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Superparamagnetic iron oxide nanoparticles functionalized by peptide nucleic acids

机译：肽核酸功能化的超顺磁性氧化铁纳米颗粒

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A novel efficient method has been developed for covalently linking Peptide Nucleic Acid (PNA) oligomers and superparamagnetic iron oxide nanoparticles (SPION), to produce water soluble hybrid nanomaterials that can act as MRI contrast agents, as hyperthermia promoters and as PNA carriers. The multistep procedure involves: (i) preparation of oleate-stabilized SPION by using the thermal decomposition method, to control the size of the magnetic core (here 17 ± 2 nm, by TEM measurement); (ii) exchange of the oleate layer by dimercaptosuccinic acid (DMSA), to impart water solubility and to provide functional groups for PNA grafting; (iii) functionalization of a PNA oligomer with a terminal maleimide moiety, to allow SPION–PNA conjugation by thiol-maleimide Michael addition reaction, exploiting the SH groups of DMSA on the SPION surface. The method was tested using a model PNA decamer containing all four nucleobases (–CTAGATCACT–). SPION–PNA conjugation by SH addition was found more efficient than conjugation through amide bond between the COOH groups of DMSA and the terminal NH2 groups of PNA. Elemental analysis, UV-Vis and IR spectra, and ζ-potential measures confirmed the PNA binding (a loading of ca. 400 PNA strands per SPION was estimated, molar ratio ca. 1?:?15 with respect to DMSA). A detailed characterization of the morphology, relaxivity and magnetic properties of the SPION used for PNA binding is reported, and compared to the one relative to the SPION–PNA conjugate. The analysis of the magnetic behaviour showed that the nanoparticles are in the superparamagnetic regime at room temperature, and have a considerably high saturation magnetization (85 emu g?1). Magnetic hyperthermia measurements gave a remarkable value of the heat released (SAR = 65 W g?1), which makes these SPION suitable for magnetic hyperthermia treatments. The SPION were also able to effectively shorten both longitudinal and transverse relaxation times of water, with r1 and r2 values higher with respect to routinely used commercial contrast agents, at the typical fields of clinical instrumentation.
机译：已经开发出一种新颖的有效方法，用于共价连接肽核酸（PNA）寡聚物和超顺磁性氧化铁纳米颗粒（SPION），以生产可充当MRI造影剂，热疗促进剂和PNA载体的水溶性杂化纳米材料。多步骤程序涉及：（i）通过热分解方法制备油酸酯稳定的SPION，以控制磁芯的尺寸（此处为17±2 nm，通过TEM测量）; （ii）通过二巯基琥珀酸（DMSA）交换油酸酯层，以赋予水溶性并提供用于PNA接枝的官能团; （iii）利用末端马来酰亚胺部分对PNA低聚物进行功能化，以允许SPION-PNA通过硫醇-马来酰亚胺迈克尔加成反应进行偶联，从而利用SPION表面的DMSA的SH基团。使用包含所有四个核碱基（–CTAGATCACT–）的模型PNA decamer测试了该方法。发现通过SH添加的SPION–PNA共轭比通过DMSA的COOH基团与PNA的末端NH 2 基团之间的酰胺键共轭更有效。元素分析，UV-Vis和IR光谱以及ζ电位测量结果证实了PNA的结合（估计了 ca。每条SPION的400条PNA链的负载量，摩尔比< em> ca。 1？：？15（相对于DMSA）。报道了用于PNA结合的SPION的形态，弛豫性和磁性的详细表征，并与相对于SPION-PNA结合物的表征进行了比较。磁性的分析表明，纳米粒子在室温下处于超顺磁性状态，并具有相当高的饱和磁化强度（85 emu g ？1 ）。磁热疗测量显示出显着的释放热量值（SAR = 65 W g ？1 ），这使这些SPION适用于磁疗。 SPION还可以通过 r 1 和 r 有效地缩短水的纵向和横向松弛时间。在临床仪器的典型领域，相对于常规使用的商业造影剂， 2 值更高。

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