Mitomycin C as an Anti-Persister Strategy against Klebsiella pneumoniae: Toxicity and Synergy Studies
Abstract
:1. Introduction
2. Results
2.1. Synergy Studies
2.1.1. Optical Density Growth Curves and Viability Assay
2.1.2. Frequency of Phage-Resistant Mutants
2.1.3. Assessment of the Cellular Respiration In Vitro
2.2. Toxicity Studies
2.2.1. In Vitro: Metabolic Activity and Apoptosis Study
2.2.2. In Vivo Toxicity Assay: Murine Model
3. Discussion
4. Materials and Methods
4.1. Bacterial Strains, Phage and Growth Culture Conditions
4.2. Optical Density Growth Curves
4.3. Time-Kill Curves Assay
4.4. Frequency of Resistant Mutants
4.5. Respiration Assay Using the Tetrazolium Salt WST-1
4.6. Cytotoxicity Assay Using Human Chondrocytes T/C28a2 Cell Line
4.6.1. MTT-Cytotoxic Assay
4.6.2. Apoptosis Study Using Flow Cytometry
4.7. Acute and Cumulative of MMC and Lytic Bacteriophage vB_KpnM-VAC13 in Healthy Female C57BL6/J Mice
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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K. pneumoniae Clinical Strain | Sequence Type | Carbapenemase | Biological Origin | GenBank Accession Number | Reference |
---|---|---|---|---|---|
K2534 | ST437 | OXA-245 | Rectal | WRXG00000000.1 | [23] |
K3325 | ST42 | - | Blood | SAMEA3649525 | [23] |
ATCC | ST131 | - | Commercial | - | |
K. pneumoniae lytic Bacteriophage | Morphotype | Genus | Genome Size (bp) | GenBank Accession Number | Characterization (Reference) |
vB_KpnM-VAC13 | Myoviridae | Slopekvirus | 174,826 | MZ322895.1 | [23] |
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Pacios, O.; Herrera-Espejo, S.; Armán, L.; Ibarguren-Quiles, C.; Blasco, L.; Bleriot, I.; Fernández-García, L.; Ortiz-Cartagena, C.; Paniagua, M.; Barrio-Pujante, A.; et al. Mitomycin C as an Anti-Persister Strategy against Klebsiella pneumoniae: Toxicity and Synergy Studies. Antibiotics 2024, 13, 815. https://doi.org/10.3390/antibiotics13090815
Pacios O, Herrera-Espejo S, Armán L, Ibarguren-Quiles C, Blasco L, Bleriot I, Fernández-García L, Ortiz-Cartagena C, Paniagua M, Barrio-Pujante A, et al. Mitomycin C as an Anti-Persister Strategy against Klebsiella pneumoniae: Toxicity and Synergy Studies. Antibiotics. 2024; 13(9):815. https://doi.org/10.3390/antibiotics13090815
Chicago/Turabian StylePacios, Olga, Soraya Herrera-Espejo, Lucía Armán, Clara Ibarguren-Quiles, Lucía Blasco, Inés Bleriot, Laura Fernández-García, Concha Ortiz-Cartagena, María Paniagua, Antonio Barrio-Pujante, and et al. 2024. "Mitomycin C as an Anti-Persister Strategy against Klebsiella pneumoniae: Toxicity and Synergy Studies" Antibiotics 13, no. 9: 815. https://doi.org/10.3390/antibiotics13090815