Design and In Vitro Activity of Furcellaran/Chitosan Multilayer Microcapsules for the Delivery of Glutathione and Empty Model Multilayer Microcapsules Based on Polysaccharides
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials for Preparing Capsules
2.2. Preparation of Biopolymer Solution
2.3. Preparation of Capsules with Different Number of Layers Enclosed with Glutathione
2.4. Preparation of Empty Capsules with Four Layers
- -
- FUR/CHIT two-layer capsule enriched in GSH (5 mg or 25 mg/mL)—2L5 and 2L25
- -
- (FUR/CHIT)2 four-layer capsule enriched in GSH (5 mg or 25 mg/mL)—4L5 and 4L25
- -
- (CHIT/FUR)2 empty capsule—ECAPS
2.5. Particle Size and Zeta Potential
2.6. SEM
2.7. Simulated In Vitro Digestion Model of Gastrointestinal Tract
2.8. Simulated In Vitro Absorption Model of Gastrointestinal Tract
2.9. Cell Culture
2.10. Cell Treatment
2.11. Cell Proliferation Assessment
2.12. Cytotoxicity Assay
2.13. Muse Flow Cytometer Analysis
2.14. Statistical Analysis
3. Results and Discussion
3.1. Preparation of Capsules
3.2. Impact on Cell Proliferation
3.3. Cytotoxicity
3.4. PI3K/MAPK Activity Assay
3.5. Detection of Early and Late Markers of Biochemical Apoptosis
3.6. Cell Cycle Phase Distribution
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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PI3K/MAPK Activation | ||||||||
---|---|---|---|---|---|---|---|---|
UC | STS | GSH5 | GSH25 | 2L5 | 2L25 | 4L25 | ECAPS | |
MAPK activation | 0.97 c ± 0.46 | 0.05 a ± 0.07 | 0.73 b,c ± 0.25 | 0.83 c ± 0.25 | 0.83 c ± 1.21 | 0.3 a,c ± 0.35 | 0.2 a ± 0.2 | 0.5 a,b,c ± 0.17 |
PI3K activation | 25.27 a ± 1.79 | 56.85 d ± 0.64 | 45.0 b ± 1.14 | 42.87 b ± 1.1 | 42.4 b ± 0.96 | 52.07 c ± 1.99 | 54.4 c,d ± 1.47 | 53.9 c ± 2.14 |
dual pathway activation | 64.67 e ± 0.96 | 16.17 a ± 0.57 | 39.63 c ± 2.0 | 40.07 c ± 2.39 | 43.73 d ± 2.63 | 30.3 b ± 1.44 | 29.0 b ± 2.8 | 29.13 b ± 0.46 |
negative | 9.1 a ± 0.35 | 26.4 d ± 1.13 | 14.63 b,c ± 1.12 | 16.23 b,c ± 3.0 | 13.03 b ± 1.8 | 17.33 c ± 1.0 | 16.4 c ± 1.81 | 16.47 c ± 2.65 |
Apoptosis Activity | ||||||||
UC | STS | GSH5 | GSH25 | 2L5 | 2L25 | 4L25 | ECAPS | |
Live | 85.98 e ± 0.88 | 55.93 a ± 0.93 | 62.67 c ± 1.62 | 62.65 c ± 1.4 | 59.15 b ± 0.13 | 60.13 b,c ± 1.92 | 66.3 d ± 3.29 | 67.58 d ± 0.72 |
Early apoptotic | 7.78 a ± 0.43 | 22.78 c ± 1.58 | 16.98 b ± 1.19 | 14.52 b ± 1.58 | 17.95 b ± 0.75 | 15.10 b ± 3.88 | 9.9 a ± 2.08 | 10.0 a ± 1.16 |
Late apoptotic | 3.40 a ± 2.0 | 19.55 b,c ± 0.2 | 17.60 b ± 0.78 | 19.47 b,c ± 0.76 | 19.87 b,c ± 0.43 | 23.62 d ± 1.73 | 21.80 c,d ± 1.73 | 20.75 c ± 1.57 |
Dead | 1.15 a ± 0.0 | 1.4 a,b ± 0.1 | 2.75 c ± 0.41 | 3.37 d ± 0.4 | 3.03 c,d ± 0.33 | 1.15 a ± 0.52 | 2.0 b ± 0.1 | 1.67 a,b ± 0.43 |
Total apoptotic | 12.88 a ± 0.88 | 42.68 e ± 1.03 | 34.58 c ± 1.85 | 37.82 b,c ± 0.38 | 37.82 d ± 0.38 | 38.72 d ± 2.44 | 31.70 b,c ± 3.23 | 30.75 b ± 1.13 |
Caspase-3/7 Activity | ||||||||
UC | STS | GSH5 | GSH25 | 2L5 | 2L25 | 4L25 | ECAPS | |
