The Preparation And Biological Effects In Vitro Of Nanoparticles BKC And PMCM With Two-photon Excitation

Objective:Two novel nano materials with two-photon excitation were designed and synthesized,and their morphology and two-photon properties were analyzed.Cell imaging,cellular endocytosis and in vitro phototoxicity and dark toxicity under one/two-photon excitation were investigated by cell model,providing scientific basis for safety applications such as in vivo imaging and treatment of two-photon nanomaterials.Methods:1?A novel two-photon nano photosensitizer BKC was synthesized by using Fmoc coupling chemical standard solid-phase peptide technology by selecting the BP with high two-photon absorption cross section(2.8×10~5GM)and the CREKA peptide which can specifically bind to fibrin in tumor blood vessels and matrix.The nanoparticles were characterized by UV spectrophotometer,fluorescence spectrometer,nano particle size,ZETA potential analyzer,transmission electron microscope,laser confocal scanning electron microscope and other instruments.The ability of BKC nanoparticles to produce reactive oxygen species was detected using singlet oxygen capture agent(ABDA).The cytotoxicity of nanoparticles under one/two-photon irradiation was investigated.2?PMCM nanoparticles with two-photon excitation were prepared.The nanoparticles were characterized by transmission electron microscope,laser confocal microscope,nano-particle size and ZETA potential analyzer.The photothermal effect,two-photon properties and imaging ability of nanoparticles were studied.The one/two-photon toxicity of nanoparticles was investigated.Results:1.The size of BKC nanoparticles are about 5-10nm,and that of PMCM nanoparticles are about 122nm,and the morphology is regular,the scale is clear and well dispersed.2.BKC nanoparticles can produce a large amount of singlet oxygen after laser irradiation.After two-photon irradiation,the cells have significant phototoxicity and low dark toxicity.3.PMCM nanoparticles have photoacoustic and nuclear magnetic resonance imaging capabilities.4.PMCM nanoparticles have photothermal effect under one/two-photon irradiation,and the two-photon effect is more obvious.After two-photon irradiation,the cell phototoxicity is significant,but it has certain cytotoxicity.Conclusion:1.BKC nanoparticles have uniform particle size and good dispersion.Selective endocytosis and targeting.The nanoparticles can generate a large amount of active oxygen after being irradiated by one/two-photon laser,and have significant phototoxicity;the nanoparticles have low dark toxicity and good biocompatibility.2.PMCM nanoparticles have uniform particle size and good dispersion.It is easy to be swallowed by cells and has targeting property.Nanoparticles have good photothermal conversion effects under two-photon laser irradiation and can be used for PTT.The nanoparticles have a variety of imaging capabilities.The particles have significant phototoxicity under one/two-photon excitation.However,the dark toxicity of PMCM nanoparticles are high.