Mannose-Modified Chitosan Poly(lactic-co-glycolic acid) Microspheres Act as a Mannose Receptor-Mediated Delivery System Enhancing the Immune Response
Abstract
:1. Introduction
2. Materials and Methods
2.1. Reagents and Antibodies
2.2. Synthesis of Mannose-Modified Chitosan PLGA Microspheres
2.2.1. Synthesis of Mannose-Modified Chitosan
2.2.2. Physicochemical Property Analysis of MAN-CS
2.2.3. Synthesis of PLGA Microspheres
2.2.4. Determination of Drug Loading on Microspheres
2.2.5. Orthogonal Experiment Optimization
2.2.6. Synthesis of Mannose-Modified Chitosan PLGA Microspheres
2.2.7. Microsphere Surface Morphology, Particle Size, and Zeta Potential
2.3. In Vitro Experiments
2.3.1. Cytotoxicity of MAN-CS-OVA-PLGA-MPs on DCs
2.3.2. Phagocytosis of Different Microspheres by DC Cells
2.3.3. The Effect of MAN-CS-OVA-PLGA-MPs on the Delivery of OVA
2.4. In Vivo Experiments
2.4.1. Mice Grouping and Immunization
2.4.2. Immunophenotype of Spleen Lymphocytes in Mice
2.4.3. The Maturation of DCs in Mouse Spleen
2.4.4. Determination of OVA-Specific IgG and IgG Isotype in Serum of Mice
2.4.5. Detection of Cytokines in Serum of Immunized Mice by ELISA
2.5. Statistical Analysis
3. Results
3.1. Physicochemical Characteristics
3.1.1. Identification of Mannose-Modified Chitosan
3.1.2. FT-IR Spectroscopy Analysis
3.2. Orthogonal Design of PLGA Microspheres
3.3. Morphology of the Microspheres Obtained by SEM
3.4. The Particle Size and Electromotive Force of Three Types of PLGA Microspheres
3.5. In Vitro Experiments
3.5.1. Cytotoxicity of MAN-CS-OVA-PLGA-MPs towards DCs
3.5.2. Effects of MAN-CS-OVA-PLGA-MPs on the Phagocytic Activity of DCs
3.5.3. The Delivery Effect of MAN-CS-OVA-PLGA-MPs on OVA
3.6. In Vivo Experiments
3.6.1. Effect of MAN-CS-OVA-PLGA-MPs on OVA-Specific IgG and IgG Isotype Antibody Titers of Murine Serum
3.6.2. Effect of MAN-CS-OVA-PLGA-MPs on the Immunophenotype of Spleen Lymphocytes in Mice
3.6.3. Effect of MAN-CS-OVA-PLGA-MPs on the Expression of DC Surface Molecules in the Spleen
3.6.4. Effect of MAN-CS-OVA-PLGA-MPs on the Concentrations of Cytokines in the Serum of Mice
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Conflicts of Interest
References
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PLGA | PVA | OVA | |
---|---|---|---|
1 | 10 | 1 | 50 |
2 | 20 | 2 | 100 |
3 | 40 | 3 | 150 |
Chitosan | Mannose-Modified Chitosan |
---|---|
C:39.99 | C:39.26 |
H:6.42 | H:6.87 |
O:45.37 | O:45.52 |
N:7.63 | N:7.24 |
PLGA | PVA | OVA | LE(%) | |
---|---|---|---|---|
1 | 10 | 1 | 50 | 3.25 |
2 | 10 | 2 | 100 | 3.15 |
3 | 10 | 3 | 150 | 1.27 |
4 | 20 | 1 | 100 | 1.69 |
5 | 20 | 2 | 150 | 6.48 |
6 | 20 | 3 | 50 | 3.22 |
7 | 40 | 1 | 150 | 5.83 |
8 | 40 | 2 | 50 | 8.4 |
9 | 40 | 3 | 100 | 8.1 |
K1 | 2.55 | 3.59 | 4.95 | |
K2 | 3.79 | 6.01 | 4.31 | |
K3 | 7.44 | 4.19 | 4.52 | |
R | 4.88 | 2.42 | 0.43 | |
A3B2C1 |
Group | Zeta EMF (mv) | Particle Size (μm) | Drug Loading (%) |
---|---|---|---|
OVA-PLGA-MPs | −14.8 ± 1.9 a | 6.5 ± 0.4 a | 7.9 ± 0.2 a |
CS-OVA-PLGA-MPs | 31.6 ± 0.4 b | 9.0 ± 0.5 b | 6.5 ± 0.2 b |
MAN-CS-OVA-PLGA-MPs | 30.8 ± 1.1 b | 7.9 ± 0.5 c | 6.2 ± 1.2 b |
Group | IL-2 (pg/mL) | IL-4 (pg/mL) | IL-6 (pg/mL) | IFN-γ (pg/mL) |
---|---|---|---|---|
Saline | 4.295 ± 0.045 a | 11.540 ± 2.820 a | 36.835 ± 0.485 a | 27.200 ± 0.630 a |
OVA | 4.210 ± 0.580 a | 12.735 ± 0.835 a,b | 37.320 ± 1.230 a | 28.665 ± 0.845 a,b |
OVA-PLGA-MPs | 4.245 ± 0.315 a | 16.015 ± 2.845 a,b | 37.805 ± 1.275 a,b | 28.900 ± 0.430 a,b |
CS-OVA-PLGA-MPs | 4.525 ± 0.165 a | 16.255 ± 1.185 b | 39.930 ± 0.750 b | 29.565 ± 0.075 a,b |
MAN-CS-OVA-PLGA-MPs | 4.635 ± 0.045 a | 21.860 ± 3.720 c | 40.175 ± 1.975 c | 30.525 ± 1.065 b |
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Feng, H.; Yang, X.; Zhang, L.; Liu, Q.; Feng, Y.; Wu, D.; Liu, Y.; Yang, J. Mannose-Modified Chitosan Poly(lactic-co-glycolic acid) Microspheres Act as a Mannose Receptor-Mediated Delivery System Enhancing the Immune Response. Polymers 2021, 13, 2208. https://doi.org/10.3390/polym13132208
Feng H, Yang X, Zhang L, Liu Q, Feng Y, Wu D, Liu Y, Yang J. Mannose-Modified Chitosan Poly(lactic-co-glycolic acid) Microspheres Act as a Mannose Receptor-Mediated Delivery System Enhancing the Immune Response. Polymers. 2021; 13(13):2208. https://doi.org/10.3390/polym13132208
Chicago/Turabian StyleFeng, Haibo, Xiaonong Yang, Linzi Zhang, Qianqian Liu, Yangyang Feng, Daiyan Wu, Yunjie Liu, and Jie Yang. 2021. "Mannose-Modified Chitosan Poly(lactic-co-glycolic acid) Microspheres Act as a Mannose Receptor-Mediated Delivery System Enhancing the Immune Response" Polymers 13, no. 13: 2208. https://doi.org/10.3390/polym13132208