Effect of chitosan nanogels loaded with vancomycin and gamma interferon on TNF-α gene expression in macrophage cell line activated with methicillin-resistant Staphylococcus aureus (MRSA)
Abstract
Background and Objectives: Staphylococcus aureus is an opportunistic pathogen that frequently leads to asymptomatic infections. Methicillin-resistant strains (MRSA) pose a significant threat as they are resistant to most commonly used antibiotics, complicating treatment efforts. This study aimed to develop chitosan nanogels loaded with vancomycin and IFN-γ and to assess the expression of the TNF-α gene in a cell line infected with MRSA.
Materials and Methods: Following the synthesis and confirmation of the chitosan nanogels, vancomycin and IFN-γ were incorporated into these nanogels. The synthesis was validated using DLS, FTIR, TEM, and SEM. Subsequently, the antibacterial efficacy of the nanogels was assessed. Finally, four groups of cell lines were designed: control, MRSA, chitosan nanogels and IFN-γ-vancomycin chitosan nanogels. After infection of the groups (except control) with MRSA, 5 μg/mL of nanogels, and nanogels (drug and IFN-γ) were added to groups 3 and 4, respectively. Then the expression of TNF-α gene in each group was analyzed by RT-PCR at 6 and 24 hours.
Results: At pH 6.5 and 7.4, the MIC of 1 μg/mL was obtained for free vancomycin, whereas that of IFN-γ-vancomycin nanogels at both pHs was respectively 8 and 64 μg/mL. The IC50 of chitosan nanogels and nanogels loaded with vancomycin-IFN-γ on RAW264.7 cells were 2.37 and 4.15 μg/mL in 24 hours, respectively. In group 4 in comparison to the MRSA group, TNF-α expression decreased significantly following 24 hours.
Conclusion: Loading of vancomycin and IFN-γ in the chitosan nanogel can reduce TNF-α gene expression on MRSA infected cell lines.
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21. Faraji N, Esrafili A, Esfandiari B, Abednezhad A, Naghizadeh M, Arasteh J. Synthesis of pH-sensitive hyaluronic acid nanogels loaded with paclitaxel and interferon gamma: Characterization and effect on the A549 lung carcinoma cell line. Colloids Surf B Biointerfaces 2021; 205: 111845.
22. Pei Y, Mohamed MF, Seleem MN, Yeo Y. Particle engineering for intracellular delivery of vancomycin to methicillin-resistant Staphylococcus aureus (MRSA)-infected macrophages. J Control Release 2017; 267: 133-143.
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24. Saeb AT, Alshammari AS, Al-Brahim H, Al-Rubeaan KA. Production of silver nanoparticles with strong and stable antimicrobial activity against highly pathogenic and multidrug resistant bacteria. ScientificWorldJournal 2014; 2014: 704708.
25. Tongsai S, Koomanachai P. The safety and efficacy of high versus low vancomycin trough levels in the treatment of patients with infections caused by methicillin-resistant Staphylococcus aureus: a meta-analysis. BMC Res Notes 2016; 9: 455.
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27. Yang Z, Liu J, Gao J, Chen S, Huang G. Chitosan coated vancomycin hydrochloride liposomes: Characterizations and evaluation. Int J Pharm 2015; 495: 508-515.
28. Lee EJ, Jun SH, Kim HE, Kim HW, Koh YH, Jang JH. Silica xerogel-chitosan nano-hybrids for use as drug eluting bone replacement. J Mater Sci Mater Med 2010; 21: 207-214.
29. Karakeçili A, Topuz B, Korpayev S, Erdek M. Metal-organic frameworks for on-demand pH controlled delivery of vancomycin from chitosan scaffolds. Mater Sci Eng C Mater Biol Appl 2019; 105: 110098.
30. Chávez de Paz LE, Resin A, Howard KA, Sutherland DS, Wejse PL. Antimicrobial effect of chitosan nanoparticles on Streptococcus mutans biofilms. Appl Environ Microbiol 2011; 77: 3892-3895.
