The Mould War: Developing an Armamentarium against Fungal Pathogens Utilising Thymoquinone, Ocimene, and Miramistin within Bacterial Cellulose Matrices
Abstract
:1. Introduction
2. Materials and Methods
2.1. Media and Reagents
2.2. Microorganisms
2.3. Cell Culture
2.4. Fungal Starter Cultures
2.5. Production and Purification of Bacterial Cellulose
2.6. Preparation of Working Solutions and Loading of Antifungal Agents
2.7. Minimum Inhibition Concentration (MIC)
2.8. Minimum Fungicidal Concentration (MFC)
2.9. Quantification of Loaded Compound in BC
2.10. Zone of Inhibition (ZOI)
2.11. Cytotoxicity
2.12. Scanning Electron Microscopy (SEM)
2.13. Fourier Transform Infrared Spectroscopy
2.14. Statistical Analysis
3. Results and Discussion
3.1. Preparation of BC Hydrogels and Characterisation
3.2. Antifungal Studies
3.3. Cytotoxicity Studies
4. Conclusions and Future Prospects
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Step | Concentration (mg/L) | Source | Volume of Antifungal (μL) | Volume of Solvent (μL) | Intermediate Concentration (mg/L) | Concentration (mg/L) after 1:100 Dilution with Double Strength RPMI 2% (w/v) |
---|---|---|---|---|---|---|
1 | 3200 | Stock | 200 | 0 | 3200 | 32 |
2 | 3200 | Stock | 100 | 100 | 1600 | 16 |
3 | 3200 | Stock | 50 | 150 | 800 | 8 |
4 | 3200 | Stock | 50 | 350 | 400 | 4 |
5 | 400 | Step 4 | 100 | 100 | 200 | 2 |
6 | 400 | Step 4 | 50 | 150 | 10 | 1 |
7 | 400 | Step 4 | 50 | 350 | 50 | 0.5 |
8 | 50 | Step 7 | 100 | 100 | 25 | 0.25 |
9 | 50 | Step 7 | 50 | 150 | 12.5 | 0.125 |
10 | 50 | Step 7 | 25 | 175 | 6.25 | 0.06 |
Step | Concentration (mg/L) | Source | Volume of Antifungal (μL) | Volume of Solvent (μL) | Intermediate Concentration (mg/L) | Concentration (mg/L) after 1:100 Dilution with Double Strength RPMI 2% (w/v) |
---|---|---|---|---|---|---|
1 | 12,800 | Stock | 200 | 0 | 12,800 | 128 |
2 | 12,800 | Stock | 100 | 100 | 6400 | 64 |
3 | 12,800 | Stock | 50 | 150 | 3200 | 32 |
4 | 12,800 | Stock | 50 | 350 | 1600 | 16 |
5 | 1600 | Step 4 | 100 | 100 | 800 | 8 |
6 | 1600 | Step 4 | 50 | 150 | 400 | 4 |
7 | 1600 | Step 4 | 50 | 350 | 200 | 2 |
8 | 200 | Step 7 | 100 | 100 | 100 | 1 |
9 | 200 | Step 7 | 50 | 150 | 50 | 0.5 |
10 | 200 | Step 7 | 25 | 175 | 25 | 0.26 |
Fungi | Amphotericin B | Thymoquinone | Ocimene | Miramistin | ||||
---|---|---|---|---|---|---|---|---|
MIC90 (mg/L) | MFC (mg/L) | MIC90 (mg/L)) | MFC (mg/L) | MIC90 (mg/L) | MFC (mg/L) | MIC90 (mg/L) | MFC (mg/L) | |
A. fumigatus | 0.06–1 | 0.07–1 | 1.3–2 | 1.5–2 | 0.7–1 | 1.6–2 | 21 | 27 |
A. niger | 0.06–1 | 0.09–1 | 0.9–2 | 1–2 | 0.4–0.9 | 1–2 | 18-20 | 25 |
C. albicans | 0.03 | 0.06–1 | 4.9–5.2 | 6.2–7 | 0.3–0.9 | 0.3–0.9 | 1.3–3 | 2.6–4 |
C. auris | 0.05 | 0.08–1 | 7.9–8.2 | 8.4–9.7 | 1.2–1.9 | 2–2.5 | 1.4–2.8 | 3–5 |
Drug | Unloaded BC (mg) | Loaded BC (mg) | Average Compound Loaded (µg) |
---|---|---|---|
Thymoquinone | 1.6375 ± 0.364 | 1.678 ± 0.357 | 42.0 ± 8 |
Ocimene | 1.5500 ± 0.208 | 1.588 ± 0.207 | 37.4 ± 9 |
Miramistin | 1.5750 ± 0.175 | 1.615 ± 0.183 | 46.0 ± 8 |
Amphotericin B | 1.7000 ± 0.221 | 1.733 ± 0.220 | 32.5 ± 10 |
Control | 1.6200 ± 0.125 | 1.627 ± 0.120 | 7.00 ± 2 |
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Swingler, S.; Gupta, A.; Gibson, H.; Heaselgrave, W.; Kowalczuk, M.; Adamus, G.; Radecka, I. The Mould War: Developing an Armamentarium against Fungal Pathogens Utilising Thymoquinone, Ocimene, and Miramistin within Bacterial Cellulose Matrices. Materials 2021, 14, 2654. https://doi.org/10.3390/ma14102654
Swingler S, Gupta A, Gibson H, Heaselgrave W, Kowalczuk M, Adamus G, Radecka I. The Mould War: Developing an Armamentarium against Fungal Pathogens Utilising Thymoquinone, Ocimene, and Miramistin within Bacterial Cellulose Matrices. Materials. 2021; 14(10):2654. https://doi.org/10.3390/ma14102654
Chicago/Turabian StyleSwingler, Sam, Abhishek Gupta, Hazel Gibson, Wayne Heaselgrave, Marek Kowalczuk, Grazyna Adamus, and Iza Radecka. 2021. "The Mould War: Developing an Armamentarium against Fungal Pathogens Utilising Thymoquinone, Ocimene, and Miramistin within Bacterial Cellulose Matrices" Materials 14, no. 10: 2654. https://doi.org/10.3390/ma14102654