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Polyethyleneimine-functionalized boron nitride nanospheres as efficient carriers for enhancing the immunostimulatory effect of CpG oligodeoxynucleotides.

Zhang H, Feng S, Yan T, Zhi C, Gao XD, Hanagata N - Int J Nanomedicine (2015)

Bottom Line: Furthermore, the positively charged surface of the BNNS-PEI complexes greatly improved the loading capacity and cellular uptake efficiency of CpG ODNs.Class B CpG ODNs loaded on the BNNS-PEI complexes enhanced the production of interleukin-6 and tumor necrosis factor-α from peripheral blood mononuclear cells compared with CpG ODNs directly loaded on BNNS.Therefore, BNNS-PEI complexes can be used to enhance the immunostimulatory effect and therapeutic activity of CpG ODNs and the treatment of diseases requiring interleukin-6, tumor necrosis factor-α, and interferon-α.

View Article: PubMed Central - PubMed

Affiliation: The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, People's Republic of China.

ABSTRACT
CpG oligodeoxynucleotides (ODNs) stimulate innate and adaptive immune responses. Thus, these molecules are promising therapeutic agents and vaccine adjuvants against various diseases. In this study, we developed a novel CpG ODNs delivery system based on polyethyleneimine (PEI)-functionalized boron nitride nanospheres (BNNS). PEI was coated on the surface of BNNS via electrostatic interactions. The prepared BNNS-PEI complexes had positive zeta potential and exhibited enhanced dispersity and stability in aqueous solution. In vitro cytotoxicity assays revealed that the BNNS-PEI complexes with concentrations up to 100 μg/mL exhibited no obvious cytotoxicity. Furthermore, the positively charged surface of the BNNS-PEI complexes greatly improved the loading capacity and cellular uptake efficiency of CpG ODNs. Class B CpG ODNs loaded on the BNNS-PEI complexes enhanced the production of interleukin-6 and tumor necrosis factor-α from peripheral blood mononuclear cells compared with CpG ODNs directly loaded on BNNS. Contrary to the free CpG ODNs or CpG ODNs directly loaded on BNNS, class B CpG ODNs loaded on the BNNS-PEI complexes induced interferon-α simultaneously. PEI coating may have changed the physical form of class B CpG ODNs on BNNS, which further affected their interaction with Toll-like receptor 9 and induced interferon-α. Therefore, BNNS-PEI complexes can be used to enhance the immunostimulatory effect and therapeutic activity of CpG ODNs and the treatment of diseases requiring interleukin-6, tumor necrosis factor-α, and interferon-α.

No MeSH data available.


Loading of CpG ODNs on BNNS–PEI complexes.Notes: (A) Gel electrophoresis image of the supernatant after interaction of CpG ODNs with BNNS–PEI at increasing weight ratios. (B) UV-vis spectra of CpG ODNs, BNNS–PEI, and BNNS–PEI/CpG ODNs complexes. (C) Loading capacity of CpG ODNs on BNNS and BNNS-PEI complexes with increasing CpG ODNs concentrations, denoted as μg CpG ODNs loaded on 1 mg BNNS. Data are presented as mean ± standard deviation (n=3).Abbreviations: BNNS, boron nitride nanospheres; ODN, oligodeoxynucleotide; PEI, polyethyleneimine.
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f6-ijn-10-5343: Loading of CpG ODNs on BNNS–PEI complexes.Notes: (A) Gel electrophoresis image of the supernatant after interaction of CpG ODNs with BNNS–PEI at increasing weight ratios. (B) UV-vis spectra of CpG ODNs, BNNS–PEI, and BNNS–PEI/CpG ODNs complexes. (C) Loading capacity of CpG ODNs on BNNS and BNNS-PEI complexes with increasing CpG ODNs concentrations, denoted as μg CpG ODNs loaded on 1 mg BNNS. Data are presented as mean ± standard deviation (n=3).Abbreviations: BNNS, boron nitride nanospheres; ODN, oligodeoxynucleotide; PEI, polyethyleneimine.

Mentions: The capability of the BNNS–PEI complexes as carriers for CpG ODNs delivery was investigated by incubating the complexes with CpG ODNs. Loading of CpG ODNs onto the BNNS–PEI complexes was initially confirmed through gel electrophoresis. The band of CpG ODNs from the supernatant became weaker when the BNNS–PEI complexes were mixed with CpG ODNs at a high weight ratio (BNNS–PEI/CpG ODNs) and disappeared at the ratio of 50 (Figure 6A). This result indicates the electrostatic binding of the negatively charged CpG ODNs to the positively charged BNNS–PEI complexes, and all CpG ODNs could be bound by the BNNS–PEI complexes at the weight ratio of 50. The characteristic adsorption peak at 260 nm for CpG ODNs appeared in the UV-vis spectra of the BNNS–PEI/CpG ODN complexes, which also confirmed the binding of CpG ODNs to the BNNS–PEI complexes (Figure 6B). Meanwhile, the loading capacity of the CpG ODNs on BNNS and the BNNS–PEI complexes increased with increasing CpG ODNs concentration and then reached a plateau. The BNNS–PEI complexes possessed eightfold higher capacity to load CpG ODNs (24 μg/mg nanoparticles) than BNNS (Figure 6C). This result can be attributed to the strong positive surface charge of the BNNS–PEI complexes, which enhanced the loading of the CpG ODNs via electrostatic interactions. The release of CpG ODNs from the BNNS–PEI/CpG ODN complexes was further examined under conditions that corresponded to the physiological environment in TLR-9-localized endolysosomes. However, almost no CpG ODNs were released. This result suggests that the BNNS–PEI/CpG ODN complexes are stable and that CpG ODNs cannot be easily released from the BNNS–PEI complexes because of the strong binding.


