非病毒性基因载体的合成研究

基因治疗是一种有效的治疗先天性遗传性疾病以及后天获得性疾病的手段。它通过激发细胞的生物活性或者抑制细胞非正常的功能来治疗或者预防疾病的发生,例如细胞的基因紊乱,细胞的无序增殖。目前基因治疗所面临的问题是缺乏有效的基因递送载体。基因载体主要分为病毒性基因载体和非病毒性基因载体。与病毒性基因载体相比,非病毒性基因载体具有毒性小、安全性高、易于制备、能够荷载分子量大的DNA等优点。本文综述了非病毒性基因载体的合成研究进展。     

壳聚糖及其衍生物基因载体的研究进展

壳聚糖是一种天然的生物可降解性,生物相容性好而且安全无毒的多糖,因而它成为基因治疗载体研究的热点。本文就近年来壳聚糖及其衍生物作为基因载体转染的研究进展和现状作简要的综述,并对转染率的影响因素如壳聚糖的分子量、粒径、脱乙酰度等进行着重介绍。     

PEG化壳聚糖/DNA自组装复合物的制备、表征和体外Hela细胞转染研究

通过有机合成和高分子聚合等方法将亲水性的聚乙二醇接枝到壳聚糖的氨基侧链上，得到了改性的壳聚糖—聚乙二醇接枝共聚物，应用现代波谱等技术对中间产物和最终产物进行了表征，采用绿色荧光蛋白基因质粒pEGFP—N1为DNA模型，在溶液中通过自动(静电)吸附得到PEG化的壳聚糖／DNA自组装复合物，初步研究了该自组装复合物对Hela细胞的体外转染效率。结果表明，活化的聚乙二醇被成功地接枝到壳聚糖上，使不溶于水的壳聚糖改性为水溶性的PEG化的壳聚糖。PEG化壳聚糖／DNA自组装复合物在Hela细胞体外转染率达到81％。因此，PEG化的壳聚糖有可能成为基因转染的非病毒载体。     

叶酸和聚乙二醇接枝作基因载体用壳聚糖的合成与表征

本研究将叶酸和聚乙二醇接枝到四种不同分子量的壳聚糖氨基侧链上,以改善壳聚糖的靶向性和水溶性作基因载体。用FTIE、1HNMR、UV-Vis、DSC和TEM对产物进行了表征,结果表明,叶酸和聚乙二醇被成功地接枝到壳聚糖上,所制得的载体有望作为潜在的肿瘤细胞靶向基因载体。     

pH敏感基因载体的研究现状

基因治疗正成为遗传病、癌症等疾病的有效治疗方法. 基因治疗实现的最大挑战是开发安全有效的基因运载载体以将目的基因从血液运送到细胞质或细胞核. 目前, 常用的基因治疗载体有病毒载体和非病毒载体. 非病毒载体由于安全性好, 易于合成, 易于修饰而得到了更为广泛的研究. 其中pH敏感载体作为功能性非病毒载体, 不仅安全性高, 而且有更好的体内基因转染效率, 为非病毒性载体在临床上的应用开辟了广阔的前景. 本文主要从pH敏感脂质和pH敏感聚合物两方面对pH敏感基因载体进行简要综述, 介绍了该两类载体的构建方法及其对基因的运载机制.     

壳聚糖功能化改性及其在药物载体中的应用

壳聚糖为天然多糖甲壳素脱除部分乙酰基的产物,因其优异的理化性质和生物学活性,成为生物医学应用中最有前景的聚合物之一。由于壳聚糖在中性和碱性溶液中溶解性较差,其应用受到了一定限制,通常对其进行功能化改性,生成一系列壳聚糖衍生物。壳聚糖衍生物可改善壳聚糖的水溶性、生物活力及力学性能,从而提高其利用价值,扩大其应用领域。本文综述了几种壳聚糖常用的功能化改性方法,包括接枝共聚、交联、羧甲基化、酰基化、烷基化。并介绍了改性壳聚糖作为药物载体应用的研究进展,旨在为改性壳聚糖的研究提供一定理论基础。     

