In Vitro Study Of Cypate-Silk Nano-drug In The Treatment Of Colon Cancer

Objective:This paper is to study the preparation and application of the nano-drug loaded with cypate.Silk fibroin is a natural protein with excellent properties,such as good biocompatibility,biodegradability,non-toxic,no stimulation and easy to be made into the film,fiber,hydrogel,nanoparticles,porous materials and other forms,it has broad application prospects in the field of drug delivery.Among them,the nano carrier,due to its high specific surface area,surface energy,easy diffusion and quick response to external stimulation,aroused the attention of researchers.The prepared nanoparticles loaded with cypate-silk remained in the tumor region to enrichment,infiltration and retention.Furthermore,the near infrared fluorescence and photothermal effects of cypate could increase the temperature of tumor tissues and induce the death of tumor cells,so as to achieve the effect of killing tumor.Methods:?1?silk fiber was treated with CaCl₂-ethanol water?molar ratio 1:2:8?.After dialysis,the liquid was made and quantified by weighing method.?2?we adopt the solvent dialysis and stirring methods to prepare the nano-drug that materials were loaded with cypate-silk,and we evaluated the prepared nano-drug by measuring the particle size and particle size distribution?PDI?and other indicators,finally finding the suitable conditions.The morphology and size of the nano-drug were observed by transmission electron microscopy?TEM?.The particle size distribution of the nano-drug loaded with cypate-silk was examined by DLS,and the in vitro photothermal effect of it was also investigated.?3?The toxicity of nano-drugloaded with cypate-silk to Lovo cells was compared by MTT method with different concentrations,and the thermal toxicity of it was also investigated.?4?Live-dead staining was used to observe the survival rate of the cells under different conditions,so as to evaluate the effect of the nano-drug.?5?The cell viability was further quantified by flow cytometry.Result:Cypate-silk nanoparticles were prepared by the agitation and dialysis methods,with a particle size of?126.5+9.5 nm?.Cypate has obvious photothermal effect in vitro under near infrared light,nano-drug loaded with cypate-silk can increase the temperature above 40 within 5min at the concentration of 20g/ml;The survival rate of cells was over90%after treated with different concentrations of cypate-silk nanoparticles on Lo Vo cells for 24 h,indicating that nanoparticles loaded with cypate-silk have less toxicity and good biocompatibility;but at 808 nm of illumination?1.5W/cm²?,cypate-silk nanoparticles significantly increased cytotoxicity against the LoVo cells.Conclusion:This paper introduces the preparation of cypate silk fibroin nanometer drug with silk fibroin as the carrier,which has the advantages of proper particle size,uniform distribution and low toxicity.After the light-exposed treatment,cypate-silk nano-drug can effectively kill tumor cells.After the above studies,we found that the cypate-silk nano-drug has low cytotoxicity,good histocompatibility and photothermal effect,so it can be used as a target drug to kill tumor..