Preparation of hyaluronic acid modified CS-g-PEI/pDNA nanoparticles and mediated gene transfection against chondrocytes in vitro
CHEN Mingwei1 FAN Binbin1 LU Huading2
1.Department of Orthopaedics, Third Affiliated Hospital of Sun Yat-san University, Guangdong Province, Guangzhou 510630, China;
2.Department of Surgical Trauma and Joints, Fifth Affiliated Hospital of Sun Yat-san University, Guangdong Province, Guangzhou 519000, China
Abstract：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 pEGFP 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. Results ①TEM 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.
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2017, Vol. 14 
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