聚乙二醇改性壳聚糖作为小分子药物载体的研究进展

壳聚糖具有抗菌、抗氧化、增强胶凝特性以及可作为生物活性分子的微型或纳米载体等优点,其化学改性和应用近年来受到广泛关注。然而,壳聚糖既不溶于有机溶剂也不溶于水,极大地限制了它的应用。在改性的壳聚糖中,聚乙二醇化壳聚糖不仅能保持壳聚糖的优点,还能提高水溶性,并能有效运输生物活性分子。本文综述了近5年间聚乙二醇化壳聚糖作为紫杉醇、阿霉素、5-氟尿嘧啶等小分子载体的研究进展,为今后的研究提供有益参考和理论依据。     

聚乙二醇改性壳聚糖作为小分子药物载体的研究进展

壳聚糖具有抗菌、抗氧化、增强胶凝特性以及可作为生物活性分子的微型或纳米载体等优点,因此其化学改性和应用近年来受到广泛关注。然而,壳聚糖既不溶于有机溶剂也不溶于水,极大地限制了它的应用。在改性的壳聚糖中,聚乙二醇化壳聚糖不仅能保持壳聚糖的优点,还能提高水溶性,并能有效运输生物活性分子。因此,本文总结了2008-2012年聚乙二醇化壳聚糖作为紫杉醇、阿霉素、5-氟尿嘧啶等小分子载体的最新进展,为今后聚乙二醇化壳聚糖的研究提供有益参考和理论依据。     

负载壳聚糖及其衍生物的材料表面血液相容性研究进展

壳聚糖及其衍生物具有多种生物活性,基于壳聚糖及其衍生物的材料表面改性是获取各种生物活性表面的重要手段,在生物材料领域显示出广阔的应用前景。为了获得血液相容性良好的壳聚糖改性生物材料表面,可通过引入具备抗凝活性的壳聚糖衍生物或壳聚糖/抗凝剂复合物来抑制壳聚糖固有的促凝作用。本文综述了负载有壳聚糖或其衍生物的材料表面的血液相容性改性方法方面的进展,并根据表面改性方法的不同按照物理改性和化学改性分别对其进行了阐述。     

基于树枝状聚合物构建的核酸载体研究进展

安全有效的基因载体对于基因治疗有着重要的应用价值.相对于病毒性基因载体,非病毒基因载体具有低免疫原性、易于大规模生产和成本较低的特性,因而受到越来越多的关注,但是非病毒基因载体在转染效率和选择性方面有一定的限制性.综述了基于树枝状聚合物构建的基因载体的研究进展,包括:(i)树枝状及其衍生材料;(ii)与超分子母体材料,如环糊精、葫芦脲结合的的树枝状聚合物基因载体;(iii)与无机材料结合的树枝状聚合物载体,如金纳米粒子、氧化铁纳米粒子以及石墨烯等结合的树枝状聚合物构建的基因载体;(iv)功能响应性的树枝状聚合物载体,如p H响应型、二硫键响应型、氧化还原型以及氟代化树枝状基因载体.     

Chemo-physical and biological evaluation of poly(L-lysine)-grafted chitosan copolymers used for highly efficient gene delivery

For the success of non-viral gene delivery, it is of great importance to develop gene vectors with high efficiency but low toxicity. We demonstrate that PLL-grafted chitosan copolymers combine the advantages of PLL with its good pDNA-binding ability and of chitosan with its good biocompatibility. The chemo-physical properties of the prepared Chi-g-PLL copolymers are thoroughly characterized. The in vitro transfection study shows that the copolymers have a much higher gene transfer ability than the starting materials chitosan and PLL. A positive correlation between PLL chain lengths and transfection efficiency of the copolymers is found. Our results suggest that these novel Chi-g-PLL copolymers are good candidates for gene delivery in vivo.     

