载阿霉素自组装多肽水凝胶的制备及体外评价  

Preparation and in vitro evaluation of self-assembling peptide hydrogel loaded with doxorubicin

作　　者：张馨 陆莹[1] 姚庆强[2] 朱颐申[1] Zhang Xin;Lu Ying;Yao Qingqiang;Zhu Yishen(Nanjing Tech University,Nanjing 211816,Jiangsu Province,China;Department of Orthopedic Surgery,Nanjing First Hospital,Nanjing Medical University,Nanjing 210006,Jiangsu Province,China)

机构地区：[1]南京工业大学,江苏省南京市211816 [2]南京医科大学附属南京医院骨科,江苏省南京市210006

出　　处：《中国组织工程研究》2021年第16期2488-2493,共6页Chinese Journal of Tissue Engineering Research

基　　金：江苏省社会发展重点项目(BE2019736),项目负责人:朱颐申。

摘　　要：背景:化疗是治疗骨肉瘤的重要手段,但传统的小分子疗法临床疗效不理想,不良反应严重,因此需要研发一种治疗骨肉瘤的新治疗方法,能够低剂量下抗肿瘤并降低不良反应。目的:构建载阿霉素的自组装多肽水凝胶体系(doxorubicin self-assembling peptide hydrogel,DOX-SAPH),分析其体外药物释放性能与生物相容性。方法:将阿霉素引入到由FEFEFKFK(F为苯丙氨酸,E为谷氨酸,K为赖氨酸)构成的自组装多肽水凝胶中,制备含20,30,40 g/L FEFEFKFK的DOX-SAPH,该体系中阿霉素质量浓度为2 g/L。将3种DOX-SAPH置于PBS中,定时检测阿霉素释放量。将3种DOX-SAPH浸提液分别与兔骨髓间充质干细胞共培养,CCK-8法检测细胞存活率。将不同质量浓度的阿霉素溶液与含不同质量浓度阿霉素的DOX-SAPH浸提液(含FEFEFKFK 30 g/L)分别与MG63人骨肉瘤细胞共培养,采用CCK-8法检测细胞存活率。结果与结论:①随着水凝胶中FEFEFKFK质量浓度的增加,DOX-SAPH的缓释作用增强,体外第168小时,20,30,40 g/L DOX-SAPH的药物累计释放率分别为88%,83%,80%;②体外共培养1,3,5 d时,20 g/L DOX-SAPH浸提液组的细胞增殖快于30,40 g/L DOX-SAPH浸提液组(P<0.05),其中40 g/L DOX-SAPH浸提液组于第1天时有一定的细胞毒性;③阿霉素溶液与DOX-SAPH浸提液呈浓度依赖性抑制MG63细胞的增殖,当阿霉素溶液与DOX-SAPH浸提液中阿霉素质量浓度<12.5 mg/L时,二者对MG63细胞的毒性无明显差异(P>0.05);当阿霉素质量浓度≥12.5 mg/L时,阿霉素溶液的细胞毒性大于DOX-SAPH浸提液(P<0.05);④结果表明,DOX-SAPH具有良好的药物缓释作用,可抑制MG63骨肉瘤细胞的生长。BACKGROUND:Chemotherapy is an important method for treating osteosarcoma,but traditional small molecule therapy is not ideal for clinical efficacy and has serious adverse reactions.Therefore,it is necessary to develop a new method for osteosarcoma,which can anti-tumor at low doses and reduce adverse reactions.OBJECTIVE:To construct a self-assembling peptide hydrogel(SAPH)loaded with doxorubicin(DOX)and analyze its drug release performance and biocompatibility in vitro.METHODS:DOX was introduced into SAPH composed of FEFEFKFK(F,phenylalanine;E,glutamic acid;K,lysine),prepared to contain 20,30,and 40 g/L DOXSAPH of FEFEFKFK;the DOX mass concentration in this system was 2 g/L.Three kinds of DOX-SAPH were placed in PBS,and the amount of DOX released was regularly detected.Three DOX-SAPH extracts were co-cultured with rabbit bone marrow mesenchymal stem cells,and the cell survival rate was detected by CCK-8 method.The DOX solution containing different concentrations of DOX and the DOX-SAPH extract(containing FEFEFKFK 30 g/L)were co-cultured with MG63 human osteosarcoma cells,and the cell survival rate was detected by CCK-8 method.RESULTS AND CONCLUSION:(1)With the increase of the concentration of FEFEFKFK in the hydrogel,the sustained release effect of DOX-SAPH was enhanced.The cumulative drug release rate of DOX-SAPH at 20,30,and 40 g/L at the 168 th hour in vitro was 88%,83%,and 80%.(2)The cells in 20 g/L DOX-SAPH extract group proliferated faster than 30 and 40 g/L DOX-SAPH extract group at 1,3,and 5 days in vitro(P<0.05),in which the 40 g/L DOX-SAPH extract group had certain cytotoxicity on the first day.(3)DOX solution and DOX-SAPH extract showed concentration-dependent inhibition of MG63 cell proliferation,when the concentration of DOX in the DOX solution and the DOX-SAPH extract was lower than 12.5 mg/L,there was no significant difference in the toxicity of the two to MG63 cells(P>0.05).When the concentration of DOX was higher than 12.5 mg/L,the cell toxicity of the DOX solution was greater than that of DOX-SAP

关 键 词：骨 材料 水凝胶 肿瘤 阿霉素 体外评价 骨肉瘤 组织工程 

分 类 号：R459.9[医药卫生—治疗学] R73-34[医药卫生—临床医学]