Roacho-Pérez Jorge A, Rodríguez-Aguillón Kassandra O, Gallardo-Blanco Hugo L, Velazco-Campos María R, Sosa-Cruz Karla V, García-Casillas Perla E, Rojas-Patlán Luz, Sánchez-Domínguez Margarita, Rivas-Estilla Ana M, Gómez-Flores Víctor, Chapa-Gonzalez Christian, Sánchez-Domínguez Celia N
Departamento de Bioquímica y Medicina Molecular, Facultad de Medicina, Universidad Autónoma de Nuevo León, Monterrey 64460, Mexico.
Departamento de Genética, Facultad de Medicina, Universidad Autónoma de Nuevo León, Monterrey 64460, Mexico.
Polymers (Basel). 2021 Jan 27;13(3):400. doi: 10.3390/polym13030400.
Microspheres have been proposed for different medical applications, such as the delivery of therapeutic proteins. The first step, before evaluating the functionality of a protein delivery system, is to evaluate their biological safety. In this work, we developed chitosan/Tween 80 microspheres loaded with magnetite nanoparticles and evaluated cell damage. The formation and physical-chemical properties of the microspheres were determined by FT-IR, Raman, thermogravimetric analysis (TGA), energy-dispersive X-ray spectroscopy (EDS), dynamic light scattering (DLS), and SEM. Cell damage was evaluated by a full set of in vitro assays using a non-cancerous cell line, human erythrocytes, and human lymphocytes. At the same time, to know if these microspheres can load proteins over their surface, bovine serum albumin (BSA) immobilization was measured. Results showed 7 nm magnetite nanoparticles loaded into chitosan/Tween 80 microspheres with average sizes of 1.431 µm. At concentrations from 1 to 100 µg/mL, there was no evidence of changes in mitochondrial metabolism, cell morphology, membrane rupture, cell cycle, nor sister chromatid exchange formation. For each microgram of microspheres 1.8 µg of BSA was immobilized. The result provides the fundamental understanding of the in vitro biological behavior, and safety, of developed microspheres. Additionally, this set of assays can be helpful for researchers to evaluate different nano and microparticles.
微球已被应用于不同的医学领域，如治疗性蛋白质的递送。在评估蛋白质递送系统的功能之前，第一步是评估其生物安全性。在这项工作中，我们制备了负载磁铁矿纳米颗粒的壳聚糖/吐温80微球，并评估了细胞损伤情况。通过傅里叶变换红外光谱（FT-IR）、拉曼光谱、热重分析（TGA）、能量色散X射线光谱（EDS）、动态光散射（DLS）和扫描电子显微镜（SEM）来确定微球的形成及其物理化学性质。使用非癌细胞系、人类红细胞和人类淋巴细胞，通过一整套体外试验来评估细胞损伤。同时，为了了解这些微球是否能在其表面负载蛋白质，我们测定了牛血清白蛋白（BSA）的固定化情况。结果显示，负载于壳聚糖/吐温80微球中的磁铁矿纳米颗粒大小为7 nm，微球平均尺寸为1.431 µm。在浓度为1至100 µg/mL时，没有证据表明线粒体代谢、细胞形态、膜破裂、细胞周期或姐妹染色单体交换形成发生变化。每微克微球可固定1.8 µg BSA。该结果为所制备微球的体外生物学行为和安全性提供了基本认识。此外，这一系列试验有助于研究人员评估不同的纳米颗粒和微粒。
