Collagen and α-Tubulin of Mouse Tympanic Membrane Fibroblasts Treated with Quinolones and Aminoglycosides
Objective To assess collagen and α-tubulin levels of mouse tympanic membrane fibroblasts treated with quinolone and aminoglycoside antibiotics at concentrations found in eardrops. Study Design Prospective controlled cell culture study. Setting Academic tertiary medical center. Subjects Mouse tympani...
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| Veröffentlicht in: | Otolaryngology-head and neck surgery 2017-02, Vol.156 (2), p.341-349 |
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| creator | Milne-Davies, Bailey A. Antonelli, Patrick J. Orobello, Nicklas C. Dirain, Carolyn O. |
| description | Objective
To assess collagen and α-tubulin levels of mouse tympanic membrane fibroblasts treated with quinolone and aminoglycoside antibiotics at concentrations found in eardrops.
Study Design
Prospective controlled cell culture study.
Setting
Academic tertiary medical center.
Subjects
Mouse tympanic membrane fibroblasts.
Methods
In experiment 1, fibroblasts were treated with the following for 24 or 48 hours: phosphate-buffered saline (negative control), dilute hydrochloric acid (positive control), 0.5% gatifloxacin, or commercially available 0.3% ciprofloxacin, 0.3% ciprofloxacin + 0.1% dexamethasone, 0.3% ofloxacin, 0.5% moxifloxacin, 0.3% gentamicin, or 3.5 mg/mL of neomycin + polymyxin B sulfate + hydrocortisone. In experiment 2, cells were treated with the pure form of gatifloxacin, gentamicin, ofloxacin, or ciprofloxacin. Cells were observed with phase-contrast microscope until harvested. Proteins were extracted for Western blotting with antibodies against collagen α1 type I (collagen 1A1) and α-tubulin, and for densitometry to quantify levels.
Results
Collagen and tubulin levels in fibroblasts treated with ofloxacin, moxifloxacin, gatifloxacin, or gentamicin for 24 hours were not different from the saline control. Fibroblasts treated with neomycin + polymyxin B + hydrocortisone, ciprofloxacin + dexamethasone, or ciprofloxacin for 24 hours had lower collagen 1A1 and α-tubulin levels (all P < .001) than the negative control. After 48 hours, fibroblasts treated with neomycin + polymyxin B sulfate + hydrocortisone, ciprofloxacin + dexamethasone, ciprofloxacin, or moxifloxacin had lower collagen 1A1 (P ≤ .007) and α-tubulin (P < .0001; except ciprofloxacin, P = .033) as compared with control. In experiment 2, only cells treated with ciprofloxacin had lower collagen 1A1 and α-tubulin levels and cell viability (all P < .0001) than control. Cytotoxicity assay and phase-contrast images mirrored the protein findings.
Conclusion
The adverse impact of topical antibiotic exposure on tympanic membrane collagen and tubulin protein levels is drug specific. This may be important for selection of ototopical therapy. |
| doi_str_mv | 10.1177/0194599816672627 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1859726489</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sage_id>10.1177_0194599816672627</sage_id><sourcerecordid>1859726489</sourcerecordid><originalsourceid>FETCH-LOGICAL-c3866-f74ca7852f2dd320d19932487842661c93ac27bea7fe84ac2bd5d88defb726393</originalsourceid><addsrcrecordid>eNqFkc9O4zAQxq3VoqWwe9_Tyse9BGwn8Z9jqSggURBSOUdOPOm6SuxiN0J9LF6EZ8KhZQ8rrTjNjOb3ffJ8RugnJWeUCnFOqCpKpSTlXDDOxBc0oUSJjEsqvqLJuM7G_TE6iXFNCEmc-IaOmUi1VHyC1jPfdXoFDmtn8OtLthzqobMO-xYv_BABL3f9Rjvb4AX0ddAO8NzWwdedjtuIlwH0Fgx-tts_-GGwznfeQXx3m_ZpXHW7xkdrIH5HR63uIvw41FP0OL9czq6z2_urm9n0NmtyyXnWiqLRQpasZcbkjBiqVM4KKWTBOKeNynXDRA1atCCL1NemNFIaaOuUQa7yU_R777sJ_mmAuK16GxtIZzpIF1VUliqRhRxRskeb4GMM0FabYHsddhUl1Zhw9W_CSfLr4D7UPZi_go9IEyD3wLPtYPepYXV_fXcxJ5LQUZrtpTH9SLX2Q3ApqP-_5Q0gqpS7</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1859726489</pqid></control><display><type>article</type><title>Collagen and α-Tubulin of Mouse Tympanic Membrane Fibroblasts Treated with Quinolones and Aminoglycosides</title><source>MEDLINE</source><source>Wiley Online Library Journals Frontfile Complete</source><source>SAGE Complete</source><creator>Milne-Davies, Bailey A. ; Antonelli, Patrick J. ; Orobello, Nicklas C. ; Dirain, Carolyn O.</creator><creatorcontrib>Milne-Davies, Bailey A. ; Antonelli, Patrick J. ; Orobello, Nicklas C. ; Dirain, Carolyn O.</creatorcontrib><description>Objective
To assess collagen and α-tubulin levels of mouse tympanic membrane fibroblasts treated with quinolone and aminoglycoside antibiotics at concentrations found in eardrops.
