Salmonella-innovative targeting carrier: Loading with doxorubicin for cancer treatment

Abstract

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.

Bacterial Ghosts-Based Vaccine and Drug Delivery Systems

“…BGs that are specific to cancer cells, such as ECN ghost, have been developed [99]. Experiments on several in vitro tumor cells (hepatoma carcinoma cell and colorectal adenocarcinoma cells) showed that chemotherapeutics@BGs could adhere to tumor cells and exhibited stronger cytotoxicities than free chemotherapeutics, thereby reducing the doses of chemotherapeutic drugs [85,87,100]. Chemotherapeutics@BGs can achieve immunotherapy against tumors in animal experiments.…”

Construction and In Vitro Evaluation of a Tumor Acidic pH-Targeting Drug Delivery System Based on Escherichia coli Nissle 1917 Bacterial Ghosts

“…To evaluate drug packaging into the A-BGs, we loaded the lyophilized A-BGs with DOX using a modified published protocol [ 34 ]. We collected the A-BGs by centrifugation at 4000 rpm for 15 min and washed them three times with PBS.…”

Lactobacillus casei Ghosts as a Vehicle for the Delivery of DNA Vaccines Mediate Immune Responses

“…Bacterial ghosts (BGs) bacterial shells without contents that preserve the entire surface structure of bacteria, including the outer membrane proteins, and show potential as vaccine candidates ( 4 ). In addition, BGs can be loaded with DNA, proteins, and drugs and exploited as delivery systems ( 5 – 7 ). Because BGs contain many Toll-like receptor (TLR) agonists, they can effectively target APCs and elicit potent immune responses against BG-delivered antigens ( 8 – 10 ).…”