作　　者： ; ; ;

机构地区： 中山大学附属第三医院

出　　处： 《中国医药导报》 2017年第21期4-9,F0003,共7页

摘　　要： 目的探讨新型透明质酸改性的聚乙烯亚胺-壳聚糖/DNA(CS-g-PEI/pDNA)纳米粒作为治疗骨关节炎(OA)的可行性。方法通过复凝聚法制备成CP/pDNA纳米粒,然后通过巯基的原位交联与巯基化的透明质酸(HASH)形成交联网络,从而合成HA-CP/pDNA纳米粒;透射电镜观察纳米粒形貌,马尔文粒度分析仪分析其不同构成比(HA-SH∶CP)时的粒径、表面电位等;凝胶电泳阻滞实验检测CP/pDNA的结合力;CCK-8细胞毒性实验检测该基因载体的细胞毒性;以HA-CP/pDNA纳米粒、裸pDNA、CS/pDNA、CP/pDNA纳米粒、LipofectamineTM2000转染软骨细胞,荧光显微镜、流式细胞仪检测基因转染效率。结果 (1)HA-CP/pDNA纳米粒呈较均一的球形,并随HA-SH∶CP质量比的增加,粒径先减小后增大,表面电位电荷逐渐降低。(2)HA-CP/pDNA纳米粒对软骨细胞毒性远低于LipofectamineTM2000(P<0.05)。(3)HA-CP/pDNA纳米粒对软骨细胞转染率较CP/pDNA、CS/pDNA纳米粒大大提高(P<0.05)。(4)大量游离HA导致HA-CP/pDNA纳米粒对软骨细胞的转染效率下降。结论 HA-CP/pDNA纳米粒具有更强的DNA保护能力,对软骨细胞毒性小,转染效率高,并且对软骨细胞具有一定的靶向性。 Objective To investigate the feasibility of hyaluronic acid modified CS-g-PEI/pDNA nanoparticles and mediated gene transfection against osteoarthritis. Methods CP was engaged with p EGFP into nanopartricles by using complex coaceration method and then covering the CP/pDNA with HA-SH, HA-SH forming disulfide cross-linked gene complex,and on the surface of the CP/pDNA form cross-linked network with HA, synthesis HA-CP/pDNA; the morphy of nanoparticle was obtained by TEM, the particle size and surface potential were measured by Malvern particle size analyzer; The pDNA binding ability with CP and the influence of HA-SH on CP were evaluated by gel retardation assay; the cytotoxicity of HA-CP/pDNA nanoparticles was evaluated by CCK-8 assays; HA-CP/pDNA nanoparticles, nake pDNA, CS/pDNA, CP/pDNA nanoparticles and lipofectamineTM2000 transfected chondrocytes, the transfection efficiency was measured by flurescence microscope, flow cytometry. Results①TEM showed that the nanoparticles were well-formed spherical shapes with compact structure, and increased with HA-SH∶CP weight ratio increasing, nanoparticle sizes decreased firstly then increased, the surface potentials gradually decrease.②The cytotoxicity of HA-CP/pDNA nanoparticles had lower cellular toxicity than LipofectamineTM2000（P〈0.05）.③The HA-CP/pDNA nanoparticles transfection efficiency for chondrocytes was highly improved compared to CS/pDNA, CP/pDNA nanoparticles（P〈0.05）.④Free HA decrease HA-CP/pDNA nanoparticles transfection efficiency notably. Conclusion HA-CP/pDNA has lower cytotoxicity and higher transfection efficiency, and it has the ability of targeting chondrocytes.

关 键 词： 透明质酸 纳米粒 基因治疗 软骨细胞

领　　域： []