Anti-melanoma Activity Of Magnetic Nanoparticles Loaded With Melittin And Photosensitizer

BackgroundMelanomas arise from the production of melanocytes,which is the most aggressive type of tumor in skin cancer Although the number of melanoma patients is low in the total number of skin cancer patients,most of the patients who die from skin cancer are melanoma patients.For metastatic patients with melanoma who cannot be removed,other conventional methods,including chemotherapy,targeted molecular drug therapy,immunotherapy and radiotherapy,need to be used.However,these methods often result in poor therapeutic effects(less than 15% for three years)and poor clinical response.Therefore,it is particularly important to develop effective methods for early and efficient diagnosis of melanoma and for effective treatment of metastatic melanoma.ObjectiveThe self-assembly build super nanoparticles,which load the photosensitizer and targeted destruction peptide(alpha melittin),dual mode on melanoma for imaging(NIR/MRI)and has targeted the dual mode of therapy(photodynamic therapy/alpha melittin).To achieve early detection and control of tumor growth and recurrence.Methods 1.Preparation and characterization of magnetic nanoparticles(alpha-melittin-pro-NP)loaded with killing peptide and photosensitizer.The iron oxide nanoparticles,phospholipids and photosensitizers are dissolved in chloroform and mixed.The self-assembled spherical nanostructure was prepared by hydration emulsification.After cooling,the improved melittin(alpha-melittin)was added to the ice box,shaking the 3H to form an alpha-melittin-pro-NP.The prepared alpha-melittin-pro-NP was measured by the electron microscope and the hydrous particle diameter of the alpha-melittin-pro-NP diameter and the UV spectrum instrument to determine its UV absorption.2.Detection of biological safety of alpha-melittin-pro-NP and targeted validation of SR-B1 receptor Characterization of nanomaterials.The biosafety of nanomaterials was detected by RBC hemolysis test.The uptake and uptake of alpha-melittin-pro-NP and Western Blot test were detected by cell flow cytometry,which verified the SR-B1 targeting of nanomaterials.3.Cell experimentThe distribution of drug in the cells was observed by confocal laser scanning;the cytotoxicity of ?-melittin-pro-NP was detected by MTT assay;the effect of ?-melittin-pro-NP on apoptosis was detected by flow cytometry;Dendritic(DC)cells,co-incubated with alpha-melittin-pro-NP and B16-F10 cells,can activate DC cells,activate immune responses,and inhibit and kill tumor cells.Results1.The prepared ?-melittin-pro-NP solution is uniform,without precipitation,and has good stability;the average particle size of the nanomaterials detected by transmission electron microscopy and hydration particle size is about 15 nm;UV spectral scanning results of prolipid and ?-Melittin-pro-NP has characteristic absorption at 670 nm,which proves that photosensitizer prolipid has been modified to ?-melittin-NP surface.2.The diameter of ?-melittin-pro-NP was measured by transmission electron microscopy and water particle diameter respectively.2.When the concentration is 10 ?M,the hemolysis rate of melittin is close to 100%,while the hemolysis rate of ?-melittin-pro-NP is about 20%;by comparing different cellular uptake nanomaterials and Western Blot results,?-melittin-pro-NP efficiently targets the SR-B1 receptor in B16-F10 cells.3.By laser confocal observation of B16-F10 cells through endocytosis,?-melittin-pro-NP uptake into the cytoplasm;MTT experiment results show that when the ?-melittin-pro-NP concentration of 1?M,non-light group The photoinhibition rates of B16-F10 cells were 30% and 90%,respectively.The results of apoptosis test showed that cells in the PBS group,?-melittin-pro-NP group,and ?-melittin-pro-NP+ light group had cell withering.The mortality rates were 3.18%,47.56%,and 90.8%,respectively;in vitro activation of DC cells showed that when the concentration of ?-melittin-pro-NP was 10 ?M,the proportion of DC cell activation was 37% higher than that of the PBS group.ConclusionThe success of the project can be synthesized at the same time carrying melittin alpha melittin and photosensitizer prolipid new nanomaterials alpha melittin-pro-NP,the nano meter material has good biocompatibility,tumor size,location,and can be observed near in frared imaging can also through the light power and alpha-destruction of melittin synergy to perform good activity of melanoma.