Department of Biology, Faculty of Science, University of Mohaghegh Ardabili, Daneshgah St, Ardabil, 11367-56199, Iran.
Department of Inorganic Chemistry, Chemistry and Chemical Engineering Research Center of Iran, Tehran, 14335-186, Iran.
AAPS PharmSciTech. 2018 May;19(4):1554-1566. doi: 10.1208/s12249-018-0969-4. Epub 2018 Feb 22.
The aim of this study was an in vitro evaluation and comparison of the cytotoxic effects of free nisin and nisin-loaded PLA-PEG-PLA nanoparticles on gastrointestinal (AGS and KYSE-30), hepatic (HepG2), and blood (K562) cancer cell lines. To create this novel anti-cancer drug delivery system, the nanoparticles were synthesized and then loaded with nisin. Subsequently, their biocompatibility, ability to enter cells, and physicochemical properties, including formation, size, and shape, were studied using hemolysis, fluorescein isothiocyanate (FITC), Fourier transform infrared (FTIR) spectroscopy, dynamic light scattering (DLS), and scanning electron microscopy (SEM), respectively. Then, its loading efficiency and release kinetics were examined to assess the potential impact of this formulation for the nanoparticle carrier candidacy. The cytotoxicities of nisin and nisin-loaded nanoparticles were evaluated by using the MTT and Neutral Red (NR) uptake assays. Detections of the apoptotic cells were done via Ethidium Bromide (EB)/Acridine Orange (AO) staining. The FTIR spectra, SEM images, and DLS graph confirmed the formations of the nanoparticles and nisin-loaded nanoparticles with spherical, distinct, and smooth surfaces and average sizes of 100 and 200 nm, respectively. The loading efficiency of the latter nanoparticles was about 85-90%. The hemolysis test represented their non-cytotoxicities and the FITC images indicated their entrance inside the cells. An increase in the percentage of apoptotic cells was observed through EB/AO staining. These results demonstrated that nisin had a cytotoxic effect on AGS, KYSE-30, HepG2, and K562 cancer cell lines, while the cytotoxicity of nisin-loaded nanoparticles was more than that of the free nisin.
本研究旨在体外评估和比较游离乳链菌肽(nisin)和载 nisin 的 PLA-PEG-PLA 纳米粒对胃肠道(AGS 和 KYSE-30)、肝(HepG2)和血(K562)癌细胞系的细胞毒性作用。为了构建这种新型抗癌药物递送系统,合成了纳米粒并负载 nisin,然后分别通过溶血实验、荧光素异硫氰酸酯(FITC)、傅里叶变换红外(FTIR)光谱、动态光散射(DLS)和扫描电子显微镜(SEM)研究了其生物相容性、进入细胞的能力以及理化性质,包括形成、大小和形状。然后,通过测定载药效率和释放动力学来评估该制剂作为纳米载体候选物的潜力。采用 MTT 和中性红(NR)摄取试验评估 nisin 和载 nisin 纳米粒的细胞毒性。通过溴化乙锭(EB)/吖啶橙(AO)染色检测凋亡细胞。FTIR 光谱、SEM 图像和 DLS 图证实了纳米粒和载 nisin 纳米粒的形成,其具有球形、分明和光滑的表面,平均粒径分别为 100nm 和 200nm。后者的载药效率约为 85-90%。溶血试验表明其无细胞毒性,FITC 图像表明其进入细胞内。EB/AO 染色观察到凋亡细胞的百分比增加。这些结果表明 nisin 对 AGS、KYSE-30、HepG2 和 K562 癌细胞系具有细胞毒性作用,而载 nisin 纳米粒的细胞毒性作用强于游离 nisin。