Live | 89.8 e ± 0.35 | 46.47 b,c ± 2.8 | 43.17 b ± 9.56 | 48.28 b,c ± 1.51 | 36.50 a ± 1.26 | 41.78 a,b ± 1.08 | 51.44 c,d ± 0.72 | 57.37 d ± 0.86 |
Early apoptotic | 2.93 a ± 0.53 | 31.87 e ± 4.59 | 19.58 c,d ± 5.25 | 7.18 a ± 0.35 | 21.15 d ± 1.26 | 16.32 b,c ± 0.85 | 17.16 b,c,d ± 0.96 | 14.27 b ± 0.15 |
Late apoptotic | 7.13 a ± 0.68 | 21.55 b ± 2.07 | 30.47 c ± 7.57 | 39.43 d ± 2.32 | 39.83 d ± 1.89 | 40.05 d ± 1.03 | 29.31 c ± 0.7 | 26.43 b,c ± 1.05 |
Dead | 0.13 a ± 0.14 | 0.12 a ± 0.08 | 6.78 b ± 7.73 | 4.43 a,b ± 0.51 | 2.52 a,b ± 0.21 | 1.85 a,b ± 0.31 | 2.09 a,b ± 0.14 | 1.99 a,b ± 0.15 |
Total apoptotic | 10.07 a ± 0.3 | 53.45 d,e ± 2.70 | 50.05 c,d ± 6.08 | 46.62 c ± 2.16 | 60.68 f ± 1.05 | 56.37 e ± 0.8 | 46.47 c ± 0.64 | 40.69 b ± 0.99 |
BCL-2 Activation | ||||||||
UC | STS | GSH5 | GSH25 | 2L5 | 2L25 | 4L25 | ECAPS | |
Activated | 82.6 d ± 1.51 | 7.53 a ± 0.84 | 14.03 a,b ± 1.89 | 12.9 a,b ± 1.68 | 15.3 b ± 2.19 | 15.47 b ± 1.81 | 26.8 c ± 9.07 | 24.72 c ± 3.51 |
Inactivated | 17.3 a ± 1.51 | 91.87 d ± 0.67 | 85.2 c,d ± 1.65 | 86.2 c,d ± 1.9 | 83.5 c ± 2.29 | 83.5 c ± 1.61 | 72.23 b ± 9.26 | 74.92 b ± 3.42 |
Non-expressing | 0.1 a ± 0.0 | 0.53 a,b,c ± 0.21 | 0.67 b,c,d ± 0.38 | 0.83 c,d ±0.31 | 1.13 d ± 0.5 | 0.93 c,d ± 0.31 | 0.63 a,b,c,d ± 0.15 | 0.21 a,b ± 0.1 |
Cell Cycle Phase Distribution | ||||||||
---|---|---|---|---|---|---|---|---|
UC | STS | GSH5 | GSH25 | 2L5 | 2L25 | 4L25 | ECAPS | |
G0/G1 | 41.80 c ± 0.79 | 26.7 a ± 2.19 | 42.47 c ± 3.26 | 36.67 b ± 1.72 | 37.3 b ± 1.68 | 42.17 c ± 1.95 | 39.37 b,c ± 2.55 | 37.57 b ± 2.22 |
S | 29.33 c ± 0.12 | 24.33 b ± 1.46 | 21.27 a ± 1.0 | 22.1 a,b ± 0.87 | 22.77 a,b ± 0.72 | 21.6 a ± 0.0 | 24.33 b ± 2.15 | 23.70 a,b ± 2.36 |
G2/M | 15.73 a ± 0.86 | 29.03 b ± 0.32 | 30.6 b,c ± 1.76 | 34.33 e ± 1.38 | 32.8 d,e ± 0.95 | 30.43 b,c ± 1.43 | 30.67 b,c ± 0.93 | 31.9 c,d ± 0.56 |
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Drozdowska, M.; Piasna-Słupecka, E.; Such, A.; Dziadek, K.; Krzyściak, P.; Kruk, T.; Duraczyńska, D.; Morawska-Tota, M.; Jamróz, E. Design and In Vitro Activity of Furcellaran/Chitosan Multilayer Microcapsules for the Delivery of Glutathione and Empty Model Multilayer Microcapsules Based on Polysaccharides. Materials 2024, 17, 2047. https://doi.org/10.3390/ma17092047
Drozdowska M, Piasna-Słupecka E, Such A, Dziadek K, Krzyściak P, Kruk T, Duraczyńska D, Morawska-Tota M, Jamróz E. Design and In Vitro Activity of Furcellaran/Chitosan Multilayer Microcapsules for the Delivery of Glutathione and Empty Model Multilayer Microcapsules Based on Polysaccharides. Materials. 2024; 17(9):2047. https://doi.org/10.3390/ma17092047
Chicago/Turabian StyleDrozdowska, Mariola, Ewelina Piasna-Słupecka, Aleksandra Such, Kinga Dziadek, Paweł Krzyściak, Tomasz Kruk, Dorota Duraczyńska, Małgorzata Morawska-Tota, and Ewelina Jamróz. 2024. "Design and In Vitro Activity of Furcellaran/Chitosan Multilayer Microcapsules for the Delivery of Glutathione and Empty Model Multilayer Microcapsules Based on Polysaccharides" Materials 17, no. 9: 2047. https://doi.org/10.3390/ma17092047