壳聚糖及其衍生物递送miRNA/siRNA的研究进展

微小RNA(microRNA,miRNA)和短链干扰RNA (small interfering RNA,siRNA)是两类具有调节基因表达功能的内源性非编码性小RNA分子.它们已成为多种疾病的潜在治疗药物,逐渐被应用于基因治疗中,而将小RNA应用于基因治疗亟需一种安全高效的递送载体.壳聚糖及其衍生物作为一种可降解、低...     

纳米阳离子多聚物在基因载体系统的应用

阳离子多聚物能与DNA通过静电吸附作用而自组装成纳米微粒,防止DNA被核酸酶降解.阳离子多聚物由于具备合成简便、储存稳定、基因荷载率高、靶向性强、免疫原性低等优点而被用作基因载体.阳离子多聚物按特性可分为两类：合成型和天然型.经典的人工合成型阳离子多聚物基因载体主要有：多聚乙烯亚胺、多聚左旋赖氨酸和树状大分子等;天然生物型阳离子多聚物基因载体主要有壳聚糖及其衍生物和明胶等.本文详细论述了各种阳离子聚合物用作基因载体的性能特点、自身缺陷、介导基因进入细胞的机理和靶向性策略,并对非病毒基因载体的发展作出展望.     

Lanthanide-doped chitosan nanospheres as cell nuclei illuminator and fluorescent nonviral vector for plasmid DNA delivery

Lanthanide-doped chitosan nanospheres (LDCNs) and lanthanide-Fe(3)O(4)-doped chitosan nanospheres (Fe(3)O(4)-LDCNs) are fabricated and show fluorescence, MRI effectiveness and desirable biocompatibility. Superior to most nanoparticles that were found retained in cytoplasmic organelles rather than the nucleus, the prepared chitosan nanospheres preferentially enter and illuminate the cell nuclei. Complexation of plasmid DNA (pDNA) to the nanospheres was accomplished via electrostatic forces between positively charged chitosan and negatively charged pDNA. Satisfactory results of the complexation indicate that the prepared chitosan nanospheres can serve as a potential fluorescent nonviral vector for pDNA delivery that can fulfill gene delivery and transfer efficiency assessment simultaneously, without an additional step of tagging fluorophores to the vectors carried out in fabrications of currently available pDNA delivery vectors.     

Recent progress in cationic polymeric gene carriers for cancer therapy

In recent years,various carriers for gene delivery nave been developed for biomedical applications.Among all kinds of gene carriers,cationic polymeric carriers for delivery therapeutic gene as non-viral carriers have received growing interests due to their improved high transfection efficiency with the relative safety.In particular,the advancement of novel polymeric gene carriers has gained much progress in the development of effective anticancer therapy.Herein,this review focused on the development of cationic polymeric carriers for cancer therapy,including polyethylenimine(PEI),polyamidoamine(PAMAM) dendrimers,polylysine(PLL),chitosan and modified cationic polymers.And recent progresses in the development of novel polymeric carriers for gene delivery,such as targeted gene carriers,responsive gene carriers and multifunctional gene carriers,were summarized.Finally,the future perspectives in the development of novel polymeric carriers for delivery gene were presented.     

Dendrimers as Non-Viral Vectors in Gene-Directed Enzyme Prodrug Therapy

Gene-directed enzyme prodrug therapy (GDEPT) has been intensively studied as a promising new strategy of prodrug delivery, with its main advantages being represented by an enhanced efficacy and a reduced off-target toxicity of the active drug. In recent years, numerous therapeutic systems based on GDEPT strategy have entered clinical trials. In order to deliver the desired gene at a specific site of action, this therapeutic approach uses vectors divided in two major categories, viral vectors and non-viral vectors, with the latter being represented by chemical delivery agents. There is considerable interest in the development of non-viral vectors due to their decreased immunogenicity, higher specificity, ease of synthesis and greater flexibility for subsequent modulations. Dendrimers used as delivery vehicles offer many advantages, such as: nanoscale size, precise molecular weight, increased solubility, high load capacity, high bioavailability and low immunogenicity. The aim of the present work was to provide a comprehensive overview of the recent advances regarding the use of dendrimers as non-viral carriers in the GDEPT therapy.     

