Preparation Of DOX@NGO-PEG-HN-1 Complex AndIn Vitro Study Of Targeted Therapy For Oral Squamous Cell Carcinoma

Objective:In order to realize the release process of tumor-targeted drugs,promote the absorption and accumulation of chemotherapy drugs by tumor cells,and reduce the toxic effects of chemotherapy drugs on normal cells,we constructed nano-graphene oxide(NGO)with peptides which have target effects as drug delivery system.Methods:1.Connect HN-1 and chemotherapeutic drug doxorubicin(DOX)with nano-graphene oxide with hydrogen-targeting and π-π bonds to oral cell-like cell carcinoma,and through Fourier Transform Infrared Spectrometer,Scanning Electron Microscope,Dynamic Light Scattering and UV fluorescence spectrophotometer characterize the synthesized materials.2.Study the biocompatibility of the nano drug-loading system and observe the stability of the nano drug-loading system in PBS,physiological saline,FBS,cell culture fluid,and ultrapure water,respectively.3.Observe the release of drugs in PBS solutions of different PHs.Tumor tissues are generally acidic,while normal tissues are neutral.Therefore,the release of drugs in different tissues was simulated through a second experiment.4.To verify the characteristics of NGO’s high loading rate of the anti-tumor drug doxorubicin(DOX).5.The toxicity and targeting of the nano drug delivery system were verified in vitro.Using oral squamous cell carcinoma cells CAL-27 and SCC-25 and normal oral keratinocytes HOK for cytotoxicity test,cell internalization and uptake test.Results:1.The appearance of characteristic peaks in Fourier infrared and ultraviolet spectra shows that DOX and HN-1 are successfully combined with NGO.The scanning electron micrograph of DOX@NGO-PEG-HN-1 shows that the distance between NGO slices increases and wrinkles and curls appear.Those are caused by the introduction of DOX and PEG modification.2.Stability experiment results show that the graphene oxide modified by PEG is evenly dispersed in five different solutions without precipitation,while NGO have obvious precipitation in all five solutions.It shows that the stability and biocompatibility of NGO modified by PEG are greatly improved.3.In different PH drug release experiments,we observed that when PH=5.6,the drug release rate and amount were significantly higher than those of the other two groups(PH=6.6,7.4).And in the first 24 hours,the release rate of the drug is relatively faster,and the release rate tends to be stable after 24 hours.In the PH=5.6 PBS solution,the drug release rate reached 70%.4.The experimental results further verify that NGO has a high load rate for DOX.It can be used as a good anti-tumor drug and a carrier for targeting peptides.5.In OSCC cells(CAL-27 and SCC-25),the cell uptake and cytotoxicity of DOX@ NGO-PEG-HN-1 is significantly higher than that of free DOX.In addition,HN-1showed significant tumor targeting and competitive inhibition.The appearance of characteristic peaks in the Fourier infrared and ultraviolet spectra shows that DOX and NH-1 successfully combined with NGO.Conclusion:1.In this experiment,a tumor targeting DOX@NGO-PEG-HN-1 drug delivery system was successfully constructed.2.DOX@NGO-PEG-HN-1 is more conducive to the release of drugs in an acidic environment and reduces the toxic and side effects on normal tissues.3.After a series of in vitro experiments,DOX@NGO-PEG-HN-1 has the characteristics of targeting and PH-sensitive drug release.Our study demonstrates a good strategy for the construction of OSCC-targeted delivery of nanoparticles to improve the cancer targeting efficiency of anticancer medicines.