparametric approach for the evaluation of lipid nanoparticles (LNPs) to identify relationships between structure, biological function, and biological activity. Our results indicate that evaluation of multiple parameters associated with barriers to delivery such as siRNA entrapment, pK a , LNP stability, and cell uptake as a collective may serve as a useful prescreening tool for the advancement of LNPs in vivo. This multiparametric approach complements the use of in vitro efficacy results alone for prescreening and improves in vitro-in vivo translation by minimizing false negatives. For the LNPs used in this work, the evaluation of multiple parameters enabled the identification of LNP pK a as one of the key determinants of LNP function and activity both in vitro and in vivo. It is anticipated that this type of analysis can aid in the identification of meaningful structure-functionactivity relationships, improve the in vitro screening process of nanoparticles before in vivo use, and facilitate the future design of potent nanocarriers.RNAi | thiol-yne | gene silencing | drug carrier

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A fast and sensitive method for measuring the integrity of siRNA-carrier complexes in full human serum

Cited by 119 publications

References 21 publications

Colloidal stability of nano-sized particles in the peritoneal fluid: Towards optimizing drug delivery systems for intraperitoneal therapy

Colloidal stability of nano-sized particles in the peritoneal fluid: Towards optimizing drug delivery systems for intraperitoneal therapy

Ultrasound assisted siRNA delivery using PEG-siPlex loaded microbubbles

Multiparametric approach for the evaluation of lipid nanoparticles for siRNA delivery

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