Caso Maria Federica, Carotenuto Felicia, Di Nardo Paolo, Migliore Alberto, Aguilera Ana, Lopez Cruz Matilde, Venanzi Mariano, Cavalieri Francesca, Rinaldi Antonio
NANOFABER srl, 00123 Rome, Italy.
Center of Regenerative Medicine, University of Rome "Tor Vergata", 00133 Rome, Italy.
Nanomaterials (Basel). 2020 May 31;10(6):1075. doi: 10.3390/nano10061075.
Different polysaccharides-namely dextran, carboxymethyl dextran, alginate, and hyaluronic acid-were compared for the synthesis of nanoporous microsponges particles (NMPs) obtained from a one-pot self-precipitation/cross-linking process. The morphologies and sizes of the NMPs were evaluated comparatively with respect to polymer-to-polymer and cross-linker solvents (water-based vs. DMSO). We found that the radial distribution of the polymer in the near-spherical NMPs was found to peak either at the core or in the corona of the particle, depending both on the specific polymer or the solvent used for the formation of NMPs. The NMP porosity and the swelling capability were evaluated via scanning electron microscopy (SEM). The degradation study indicated that after 10 h incubation with a reducing agent, approximately 80% of the NMPs were disassembled into soluble polysaccharide chains. The adsorption and release capacity of each type of NMP were evaluated using fluorescently labeled bovine serum albumin and lysozyme as model proteins, highlighting a release time typically much longer than the corresponding adsorption time. The dependence of the adsorption-release performance on pH was demonstrated as well. Confocal microscopy images allowed us to probe the different distribution of labeled proteins inside the NMP. The safety and non-cytotoxicity of NMPs were evaluated after incubation with fibroblast 3T3 cells and showed that all types of NMPs did not adversely affect the cell viability for concentrations up to 2.25 μg/mL and an exposure time up to 120 h. Confocal microscopy imaging revealed also the effective interaction between NMPs and fibroblast 3T3 cells. Overall, this study describes a rapid, versatile, and facile approach for preparing a universal non-toxic, nanoporous carrier for protein delivery under physiological conditions.
比较了不同的多糖,即葡聚糖、羧甲基葡聚糖、藻酸盐和透明质酸,用于通过一锅法自沉淀/交联过程合成纳米多孔微海绵颗粒(NMPs)。相对于聚合物与聚合物以及交联剂溶剂(水基与二甲基亚砜),对NMPs的形态和尺寸进行了比较评估。我们发现,近球形NMPs中聚合物的径向分布在颗粒的核心或冠层达到峰值,这取决于用于形成NMPs的特定聚合物或溶剂。通过扫描电子显微镜(SEM)评估了NMP的孔隙率和溶胀能力。降解研究表明,在用还原剂孵育10小时后,大约80%的NMPs分解为可溶性多糖链。使用荧光标记的牛血清白蛋白和溶菌酶作为模型蛋白评估了每种类型NMP的吸附和释放能力,突出显示释放时间通常比相应的吸附时间长得多。还证明了吸附-释放性能对pH的依赖性。共聚焦显微镜图像使我们能够探测标记蛋白在NMP内部的不同分布。在用成纤维细胞3T3孵育后评估了NMPs的安全性和无细胞毒性,结果表明,对于浓度高达2.25μg/mL和暴露时间长达120小时的情况,所有类型的NMPs均未对细胞活力产生不利影响。共聚焦显微镜成像还揭示了NMPs与成纤维细胞3T3之间的有效相互作用。总体而言,本研究描述了一种快速、通用且简便的方法,用于在生理条件下制备用于蛋白质递送的通用无毒纳米多孔载体。