Chitosan-based nanocomposites for medical applications

Chitosan as a biobased polymer is gaining increasing attention due to its extraordinary physico-chemical characteristics and properties. While a primary use of chitosan has been in horticultural and agricultural applications for plant defense and to increase crop yield, recent research reports display various new utilizations in the field of advanced biomedical devices, targeted drug delivery, and as bioimaging sensors. Chitosan possesses multiple characteristics such as antimicrobial properties, stimuli-responsiveness, tunable mechanical strength, biocompatibility, biodegradability, and water-solubility. Further, chitosan can be processed into nanoparticles, nano-vehicles, nanocapsules, scaffolds, fiber meshes, and 3D printed scaffolds for a variety of applications. In recent times, nanoparticles incorporated in chitosan matrices have been identified to show superior biological activity, as cells tend to proliferate/differentiate faster when they interact with nanocomposites rather than bulk or micron size substrates/scaffolds. The present article intents to cover chitosan-based nanocomposites used for regenerative medicine, wound dressings, drug delivery, and biosensing applications.     

Modulation of surface charge, particle size and morphological properties of chitosan-TPP nanoparticles intended for gene delivery

This work investigates the polyanion initiated gelation process in fabricating chitosan-TPP (tripolyphosphate) nanoparticles in the size range of 100-250 nm intended to be used as carriers for the delivery of gene or protein macromolecules. It demonstrates that ionic gelation of cationic chitosan molecules offers a flexible and easily controllable process for systematically and predictably manipulating particle size and surface charge which are important properties in determining gene transfection efficacy if the nanoparticles are used as non-viral vectors for gene delivery, or as delivery carriers for protein molecules. Variations in chitosan molecular weight, chitosan concentration, chitosan to TPP weight ratio and solution pH value were examined systematically for their effects on nanoparticle size, intensity of surface charge, and tendency of particle aggregation so as to enable speedy fabrication of chitosan nanoparticles with predetermined properties. The chitosan-TPP nanoparticles exhibited a high positive surface charge across a wide pH range, and the isoelectric point (IEP) of the nanoparticles was found to be at pH 9.0. Detailed imaging analysis of the particle morphology revealed that the nanoparticles possess typical shapes of polyhedrons (e.g., pentagon and hexagon), indicating a similar crystallisation mechanism during the particle formation and growth process. This study demonstrates that systematic design and modulation of the surface charge and particle size of chitosan-TPP nanoparticles can be readily achieved with the right control of critical processing parameters, especially the chitosan to TPP weight ratio.     

Chitosan nanoparticles: a promising system in novel drug delivery

The ability of nanoparticles to manipulate the molecules and their structures has revolutionized the conventional drug delivery system. The chitosan nanoparticles, because of their biodegradability, biocompatibility, better stability, low toxicity, simple and mild preparation methods, offer a valuable tool to novel drug delivery systems in the present scenario. Besides ionotropic gelation method, other methods such as microemulsion method, emulsification solvent diffusion method, polyelectrolyte complex method, emulsification cross-linking method, complex coacervation method and solvent evaporation method are also in use. The chitosan nanoparticles have also been reported to have key applications in parentral drug delivery, per-oral administration of drugs, in non-viral gene delivery, in vaccine delivery, in ocular drug delivery, in electrodeposition, in brain targeting drug delivery, in stability improvement, in mucosal drug delivery in controlled drug delivery of drugs, in tissue engineering and in the effective delivery of insulin. The present review describes origin and properties of chitosan and its nanoparticles along with the different methods of its preparation and the various areas of novel drug delivery where it has got its application.