Novel In Vivo Assessment of Antimicrobial Efficacy of Ciprofloxacin Loaded Mesoporous Silica Nanoparticles against Salmonella typhimurium Infection
Abstract
:1. Introduction
2. Results
2.1. Characterization of Mesoporous Silica Nanoparticles (MSNs), Ciprofloxacin Loading, and Release
2.2. In Vitro Antibacterial Activity of CIP–MSN against Decreased CIP Susceptible Salmonella typhimurium
2.3. Biofilm Inhibition and Transcriptional Modulatory Effect of CIP–MSN
2.4. Hematological, Biochemical, Antioxidant, and Immunological Effect of CIP–MSN on Blood, and Serum Constituents
2.5. Inhibitory Effect of CIP–MSN on Hepatic Salmonella typhimurium Load
2.6. Pro-Inflammatory Cytokines Transcriptional Modulatory Effect of CIP–MSN
2.7. Modulation of Pro-Apoptoticgenes Expression
2.8. Histopathological Evaluation
3. Discussion
4. Materials and Methods
4.1. Synthesis and Characterization of Mesoporous Silica Nanoparticles (MSNs) and Ciprofloxacin Loading
4.2. Antibacterial Effect of Ciprofloxacin—Loaded Mesoporous Silica Nanoparticles
4.2.1. Agar Well Diffusion Assay
4.2.2. Minimum Inhibitory Concentration
4.3. Ciprofloxacin Loaded Mesoporous Silica Nanoparticles Effect on Biofilm Formation
4.4. Expression of Genes Associated Virulence in Biofilm Culture
4.5. Experimental Design, and Oral Challenging with Salmonella typhimurium
4.6. Hematological, Biochemical, Oxidative Stress Mediators, and Immunological Measurements
4.7. Quantification of S. typhimurium DNA Copies
4.8. Pro-Inflammatory Cytokines, and Pro-Apoptotic Gene Expression Analysis by Real-Time PCR
4.9. Histopathologic Evaluation
4.10. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Zone of Inhibition (mm) | MIC (mg/L) | MBC (mg/L) | |
---|---|---|---|
CIP | 8.7 b ± 0.2 | 1.0 | 2.0 |
MSN | 3.4 c ± 0.37 | ND | ND |
CIP–MSN | 40.5 a ± 0.4 | 0.03125 | 0.0625 |
At 7 Days Postinfection | At 14 Days Postinfection | |||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Groups | NC | PC | CIP | MSN | CIP@ MSN | p Value | SEM | NC | PC | CIP | MSN | CIP–MSN | p Value | SEM |
RBCs | 12.5 a | 7.96 b | 9.17 ab | 8.23 b | 11.19 a | <0.001 | 0.09 | 12.63 a | 9.66 c | 10.30 b | 10.20 b | 11.60 ab | 0.09 | 12.63 a |
Hb | 12.9 a | 6.3 b | 11.2 a | 7.10 b | 11.56 a | <0.001 | 0.07 | 12.80 a | 8.80 c | 11.98 a | 10.26 b | 12.00 a | 0.12 | 12.80 a |
PCV | 41.4 a | 24.5 c | 37.43 b | 26.60 c | 39.5 ab | 0.03 | 0.14 | 42.30 a | 29.69 c | 38.6 ab | 37.75 b | 39.5 a | 0.13 | 42.30 a |
Biochemical biomarkers for tissue injury analysis | ||||||||||||||
ALT (U/L) | 48.3 c | 74.7 a | 58.7 b | 65.63 ab | 50.86 c | <0.001 | 0.21 | 46.7 d | 85.1 a | 52.36 c | 57.99 b | 48.36 cd | 0.25 | 48.3 c |
AST (U/L) | 25.2 b | 42.3 a | 27.6 b | 26.30 b | 25.53 b | <0.001 | 0.19 | 24.7 c | 48.4 a | 25.8 c | 34.96 b | 23.46 c | 0.16 | 25.2 b |
Urea (μmol/L) | 32.1 c | 52.3 a | 38.76 b | 50.90 a | 35.53 bc | <0.001 | 0.12 | 31.6 c | 59.8 a | 32.6 c | 41.90 b | 32.1 c | 0.14 | 32.1 c |
Creatinine (mg/dL) | 1.33 b | 2.73 a | 1.45 b | 1.95 ab | 1.41 b | <0.001 | 0.09 | 1.34 c | 3.07 a | 1.38 c | 2.35 b | 1.33 c | 0.08 | 1.33 b |
Oxidative stress mediators analysis | ||||||||||||||
NO | 153.8 d | 532.60 a | 246.40 b | 480.60 | 185.36 c | 0.027 | 0.25 | 150.5 e | 559.1 a | 202.16 d | 140.22 b | 177.66 c | 0.31 | 153.8 d |
