Abstract
Breast cancer is a type of cancer that can metastasize to distant organs of the body if left untreated, causing significant morbidity. The use of chemotherapeutic agents, one of the most employed approaches, possesses several shortcomings, including low tissue availability, short circulation time, and toxicity to healthy cells. To overcome these shortcomings, this study aimed to develop and characterize nanovesicles loaded with doxorubicin hydrochloride and paclitaxel, hypothesizing that combining both drugs in a single nanovesicle would enhance targeting to tumor cells, reduce drug dosing and toxicity resulting in improved patient compliance. The study developed and validated an UPLC method for the simultaneous detection and quantitation of both drugs in aqueous solutions. The UPLC method was validated for linearity, precision, sensitivity, and accuracy. Standard curves developed for both drugs were linear over a concentration range of 3.13 µg/mL through 50 µg/mL. The retention time of doxorubicin hydrochloride and paclitaxel were 1.53 minutes, and 4.06 minutes, respectively.
Thin-film hydration method was used to formulate blank, drug-loaded liposomes, transfersomes, and niosomes. The nanovesicles were physiochemically characterized for particle size, polydispersity index, zeta potential, drug loading, encapsulation efficiency, and drug release. The particle size for blank and drug-loaded nanovesicles ranged from 150 nm-250 nm, and PDI and zeta potential varied from 0.14-0.20 and -0.56 mV - +0.54 mV, respectively. The results indicated that 2.0-2.5% of doxorubicin hydrochloride and 7.0-7.5% of paclitaxel were successfully loaded in the formulations, with high encapsulation efficiency observed for both drugs.
Drug release studies using dialysis membranes with a molecular weight cut-off of 10,000 showed sustained release of drugs from the nano vesicular drug delivery systems for up to 72 hours. Both blank and drug-loaded nanovesicles were physically stable for a minimum of seven days when stored at room temperature and 4⁰C. The cytotoxicity of both drugs, blank nanovesicles, as well as drug-loaded nanovesicles, was investigated through MTT cytotoxicity assay and apoptosis study using flow cytometry. The liposomes, transfersomes, and niosomes developed in the study demonstrated potent and efficient cytotoxicity and apoptotic cell death against the MDA-MB 231 breast cancer cell line, requiring lower doses than their individual drug solutions.
In conclusion, the study developed and characterized three nanovesicles loaded with doxorubicin hydrochloride and paclitaxel, demonstrating their potential as an effective drug delivery system for the treatment of breast cancer.