Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT), Argentina; Área de Biocoloides y Nanotecnología, Instituto de Tecnología de Alimentos, Facultad de Ingeniería Química, Universidad Nacional del Litoral, 1 de Mayo 3250, Santa Fe, 3000, Argentina.
Área de Biocoloides y Nanotecnología, Instituto de Tecnología de Alimentos, Facultad de Ingeniería Química, Universidad Nacional del Litoral, 1 de Mayo 3250, Santa Fe, 3000, Argentina.
Colloids Surf B Biointerfaces. 2019 Jan 1;173:43-51. doi: 10.1016/j.colsurfb.2018.09.046. Epub 2018 Sep 21.
Chrysin (5,7-dihydroxyflavone) (Chrys) is a natural flavone extracted from many plants, and it has been proposed as a bioactive agent for cancer therapy. Nevertheless, its use is limited mainly due to its poor water solubility. Bovine serum albumin (BSA) is a water soluble, biocompatible and non-toxic protein with a promising application in lipophilic bioactive compound delivery. Moreover, BSA is heat sensitive, feature that could be used for producing self-assembled nanoparticle with tailor-made properties. In this contribution, we studied the formation of BSA nanoparticles (BSAnp) by thermal treatment at different conditions of temperature (70 °C/5 min and 85 °C/5 min), protein concentration (1.0-4.0%wt.) and aqueous medium pH values (9.0 and 11.0) in which it is known that BSA is found in different unfolded conformations. Binding of Chrys dissolved in dimethyl sulfoxide (DMSO) was studied by fluorescence titration experiments. Characterization of Chrys-loaded and unloaded BSAnp was performed in phosphate buffered saline (PBS) pH 7.4 by applying a set of complementary techniques: dynamic light scattering (DLS), size exclusion fast protein liquid chromatography (SEC-FPLC) and transmission electron microscopy (TEM). Different populations of BSAnp were obtained, which showed different diameters in the range of 1328 nm, ζ potentials around -10.0 mV, molecular weight in the range of 400-1000 kDa and spherical shape. Chrys encapsulation efficiency (EE. %) was also determined, and values between 44-84% were obtained, which mainly depended on the mode of Chrys binding and physicochemical BSAnp properties. Results highlight the ability of self-assembled BSAnp for Chrys vehiculization in an aqueous medium which could found potential application in antitumor therapies.
白杨素（5,7-二羟基黄酮）（Chrys）是一种从多种植物中提取的天然黄酮类化合物，它已被提议作为癌症治疗的生物活性药物。然而，其应用主要受到其较差的水溶性的限制。牛血清白蛋白（BSA）是一种水溶性、生物相容和无毒的蛋白质，在亲脂性生物活性化合物传递方面具有广阔的应用前景。此外，BSA对热敏感，这一特性可用于制备具有定制特性的自组装纳米颗粒。在本研究中，我们研究了在不同条件下（70°C/5min 和 85°C/5min 的温度、1.0-4.0%wt. 的蛋白质浓度和 9.0 和 11.0 的水介质 pH 值）通过热处理形成 BSA 纳米颗粒（BSAnp），在这些条件下，BSA 以不同的未折叠构象存在。通过荧光滴定实验研究了 Chrys 在二甲基亚砜（DMSO）中的溶解情况。在 pH 为 7.4 的磷酸盐缓冲盐水（PBS）中，通过应用一系列互补技术对 Chrys 负载和未负载的 BSAnp 进行了表征：动态光散射（DLS）、尺寸排阻快速蛋白液相色谱（SEC-FPLC）和透射电子显微镜（TEM）。得到了不同粒径范围在 1328nm、ζ 电位约-10.0mV、分子量在 400-1000kDa 范围内和球形的 BSAnp 不同群体。还确定了 Chrys 包封效率（EE. %），得到了 44-84%的包封效率，这主要取决于 Chrys 的结合方式和 BSAnp 的物理化学性质。研究结果突出了自组装 BSAnp 对 Chry 在水介质中载体化的能力，这可能在抗肿瘤治疗中具有潜在的应用。
