Efficient and low-risk treatment of skin cancer using antibacterial nanoparticle loaded film containing anti-cancer drug and expression changes of apoptotic genes

Skin cancer is the out-of-control growth of abnormal cells in the epidermis, caused by unrepaired DNA damage that triggers mutations. These mutations lead the skin cells to multiply rapidly and form malignant tumors.The main types of skin cancer are basal cell carcinoma (BCC), squamous cell carcinoma (SCC), melanoma and Merkel cell carcinoma (MCC). Melanoma rates have been rising rapidly over the past ۳۰ years. Despite advances in skin cancer treatment methods, complications such as bacterial infections after surgery are one of the problems of this treatment method. Nano technology provides solutions for this problem. This study presented a new antibacterial wound dressing film infused with nanoparticles containing anti-cancer drug dacarbazine. This anti-cancer and anti-bacterial film offers a strategy in the path of efficient and less risky treatment, and improve patient outcomes. Methods: ۱-Nanoparticles synthesis: In order to generate ZnAl-LDH/ZIF-۸-MOF, ZnAl-LDH was set up using the coprecipitation method then, we added ۲-methylimidazole and zinc nitrate hexahydrate to the ZnAl-LDH and ethanol (AR) solution at room temperature and stirred to obtain a white wet product. After several times of centrifuging and washing, the white product dried and prepared. ۲-In vitro tests: ۲-۱- Release profile: The film was immersed in ۱۰ mL PBS, pH ۷.۴, and shaken at ۳۷ °C. At set intervals, ۳ mL of the medium was taken out and replaced with fresh PBS. Released ZnAl-LDH/ZIF-۸-MOF was quantified using a UV–visible spectrophotometer ۲-۲- Bactericidal test: The antibacterial activities of the film were tested against pathogens S. aureus and P. aeruginosa. ۲-۳- Cell viability: The cytotoxic effects of drug-encapsulated film on A۳۷۴ skin cancer cells were evaluated by MTT assay. ۲-۴- Gene Expression analysis: The in vitro effectiveness of drug-loaded film to alter the expression of apoptotic genes (bcl-۲, bax) was analyzed using real-time PCR. Results: Release profile: The in vitro release profile of ZnAl-LDH/ZIF-۸-MOF from the film mats, demonstrates a sustained release pattern. Antibacterial assays of nanoparticle-loaded films against mentioned bacteria revealed that the biopolymers utilized in film preparation create an antibacterial zone around wounds. Cell viability:While the control cells do not show any signs of toxicity, the skin cancer cells treated with the film have changed color and show toxicity.The drug-loaded film demonstrated an inhibitory effect on skin cancer cell viability after ۷۲ hours, confirming its retained biological activity. Gene expression: Firstly, the expression levels of all genes were normalized with the expression level of the housekeeping gene β-actin. Results showed that the mRNA expression levels of bax and bak genes as a apoptos promoting gene were significantly up-regulated . Also, it has been revealed that the mRNA levels of anti-apoptotic gene, bcl-۲, significantly down-regulated in the cells treated with nanoparticle-loaded films And an increase in cell death. Conclusion: The developed nanoparticle-loaded wound dressing film effectively showed anti-cancer and anti-bacterial properties for the treatment of skin cancer. The sustained release of dacarbazine,with the antibacterial properties of the nanoparticles, demonstrates promising potential for enhancing post-surgery wound healing . The film's biocompatibility and in vitro efficacy in inhibiting skin cancer cell viability and modulating apoptotic genes suggest its potential for clinical translation.

کلیدواژه ها:

squamous cell carcinoma ، dacarbazine ، Nanoparticles ، ZnAl-LDH/ZIF-۸-MOF