Nova nanosem 230

Crystallographic phases of all the products were investigated by powder X-ray diffraction (XRD, Rigaku D/max2500) with Cu Kα (λ = 1.5406 Å) radiation. The morphologies of the samples were examined by field-emission scanning electron microscopy (SEM, FEI Nova NanoSEM 230) and transmission electron microscopy (TEM; JEOL-JEM-2100F transmission electron microscope). A combined Differential Scanning Calorimetry (DSC)/Thermogravimetric Analysis (TGA) instrument (Netzsch STA449C, Germany) was used to study the reactions during the annealing process and measure the carbon content in CW-LVP. Raman spectra were obtained using a Renishaw INVIA micro-Raman spectroscopy system. Nitrogen adsorption-desorption measurements were conducted at 77K (NOVA 4200e, Quantachrome Instruments).

The pDNA‐NPs were prepared by the double emulsion solvent evaporation method (w/o/w). Briefly, 400 μl of diethylpyrocarbonate (DEPC) water containing 100 μg of plasmid DNA was added to 1 ml of dichloromethane containing PEI₂₅₀₀₀‐C₁₄ (2 mg) and PLGA (10 mg). The above mixture was sonicated for 2 min at 60 W in an ice bath to form a primary W/O emulsion. Then, 3 ml DEPC water was added to the primary emulsion and further emulsified by ultrasound for 2 min at 60 W to form a secondary W/O/W emulsion. Subsequently, the dichloromethane was removed by a rotary evaporator (RE‐52AA, Shanghai), and the pDNA‐NPs were finally obtained. The morphology and size of the nanoparticles were characterized by scanning electron microscopy (SEM, Nova NanoSEM 230, USA) and transmission electron microscopy (TEM, JEOL, Japan). Nano ZS (Malvern Instruments, Malvern, UK) was used to investigate the hydrodynamic size (DLS) and zeta potential of pDNA‐NPs.

The crystal structure and the composition of the samples were investigated by X-ray diffraction (XRD, Rigaku Dmax/2550VB + 18 kW) and energy dispersive X-ray spectroscopy (EDX, FEI Nova Nano SEM230). The morphology and microstructure of samples was observed through scanning electron microscopy (SEM, FEI Nova Nano SEM230) and transmission electron microscopy (TEM, JEOL JEM-2010). The thermogravimetric analysis (TGA) was tested in air at a ramp rate of 10°C min⁻¹. The Raman spectrum test was conducted on a Renishaw in Via 2000. The specific surface areas and the pore size distribution were measured by the Brunauer-Emmett-Teller (BET) test and Barrett-Joyner-Halenda (BJH) method, respectively. The surface chemical composition of the sample was tested by the X-ray photoelectron spectroscopy (XPS, Thermo Scientific ESCALAB 250XI).

The crystalline structure was tested using the X-ray diffractometer (XRD, Rigaku3014) with Cu-Kα1 radiation (λ = 0.15418 nm). The morphologies of the samples were observed by using a Nova NanoSEM 230 and a JEOL JEM-2100 TEM. The chemical composition of the samples was investigated by using X-ray photoelectron spectroscopy (XPS, Thermo ESCALAB 250) with an Al-Kα radiation. The Brunauer–Emmett–Teller (BET) surface area was tested by V-Sorb 2800P in an N₂ atmosphere.