Cellular Uptake and Distribution of Gemini Surfactant Nanoparticles Used as Gene Delivery Agents

Wei Jin; Mays Al-Dulaymi; Ildiko Badea; Scot C Leary; Jeveria Rehman; Anas El-Aneed

doi:10.1208/s12248-019-0367-1

Cellular Uptake and Distribution of Gemini Surfactant Nanoparticles Used as Gene Delivery Agents

AAPS J. 2019 Aug 6;21(5):98. doi: 10.1208/s12248-019-0367-1.

Authors

Wei Jin¹, Mays Al-Dulaymi¹, Ildiko Badea¹, Scot C Leary², Jeveria Rehman³, Anas El-Aneed⁴

Affiliations

¹ Drug Design & Discovery Group, College of Pharmacy and Nutrition, University of Saskatchewan, 107 Wiggins Road, Saskatoon, Saskatchewan, S7N 5E5, Canada.
² Department of Biochemistry, Microbiology and Immunology, University of Saskatchewan, 107 Wiggins Road, Saskatoon, Saskatchewan, S7N 5E5, Canada.
³ Department of Chemistry, University of Saskatchewan, 110 Science Place, Saskatoon, Saskatchewan, S7N 5C9, Canada.
⁴ Drug Design & Discovery Group, College of Pharmacy and Nutrition, University of Saskatchewan, 107 Wiggins Road, Saskatoon, Saskatchewan, S7N 5E5, Canada. anas.el-aneed@usask.ca.

PMID: 31388860
DOI: 10.1208/s12248-019-0367-1

Abstract

Gemini surfactants are promising molecules utilized as non-viral gene delivery vectors. However, little is known about their cellular uptake and distribution after they release their therapeutic cargo. Therefore, we quantitatively evaluated the cellular uptake and distribution of three gemini surfactants: unsubstituted (16-3-16), with pyridinium head groups (16(Py)-S-2-S-16(Py)) and substituted with a glycyl-lysine di-peptide (16-7N(GK)-16). We also assessed the relationship between cellular uptake and distribution of each gemini surfactant and its overall efficiency and toxicity. Epidermal keratinocytes PAM 212 were treated with gemini surfactant nanoparticles formulated with plasmid DNA and harvested at various time points to collect the enriched nuclear, mitochondrial, plasma membrane, and cytosolic fractions. Gemini surfactants were then extracted from each subcellular fraction and quantified using a validated flow injection analysis-tandem mass spectrometry (FIA-MS/MS) method. Mass spectrometry is superior to the use of fluorescent tags that alter the physicochemical properties and pharmacokinetics of the nanoparticles and can be cleaved from the gemini surfactant molecules within biological systems. Overall, a significantly higher cellular uptake was observed for 16-7N(GK)-16 (17.0%) compared with 16-3-6 (3.6%) and 16(Py)-S-2-S-16(Py) (1.4%), which explained the relatively higher transfection efficiency of 16-7N(GK)-16. Gemini surfactants 16-3-16 and 16(Py)-S-2-S-16(Py) displayed similar subcellular distribution patterns, with major accumulation in the nucleus, followed by the mitochondrion, cytosol, and plasma membrane. In contrast, 16-7N(GK)-16 was relatively evenly distributed across all four subcellular fractions. However, accumulation within the nucleus after 5 h of treatment was the highest for 16(Py)-S-2-S-16(Py) (50.3%), followed by 16-3-16 (41.8%) and then 16-7N(GK)-16 (33.4%), possibly leading to its relatively higher toxicity. Graphical Abstract.

Keywords: FIA-MS/MS; gemini surfactants; gene delivery; subcellular distribution; toxicity; transfection.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line
Flow Injection Analysis
Gene Transfer Techniques*
Keratinocytes / cytology
Keratinocytes / metabolism*
Mice
Nanoparticles*
Surface-Active Agents / chemistry*
Tandem Mass Spectrometry
Time Factors
Transfection

Substances

Surface-Active Agents