Ayad Camille, Yavuz Altan, Salvi Jean-Paul, Libeau Pierre, Exposito Jean-Yves, Ginet Valentine, Monge Claire, Verrier Bernard, Arruda Danielle Campiol
UMR 5305: Laboratoire de Biologie Tissulaire et d'Ingénierie Thérapeutique, Institut de Biologie et Chimie des Protéines, CNRS/Université Claude Bernard Lyon 1, 7 Passage du Vercors, CEDEX 07, 69367 Lyon, France.
Pharmaceutics. 2022 Jun 18;14(6):1297. doi: 10.3390/pharmaceutics14061297.
Polymeric and/or lipid platforms are promising tools for nucleic acid delivery into cells. We previously reported a lipid-polymer nanocarrier, named LipoParticles, consisting of polylactic acid nanoparticles surrounded by cationic lipids, and allowing the addition of mRNA and cationic LAH4-1 peptide at their surface. Although this mRNA platform has shown promising results in vitro in terms of mRNA delivery and translation, the bulk method used to prepare LipoParticles relies on a multistep and time-consuming procedure. Here, we developed an automated process using a microfluidic system to prepare LipoParticles, and we compared it to the bulk method in terms of morphology, physicochemical properties, and ability to vectorize and deliver mRNA in vitro. LipoParticles prepared by microfluidic presented a smaller size and more regular spherical shape than bulk method ones. In addition, we showed that the total lipid content in LipoParticles was dependent on the method of preparation, influencing their ability to complex mRNA. LipoParticles decorated with two mRNA/LAHA-L1 ratios (1/20, 1/5) could efficiently transfect mouse DC2.4 cells except for the automated 1/5 assay. Moreover, the 1/5 mRNA/LAHA-L1 ratio drastically reduced cell toxicity observed in 1/20 ratio assays. Altogether, this study showed that homogeneous LipoParticles can be produced by microfluidics, which represents a promising platform to transport functional mRNA into cells.
聚合物和/或脂质平台是将核酸递送至细胞的有前景的工具。我们之前报道了一种脂质-聚合物纳米载体，名为脂质颗粒（LipoParticles），它由被阳离子脂质包围的聚乳酸纳米颗粒组成，并允许在其表面添加mRNA和阳离子LAH4-1肽。尽管这个mRNA平台在mRNA递送和翻译方面已在体外显示出有前景的结果，但用于制备脂质颗粒的批量方法依赖于多步骤且耗时的程序。在此，我们开发了一种使用微流控系统制备脂质颗粒的自动化方法，并在形态、物理化学性质以及体外将mRNA载体化和递送的能力方面将其与批量方法进行了比较。通过微流控制备的脂质颗粒比通过批量方法制备的脂质颗粒尺寸更小且呈更规则的球形。此外，我们表明脂质颗粒中的总脂质含量取决于制备方法，这会影响它们与mRNA复合的能力。用两种mRNA/LAHA-L1比例（1/20、1/5）修饰的脂质颗粒能够有效转染小鼠DC2.4细胞，但自动化的1/5检测除外。此外，1/5的mRNA/LAHA-L1比例显著降低了在1/20比例检测中观察到的细胞毒性。总之，这项研究表明可以通过微流控制备均匀的脂质颗粒，这代表了一个将功能性mRNA转运到细胞中的有前景的平台。