Study Design
Prospective controlled cell culture study.
Setting
Academic tertiary medical center.
Subjects
Mouse tympanic membrane fibroblasts.
Methods
In experiment 1, fibroblasts were treated with the following for 24 or 48 hours: phosphate-buffered saline (negative control), dilute hydrochloric acid (positive control), 0.5% gatifloxacin, or commercially available 0.3% ciprofloxacin, 0.3% ciprofloxacin + 0.1% dexamethasone, 0.3% ofloxacin, 0.5% moxifloxacin, 0.3% gentamicin, or 3.5 mg/mL of neomycin + polymyxin B sulfate + hydrocortisone. In experiment 2, cells were treated with the pure form of gatifloxacin, gentamicin, ofloxacin, or ciprofloxacin. Cells were observed with phase-contrast microscope until harvested. Proteins were extracted for Western blotting with antibodies against collagen α1 type I (collagen 1A1) and α-tubulin, and for densitometry to quantify levels.
Results
Collagen and tubulin levels in fibroblasts treated with ofloxacin, moxifloxacin, gatifloxacin, or gentamicin for 24 hours were not different from the saline control. Fibroblasts treated with neomycin + polymyxin B + hydrocortisone, ciprofloxacin + dexamethasone, or ciprofloxacin for 24 hours had lower collagen 1A1 and α-tubulin levels (all P < .001) than the negative control. After 48 hours, fibroblasts treated with neomycin + polymyxin B sulfate + hydrocortisone, ciprofloxacin + dexamethasone, ciprofloxacin, or moxifloxacin had lower collagen 1A1 (P ≤ .007) and α-tubulin (P < .0001; except ciprofloxacin, P = .033) as compared with control. In experiment 2, only cells treated with ciprofloxacin had lower collagen 1A1 and α-tubulin levels and cell viability (all P < .0001) than control. Cytotoxicity assay and phase-contrast images mirrored the protein findings.
Conclusion
The adverse impact of topical antibiotic exposure on tympanic membrane collagen and tubulin protein levels is drug specific. This may be important for selection of ototopical therapy.</description><identifier>ISSN: 0194-5998</identifier><identifier>EISSN: 1097-6817</identifier><identifier>DOI: 10.1177/0194599816672627</identifier><identifier>PMID: 27677596</identifier><language>eng</language><publisher>Los Angeles, CA: SAGE Publications</publisher><subject>aminoglycoside ; Aminoglycosides - pharmacology ; Animals ; Blotting, Western ; Cells, Cultured ; collagen ; Collagen - drug effects ; eardrop ; fibroblast ; Fibroblasts - drug effects ; Mice ; Mice, Transgenic ; Prospective Studies ; quinolone ; Quinolones - pharmacology ; Tubulin - drug effects ; Tympanic Membrane - cytology</subject><ispartof>Otolaryngology-head and neck surgery, 2017-02, Vol.156 (2), p.341-349</ispartof><rights>American Academy of Otolaryngology—Head and Neck Surgery Foundation 2016</rights><rights>2017 American Association of Otolaryngology‐Head and Neck Surgery Foundation (AAO‐HNSF)</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3866-f74ca7852f2dd320d19932487842661c93ac27bea7fe84ac2bd5d88defb726393</citedby><cites>FETCH-LOGICAL-c3866-f74ca7852f2dd320d19932487842661c93ac27bea7fe84ac2bd5d88defb726393</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://journals.sagepub.com/doi/pdf/10.1177/0194599816672627$$EPDF$$P50$$Gsage$$H</linktopdf><linktohtml>$$Uhttps://journals.sagepub.com/doi/10.1177/0194599816672627$$EHTML$$P50$$Gsage$$H</linktohtml><link.rule.ids>314,777,781,1412,21800,27905,27906,43602,43603,45555,45556</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27677596$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Milne-Davies, Bailey A.</creatorcontrib><creatorcontrib>Antonelli, Patrick J.</creatorcontrib><creatorcontrib>Orobello, Nicklas C.</creatorcontrib><creatorcontrib>Dirain, Carolyn O.</creatorcontrib><title>Collagen and α-Tubulin of Mouse Tympanic Membrane Fibroblasts Treated with Quinolones and Aminoglycosides</title><title>Otolaryngology-head and neck surgery</title><addtitle>Otolaryngol Head Neck Surg</addtitle><description>Objective
To assess collagen and α-tubulin levels of mouse tympanic membrane fibroblasts treated with quinolone and aminoglycoside antibiotics at concentrations found in eardrops.
