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Biodistribution and Toxicity Studies of PRINT Hydrogel Nanoparticles in Mosquito Larvae and Cells.

Phanse Y, Dunphy BM, Perry JL, Airs PM, Paquette CC, Carlson JO, Xu J, Luft JC, DeSimone JM, Beaty BJ, Bartholomay LC - PLoS Negl Trop Dis (2015)

Bottom Line: Positively charged NPs were more associated with the gastric caeca in the gastrointestinal tract.Positively charged NPs trafficked to the cytosol, but negatively charged NPs co-localized with lysosomes.Following in vitro and in vivo challenge, none of the NPs tested induced any cytotoxic effects.

View Article: PubMed Central - PubMed

Affiliation: Department of Entomology, Iowa State University, Ames, Iowa, United States of America.

ABSTRACT
Mosquito-borne diseases continue to remain major threats to human and animal health and impediments to socioeconomic development. Increasing mosquito resistance to chemical insecticides is a great public health concern, and new strategies/technologies are necessary to develop the next-generation of vector control tools. We propose to develop a novel method for mosquito control that employs nanoparticles (NPs) as a platform for delivery of mosquitocidal dsRNA molecules to silence mosquito genes and cause vector lethality. Identifying optimal NP chemistry and morphology is imperative for efficient mosquitocide delivery. Toward this end, fluorescently labeled polyethylene glycol NPs of specific sizes, shapes (80 nm x 320 nm, 80 nm x 5000 nm, 200 nm x 200 nm, and 1000 nm x 1000 nm) and charges (negative and positive) were fabricated by Particle Replication in Non-Wetting Templates (PRINT) technology. Biodistribution, persistence, and toxicity of PRINT NPs were evaluated in vitro in mosquito cell culture and in vivo in Anopheles gambiae larvae following parenteral and oral challenge. Following parenteral challenge, the biodistribution of the positively and negatively charged NPs of each size and shape was similar; intense fluorescence was observed in thoracic and abdominal regions of the larval body. Positively charged NPs were more associated with the gastric caeca in the gastrointestinal tract. Negatively charged NPs persisted through metamorphosis and were observed in head, body and ovaries of adults. Following oral challenge, NPs were detected in the larval mid- and hindgut. Positively charged NPs were more efficiently internalized in vitro than negatively charged NPs. Positively charged NPs trafficked to the cytosol, but negatively charged NPs co-localized with lysosomes. Following in vitro and in vivo challenge, none of the NPs tested induced any cytotoxic effects.

No MeSH data available.


Related in: MedlinePlus

In vivo fluorescence quantification of particles.Fourth instar larvae were injected with 45 nL (235 ng) (A) 80 nm x 320 nm (B) 200 nm x 200 nm and (C) 80 nm x 5000 nm particles. At indicated time points, larvae were dissected and fluorescence from thorax and abdomen quantified. Data are expressed as the mean ± SEM of three independent experiments. n = 10 larvae/group/time point. Statistical difference (p<0.05) from controls and positives is indicated by * and #, respectively.
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pntd.0003735.g005: In vivo fluorescence quantification of particles.Fourth instar larvae were injected with 45 nL (235 ng) (A) 80 nm x 320 nm (B) 200 nm x 200 nm and (C) 80 nm x 5000 nm particles. At indicated time points, larvae were dissected and fluorescence from thorax and abdomen quantified. Data are expressed as the mean ± SEM of three independent experiments. n = 10 larvae/group/time point. Statistical difference (p<0.05) from controls and positives is indicated by * and #, respectively.

Mentions: For efficient, systemic RNAi, dsRNA should reach and persist in the hemolymph where it will be distributed throughout the body by virtue of circulation through the open circulatory system [35]. Studies have shown that dsRNA is enzymatically degraded in hemolymph of some insects e.g., Manduca sexta [11]. Thus, to increase hemolymph delivery, the NP vehicle should protect the dsRNA when trafficking in the larval body. In this study, in vivo imaging was employed to test the persistence of hydrogel NPs in larval thoraces and abdomens. All of the particle groups tested persisted through 3 days p.i. (Fig 5). Interestingly, the 80 nm x 320 nm and 200 nm x 200 nm negatively charged NPs were more abundantly detected at each time point than positively charged NPs. This difference was not as great with the 80 nm x 5000 nm NPs. This may be attributable to more efficient internalization of the positively charged NPs in mosquito cells in vivo as was shown in vitro (Fig 1).


