N-Acetylcysteine Protects Bladder Epithelial Cells from Bacterial Invasion and Displays Antibiofilm Activity against Urinary Tract Bacterial Pathogens
<b>Introduction:</b> Urinary tract infections (UTIs) affect more than 150 million individuals annually. A strong correlation exists between bladder epithelia invasion by uropathogenic bacteria and patients with recurrent UTIs. Intracellular bacteria often recolonise epithelial cells post...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-07-01
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| Series: | Antibiotics |
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| Online Access: | https://www.mdpi.com/2079-6382/10/8/900 |
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| author | Arthika Manoharan Samantha Ognenovska Denis Paino Greg Whiteley Trevor Glasbey Frederik H. Kriel Jessica Farrell Kate H. Moore Jim Manos Theerthankar Das |
| author_facet | Arthika Manoharan Samantha Ognenovska Denis Paino Greg Whiteley Trevor Glasbey Frederik H. Kriel Jessica Farrell Kate H. Moore Jim Manos Theerthankar Das |
| author_sort | Arthika Manoharan |
| collection | DOAJ |
| description | <b>Introduction:</b> Urinary tract infections (UTIs) affect more than 150 million individuals annually. A strong correlation exists between bladder epithelia invasion by uropathogenic bacteria and patients with recurrent UTIs. Intracellular bacteria often recolonise epithelial cells post-antibiotic treatment. We investigated whether N-acetylcysteine (NAC) could prevent uropathogenic <i>E. coli</i> and <i>E. faecalis</i> bladder cell invasion, in addition to its effect on uropathogens when used alone or in combination with ciprofloxacin. <b>Methods:</b> An invasion assay was performed in which bacteria were added to bladder epithelial cells (BECs) in presence of NAC and invasion was allowed to occur. Cells were washed with gentamicin, lysed, and plated for enumeration of the intracellular bacterial load. Cytotoxicity was evaluated by exposing BECs to various concentrations of NAC and quantifying the metabolic activity using resazurin at different exposure times. The effect of NAC on the preformed biofilms was also investigated by treating 48 h biofilms for 24 h and enumerating colony counts. Bacteria were stained with propidium iodide (PI) to measure membrane damage. <b>Results:</b> NAC completely inhibited BEC invasion by multiple <i>E. coli</i> and <i>E. faecalis</i> clinical strains in a dose-dependent manner (<i>p</i> < 0.01). This was also evident when bacterial invasion was visualised using GFP-tagged <i>E. coli</i>. NAC displayed no cytotoxicity against BECs despite its intrinsic acidity (pH ~2.6), with >90% cellular viability 48 h post-exposure. NAC also prevented biofilm formation by <i>E. coli</i> and <i>E. faecalis</i> and significantly reduced bacterial loads in 48 h biofilms when combined with ciprofloxacin. NAC visibly damaged <i>E. coli</i> and <i>E. faecalis</i> bacterial membranes, with a threefold increase in propidium iodide-stained cells following treatment (<i>p</i> < 0.05). <b>Conclusions:</b> NAC is a non-toxic, antibiofilm agent in vitro and can prevent cell invasion and IBC formation by uropathogens, thus providing a potentially novel and efficacious treatment for UTIs. When combined with an antibiotic, it may disrupt bacterial biofilms and eliminate residual bacteria. |
| first_indexed | 2024-03-10T09:03:59Z |
| format | Article |
| id | doaj.art-1128c22280c74da4a94c96a2ac1029c5 |
| institution | Directory Open Access Journal |
| issn | 2079-6382 |
| language | English |
| last_indexed | 2024-03-10T09:03:59Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibiotics |
| spelling | doaj.art-1128c22280c74da4a94c96a2ac1029c52023-11-22T06:32:58ZengMDPI AGAntibiotics2079-63822021-07-0110890010.3390/antibiotics10080900N-Acetylcysteine Protects Bladder Epithelial Cells from Bacterial Invasion and Displays Antibiofilm Activity against Urinary Tract Bacterial PathogensArthika Manoharan0Samantha Ognenovska1Denis Paino2Greg Whiteley3Trevor Glasbey4Frederik H. Kriel5Jessica Farrell6Kate H. Moore7Jim Manos8Theerthankar Das9Department of Infectious Diseases and Immunology, School of Medical Sciences, Charles Perkins Centre, The University of Sydney, Sydney, NSW 2006, AustraliaDepartment of Urogynaecology, St George Hospital, University of New South Wales, Sydney, NSW 2052, AustraliaDepartment of Infectious Diseases and Immunology, School of Medical Sciences, Charles Perkins Centre, The University of Sydney, Sydney, NSW 2006, AustraliaWhiteley Corporation, 19–23 Laverick Avenue, Tomago, NSW 2319, AustraliaWhiteley Corporation, 