载阿霉素金纳米粒的制备和细胞毒性研究

Preparation and cytotoxicity of doxorubicin-containing gold nanoparticles

目的构建载阿霉素(DOX)的甲氧基聚乙二醇(mPEG)修饰的金纳米粒AuNPs-mPEG@DOX,以降低DOX的毒副作用。方法制备AuNPs-mPEG@DOX,通过粒径、电位和紫外可见光吸收光谱(UV-Vis)进行表征。考察连接巯基的DOX(HS-DOX)投药浓度对AuNPs-mPEG@DOX吸附率和载药量的影响。建立未吸附HS-DOX含量测定的高效液相色谱法(HPLC),对专属性、线性、精密度、稳定性和加样回收率进行考察。采用CCK-8法检测AuNPs-mPEG@DOX对MCF-10A和MCF-7细胞的毒性作用。结果成功制备了AuNPs-mPEG@DOX,粒径为(46.12±0.49)nm,电位为(18.60±1.51)mV,最大吸收波长为530 nm。建立了可用于检测AuNPs-mPEG@DOX未吸附HS-DOX含量的HPLC方法,测定最佳投药浓度11.18μg/ml,HS-DOX条件下的吸附率为(9.21±2.88)%,载药量为(2.01±0.62)%。细胞毒性实验表明AuNPs-mPEG@DOX可明显降低DOX对正常乳腺细胞的毒副作用;DOX在≥4.75μmol/L时,AuNPs-mPEG@DOX与游离DOX对乳腺肿瘤细胞的细胞毒性作用一致。结论AuNPs-mPEG@DOX可有效降低DOX的毒副作用,为后续AuNPs连接药物降低其毒副作用的研究提供参考。

Objective To construct methoxy polyethylene glycol(mPEG)modified gold nanoparticles(AuNPs)loaded with doxorubicin(DOX)AuNPs-mPEG@DOX in order to reduce the toxicity and side effects of DOX.Methods AuNPs-mPEG@DOX was prepared and characterized by Z-Average,Zeta potential and UV-Vis spectroscopy.The impact of thiol-linked DOX(HS-DOX)at various dosage concentrations on the drug adsorption rate and drug loading of AuNPs-mPEG@DOX was investigated.Furthermore,a HPLC method was developed to accurately determine the content of unadsorbed HS-DOX in AuNPs-mPEG@DOX.The specificity,linearity,precision,stability and average recovery of this method were thoroughly investigated.The cytotoxic effect of AuNPs-mPEG@DOX on MCF-10A and MCF-7 cells was evaluated using a CCK-8 assay.Results AuNPs-mPEG@DOX was successfully prepared with Z-Average of(46.12±0.49)nm,Zeta potential of(18.60±1.51)nm and the maximum absorption wavelength of 530 nm.An efficient HPLC method for the detection of unadsorbed HS-DOX in AuNPs-mPEG@DOX was devised.The optimal dosage concentration of HS-DOX for AuNPs-mPEG@DOX was determined to be 11.18μg/ml,resulting in a drug adsorption rate of(9.21±2.88)%and a drug loading rate of(2.01±0.62)%.Cytotoxicity experiments demonstrated that AuNPs-mPEG@DOX significantly reduced the toxic and side effects of DOX on normal breast cells.Additionally,AuNPs-mPEG@DOX and free DOX exhibited comparable cytotoxic effects on breast tumor cells when DOX concentration was equal to or greater than 4.75μmol/L.Conclusion AuNPs-mPEG@DOX effectively reduce the toxicity of DOX,providing a reference for future research on reducing the toxicity of AuNPs-linked drugs.