Polydopamine Nanoparticle As A Multifunctional Nanocarrierfor Combined Radiophotodynamic Therapy Of Cancer

Part?: Synthesis and characterization of poly-dopamine nanoparticlesObiective: Preparation and representation of multifunctional poly-dopamine nanoparticleMethods: 1)Polydopamine(PDA)multifunctional nanoparticles were prepared by polymerization,centrifugation and washing with dopamine hydrochloride as raw materials,and their characterization was studied.2)PEG nanoparticles modified with polyethylene glycol(PEG)to increase their water solubility and reduce toxicity.3)PDA-PEG nanoparticles were mixed with different concentrations of curcumin(Cur)and chlorin e6(Ce6)solution to explore the loading rate of curcumin(Cur)and chlorin e6(Ce6).4)PDA-PEG/Cur/Ce6 multifunctional nanomaterials were prepared by mixing PDA-PEG nanoparticles,curcumin and Ce6 solution.5)The dialysis bags containing PDA-PEG/Cur/Ce6 solution were placed in different p H buffer,and the temperature was kept at 37 ?.The buffer was taken at different time points and analyzed the concentration of curcumin and Ce6 in the buffer at different time points by high performance liquid chromatography(HPLC).Results: 1)Scanning electron microscopy(SEM)showed that poly-dopamine nanoparticles(PDA)were spherical nanoparticles with an average diameter of about 100 nm.2)After PEG(PEG)modification,PDA-PEG has good water solubility and can be stably present in water,sodium chloride solution and medium.3)The loading curve shows that curcumin(Cur)and chlorin e6(Ce6)can be loaded on PDA-PEG particles at a loading rate of 15% and 30%,respectively.4)The diameter of PDA-PEG/Cur/Ce6 nanoparticles is about 260 nm,and it has good biostability and can be used in biological experiments.5)The drug release rates of curcumin and Ce6 were all p H-dependent,and both drugs were reduced under acidic conditions.Conclusion: The synthesis of PDA-PEG/Cur/Ce6 nanocomposites is relatively simple and the raw materials are easy to obtained.PDA-PEG/Cur/Ce6 nanomaterials with good water solubility could be stable in a variety of solvents for a long time and,could be used for biological experiments.Part ?: Biological toxicity of poly-dopamine nanomaterials and preliminary study on the therapeutic effect of cancertherapyObjective: To investigate the cytotoxicity of PDA-PEG nano-materials,and to observe the effect of PDA-PEG / Cur / Ce6 on tumor combination therapy.Methods: 1)Human lung cancer(A549)cells were incubated in different concentrations of PDA-PEG for 24 hours,and the in vitro toxicity of PDA-PEG nanoparticles was detected by toxicity test(MTT).2)After incubation of human lung cancer(A549)with PDA-PEG/Cur/Ce6 for 6 hours,the nuclei of DAPI cells were used to observe the expression of PDA-PEG/Cur/Ce6 in fluorescence cells according to the characteristics of Cur and Ce6 autofluorescence Intake situation.3)A549 cells were incubated for 24 hours in cell medium containing different concentrations of PDA-PEG/Cur/Ce6,divided into two groups,one group at 6Gy dose X-ray and the other group untreated.After 48 hours of incubation,cell viability was assessed by MTT assay and radiotherapy was evaluated.4)A549 cells were incubated for 24 hours in a medium containing different concentrations of PDA-PEG/Cur/Ce6,divided into two groups,one group irradiated with 660 nm wavelength laser(power 5 m W / cm2)for 30 minutes,and the other Do not deal with it.After 12 hours,the cell viability was measured by MTT assay and the photodynamic therapy was evaluated.Results: 1)PDA-PEG nanomaterials did not have cytotoxicity.2)PDA-PEG/Cur/Ce6 can well accumulate in cells.3)without the X-ray irradiation group,the cells survived well,by X-ray irradiation group,the cell survival rate decreased with the Cur concentration gradually decreased,and the trend was obvious.4)without the 660 nm laser irradiation group,the cells survived well,by 660 nm laser irradiation group,with the Ce6 concentration gradually increased cell survival rate gradually decreased,and the trend is obvious.Conclusion: PDA-PEG nanomaterials have good biocompatibility,no cytotoxicity,is a good drug carrier,can be used to load drugs and the combination of tumor therapy.Part ?: PDA-PEG/Cur/Ce6 nanomaterials for the combination of radiotherapy and photodynamic therapyObjective:To investigate the effect of PDA-PEG/Cur/Ce6 nanomaterials on radiotherapy and photodynamic therapy.Methods: 1)After 24 hours of incubation,A549 cells were divided into 6 groups and numbered.Group 1 were untreated,group 2treated with PDA-PEG/Cur/Ce6(NPs),group 3 were treated with 6Gy X-ray,group 4 were incubated with PDA-PEG/Cur/Ce6,and after for 12 hours,6Gy X-ray were treated(NPs + X group),group 5 were incubated with PDA-PEG/Cur/Ce6,and after 12 hours,irradiated with 660 nm laser for 30 minutes(NPs + PDT group),group 6 were incubated with PDA-PEG/Cur/Ce6 for 12 hours,then irradiated 6Gy X-ray,and then 660 nm laser irradiation for 30 minutes(combined treatment group),the concentration of PDA-PEG/Cur/Ce6 were all identical in 6 groups.2)A549 cells were injected subcutaneously in the back of mice to establish tumor-bearing mice model.Tumor-bearing mice were randomly divided into 6 groups(treatment were the same as cells above).3)After treatment,tumor size and weight were measured every two days,to assess the treatment effect.During the course of the experiment,animal activity,eating,defecation and mental state were observed daily.On the 26 th day after treatment,the main organs of each group were taken and the histological examination was performed by H & E staining to evaluate the long-term organ toxicity of nanomaterials.Results: 1)The cell viability of group 1 and group 2 was almost no difference,the activity of group 4 was lower than that of group 2 and group 3,the activity of group 5 was lower than that of group 2,and the activity of group 6 was the lowest.2)Resume tumor-bearing model completed in 20 days,before treatment,tumor size difference.3)No death was observed in each group of mice.The tumor group was the most obvious in the blank group.The tumor growth was the slowest in the combined group,and there was no significant difference between the two groups.Conclusion: PDA-PEG/Cur/Ce6 nanocomposites show good therapeutic effect in cell test.In animal experiments,PDA-PEG/Cur/Ce6 did not have long-term organ toxicity and showed good radiological and photodynamic therapy.PDA-PEG/Cur/Ce6 can be used to achieve the combined treatment of cancer,opened a new chapter in cancer treatment.