19–23 Laverick Avenue, Tomago, NSW 2319, AustraliaWhiteley Corporation, 19–23 Laverick Avenue, Tomago, NSW 2319, AustraliaDepartment of Infectious Diseases and Immunology, School of Medical Sciences, Charles Perkins Centre, The University of Sydney, Sydney, NSW 2006, AustraliaDepartment of Urogynaecology, St George Hospital, University of New South Wales, Sydney, NSW 2052, AustraliaDepartment of Infectious Diseases and Immunology, School of Medical Sciences, Charles Perkins Centre, The University of Sydney, Sydney, NSW 2006, AustraliaDepartment of Infectious Diseases and Immunology, School of Medical Sciences, Charles Perkins Centre, The University of Sydney, Sydney, NSW 2006, Australia<b>Introduction:</b> Urinary tract infections (UTIs) affect more than 150 million individuals annually. A strong correlation exists between bladder epithelia invasion by uropathogenic bacteria and patients with recurrent UTIs. Intracellular bacteria often recolonise epithelial cells post-antibiotic treatment. We investigated whether N-acetylcysteine (NAC) could prevent uropathogenic <i>E. coli</i> and <i>E. faecalis</i> bladder cell invasion, in addition to its effect on uropathogens when used alone or in combination with ciprofloxacin. <b>Methods:</b> An invasion assay was performed in which bacteria were added to bladder epithelial cells (BECs) in presence of NAC and invasion was allowed to occur. Cells were washed with gentamicin, lysed, and plated for enumeration of the intracellular bacterial load. Cytotoxicity was evaluated by exposing BECs to various concentrations of NAC and quantifying the metabolic activity using resazurin at different exposure times. The effect of NAC on the preformed biofilms was also investigated by treating 48 h biofilms for 24 h and enumerating colony counts. Bacteria were stained with propidium iodide (PI) to measure membrane damage. <b>Results:</b> NAC completely inhibited BEC invasion by multiple <i>E. coli</i> and <i>E. faecalis</i> clinical strains in a dose-dependent manner (<i>p</i> < 0.01). This was also evident when bacterial invasion was visualised using GFP-tagged <i>E. coli</i>. NAC displayed no cytotoxicity against BECs despite its intrinsic acidity (pH ~2.6), with >90% cellular viability 48 h post-exposure. NAC also prevented biofilm formation by <i>E. coli</i> and <i>E. faecalis</i> and significantly reduced bacterial loads in 48 h biofilms when combined with ciprofloxacin. NAC visibly damaged <i>E. coli</i> and <i>E. faecalis</i> bacterial membranes, with a threefold increase in propidium iodide-stained cells following treatment (<i>p</i> < 0.05). <b>Conclusions:</b> NAC is a non-toxic, antibiofilm agent in vitro and can prevent cell invasion and IBC formation by uropathogens, thus providing a potentially novel and efficacious treatment for UTIs. When combined with an antibiotic, it may disrupt bacterial biofilms and eliminate residual bacteria.https://www.mdpi.com/2079-6382/10/8/900NAC<i>E. coli</i>UPEC<i>E. faecalis</i>biofilmsantibiotic resistance |
| spellingShingle | Arthika Manoharan Samantha Ognenovska Denis Paino Greg Whiteley Trevor Glasbey Frederik H. Kriel Jessica Farrell Kate H. Moore Jim Manos Theerthankar Das N-Acetylcysteine Protects Bladder Epithelial Cells from Bacterial Invasion and Displays Antibiofilm Activity against Urinary Tract Bacterial Pathogens Antibiotics NAC <i>E. coli</i> UPEC <i>E. faecalis</i> biofilms antibiotic resistance |
| title | N-Acetylcysteine Protects Bladder Epithelial Cells from Bacterial Invasion and Displays Antibiofilm Activity against Urinary Tract Bacterial Pathogens |
| title_full | N-Acetylcysteine Protects Bladder Epithelial Cells from Bacterial Invasion and Displays Antibiofilm Activity against Urinary Tract Bacterial Pathogens |
| title_fullStr | N-Acetylcysteine Protects Bladder Epithelial Cells from Bacterial Invasion and Displays Antibiofilm Activity against Urinary Tract Bacterial Pathogens |
| title_full_unstemmed | N-Acetylcysteine Protects Bladder Epithelial Cells from Bacterial Invasion and Displays Antibiofilm Activity against Urinary Tract Bacterial Pathogens |
| title_short | N-Acetylcysteine Protects Bladder Epithelial Cells from Bacterial Invasion and Displays Antibiofilm Activity against Urinary Tract Bacterial Pathogens |
| title_sort | n acetylcysteine protects bladder epithelial cells from bacterial invasion and displays antibiofilm activity against urinary tract bacterial pathogens |
| topic | NAC <i>E. coli</i> UPEC <i>E. faecalis</i> biofilms antibiotic resistance |
| url | https://www.mdpi.com/2079-6382/10/8/900 |
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