RESUMO
Iron oxide nanoparticles (IONPs) synthesized via thermal decomposition find diverse applications in biomedicine owing to precise control of their physico-chemical properties. However, use in such applications requires phase transfer from organic solvent to water, which remains a bottleneck. Through the thermal decomposition of iron oleate (FeOl), we systematically investigate the impact of synthesis conditions such as oleic acid (OA) amount, temperature increase rate, dwell time, and solvent on the size, magnetic saturation, and crystallinity of IONPs. Solvent choice significantly influences these properties, manipulating which, synthesis of monodisperse IONPs within a tunable size range (10-30 nm) and magnetic properties (75 to 42 Am2Kg-1) is obtained. To enable phase transfer of IONPs, we employ flash nanoprecipitation (FNP) for the first time as a method for scalable and precise size control, demonstrating its potential over conventional methods. Poly(lactic-co-glycolic acid) (PLGA)-coated IONPs with hydrodynamic diameter (Hd) in the range of 250 nm, high colloidal stability and high IONPs loadings up to 43% were obtained, such physicochemical properties being tuned exclusively by the size and hydrophobicity of starting IONPs. They showed no discernible cytotoxicity in human dermal fibroblasts, highlighting the applicability of FNP as a novel method for the functionalization of hydrophobic IONPs for biomedicine.
RESUMO
Solid-phase reversible immobilization (SPRI) bead technology is widely used in molecular biology for convenient DNA manipulation. However, commercial SPRI bead kits lack cost advantages and flexibility. It is, therefore, necessary to develop new and alternative cost-effective methods of on-par or better quality. Herein, an easy and cost-effective method is proposed for synthesizing polyacrylic acid-coated iron oxide nanoparticles (PAA-IONPs) through in situ polymerization at lab scale for high-efficiency nucleic acid extraction and size selection. A design of experiment (DoE) approach was used to investigate the influence of iron oxide nanoparticles (IONPs), acrylic acid (AA) monomer, and sodium dodecyl sulfate (SDS) surfactant amounts on the sizes and carboxyl group densities of PAA-IONPs. Thorough characterization by thermogravimetric analysis (TGA), attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR) and vibrating sample magnetometry (VSM) highlights the importance of a low starting pH achieved by a high ratio of AA/IONPs, to yield the largest sizes (554 nm) and highest carboxyl group densities (2.13 mmol g-1) obtained in this study. An efficient DNA purification strategy is then presented using homemade beads-suspension buffer and optimized bead concentrations (17% PEG 8000, 2.5 M NaCl, and 3 mg mL-1 PAA-IONPs). This method shows comparable performance to the control (AMPure XP beads) for DNA recovery. An adjustable PAA-IONPs DNA purification system was also developed to be used for DNA-size selection at low DNA amounts (50-100 ng) with a high degree of resolution and recovery. In conclusion, this work offers an optimized PAA-IONPs synthesis protocol and a flexible DNA purification approach that will enable researchers to manipulate DNA under various conditions, holding the significant potential to benefit future molecular biology research and diagnostics.