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Intranasal, siRNA Delivery to the Brain by TAT/MGF Tagged PEGylated Chitosan Nanoparticles.

Malhotra M, Tomaro-Duchesneau C, Saha S, Prakash S - J Pharm (Cairo) (2013)

Bottom Line: The currently used delivery strategies such as implantation of catheters, intracarotid infusions, surgeries, and chemotherapies are invasive in nature and pose a greater risk of postsurgical complications, which can have fatal side effects.The results indicate that 0.5 mg/kg of siRNA is delivered successfully to the hippocampus, thalamus, hypothalamus, and Purkinje cells in the cerebellum after 4 hrs of post intranasal delivery.The results indicate maximum delivery to the brain in comparison to other tissues with no cellular toxic effects.

View Article: PubMed Central - PubMed

Affiliation: Biomedical Technology and Cell Therapy Research Laboratory, Departments of Biomedical Engineering, Faculty of Medicine, McGill University, 3775 University Street, Room 311, Lyman Duff Medical Building, Montreal, QC, Canada H3A 2B4.

ABSTRACT
Neurodegeneration is characterized by progressive loss of structure and function of neurons. Several therapeutic methods and drugs are available to alleviate the symptoms of these diseases. The currently used delivery strategies such as implantation of catheters, intracarotid infusions, surgeries, and chemotherapies are invasive in nature and pose a greater risk of postsurgical complications, which can have fatal side effects. The current study utilizes a peptide (TAT and MGF) tagged PEGylated chitosan nanoparticle formulation for siRNA delivery, administered intranasally, which can bypass the blood brain barrier. The study investigates the optimal dose, duration, biodistribution, and toxicity, of the nanoparticle-siRNA formulation, in-vivo. The results indicate that 0.5 mg/kg of siRNA is delivered successfully to the hippocampus, thalamus, hypothalamus, and Purkinje cells in the cerebellum after 4 hrs of post intranasal delivery. The results indicate maximum delivery to the brain in comparison to other tissues with no cellular toxic effects. This study shows the potential of peptide-tagged PEGylated chitosan nanoparticles to be delivered intranasally and target brain tissue for the treatment of neurological disorders.

No MeSH data available.


Related in: MedlinePlus

(a) Histopathological images of brain tissue (cerebral cortex and cerebellum) 4 hrs after receiving the nanoparticle formulation carrying doses of biotin-siRNA: (A) 0.25 mg/kg, (B) 0.5 mg/kg, (C) 1 mg/kg, (D) 2 mg/kg, and (E) control. (b) Quantitative analysis of the stained area in tissues using Image J. This study proved that the novel nanoparticle formulation successfully delivered the biotin-siRNA with high efficiency and selective targeting. The optimal dose of siRNA delivered via nanoparticles was determined to be 0.5 mg/kg. (c) Histopathological images of the cerebral cortex and cerebellum with nanoparticles carrying 0.5 mg/kg of biotin-siRNA at different time points, (A) 4 hrs, (B) 16 hrs, (C) 24 hrs, and (D) 48 hrs. (d) Quantitative analysis of the stained area in tissues using Image J. This study confirmed successful delivery of biotin-siRNA to the brain within 4 hrs of intranasal administration, with its clearance after 16 h. The graph shows a representative result of independent readings from two animals in each group (n = 2) mean ± s.d. ∗∗∗P < 0.01 was considered highly significant based on Tukey's post hoc analysis, when compared with other groups.
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fig3: (a) Histopathological images of brain tissue (cerebral cortex and cerebellum) 4 hrs after receiving the nanoparticle formulation carrying doses of biotin-siRNA: (A) 0.25 mg/kg, (B) 0.5 mg/kg, (C) 1 mg/kg, (D) 2 mg/kg, and (E) control. (b) Quantitative analysis of the stained area in tissues using Image J. This study proved that the novel nanoparticle formulation successfully delivered the biotin-siRNA with high efficiency and selective targeting. The optimal dose of siRNA delivered via nanoparticles was determined to be 0.5 mg/kg. (c) Histopathological images of the cerebral cortex and cerebellum with nanoparticles carrying 0.5 mg/kg of biotin-siRNA at different time points, (A) 4 hrs, (B) 16 hrs, (C) 24 hrs, and (D) 48 hrs. (d) Quantitative analysis of the stained area in tissues using Image J. This study confirmed successful delivery of biotin-siRNA to the brain within 4 hrs of intranasal administration, with its clearance after 16 h. The graph shows a representative result of independent readings from two animals in each group (n = 2) mean ± s.d. ∗∗∗P < 0.01 was considered highly significant based on Tukey's post hoc analysis, when compared with other groups.

Mentions: The optimal dose to be delivered to the four-week old C57BL/6J male mice was determined by administering the animals with different doses of biotin-tagged scrambled siRNA. Figure 3(a) represents histopathological sections of the cerebral cortex and cerebellum from animals receiving different concentrations of nanoparticle-siRNA formulations. The animals were sacrificed after 4 h. The dark brown stained pyramidal neuronal cells obtained with 0.5 mg/kg of scrambled biotin-siRNA complexed nanoparticles ensured the delivery of siRNA in the neuronal cells of cerebral cortex (P = 0.0001) and in the Purkinje cells of cerebellum (P = 0.0001) as compared to the untreated control. Other animals that received 0.25 mg/kg of scrambled biotin-siRNA showed faint staining in the neuronal cells of cerebral cortex (P = 0.006); whereas, animals that received 1 and 2 mg/kg of scrambled biotin-siRNA dose did not show any staining in the tissue. Figure 3(b) represents the quantitative analysis of the tissues using Image J software (NIH, USA), which calculates the mean percentage area of the dark brown stained cells.