Polyethyleneimine-functionalized boron nitride nanospheres as efficient carriers for enhancing the immunostimulatory effect of CpG oligodeoxynucleotides.

Zhang H, Feng S, Yan T, Zhi C, Gao XD, Hanagata N - Int J Nanomedicine (2015)

Loading of CpG ODNs on BNNS–PEI complexes.Notes: (A) Gel electrophoresis image of the supernatant after interaction of CpG ODNs with BNNS–PEI at increasing weight ratios. (B) UV-vis spectra of CpG ODNs, BNNS–PEI, and BNNS–PEI/CpG ODNs complexes. (C) Loading capacity of CpG ODNs on BNNS and BNNS-PEI complexes with increasing CpG ODNs concentrations, denoted as μg CpG ODNs loaded on 1 mg BNNS. Data are presented as mean ± standard deviation (n=3).Abbreviations: BNNS, boron nitride nanospheres; ODN, oligodeoxynucleotide; PEI, polyethyleneimine.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4554408&req=5

f6-ijn-10-5343: Loading of CpG ODNs on BNNS–PEI complexes.Notes: (A) Gel electrophoresis image of the supernatant after interaction of CpG ODNs with BNNS–PEI at increasing weight ratios. (B) UV-vis spectra of CpG ODNs, BNNS–PEI, and BNNS–PEI/CpG ODNs complexes. (C) Loading capacity of CpG ODNs on BNNS and BNNS-PEI complexes with increasing CpG ODNs concentrations, denoted as μg CpG ODNs loaded on 1 mg BNNS. Data are presented as mean ± standard deviation (n=3).Abbreviations: BNNS, boron nitride nanospheres; ODN, oligodeoxynucleotide; PEI, polyethyleneimine.
Mentions: The capability of the BNNS–PEI complexes as carriers for CpG ODNs delivery was investigated by incubating the complexes with CpG ODNs. Loading of CpG ODNs onto the BNNS–PEI complexes was initially confirmed through gel electrophoresis. The band of CpG ODNs from the supernatant became weaker when the BNNS–PEI complexes were mixed with CpG ODNs at a high weight ratio (BNNS–PEI/CpG ODNs) and disappeared at the ratio of 50 (Figure 6A). This result indicates the electrostatic binding of the negatively charged CpG ODNs to the positively charged BNNS–PEI complexes, and all CpG ODNs could be bound by the BNNS–PEI complexes at the weight ratio of 50. The characteristic adsorption peak at 260 nm for CpG ODNs appeared in the UV-vis spectra of the BNNS–PEI/CpG ODN complexes, which also confirmed the binding of CpG ODNs to the BNNS–PEI complexes (Figure 6B). Meanwhile, the loading capacity of the CpG ODNs on BNNS and the BNNS–PEI complexes increased with increasing CpG ODNs concentration and then reached a plateau. The BNNS–PEI complexes possessed eightfold higher capacity to load CpG ODNs (24 μg/mg nanoparticles) than BNNS (Figure 6C). This result can be attributed to the strong positive surface charge of the BNNS–PEI complexes, which enhanced the loading of the CpG ODNs via electrostatic interactions. The release of CpG ODNs from the BNNS–PEI/CpG ODN complexes was further examined under conditions that corresponded to the physiological environment in TLR-9-localized endolysosomes. However, almost no CpG ODNs were released. This result suggests that the BNNS–PEI/CpG ODN complexes are stable and that CpG ODNs cannot be easily released from the BNNS–PEI complexes because of the strong binding.

Bottom Line: Furthermore, the positively charged surface of the BNNS-PEI complexes greatly improved the loading capacity and cellular uptake efficiency of CpG ODNs.Class B CpG ODNs loaded on the BNNS-PEI complexes enhanced the production of interleukin-6 and tumor necrosis factor-α from peripheral blood mononuclear cells compared with CpG ODNs directly loaded on BNNS.Therefore, BNNS-PEI complexes can be used to enhance the immunostimulatory effect and therapeutic activity of CpG ODNs and the treatment of diseases requiring interleukin-6, tumor necrosis factor-α, and interferon-α.

View Article: PubMed Central - PubMed

Affiliation: The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, People's Republic of China.

ABSTRACT
CpG oligodeoxynucleotides (ODNs) stimulate innate and adaptive immune responses. Thus, these molecules are promising therapeutic agents and vaccine adjuvants against various diseases. In this study, we developed a novel CpG ODNs delivery system based on polyethyleneimine (PEI)-functionalized boron nitride nanospheres (BNNS). PEI was coated on the surface of BNNS via electrostatic interactions. The prepared BNNS-PEI complexes had positive zeta potential and exhibited enhanced dispersity and stability in aqueous solution. In vitro cytotoxicity assays revealed that the BNNS-PEI complexes with concentrations up to 100 μg/mL exhibited no obvious cytotoxicity. Furthermore, the positively charged surface of the BNNS-PEI complexes greatly improved the loading capacity and cellular uptake efficiency of CpG ODNs. Class B CpG ODNs loaded on the BNNS-PEI complexes enhanced the production of interleukin-6 and tumor necrosis factor-α from peripheral blood mononuclear cells compared with CpG ODNs directly loaded on BNNS. Contrary to the free CpG ODNs or CpG ODNs directly loaded on BNNS, class B CpG ODNs loaded on the BNNS-PEI complexes induced interferon-α simultaneously. PEI coating may have changed the physical form of class B CpG ODNs on BNNS, which further affected their interaction with Toll-like receptor 9 and induced interferon-α. Therefore, BNNS-PEI complexes can be used to enhance the immunostimulatory effect and therapeutic activity of CpG ODNs and the treatment of diseases requiring interleukin-6, tumor necrosis factor-α, and interferon-α.

No MeSH data available.