MPO | 2.21 e | 10.80 a | 7.62 c | 8.65 b | 6.11 d | 0.03 | 0.09 | 2.16 c | 10.08 a | 6.76 b | 3.57 c | 0.10 | 2.21 c | |
CRP | 1.12 e | 54.30 a | 24.4 c | 50.36 b | 16.26 d | 0.14 | 0.14 | 1.16 e | 53.3 a | 17.4 c | 44.30 b | 9.2 d | <0.001 | 0.08 |
Chemokines and pro-inflammatory cytokines analysis | ||||||||||||||
CXCL10 | 156.00 e | 393.70 a | 224.80 c | 385.64 b | 181.6 d | 0.03 | 0.24 | 152.6 d | 416.3 a | 197.9 b | 400.02 a | 161.83 c | 0.02 | 0.24 |
CXCL11 | 108.86 e | 238.70 a | 161.30 c | 220.30 b | 130.53 d | 0.02 | 0.29 | 108.5 d | 243.4 a | 140.6 b | 239.23 a | 113.16 c | 0.01 | 0.31 |
IFN- γ | 38.36 e | 99.50 a | 53.26 c | 82.23 b | 43.00 d | <0.001 | 0.11 | 38.7 c | 104.4 a | 46.4 b | 100.23 a | 38.36 c | 0.03 | 0.23 |
IL-6 | 16.28 c | 47.36 a | 47.93 a | 45.636 a | 34.86 b | <0.001 | 0.13 | 15.86 c | 47.56 a | 34.56 b | 49.36 a | 16.59 c | <0.001 | 0.14 |
TNF-α | 11.5 c | 23.61 a | 16.50 b | 20.36 a | 16.4 b | <0.001 | 0.10 | 11.7 c | 21.96 a | 15.1 b | 22.90 a | 13.2 c | <0.001 | 0.06 |
Target Gene | Primer Sequence (5′-3′) | Accession No./Reference |
---|---|---|
iNOS | F-ACCTTCCGGGCAGCCTGTGA R-CAAGGAGGGTGGTGCGGCTG-3′ | NM_012611 |
COX-2 | F-GCTCAGCC ATACAGCAAATCC R-GGGAGTCGGGCAAT CATCAG | NM_017232 |
Caspase-3 | F-GCAGCTAACCTCAGAGAGACATTC R-ACGAGTAAGGTCATTTTTATTCCTGACTT | NM_012922 |
Bcl-2 | F-TGCGCTCAGCCCTGTG R-GGTAGCGACGAGAGAAGTCATC | NM_016993 |
BAX | F-CAAGAAGCTGAGCGAGTGTCT R-CAATCATCCTCTGCAGCTCCATATT | NM_017059 |
Cytochrome C | F-TTTGAATTCCTCATTAGTAGCTTTTTTGG R-CCATCCCTACGCATCCTTTAC | NM_012839 |
IL-1β | F-TGACAGACCCCAAAAGATTAAGG R-CTCATCTGGACAGCCCAAGTC | NM_031512.2 |
IL-6 | F-CCACCAGGAACGAAAGTCAAC R-TTGCGGAGAGAAACTTCATAGCT | NM_012589.2 |
TNF-α | F-CAGCCGATTTGCCATTTCA R-AGGGCTCTTGATGGCAGAGA | L19123.1 |
β-actin | F-CGCAGTTGGTTGGAGCAAA R-ACAATCAAAGTCCTCAGCCACAT | V01217.1 |
GAPDH | F-TGCTGGTGCTGAGTATGTCG-3′ R-TTGAGAGCAATGCCAGCC-3′ | NM_017008 |
invA. | F-ACAGTGCTCGTTTACGACCTGAAT R-AGACGACTGGTACTGATCGATAAT | [55] |
FimA | F-TTGCGAGTCTGATGTTTGTCG 62 R-CACGCTCACCGGAGTAGGAT | [55] |
16S rRNA. | F-AGGCCTTCGGGTTGTAAAGT R-GTTAGCCGGTGCTTCTTCTG | [55] |
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Alandiyjany, M.N.; Abdelaziz, A.S.; Abdelfattah-Hassan, A.; Hegazy, W.A.H.; Hassan, A.A.; Elazab, S.T.; Mohamed, E.A.A.; El-Shetry, E.S.; Saleh, A.A.; ElSawy, N.A.; et al. Novel In Vivo Assessment of Antimicrobial Efficacy of Ciprofloxacin Loaded Mesoporous Silica Nanoparticles against Salmonella typhimurium Infection. Pharmaceuticals 2022, 15, 357. https://doi.org/10.3390/ph15030357
Alandiyjany MN, Abdelaziz AS, Abdelfattah-Hassan A, Hegazy WAH, Hassan AA, Elazab ST, Mohamed EAA, El-Shetry ES, Saleh AA, ElSawy NA, et al. Novel In Vivo Assessment of Antimicrobial Efficacy of Ciprofloxacin Loaded Mesoporous Silica Nanoparticles against Salmonella typhimurium Infection. Pharmaceuticals. 2022; 15(3):357. https://doi.org/10.3390/ph15030357
Chicago/Turabian StyleAlandiyjany, Maher N., Ahmed S. Abdelaziz, Ahmed Abdelfattah-Hassan, Wael A. H. Hegazy, Arwa A. Hassan, Sara T. Elazab, Eman A. A. Mohamed, Eman S. El-Shetry, Ayman A. Saleh, Naser A. ElSawy, and et al. 2022. "Novel In Vivo Assessment of Antimicrobial Efficacy of Ciprofloxacin Loaded Mesoporous Silica Nanoparticles against Salmonella typhimurium Infection" Pharmaceuticals 15, no. 3: 357. https://doi.org/10.3390/ph15030357