CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES >
Preparation of Echinococcus granulosus peptide embedded in chitosan quaternary ammonium salt nanoparticles
Received date: 2022-11-10
Revised date: 2023-03-26
Online published: 2023-05-30
Supported by
Shanghai Natural Science Foundation(19ZR1462600);National Nature Science Foundation of China(81971969);National Nature Science Foundation of China(82272369)
Objective To prepare nanoparticles (NPs) containing the epitope peptide FL46 of Echinococcus granulosus using hydroxypropyltrimethyl ammonium chloride chitosan (HACC) and sodium tripolyphosphate (TPP) as carriers, to screen the optimal preparation conditions, and evaluate the encapsulation effect and morphological characterisation. Methods The FL46-HACC-NPs suspension was prepared by slowly adding FL46 solution dropwise to HACC solution using the ionic cross-linking method, stirring at 800 r/min and adding TPP solution dropwise. The average particle size, polydispersity index (PDI) and surface potential (Zeta) were measured using a nanoparticle size potentiometer, the microscopic morphology was observed under the transmission electron microscope and the encapsulation rate was calculated by BCA protein concentration measurement assay kit. A three-factor, four-level orthogonal experiment was conducted with HACC concentration (0.25, 0.50, 0.75, 1.00 mg/ml), TPP concentration (0.25, 0.50, 0.75, 1.00 mg/ml), and HACC to TPP mass ratio (8∶1, 9∶1, 10∶1, 11∶1), with single factor method to examine the effect on the characterisation of FL46-HACC-NPs. Results FL46-HACC-NPs solution can be clarified, milky and precipitated at different mass concentrations and mass ratios of HACC and TPP. After single-factor experiments, the optimal preparation conditions were screened for small particle size and high encapsulation rate. The optimal preparation conditions were HACC concentration of 0.25 mg/ml, TPP concentration of 0.50 mg/ml, and the mass ratio of HACC to TPP 9∶1. Under the optimal conditions, spherical or polygonal FL46-HACC-NPs with the particle size of (307.37 ± 2.17) nm, PDI of (0.228 ± 0.008), Zeta potential of (29.77 ± 0.46) mV and encapsulation rate of (75.95 ± 1.60) % can be prepared. Transmission electron microscopy shows that the FL46-HACC-NPs have a homogeneous morphological structure and are sphere-like or polygonal in shape. Conclusion Optimised FL46-HACC-NPs can be effectively prepared by the ion cross-linking method, which can provide basis for the study of the protective effect of E. granulosus polypeptide vaccine.
WU Xiaoying , HU Yuan , CAO Jianping . Preparation of Echinococcus granulosus peptide embedded in chitosan quaternary ammonium salt nanoparticles[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2023 , 41(3) : 300 -305 . DOI: 10.12140/j.issn.1000-7423.2023.03.006
| [1] | Anvari D, Rezaei F, Ashouri A, et al. Current situation and future prospects of Echinococcus granulosus vaccine candidates: a systematic review[J]. Transbound Emerg Dis, 2021, 68(3): 1080-1096. |
| [2] | Pourseif MM, Moghaddam G, Saeedi N, et al. Current status and future prospective of vaccine development against Echinococcus granulosus[J]. Biologicals, 2018, 51: 1-11. |
| [3] | Zhang F, Li S, Zhu Y, et al. Immunization of mice with EGG1Y162-1/2 provides protection against Echinococcus granulosus infection in BALB/c mice[J]. Mol Immunol, 2018, 94: 183-189. |
| [4] | Zhu GQ, Yan HB, Li L, et al. Advances in research on vaccines against echinococcosis[J]. Chin J Zoonoses, 2019, 35(1): 59-65. (in Chinese) |
| (朱国强, 闰鸿斌, 李立, 等. 棘球蚴(包虫)病疫苗研究进展[J]. 中国人兽共患病学报, 2019, 35(1): 59-65.) | |
| [5] | Xu T, Liu L, Shi C, et al. A recombinant rabies virus expressing Echinococcus granulosus EG95 induces protective immunity in mice[J]. Transbound Emerg Dis, 2022, 69(4): e254-e266. |
| [6] | Parvizpour S, Pourseif MM, Razmara J, et al. Epitope-based vaccine design: a comprehensive overview of bioinformatics approaches[J]. Drug Discov Today, 2020, 25(6): 1034-1042. |
| [7] | Negahdaripour M, Golkar N, Hajighahramani N, et al. Harnessing self-assembled peptide nanoparticles in epitope vaccine design[J]. Biotechnol Adv, 2017, 35(5): 575-596. |
| [8] | Ahmed TA, Aljaeid BM. Preparation, characterization, and potential application of chitosan, chitosan derivatives, and chitosan metal nanoparticles in pharmaceutical drug delivery[J]. Drug Des Dev Ther, 2016, 10: 483-507. |
| [9] | Sivanesan I, Gopal J, Muthu M, et al. Green synthesized chitosan/chitosan nanoforms/nanocomposites for drug delivery applications[J]. Polymers, 2021, 13(14): 2256. |
| [10] | Chae KS, Shin CS, Shin WS. Characteristics of cricket (Gryllus bimaculatus) chitosan and chitosan-based nanoparticles[J]. Food Sci Biotechnol, 2018, 27(3): 631-639. |
