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. Author manuscript; available in PMC: 2015 Nov 1.
Published in final edited form as: Biomaterials. 2014 Aug 8;35(34):9343–9354. doi: 10.1016/j.biomaterials.2014.07.043

Figure 3. Functional evaluation of MLNPs for transfection and cytotoxicity.

Figure 3

Population transfection optimization on 293T cells. MLNPs were loaded with pLuc. A. Toxicity of MLNPs after surface modification. B. The effect of surface modification on particle transfection. All MLNPs were synthesized with DCM. C. The effect of utilizing a smaller PEG1 linker, changing the organic solvent during fabrication to EA, and encapsulating TCHD within the PLGA core on particle transfection. D. The effect of initial seeding density on particle transfection. E. Percent cell transfection. FACS of 293T cells transfected with MLNPs or lipofectamine particles carrying pGFP. Fluorescence measured via the FL1-H channel detected transfected cells. After 72 h, approximately 37% of cells were transfected by MLNPs and 58% of cells were transfected by lipofectamine. The dotted green line indicates a gate containing at least 99% of the control population. *P<0.05. DCM – dichloromethane, EA – ethyl acetate, FACS - fluorescence-activated cell sorting, PEG - polyethylene glycol, PEG1 – 1 kDa PEG, PEG5 – 5 kDa PEG, PLGA - poly(lactic-co-glycolic) acid, PD - PLGA nanoparticles with surface polyethyleneimine (PEI) and plasmid DNA, PDP - PD particles modified with a second layer of PEI, PDP-PEG-mAP - PDP particles modified with a heterobifunctional PEG linker and modified antennapedia. MLNP – multi-layered nanoparticles, pLuc – luciferase encoding plasmid, pGFP – green fluorescent protein encoding plasmid, TCHD - trans-1,2-Cyclohexanediol.