Cell- based targeted delivery is recently gain attention as a promising platform for delivery of anticancer drug in selective and efficient manner. As a new biotechnology platform, bacterial ghosts (BGs) have novel biomedical application as targeted drug delivery system (TDDS). In the current work, Salmonellas’ BGs was utilized for the first time as hepatocellular cancer (HCC) in-vitro targeted delivery system. Successful BGs loading and accurate analysis of doxorubicin (DOX) were necessary steps for testing the applicability of DOX loaded BGs in targeting the liver cancer cells. Loading capacity was maximized to reach 27.5 µg/mg (27.5% encapsulation efficiency), by incubation of 10 mg BGs with 1 mg DOX at pH 9 in constant temperature (25 °C) for 10 min. In-vitro release study of DOX loaded BGs showed a sustained release (182 h) obeying Higuchi sustained kinetic release model. The death rate (tested by MTT assay) of HepG2 reached to 64.5% by using of 4 μg/ml, while it was about 51% using the same concentration of the free DOX (P value < 0.0001 One-way ANOVA analysis). The proliferative inhibitory concentration (IC50) of the DOX combined formula was 1.328 µg/ml that was about one third of the IC50 of the free DOX (3.374 μg/ml). Apoptosis analysis (tested by flow-cytometry) showed more accumulation in early apoptosis (8.3%) and late apoptosis/necrosis (91%) by applying 1 μg/ml BGs combined DOX, while 1 μg/ml free DOX showed 33.4% of cells in early apoptosis and 39.3% in late apoptosis/necrosis, (P value˃ 0.05: one-way ANOVA). In conclusion, DOX loaded Salmonellas’ BGs are successfully prepared and tested in vivo with promising potential as hepatocellular cancer (HCC) targeted delivery system.

基于细胞的靶向递送作为一种以选择性和有效方式递送抗癌药物的有前途的平台最近受到关注。作为一种新的生物技术平台，细菌幽灵（BG）具有新型的生物医学应用作为靶向药物递送系统（TDDS）。在当前的工作中，沙门氏菌的BG首次用作肝细胞癌（HCC）体外靶向递送系统。成功加载BG和准确分析阿霉素（DOX）是测试加载DOX的BG在靶向肝癌细胞中的适用性的必要步骤。通过将10 mg BG与1 mg DOX在pH值为9的恒温条件下（25°C）孵育10分钟，最大可以使装载量达到27.5 µg / mg（27.5％的封装效率）。体外载有DOX的BG的释放研究表明，服从Higuchi持续动力学释放模型的持续释放（182 h）。使用4μg/ ml，HepG2的死亡率（通过MTT分析测试）达到64.5％，而使用相同浓度的游离DOX，其死亡率约为51％（P值 <0.0001单向ANOVA分析）。DOX组合配方的增殖抑制浓度（IC50）为1.328 µg / ml，约为游离DOX（3.374μg/ ml）的IC50的三分之一。凋亡分析（通过流式细胞仪测试）显示，通过应用1μg/ ml BG联合DOX，早期凋亡（8.3％）和晚期凋亡/坏死（91％）中的积累更多，而1μg/ ml游离DOX显示33.4％的细胞在早期凋亡中占39.3％，在晚期凋亡/坏死中占39.3％（P值˃0.05：单向方差分析。总之，已成功制备并加载了DOX的沙门氏菌BG，并在体内进行了测试，具有作为肝细胞癌（HCC）靶向递送系统的潜力。