| [11] | Shariatinia Z. Pharmaceutical applications of chitosan[J]. Adv Colloid Interface Sci, 2019, 263: 131-194. |
| [12] | Balasubramaniyan M, Santhanam M, Vinayagam V, et al. Immunomodulatory effects of chitosan nanoparticles as vaccine delivery agent against lymphatic filariasis through mucosalimmunization[J]. Int J Biol Macromol, 2022, 222: 2392-2398. |
| [13] | Li YJ, Zhu YJ, Sha T, et al. A multi-epitope chitosan nanoparticles vaccine of canine against Echinococcus granulosus[J]. J Biomed Nanotechnol, 2021, 17(5): 910-920. |
| [14] | Xu Y, Zhou Z, Brooks B, et al. Layer-by-layer delivery of multiple antigens using trimethylchitosan nanoparticles as a malaria vaccine candidate[J]. Front Immunol, 2022, 13: 900080. |
| [15] | Tamara FR, Lin C, Mi FL, et al. Antibacterial effects of chitosan/cationic peptide nanoparticles[J]. Nanomaterials (Basel), 2018, 8(2): 88. |
| [16] | Chen KY, Zeng SY. Fabrication of quaternized chitosan nanoparticles using tripolyphosphate/genipin dual cross-linkers as a protein delivery system[J]. Polymers, 2018, 10(11): 1226. |
| [17] | Gao Y, Gong X, Yu S, et al. Immune enhancement of N-2-hydroxypropyl trimethyl ammonium chloride chitosan/carboxymethyl chitosan nanoparticles vaccine[J]. Int J Biol Macromol, 2022, 220: 183-192. |
| [18] | Rodrigues Dos Santos Junior S, Kelley Lopes da Silva F, Santos Dias L, et al. Intranasal vaccine using P10 peptide complexed within chitosan polymeric nanoparticles as experimental therapy for paracoccidioidomycosis in murine model[J]. J Fungi (Basel), 2020, 6(3): 160. |
| [19] | Ahmad Mahmood M, Madni A, Rehman M, et al. Ionically cross-linked chitosan nanoparticles for sustained delivery of docetaxel: fabrication, post-formulation and acute oral toxicity evaluation[J]. Int J Nanomed, 2019, 14: 10035-10046. |
| [20] | Wang D, Jiang W. Preparation of chitosan-based nanoparticles for enzyme immobilization[J]. Int J Biol Macromol, 2019, 126: 1125-1132. |
| [21] | Rappuoli R, Serruto D. Self-assembling nanoparticles usher in a new era of vaccine design[J]. Cell, 2019, 176(6): 1245-1247. |
| [22] | Albalawi AE, Alanazi AD, Baharvand P, et al. High potency of organic and inorganic nanoparticles to treat cystic echinococcosis: an evidence-based review[J]. Nanomaterials (Basel), 2020, 10(12): 2538. |
| [23] | Lei Y, Zhao FR, Shao JJ, et al. Application of built-in adjuvants for epitope-based vaccines[J]. PeerJ, 2019, 6: e6185. |
| [24] | Guo JJ, Sun XH, Yin HQ, et al. Chitosan microsphere used as an effective system to deliver a linked antigenic peptides vaccine protect mice against acute and chronic toxoplasmosis[J]. Front Cell Infect Microbiol, 2018, 8: 163. |
| [25] | Pathak K, Misra SK, Sehgal A, et al. Biomedical applications of quaternizedchitosan[J]. Polymers, 2021, 13(15): 2514. |
| [26] | Li X, Xing R, Xu C, et al. Immunostimulatory effect of chitosan and quaternary chitosan: a review of potential vaccine adjuvants[J]. Carbohydr Polym, 2021, 264: 118050. |
| [27] | Zhao YH, Du WW, Wu HH, et al. Chitosan/sodium tripolyphosphate nanoparticles as efficient vehicles for enhancing the cellular uptake of fish-derived peptide[J]. J Food Biochem, 2019, 43(2): e12730. |
| [28] | Du Z, Liu J, Zhang T, et al. A study on the preparation of chitosa-tripolyphosphate nanoparticles and its entrapment mechanism for egg white derived peptides[J]. Food Chem, 2019, 286: 530-536. |
| [29] | Hejjaji EMA, Smith AM, Morris GA. Evaluation of the mucoadhesive properties of chitosan nanoparticles prepared using different chitosan to tripolyphosphate (CS∶TPP) ratios[J]. Int J Biol Macromol, 2018, 120: 1610-1617. |
| [30] | IranpurMobarakeh V, Modarressi MH, Rahimi P, et al. Optimization of chitosan nanoparticles as an anti-HIV siRNA delivery vehicle[J]. Int J Biol Macromol, 2019, 129: 305-315. |
| [31] | Changsan N, Sinsuebpol C. Dry powder inhalation formulation of chitosan nanoparticles for co-administration of isoniazid and pyrazinamide[J]. Pharm Dev Technol, 2021, 26(2): 181-192. |
| [32] | Aline Bruinsmann F, Pigana S, Aguirre T, et al. Chitosan-coated nanoparticles: effect of chitosan molecular weight on nasal transmucosaldelivery[J]. Pharmaceutics, 2019, 11(2): 86. |
| [33] | Omer AM, Ziora ZM, Tamer TM, et al. Formulation of quaternizedaminated chitosan nanoparticles for efficient encapsulation and slow release of curcumin[J]. Molecules, 2021, 26(2): 449. |
| [34] | AbdelAllah NH, Gaber Y, Rashed ME, et al. Alginate-coated chitosan nanoparticles act as effective adjuvant for hepatitis A vaccine in mice[J]. Int J Biol Macromol, 2020, 152: 904-912. |
| [35] | Pant A, Negi JS. Novel controlled ionic gelation strategy for chitosan nanoparticles preparation using TPP-β-CD inclusion complex[J]. Eur J Pharm Sci, 2018, 112: 180-185. |
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