Biodistribution and Toxicity Studies of PRINT Hydrogel Nanoparticles in Mosquito Larvae and Cells.

Phanse Y, Dunphy BM, Perry JL, Airs PM, Paquette CC, Carlson JO, Xu J, Luft JC, DeSimone JM, Beaty BJ, Bartholomay LC - PLoS Negl Trop Dis (2015)

In vivo fluorescence quantification of particles.Fourth instar larvae were injected with 45 nL (235 ng) (A) 80 nm x 320 nm (B) 200 nm x 200 nm and (C) 80 nm x 5000 nm particles. At indicated time points, larvae were dissected and fluorescence from thorax and abdomen quantified. Data are expressed as the mean ± SEM of three independent experiments. n = 10 larvae/group/time point. Statistical difference (p<0.05) from controls and positives is indicated by * and #, respectively.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4440723&req=5

pntd.0003735.g005: In vivo fluorescence quantification of particles.Fourth instar larvae were injected with 45 nL (235 ng) (A) 80 nm x 320 nm (B) 200 nm x 200 nm and (C) 80 nm x 5000 nm particles. At indicated time points, larvae were dissected and fluorescence from thorax and abdomen quantified. Data are expressed as the mean ± SEM of three independent experiments. n = 10 larvae/group/time point. Statistical difference (p<0.05) from controls and positives is indicated by * and #, respectively.
Mentions: For efficient, systemic RNAi, dsRNA should reach and persist in the hemolymph where it will be distributed throughout the body by virtue of circulation through the open circulatory system [35]. Studies have shown that dsRNA is enzymatically degraded in hemolymph of some insects e.g., Manduca sexta [11]. Thus, to increase hemolymph delivery, the NP vehicle should protect the dsRNA when trafficking in the larval body. In this study, in vivo imaging was employed to test the persistence of hydrogel NPs in larval thoraces and abdomens. All of the particle groups tested persisted through 3 days p.i. (Fig 5). Interestingly, the 80 nm x 320 nm and 200 nm x 200 nm negatively charged NPs were more abundantly detected at each time point than positively charged NPs. This difference was not as great with the 80 nm x 5000 nm NPs. This may be attributable to more efficient internalization of the positively charged NPs in mosquito cells in vivo as was shown in vitro (Fig 1).

Bottom Line: Positively charged NPs were more associated with the gastric caeca in the gastrointestinal tract.Positively charged NPs trafficked to the cytosol, but negatively charged NPs co-localized with lysosomes.Following in vitro and in vivo challenge, none of the NPs tested induced any cytotoxic effects.

View Article: PubMed Central - PubMed

Affiliation: Department of Entomology, Iowa State University, Ames, Iowa, United States of America.

ABSTRACT
Mosquito-borne diseases continue to remain major threats to human and animal health and impediments to socioeconomic development. Increasing mosquito resistance to chemical insecticides is a great public health concern, and new strategies/technologies are necessary to develop the next-generation of vector control tools. We propose to develop a novel method for mosquito control that employs nanoparticles (NPs) as a platform for delivery of mosquitocidal dsRNA molecules to silence mosquito genes and cause vector lethality. Identifying optimal NP chemistry and morphology is imperative for efficient mosquitocide delivery. Toward this end, fluorescently labeled polyethylene glycol NPs of specific sizes, shapes (80 nm x 320 nm, 80 nm x 5000 nm, 200 nm x 200 nm, and 1000 nm x 1000 nm) and charges (negative and positive) were fabricated by Particle Replication in Non-Wetting Templates (PRINT) technology. Biodistribution, persistence, and toxicity of PRINT NPs were evaluated in vitro in mosquito cell culture and in vivo in Anopheles gambiae larvae following parenteral and oral challenge. Following parenteral challenge, the biodistribution of the positively and negatively charged NPs of each size and shape was similar; intense fluorescence was observed in thoracic and abdominal regions of the larval body. Positively charged NPs were more associated with the gastric caeca in the gastrointestinal tract. Negatively charged NPs persisted through metamorphosis and were observed in head, body and ovaries of adults. Following oral challenge, NPs were detected in the larval mid- and hindgut. Positively charged NPs were more efficiently internalized in vitro than negatively charged NPs. Positively charged NPs trafficked to the cytosol, but negatively charged NPs co-localized with lysosomes. Following in vitro and in vivo challenge, none of the NPs tested induced any cytotoxic effects.

No MeSH data available.


Related in: MedlinePlus