Paquette Cynthia C H, Phanse Yashdeep, Perry Jillian L, Sanchez-Vargas Irma, Airs Paul M, Dunphy Brendan M, Xu Jing, Carlson Jonathan O, Luft J Christopher, DeSimone Joseph M, Bartholomay Lyric C, Beaty Barry J
Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado, United States of America.
Department of Entomology, Iowa State University, Ames, Iowa, United States of America.
PLoS Negl Trop Dis. 2015 May 21;9(5):e0003745. doi: 10.1371/journal.pntd.0003745. eCollection 2015 May.
Nanotechnology offers great potential for molecular genetic investigations and potential control of medically important arthropods. Major advances have been made in mammalian systems to define nanoparticle (NP) characteristics that condition trafficking and biodistribution of NPs in the host. Such information is critical for effective delivery of therapeutics and molecules to cells and organs, but little is known about biodistribution of NPs in mosquitoes.
METHODOLOGY/PRINCIPAL FINDINGS: PRINT technology was used to construct a library of fluorescently labeled hydrogel NPs of defined size, shape, and surface charge. The biodistribution (organ, tissue, and cell tropisms and trafficking kinetics) of positively and negatively charged 200 nm x 200 nm, 80 nm x 320 nm, and 80 nm x 5000 nm NPs was determined in adult Anopheles gambiae mosquitoes as a function of the route of challenge (ingestion, injection or contact) using whole body imaging and fluorescence microscopy. Mosquitoes readily ingested NPs in sugar solution. Whole body fluorescence imaging revealed substantial NP accumulation (load) in the alimentary tracts of the adult mosquitoes, with the greatest loads in the diverticula, cardia and foregut. Positively and negatively charged NPs differed in their biodistribution and trafficking. Following oral challenge, negatively charged NPs transited the alimentary tract more rapidly than positively charged NPs. Following contact challenge, negatively charged NPs trafficked more efficiently in alimentary tract tissues. Following parenteral challenge, positively and negatively charged NPs differed in tissue tropisms and trafficking in the hemocoel. Injected NPs were also detected in cardia/foregut, suggesting trafficking of NPs from the hemocoel into the alimentary tract.
CONCLUSIONS/SIGNIFICANCE: Herein we have developed a tool box of NPs with the biodistribution and tissue tropism characteristics for gene structure/function studies and for delivery of vector lethal cargoes for mosquito control.
纳米技术在分子遗传学研究以及对医学上重要节肢动物的潜在控制方面具有巨大潜力。在哺乳动物系统中已取得重大进展，以确定纳米颗粒（NP）的特性，这些特性决定了纳米颗粒在宿主体内的运输和生物分布。此类信息对于将治疗剂和分子有效递送至细胞和器官至关重要，但对于纳米颗粒在蚊子体内的生物分布却知之甚少。
方法/主要发现：利用PRINT技术构建了一个具有特定大小、形状和表面电荷的荧光标记水凝胶纳米颗粒文库。通过全身成像和荧光显微镜，确定了带正电和带负电的200 nm×200 nm、80 nm×320 nm和80 nm×5000 nm纳米颗粒在成年冈比亚按蚊体内的生物分布（器官、组织和细胞嗜性以及运输动力学），其取决于挑战途径（摄入、注射或接触）。蚊子很容易在糖溶液中摄入纳米颗粒。全身荧光成像显示成年蚊子的消化道中有大量纳米颗粒积累（负载），在憩室、贲门和前肠中的负载量最大。带正电和带负电的纳米颗粒在生物分布和运输方面存在差异。经口服挑战后，带负电的纳米颗粒比带正电的纳米颗粒更快地穿过消化道。经接触挑战后，带负电的纳米颗粒在消化道组织中的运输效率更高。经肠胃外挑战后，带正电和带负电的纳米颗粒在血腔中的组织嗜性和运输情况有所不同。在贲门/前肠中也检测到注射的纳米颗粒，这表明纳米颗粒从血腔运输到了消化道。
结论/意义：在此，我们开发了一个纳米颗粒工具箱，其具有生物分布和组织嗜性特征，可用于基因结构/功能研究以及为控制蚊子而递送载体致死物质。
