Designing of Chitosan Derivatives Nanoparticles with Antiangiogenic Effect for Cancer Therapy
Abstract
:1. Introduction
2. Materials and Methods
2.1. Design of Chitosan-Derived Nanoparticles
2.2. Spectral Characterization of Nanoparticles: FT-IR Analysis
2.3. Research Regarding Antioxidant Assays
2.4. In Vivo Evaluation of Angiogenic Activity Using a CAM Model
2.5. In Vitro Cytotoxicity Evaluation Using an MTT Assay
2.5.1. Cell Cultures
2.5.2. Cell Morphology Examination at 48 h of Treatment with Samples
2.6. Statistical Methods
- -
- one way ANOVA was conducted, for the variables X = blood vessels and Y = study batch, using an alpha of 0.05;
- -
- for multiple comparisons of normal distributed series of values, a post-hoc Bonferroni test was applied after one-way ANOVA in case of significance. One-Way ANOVA was significant (alpha of 0.05) and, in these conditions, Bonferroni correction (post-hoc Bonferroni) was performed to reduce the error rate when testing multiple hypotheses.
- -
- Skewness (−2 < p < 2) tests are tests of normality in frequentist statistics, available when using the distribution platform to examine a continuous variable.
3. Results
3.1. Chitosan-Derivatives Nanoparticles: DLS Measurements
3.2. FT-IR Characterization of Nanoparticles
3.3. Researches Regarding Antioxidant Assays
3.3.1. Determination of Total Antioxidant Activity (TAA)
3.3.2. Ferric Reducing Power
3.3.3. Radical Scavenging Ability
3.4. In Vivo Evaluation of Angiogenic Activity Using a CAM Model
- variations from 0 to 8 vessels/field;
- mean 3.88 ± 2.28 vessels/field;
- the median value (4 vessels/field) close to the average value and the Skewness test result <2 suggests that the series of values of the number of vessels was homogeneous, so statistical significance tests can be applied.
3.5. In Vitro Biocompatibility Evaluation Using MTT Cell Viability Assay
3.6. In Vitro MTT Assay on Human Epithelial Cervical Tumor Hep-2 Cells
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Compound | CL (chitosan) | CLA | CLB | CLC | CLD |
---|---|---|---|---|---|
Dimension (nm) | 390.2 | 622.2 | 571 | 351 | 435 |
All Groups | CL | CLA | CLB | CLC | CLD | ||
---|---|---|---|---|---|---|---|
N | 25 | 5 | 5 | 5 | 5 | 5 | |
Mean | 3.88 | 5.80 | 4.80 | 5.20 | 2.40 | 1.20 | |
p post-hoc Bonferroni | ns) | ns) ns) | b) ns) ns) | a) b) b) ns) | |||
Median | 4.00 | 6.00 | 5.00 | 5.00 | 3.00 | 1.00 | |
Std.Deviation | 2.28 | 1.79 | 1.30 | 1.92 | 1.52 | 0.84 | |
Variance | 5.19 | 3.20 | 1.70 | 3.70 | 2.30 | 0.70 | |
Skewness Test | 0.092 | 0.052 | −0.541 | 0.590 | −1.118 | −0.512 | |
Std.Err.Skewness | 0.464 | 0.913 | 0.913 | 0.913 | 0.913 | 0.913 |
Cell Culture | Time | Untreated Control | Positive Control | CL | CLA | CLB | CLC | CLD |
---|---|---|---|---|---|---|---|---|
NCTC | 24 h | 100% | 11.45% | 95.72% | 105.79% | 100.00% | 98.07% | 104.83% |
48 h | 100% | 7.31% | 89.29% | 99.28% | 101.96% | 107.52% | 107.52% | |
Hep-2 | 24 h | 100% | 13.86% | 113.69% | 85.00% | 111.96% | 99.35% | 98.15% |
48 h | 100% | 6.95% | 87.85% | 66.16% | 79.70% | 69.91% | 66.89% |
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Dragostin, O.-M.; Tatia, R.; Samal, S.K.; Oancea, A.; Zamfir, A.S.; Dragostin, I.; Lisă, E.-L.; Apetrei, C.; Zamfir, C.L. Designing of Chitosan Derivatives Nanoparticles with Antiangiogenic Effect for Cancer Therapy. Nanomaterials 2020, 10, 698. https://doi.org/10.3390/nano10040698
Dragostin O-M, Tatia R, Samal SK, Oancea A, Zamfir AS, Dragostin I, Lisă E-L, Apetrei C, Zamfir CL. Designing of Chitosan Derivatives Nanoparticles with Antiangiogenic Effect for Cancer Therapy. Nanomaterials. 2020; 10(4):698. https://doi.org/10.3390/nano10040698
Chicago/Turabian StyleDragostin, Oana-Maria, Rodica Tatia, Sangram Keshari Samal, Anca Oancea, Alexandra Simona Zamfir, Ionuț Dragostin, Elena-Lăcrămioara Lisă, Constantin Apetrei, and Carmen Lăcrămioara Zamfir. 2020. "Designing of Chitosan Derivatives Nanoparticles with Antiangiogenic Effect for Cancer Therapy" Nanomaterials 10, no. 4: 698. https://doi.org/10.3390/nano10040698