31. Divya K, Vijayan S, George TK, Jisha MS. Antimicrobial properties of chitosan nanoparticles: Mode of action and factors affecting activity. Fibers Polym 2017; 18: 221-230.
32. Costa EM, Silva S, Vicente S, Neto C, Castro PM, Veiga M, et al. Chitosan nanoparticles as alternative anti-staphylococci agents: Bactericidal, antibiofilm and antiadhesive effects. Mater Sci Eng C Mater Biol Appl 2017; 79: 221-226.
33. Dey S, Bishayi B. Killing of Staphylococcus aureus in murine macrophages by chloroquine used alone and in combination with ciprofloxacin or azithromycin. J Inflamm Res 2015; 8: 29-47.
34. Turner MD, Nedjai B, Hurst T, Pennington DJ. Cytokines and chemokines: At the crossroads of cell signalling and inflammatory disease. Biochim Biophys Acta 2014; 1843: 2563-2582.
35. Popa C, Netea MG, van Riel PL, van der Meer JW, Stalenhoef AF. The role of TNF-α in chronic inflammatory conditions, intermediary metabolism, and cardiovascular risk. J Lipid Res 2007; 48: 751-762.
36. Dasgupta A. Advances in antibiotic measurement. Adv Clin Chem 2012; 56: 75-104.
37. Nalos M, Santner-Nanan B, Parnell G, Tang B, McLean AS, Nanan R. Immune effects of interferon gamma in persistent staphylococcal sepsis. Am J Respir Crit Care Med 2012; 185: 110-112.
38. Smith RP, Baltch AL, Ritz WJ, Michelsen PB, Bopp LH. IFN-γ enhances killing of methicillin-resistant Staphylococcus aureus by human monocytes more effectively than GM-CSF in the presence of daptomycin and other antibiotics. Cytokine 2010; 51: 274-277.
39. Barin JG, Talor MV, Schaub JA, Diny NL, Hou X, Hoyer M, et al. Collaborative Interferon-γ and Interleukin-17 signaling protects the oral mucosa from Staphylococcus aureus. Am J Pathol 2016; 186: 2337-2352.
2. Yah CS, Simate GS. Nanoparticles as potential new generation broad spectrum antimicrobial agents. Daru 2015; 23: 43.
3. Lakhundi S, Zhang K. Methicillin-resistant Staphylococcus aureus: molecular characterization, evolution, and epidemiology. Clin Microbiol Rev 2018; 31(4): e00020-18.
4. Serra R, Grande R, Butrico L, Rossi A, Settimio UF, Caroleo B, et al. Chronic wound infections: The role of Pseudomonas aeruginosa and Staphylococcus aureus. Expert Rev Anti Infect Ther 2015; 13: 605-613.
5. Shalaby MW, Dokla EME, Serya RAT, Abouzid KAM. Penicillin binding protein 2a: An overview and a medicinal chemistry perspective. Eur J Med Chem 2020; 199: 112312.
6. Lee AS, de Lencastre H, Garau J, Kluytmans J, Malhotra-Kumar S, Peschel A, et al. Methicillin-resistant Staphylococcus aureus. Nat Rev Dis Primers 2018; 4: 18033.
7. Friães A, Resina C, Manuel V, Lito L, Ramirez M, Melo-Cristino J. Epidemiological survey of the first case of vancomycin-resistant Staphylococcus aureus infection in Europe. Epidemiol Infect 2015; 143: 745-748.
8. Bhutiani N, Li Q, Anderson CD, Gallagher HC, De Jesus M, Singh R, et al. Enhanced gut barrier integrity sensitizes colon cancer to immune therapy. Oncoimmunology 2018; 7(11): e1498438.
9. Ardolino M, Raulet DH. Cytokine therapy restores antitumor responses of NK cells rendered anergic in MHC I-deficient tumors. Oncoimmunology 2015; 5(1): e1002725.
