| Doxorubicin(DOX),also known as doxorubicin,is a widely used anticancer drug,which can cause heart and liver damage and myocardial disease after clinical application.In order to improve the effect of clinical use and achieve the purpose of high efficiency and low toxicity,mesoporous silica with good chemical stability and biocompatibility was selected as drug carrier after PEG modification to study drug loaded nanoparticles.The drug in MCF-7 breast cancer cells was analyzed qualitatively and quantitatively by laser confocal microscopy and HPLC.The pharmacokinetic behavior of adriamycin loaded nanoparticles was studied by the principle of pharmacokinetics.The quality analysis and data theoretical support were provided for the evaluation and development of the effectiveness of adriamycin loaded nanoparticles from the micro cell level.The research of this subject is as follows:1.Firstly,mesoporous silica nanoparticles and PEG modified mesoporous silica nanoparticles composites were prepared.The particle size and potential analysis were characterized by laser particle size analyzer(Nano-ZS90),the morphology of nanoparticles was characterized by transmission electron microscopy(TEM),and the structure of functional groups was confirmed by Fourier transform infrared spectroscopy(FT-IR).The mesoporous structure of MSN was confirmed by small angle X-ray powder diffraction curve.The specific surface area,pore volume and pore diameter of MSN were calculated by nitrogen adsorption / desorption experiment,which were 643.22 m2/g,0.576 cm3/g and 3.407 nm respectively.The specific surface area,pore volume and pore diameter of msn-peg were 587.09 m2/g,0.572 cm3/g and 3.05 nm respectively.2.The HPLC method for the determination of doxorubicin was established.This method has strong specificity,good precision and repeatability,and has a short retention time of about 2 min,which greatly improves the detection efficiency and can be used for rapid detection.3.The established standard curve has a good linear relationship within the concentration range of 5-200 ?g/m L.Adriamycin was selected as model drug to prepare drug loaded nanoparticles.Through box Behnken design(BBD)-response surface method,the main factors affecting the entrapment efficiency and drug loading were screened.The final process was determined as follows: the mass ratio of msn-peg to adriamycin was 3:1,the concentration of adriamycin was 2 mg/m L,and the stirring speed was 1000 rpm.The results showed that the average entrapment efficiency of adriamycin loaded nanoparticles was 72.22% and the average drug loading was 9.28%.The release behavior of drug loaded nanoparticles was studied.The results showed that the cumulative release of doxorubicin in PBS with p H5.5 was 88.5% in 24 hours,while that in PBS with ph6.8 and p H7.4 was 63.7% and 47.8% respectively,which indicated that the drug loaded nanoparticles could release more rapidly in acid condition.4.In vitro safety evaluation showed that the DOX@MSN and DOX@MSN-PEG which had good blood compatibility and low hemolysis rate.MTT assay was used to test the cytotoxicity of MSN and MSN-PEG.The results showed that the cell survival rate was above 80% when the concentration was below 400 ?g/m L,indicating that the two carrier materials were safe,non-toxic and compatible.Then,the cytotoxicity test of DOX@MSN and DOX@MSN-PEG and the commercially available doxorubicin preparations within 48 h after administration showed that the prepared drug-loaded nanoparticles had obvious killing effects on tumor cells.5.Taking breast cancer MCF-7 as a cell model,the uptake and elimination of drugs in cells were observed by laser confocal microscopy.The results showed that all adriamycin entered the nucleus after 12 hours of culture,and achieved the same effect in a short period of time DOX@MSN and DOX@MSN-PEG The elimination rate of doxorubicin is slower than that of doxorubicin,and the cell retention time is longer.A HPLC method was established for the determination of doxorubicin in cancer cells.According to the first-order absorption single chamber model,DOX@MSN The Ke decreased to 0.277 times,t1/2 increased to 1.82 times,AUC increased to 1.478 times;DOX@MSN-PEG The Ke decreased to 0.13 times,t1/2 increased to 2.11 times,AUC increased to 1.84 times.The data are consistent with the results of laser confocal observation and analysis.It is further proved that loading adriamycin with msn-peg at the cell level is beneficial to increase the antitumor effect and achieve the goal of high efficiency and low toxicity.Dox@MSN-PEG drug-loaded nanoparticles prepared can be absorbed and tend to be completely released,and the drug stays in cells for a longer time,which improves the drug's cytokinetic behavior and achieves the goal of high efficiency and low toxicity,and predicts and evaluates the drug's therapeutic effect quickly at the cellular level. |
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