Fabrication of drug nanocarrier system of gelatin coated graphene oxide nanosheets complexed with 5FU to study anti-cancer effects in skin cancer cells (A_375)

Abstract

Introduction: Skin cancer is one of the most common types of cancer in the world. The common methods of drug administration usually require a high dose or frequent use, which is due to low efficiency of drug delivery. Considering the good potential of nanocarriers, treating cancer could be more efficient and with fewer doses and subsequently less side effects in the body. The nanostructure of graphene oxide (GO) has attracted attention due to its numerous active functional groups that have provided the possibility of loading drugs and modifying its surface with polymers. In this study, to increase the efficiency of fluorouracil (5-FU), we developed a gelatin-coated nanocarrier (GGO) as a pH-sensitive transporter for the controlled release of chemotherapy drug to skin cancer cells (A-375). Then, effects GGO@5-FU on the cytotoxicity, the apoptosis, and the expression of genes involved in apoptosis were examined in A-375 cells. Methods: First, (GO@5-FU, GGO, GGO@5-FU) nanoparticles were synthesized and characterized by UV, XRD, DLS, SEM, EDX-MAP, AFM and FTIR techniques. The free drug fluorouracil loaded on the nanocarrier was analyzed on the A-375 cell line by MTT method and DAPI staining, respectively, to study the cytotoxicity and apoptosis, and finally the expression levels of genes involved in cell apoptosis (Bcl2 and Caspase3) was analyzed by real-time PCR. The statistical analysis of data was performed using GrapHpad Prism9 software, where the p value less than 0.05 was considered significant. Results: The characterization results showed that the synthesized nanocomposite has good physicochemical properties. The drug loading and encapsulation efficiency of 5-FU on GGO were 33% and 50%, respectively. From the release studies, the release rate of the drug after 96 hours was higher under acidic conditions compared to that of neutral pH. The toxicity and DAPI results also showed that GGO@5-FU caused dose-dependent inhibition of A-375 cells at lower concentrations compared to the free drug. Real-time PCR results revealed that GGO@5-FU treated cells showed lower Bcl2 gene expression and higher Caspase3 expression than the free drug group (P value < 0.05). Conclusion: This study showed that a gelatin-coated graphene oxide nanocarrier enhances the efficacy of the drug 5-FU at lower doses with sustained and controlled release in a dose-