10. Yang P-M, Chou C-J, Tseng S-H, Hung C-F. Bioinformatics and in vitro experimental analyses identify the selective therapeutic potential of interferon gamma and apigenin against cervical squamous cell carcinoma and adenocarcinoma. Oncotarget 2017; 8: 46145-46162.
11. McNab F, Mayer-Barber K, Sher A, Wack A, O’Garra A. Type I interferons in infectious disease. Nat Rev Immunol 2015; 15: 87-103.
12. Swindle EJ, Brown JM, Rådinger M, Deleo FR, Metcalfe DD. Interferon-γ enhances both the anti-bacterial and the pro-inflammatory response of human mast cells to Staphylococcus aureus. Immunology 2015; 146: 470-485.
13. Gao W, Chen Y, Zhang Y, Zhang Q, Zhang L. Nanoparticle-based local antimicrobial drug delivery. Adv Drug Deliv Rev 2018; 127: 46-57.
14. Kalhapure RS, Jadhav M, Rambharose S, Mocktar C, Singh S, Renukuntla J, et al. pH-responsive chitosan nanoparticles from a novel twin-chain anionic amphiphile for controlled and targeted delivery of vancomycin. Colloids Surf B Biointerfaces 2017; 158: 650-657.
15. Honary S, Ebrahimi P, Hadianamrei R. Optimization of particle size and encapsulation efficiency of vancomycin nanoparticles by response surface methodology. Pharm Dev Technol 2014; 19: 987-998.
16. Chakraborty SP, Sahu SK, Pramanik P, Roy S. In vitro antimicrobial activity of nanoconjugated vancomycin against drug resistant Staphylococcus aureus. Int J Pharm 2012; 436: 659-676.
17. Cheung RC, Ng TB, Wong JH, Chan WY. Chitosan: An update on potential biomedical and pharmaceutical applications. Mar Drugs 2015; 13: 5156-5186.
18. Spyropoulos V, Chalkias A, Georgiou G, Papalois A, Kouskouni E, Baka S, et al. Initial Immune Response in Escherichia coli, Staphylococcus aureus, and Candida albicans Bacteremia. Inflammation 2020; 43: 179-190.
19. Osuchowski MF, Craciun F, Weixelbaumer KM, Duffy ER, Remick DG. Sepsis chronically in MARS: systemic cytokine responses are always mixed regardless of the outcome, magnitude, or phase of sepsis. J Immunol 2012; 189: 4648-4656.
20. Li P, Zhao J, Chen Y, Cheng B, Yu Z, Zhao Y, et al. Preparation and characterization of chitosan physical hydrogels with enhanced mechanical and antibacterial properties. Carbohydr Polym 2017; 157: 1383-1392.
21. Faraji N, Esrafili A, Esfandiari B, Abednezhad A, Naghizadeh M, Arasteh J. Synthesis of pH-sensitive hyaluronic acid nanogels loaded with paclitaxel and interferon gamma: Characterization and effect on the A549 lung carcinoma cell line. Colloids Surf B Biointerfaces 2021; 205: 111845.
22. Pei Y, Mohamed MF, Seleem MN, Yeo Y. Particle engineering for intracellular delivery of vancomycin to methicillin-resistant Staphylococcus aureus (MRSA)-infected macrophages. J Control Release 2017; 267: 133-143.
23. Chang VS, Dhaliwal DK, Raju L, Kowalski RP. Antibiotic Resistance in the Treatment of Staphylococcus aureus Keratitis: a 20-Year Review. Cornea 2015; 34: 698-703.
24. Saeb AT, Alshammari AS, Al-Brahim H, Al-Rubeaan KA. Production of silver nanoparticles with strong and stable antimicrobial activity against highly pathogenic and multidrug resistant bacteria. ScientificWorldJournal 2014; 2014: 704708.
25. Tongsai S, Koomanachai P. The safety and efficacy of high versus low vancomycin trough levels in the treatment of patients with infections caused by methicillin-resistant Staphylococcus aureus: a meta-analysis. BMC Res Notes 2016; 9: 455.
