Self-Assembled Organic Nano-photosensitizer For Anti-HeLa Photodynamic Or Photothermal Therapy

Cervical cancer is the most common malignant tumor in gynecology,which seriously threatens women's health.Traditional treatments include surgery,radiotherapy and chemotherapy.Traditional therapy plays a leading role in the treatment of cervical cancer,but it also has its limitations.Photodynamic therapy(PDT)and photothermal therapy(PTT)are emerging therapies with their unique advantages,such as minimal trauma,local treatment,repeatable treatment and weak toxicity.The lesions of cervical cancer can be accessed from the outside through natural channel vagina,which provides great convenience for photosensitive therapy.Some limitations of traditional therapies can also be remedied by photosensitization therapy.Photosensitive therapy for cervical cancer has its unique superiority and practicability.Photodynamic therapy for cervical cancer is still in its infancy.Photothermal therapy for cervical cancer has not been reported,and is still in the basic research.Photosensitizer(PS)is one of the three elements of photodynamic.Similarly.photothermal conversion agent(PTCA)1s the core of photothermal efficiency.The study of photosensitizers and photothermal conversion agents has always been a hot spot in the basic research of phototherapies.Boron-dipyrromethene(BODIPY)is a new fluorescent dye.Modification of its parent structure will make it more suitable for bioimaging and cancer treatment.Chlorin is the second-generation photodynamic photosensitizer,but its use in photothermal therapy has rarely been reported.Based on these,the following two systems are designed and studied from the perspectives of photodynamic and photothermal.Purpose:1)To synthesize a new photosensitizer and combine it with chemotherapeutic drugs to form nanomaterials by self-assembly.To explore its anti-cervical cancer cell capacity at the cellular and animal levels.2)Comparing the photothermal properties of porphyrin and chlorin self-assembled nanoparticles in detail to explore the photothermal therapy effect of chlorin on cervical cancer cell.Method:1)Di-iodinated boron-dipyrromethene(BDP-I2)was iodized and then mixed with paclitaxel(PTX)to prepare nano-materials by nano-precipitation method.The structure and purity of the samples were verified by 1H NMR and MS.The basic properties of the material were characterized by transmission electron microscopy,dynamic light scattering,ultraviolet spectrum and fluorescence spectrum.Laser confocal microscopy was used to observe the endocytosis and co-localization of lysosomes.ICG and DCFH-DA indicators were used to monitor the photodynamic effects of drugs in vitro and in cell,respectively.The anti-HeLa ability was investigated by MTT cell activity assay and live and dead staining.The subcutaneous tumorigenesis experiment of nude mice was used to explore the anti-Hela ability at animal level.2)Porphyrin was prepared and reduced to chlorin by one step.Nano-materials were prepared by nano-precipitation method.The structure and purity of the samples were verified by 1H NMR and MS.The basic properties of the material were characterized by transmission electron microscopy,dynamic light scattering,ultraviolet spectrum and fluorescence spectrum.638 nm laser was used as light source.Thermocouple was used to monitor the difference of temperature rise in vitro.Laser confocal microscopy was used to observe the endoeytosis of drugs.The difference of anti-HeLa ability was investigated by MTT cell activity assay and live and dead staining.The Kunming mice model of tumor-bearing was established,the irradiation was given to the mice after drug treatments.The difference of tumor temperature was monitored by thermal imaging camera in real time.To explore the Anti-cervical cancer ability of drugs at animal level,further the growth of tumors were monitored.Result:1)BDP-I2 was successfully synthesized proved by H-NMR and MS.Under transmission electron microscopy(TEM),nanomaterials were novel rods.Both transmission electron microscopy(TEM)and dynamic light scattering(DLS)show that the particle size was less than 200 nm.Ultraviolet absorption and visible nano-material absorption showed blue-shift,fluorescence spectra contrast showed visible fluorescence quenching of nanorods.Both in vitro and in cell,good photodynamic effects were prestented.The endocytosis of the drug was good.The co-localization of lysosomes and lysosome escape experiments were successful.At the cellular and animal levels,the combination of paclitaxel and BDP-h was superior to paclitaxel or BDP-I2 alone in anti-Hela.2)The product was successfully synthesized peoved by H-NMR and MS.The nanomaterials were spherical under transmission electron microscopy.Both transmission electron microscopy(TEM)and dynamic light scattering(DLS)showed that the particle size was less than 200 nm.Ultraviolet absorption spectra showed that the absorption peak of chlorin at 650 nm increased obviously,and fluorescence quenching of nanoparticles was observed in fluorescence spectra.In vitro photothermal heating test showed that chlorin was superior to porphyrin in heating performance.The calculated photothermal conversion efficiency of porphyrin and chlorin were 34%and 45%,respectively.The endocytosis of the two drugs was good.After intratumoral administration and irradiation,chlorin still has excellent warming effect.Animal anti-tumor experiments showed that chlorines till had the advantage of Anti-cervical cancer in vivo.Conclusion:1)Nanorods were prepared by the co-assembly of photosensitizer BDP-I2 and chemical anticancer drug PTX.The obtained nanorods can be effectively ingested by cells,enhance the lysosome escape of PTX,and promote the cytotoxicity of cells in vitro.Animal experiments in vivo further validate the excellent therapeutic efficacy.The design of this combination therapy is simple and effective.Current work emphasizes the progress of small molecule self-assembly and helps to promote these to future clinical trials.2)Chlorin not only has excellent biocompatibility and stability of porphyrin photosensitizers,but also exhibits more excellent heating and therapeutic advantages in vitro,cell and animal levels.This article provides a powerful photothermal agent for the treatment of cervical cancer,and also looks forward to providing a convenient and effective new idea for the application of porphyrin and chlorin in the future.Innovations:1)? The self-assembly of di-iodinated boron-dipyrromethene(BDP-I2)as a photosensitive material for photodynamic therapy has not been reported.?The rod-like structure formed by self-assembly is novel and has more endocytosis advantages than spherical nanoparticles.? In view of the poor water solubility and drug resistance of paclitaxel,this paper combined paclitaxel with BDP-I2 to form nano-materials,which can achieve high drug delivery efficiency without other additional carriers,effectively combining chemotherapy and photodynamic therapy.2)Chlorin is mostly used in photodynamic therapy,but seldom reported in photothermal therapy.In this paper,638 nm laser source is selected,where chlorin shows excdlent photothermal performance.This paper eompares porphyrin with chlorin in system,which provides a powerful photothermal preparation for the treatment of cervical cancer,and suggesting a convenient and effective new idea for the application of porphyrin and chlorin in the future.