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Preparation of Conotoxin-Encapsulated Chitosan Nanoparticles and Evaluation of Their Skin Permeability

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Abstract

μ-Conotoxin CnIIIC (conotoxin, CTX)–loaded chitosan nanoparticles (CTX-NPs) were prepared using the ionic cross-linking method. The CTX-NPs were spherical and well with a polydispersity index of 0.292 ± 0.039, drug loading efficiency of 25.9 ± 1.2%, and encapsulation efficiency of 95.6 ± 1.3%. In vitro release studies showed that the release behavior of CTX-NPs in a pH 5.0 acetate buffer followed zero-order kinetics. In vitro transdermal experiments using Franz diffusion cells mounted with mouse abdominal skin demonstrated that the cumulative intradermal deposition amount of CTX per unit area in 8 h (D8) and permeability coefficient (Pf) of CTX loaded on CTX-NPs were 2.30- and 7.71-times that of the CTX solution. In vivo transdermal experiments in mice showed that the amount of CTX deposited in the skin after 8 h of CTX saline administration was significantly lower than that of CTX deposited in the skin after administration of CTX-NPs. In vitro fluorescence labeling transdermal studies through Franz diffusion cells mounted with mouse abdominal skin indicated that CTX-NPs aggregated at hair follicles. Skin irritation tests in mice indicated that the irritation due to CTX-NPs was negligible. The cytotoxicity experiment showed that the viability of Balb/c 3T3 cells with CTX-NPs containing 230 μg/mL (0.08 μM) CTX was greater than 75%. CTX-NPs increase intradermal deposition of CTX by accumulating in hair follicles, which has positive implications for transdermal penetration of CTX.

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Data Availability

Data sharing is not applicable to this article as no new data were created or analyzed in this study.

Abbreviations

CS:

Chitosan

DMEM:

Dulbecco’s modified Eagle’s medium

HPLC:

High-performance liquid chromatography

NIH:

National Institutes of Health

NLC:

Nanostructured lipid carriers

OD:

Optical density

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Acknowledgements

We would like to express our gratitude to all laboratory members for their technical advice and assistance. We also thank Renrui Biotechnology Inc. for providing the reagents.

Funding

This work was supported by grants from the 2019 Linyi Key R&D Program Project (grant number 2019ZDYF007) , Rizhao Science and Technology Innovation Project (Project Number: 2019CXZX1205) and Shandong Province Small And Medium-sized Technology-based Enterprise Innovation Ability Improvement Project (Project Number: 2021TSGC1246).

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Authors and Affiliations

  1. College of Medicine, Linyi University, Linyi, 276000, Shandong, People’s Republic of China

    Haigang Li, JiPeng Yao, Yong Guo, JingJing Huo, Haijuan Zhang & Chun Zhang

  2. Shandong Renrui Biotechnology Inc., RiZhao, Shandong, People’s Republic of China

    Zengtao Zhang & Chun Zhang

  3. Department of Thoracic Surgery, Linyi People’s Hospital, Linyi, 276000, Shandong, People’s Republic of China

    Jinlong Zhao

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  1. Haigang Li
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Contributions

J.Z. and C.Z. conceived and designed study. Y.G. and J.H. conducted experiments. H. Z and Z.Z. analyzed data. H.L. and J.Y. wrote the manuscript. All authors read and approved the manuscript.

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Correspondence to Jinlong Zhao or Chun Zhang.

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Li, H., Yao, J., Guo, Y. et al. Preparation of Conotoxin-Encapsulated Chitosan Nanoparticles and Evaluation of Their Skin Permeability. AAPS PharmSciTech 24, 53 (2023). https://doi.org/10.1208/s12249-023-02509-8

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