26. Hermsen ED, Hanson M, Sankaranarayanan J, Stoner JA, Florescu MC, Rupp ME. Clinical outcomes and nephrotoxicity associated with vancomycin trough concentrations during treatment of deep-seated infections. Expert Opin Drug Saf 2010; 9: 9-14.
27. Yang Z, Liu J, Gao J, Chen S, Huang G. Chitosan coated vancomycin hydrochloride liposomes: Characterizations and evaluation. Int J Pharm 2015; 495: 508-515.
28. Lee EJ, Jun SH, Kim HE, Kim HW, Koh YH, Jang JH. Silica xerogel-chitosan nano-hybrids for use as drug eluting bone replacement. J Mater Sci Mater Med 2010; 21: 207-214.
29. Karakeçili A, Topuz B, Korpayev S, Erdek M. Metal-organic frameworks for on-demand pH controlled delivery of vancomycin from chitosan scaffolds. Mater Sci Eng C Mater Biol Appl 2019; 105: 110098.
30. Chávez de Paz LE, Resin A, Howard KA, Sutherland DS, Wejse PL. Antimicrobial effect of chitosan nanoparticles on Streptococcus mutans biofilms. Appl Environ Microbiol 2011; 77: 3892-3895.
31. Divya K, Vijayan S, George TK, Jisha MS. Antimicrobial properties of chitosan nanoparticles: Mode of action and factors affecting activity. Fibers Polym 2017; 18: 221-230.
32. Costa EM, Silva S, Vicente S, Neto C, Castro PM, Veiga M, et al. Chitosan nanoparticles as alternative anti-staphylococci agents: Bactericidal, antibiofilm and antiadhesive effects. Mater Sci Eng C Mater Biol Appl 2017; 79: 221-226.
33. Dey S, Bishayi B. Killing of Staphylococcus aureus in murine macrophages by chloroquine used alone and in combination with ciprofloxacin or azithromycin. J Inflamm Res 2015; 8: 29-47.
34. Turner MD, Nedjai B, Hurst T, Pennington DJ. Cytokines and chemokines: At the crossroads of cell signalling and inflammatory disease. Biochim Biophys Acta 2014; 1843: 2563-2582.
35. Popa C, Netea MG, van Riel PL, van der Meer JW, Stalenhoef AF. The role of TNF-α in chronic inflammatory conditions, intermediary metabolism, and cardiovascular risk. J Lipid Res 2007; 48: 751-762.
36. Dasgupta A. Advances in antibiotic measurement. Adv Clin Chem 2012; 56: 75-104.
37. Nalos M, Santner-Nanan B, Parnell G, Tang B, McLean AS, Nanan R. Immune effects of interferon gamma in persistent staphylococcal sepsis. Am J Respir Crit Care Med 2012; 185: 110-112.
38. Smith RP, Baltch AL, Ritz WJ, Michelsen PB, Bopp LH. IFN-γ enhances killing of methicillin-resistant Staphylococcus aureus by human monocytes more effectively than GM-CSF in the presence of daptomycin and other antibiotics. Cytokine 2010; 51: 274-277.
39. Barin JG, Talor MV, Schaub JA, Diny NL, Hou X, Hoyer M, et al. Collaborative Interferon-γ and Interleukin-17 signaling protects the oral mucosa from Staphylococcus aureus. Am J Pathol 2016; 186: 2337-2352.
| Files | ||
| Issue | Vol 16 No 5 (2024) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v16i5.16794 | |
| Keywords | ||
| Tumor necrosis factor (TNF-α); Methicillin-resistant Staphylococcus aureus (MRSA); Vancomycin; Interferon-gamma (IFN-γ) chitosan nanogels | ||
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How to Cite
1.
Arkiya S, Hesampour A, Esrafili A, Arasteh J. Effect of chitosan nanogels loaded with vancomycin and gamma interferon on TNF-α gene expression in macrophage cell line activated with methicillin-resistant Staphylococcus aureus (MRSA). Iran J Microbiol. 2024;16(5):614-623.