Intranasal, siRNA Delivery to the Brain by TAT/MGF Tagged PEGylated Chitosan Nanoparticles.

Malhotra M, Tomaro-Duchesneau C, Saha S, Prakash S - J Pharm (Cairo) (2013)

(a) Histopathological images of brain tissue (cerebral cortex and cerebellum) 4 hrs after receiving the nanoparticle formulation carrying doses of biotin-siRNA: (A) 0.25 mg/kg, (B) 0.5 mg/kg, (C) 1 mg/kg, (D) 2 mg/kg, and (E) control. (b) Quantitative analysis of the stained area in tissues using Image J. This study proved that the novel nanoparticle formulation successfully delivered the biotin-siRNA with high efficiency and selective targeting. The optimal dose of siRNA delivered via nanoparticles was determined to be 0.5 mg/kg. (c) Histopathological images of the cerebral cortex and cerebellum with nanoparticles carrying 0.5 mg/kg of biotin-siRNA at different time points, (A) 4 hrs, (B) 16 hrs, (C) 24 hrs, and (D) 48 hrs. (d) Quantitative analysis of the stained area in tissues using Image J. This study confirmed successful delivery of biotin-siRNA to the brain within 4 hrs of intranasal administration, with its clearance after 16 h. The graph shows a representative result of independent readings from two animals in each group (n = 2) mean ± s.d. ∗∗∗P < 0.01 was considered highly significant based on Tukey's post hoc analysis, when compared with other groups.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig3: (a) Histopathological images of brain tissue (cerebral cortex and cerebellum) 4 hrs after receiving the nanoparticle formulation carrying doses of biotin-siRNA: (A) 0.25 mg/kg, (B) 0.5 mg/kg, (C) 1 mg/kg, (D) 2 mg/kg, and (E) control. (b) Quantitative analysis of the stained area in tissues using Image J. This study proved that the novel nanoparticle formulation successfully delivered the biotin-siRNA with high efficiency and selective targeting. The optimal dose of siRNA delivered via nanoparticles was determined to be 0.5 mg/kg. (c) Histopathological images of the cerebral cortex and cerebellum with nanoparticles carrying 0.5 mg/kg of biotin-siRNA at different time points, (A) 4 hrs, (B) 16 hrs, (C) 24 hrs, and (D) 48 hrs. (d) Quantitative analysis of the stained area in tissues using Image J. This study confirmed successful delivery of biotin-siRNA to the brain within 4 hrs of intranasal administration, with its clearance after 16 h. The graph shows a representative result of independent readings from two animals in each group (n = 2) mean ± s.d. ∗∗∗P < 0.01 was considered highly significant based on Tukey's post hoc analysis, when compared with other groups.
Mentions: The optimal dose to be delivered to the four-week old C57BL/6J male mice was determined by administering the animals with different doses of biotin-tagged scrambled siRNA. Figure 3(a) represents histopathological sections of the cerebral cortex and cerebellum from animals receiving different concentrations of nanoparticle-siRNA formulations. The animals were sacrificed after 4 h. The dark brown stained pyramidal neuronal cells obtained with 0.5 mg/kg of scrambled biotin-siRNA complexed nanoparticles ensured the delivery of siRNA in the neuronal cells of cerebral cortex (P = 0.0001) and in the Purkinje cells of cerebellum (P = 0.0001) as compared to the untreated control. Other animals that received 0.25 mg/kg of scrambled biotin-siRNA showed faint staining in the neuronal cells of cerebral cortex (P = 0.006); whereas, animals that received 1 and 2 mg/kg of scrambled biotin-siRNA dose did not show any staining in the tissue. Figure 3(b) represents the quantitative analysis of the tissues using Image J software (NIH, USA), which calculates the mean percentage area of the dark brown stained cells.

Bottom Line: The currently used delivery strategies such as implantation of catheters, intracarotid infusions, surgeries, and chemotherapies are invasive in nature and pose a greater risk of postsurgical complications, which can have fatal side effects.The results indicate that 0.5 mg/kg of siRNA is delivered successfully to the hippocampus, thalamus, hypothalamus, and Purkinje cells in the cerebellum after 4 hrs of post intranasal delivery.The results indicate maximum delivery to the brain in comparison to other tissues with no cellular toxic effects.

View Article: PubMed Central - PubMed

Affiliation: Biomedical Technology and Cell Therapy Research Laboratory, Departments of Biomedical Engineering, Faculty of Medicine, McGill University, 3775 University Street, Room 311, Lyman Duff Medical Building, Montreal, QC, Canada H3A 2B4.

ABSTRACT
Neurodegeneration is characterized by progressive loss of structure and function of neurons. Several therapeutic methods and drugs are available to alleviate the symptoms of these diseases. The currently used delivery strategies such as implantation of catheters, intracarotid infusions, surgeries, and chemotherapies are invasive in nature and pose a greater risk of postsurgical complications, which can have fatal side effects. The current study utilizes a peptide (TAT and MGF) tagged PEGylated chitosan nanoparticle formulation for siRNA delivery, administered intranasally, which can bypass the blood brain barrier. The study investigates the optimal dose, duration, biodistribution, and toxicity, of the nanoparticle-siRNA formulation, in-vivo. The results indicate that 0.5 mg/kg of siRNA is delivered successfully to the hippocampus, thalamus, hypothalamus, and Purkinje cells in the cerebellum after 4 hrs of post intranasal delivery. The results indicate maximum delivery to the brain in comparison to other tissues with no cellular toxic effects. This study shows the potential of peptide-tagged PEGylated chitosan nanoparticles to be delivered intranasally and target brain tissue for the treatment of neurological disorders.

No MeSH data available.


Related in: MedlinePlus