Preparation and characterization of protein-loaded N-trimethyl chitosan nanoparticles as nasal delivery system
Preparation and characterization of protein-loaded N-trimethyl chitosan nanoparticles as nasal delivery system
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DOI：
10.1016/j.jconrel.2005.11.014
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发表时间：
2006-03-10
影响因子：
10.8
通讯作者：
Jiskoot, W
中科院分区：
文献类型：
--
作者：
Amidi, M;Romeijn, SG;Jiskoot, W
在这项研究中，N-三甲基壳聚糖（TMC）纳米粒子作为鼻腔递送蛋白质的载体系统的潜力进行了研究。TMC纳米颗粒通过TMC溶液（有或没有卵清蛋白）与三聚磷酸盐在环境温度下在搅拌下的离子交联来制备。纳米粒子的大小，zeta电位和形态进行了研究作为一个功能的制备条件。研究了蛋白负载、蛋白完整性和蛋白释放。通过鸡胚气管纤毛搏动频率测定和体外细胞毒性试验来测试TMC纳米粒的毒性。采用激光共聚焦扫描显微镜观察大鼠鼻黏膜上皮对载FITC-白蛋白的TMC纳米粒的摄取情况。纳米颗粒具有约350 nm的平均尺寸和正ζ电位。它们显示出高达95%的负载效率和高达50%（w/w）的负载容量。包封的卵清蛋白的完整性得以保留。释放研究表明，超过70%的蛋白质在37 ℃下在PBS（pH 7.4）中孵育时保持与TMC纳米颗粒结合至少3小时。用Calu-3细胞进行的细胞毒性试验显示纳米颗粒没有毒性作用，而观察到对鸡气管纤毛搏动频率的部分可逆的纤毛抑制作用。体内摄取研究表明，FITC-白蛋白相关的TMC纳米颗粒的运输通过鼻粘膜。总之，TMC纳米颗粒是一种潜在的新的蛋白质通过鼻粘膜运输系统。(c)2005 Elsevier B. V.保留所有权利。
In this study, the potential of N-trimethyl chitosan (TMC) nanoparticles as a carrier system for the nasal delivery of proteins was investigated. TMC nanoparticles were prepared by ionic crosslinking of TMC solution (with or without ovalbumin) with tripolyphosphate, at ambient temperature while stirring. The size, zeta-potential and morphology of the nanoparticles were investigated as a function of the preparation conditions. Protein loading, protein integrity and protein release were studied. The toxicity of the TMC nanoparticles was tested by ciliary beat frequency measurements of chicken embryo trachea and in vitro cytotoxicity assays. The in vivo uptake of FITC-albumin-loaded TMC nanoparticles by nasal epithelia tissue in rats was studied by confocal laser scanning microscopy. The nanoparticles had an average size of about 350 nm and a positive zeta-potential. They showed a loading efficiency up to 95% and a loading capacity up to 50% (w/w). The integrity of the entrapped ovalbumin was preserved. Release studies showed that more than 70% of the protein remained associated with the TMC nanoparticles for at least 3 h on incubation in PBS (pH 7.4) at 37 degrees C. Cytotoxicity tests with Calu-3 cells showed no toxic effects of the nanoparticles, whereas a partially reversible cilio-inhibiting effect on the ciliary beat frequency of chicken trachea was observed. In vivo uptake studies indicated the transport of FITC-albumin-associated TMC nanoparticles across the nasal mucosa. In conclusion, TMC nanoparticles are a potential new delivery system for transport of proteins through the nasal mucosa. (c) 2005 Elsevier B.V. All rights reserved.