Study Design
Prospective controlled cell culture study.
Setting
Academic tertiary medical center.
Subjects
Mouse tympanic membrane fibroblasts.
Methods
In experiment 1, fibroblasts were treated with the following for 24 or 48 hours: phosphate-buffered saline (negative control), dilute hydrochloric acid (positive control), 0.5% gatifloxacin, or commercially available 0.3% ciprofloxacin, 0.3% ciprofloxacin + 0.1% dexamethasone, 0.3% ofloxacin, 0.5% moxifloxacin, 0.3% gentamicin, or 3.5 mg/mL of neomycin + polymyxin B sulfate + hydrocortisone. In experiment 2, cells were treated with the pure form of gatifloxacin, gentamicin, ofloxacin, or ciprofloxacin. Cells were observed with phase-contrast microscope until harvested. Proteins were extracted for Western blotting with antibodies against collagen α1 type I (collagen 1A1) and α-tubulin, and for densitometry to quantify levels.
Results
Collagen and tubulin levels in fibroblasts treated with ofloxacin, moxifloxacin, gatifloxacin, or gentamicin for 24 hours were not different from the saline control. Fibroblasts treated with neomycin + polymyxin B + hydrocortisone, ciprofloxacin + dexamethasone, or ciprofloxacin for 24 hours had lower collagen 1A1 and α-tubulin levels (all P < .001) than the negative control. After 48 hours, fibroblasts treated with neomycin + polymyxin B sulfate + hydrocortisone, ciprofloxacin + dexamethasone, ciprofloxacin, or moxifloxacin had lower collagen 1A1 (P ≤ .007) and α-tubulin (P < .0001; except ciprofloxacin, P = .033) as compared with control. In experiment 2, only cells treated with ciprofloxacin had lower collagen 1A1 and α-tubulin levels and cell viability (all P < .0001) than control. Cytotoxicity assay and phase-contrast images mirrored the protein findings.
Conclusion
The adverse impact of topical antibiotic exposure on tympanic membrane collagen and tubulin protein levels is drug specific. This may be important for selection of ototopical therapy.</description><subject>aminoglycoside</subject><subject>Aminoglycosides - pharmacology</subject><subject>Animals</subject><subject>Blotting, Western</subject><subject>Cells, Cultured</subject><subject>collagen</subject><subject>Collagen - drug effects</subject><subject>eardrop</subject><subject>fibroblast</subject><subject>Fibroblasts - drug effects</subject><subject>Mice</subject><subject>Mice, Transgenic</subject><subject>Prospective Studies</subject><subject>quinolone</subject><subject>Quinolones - pharmacology</subject><subject>Tubulin - drug effects</subject><subject>Tympanic Membrane - cytology</subject><issn>0194-5998</issn><issn>1097-6817</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc9O4zAQxq3VoqWwe9_Tyse9BGwn8Z9jqSggURBSOUdOPOm6SuxiN0J9LF6EZ8KhZQ8rrTjNjOb3ffJ8RugnJWeUCnFOqCpKpSTlXDDOxBc0oUSJjEsqvqLJuM7G_TE6iXFNCEmc-IaOmUi1VHyC1jPfdXoFDmtn8OtLthzqobMO-xYv_BABL3f9Rjvb4AX0ddAO8NzWwdedjtuIlwH0Fgx-tts_-GGwznfeQXx3m_ZpXHW7xkdrIH5HR63uIvw41FP0OL9czq6z2_urm9n0NmtyyXnWiqLRQpasZcbkjBiqVM4KKWTBOKeNynXDRA1atCCL1NemNFIaaOuUQa7yU_R777sJ_mmAuK16GxtIZzpIF1VUliqRhRxRskeb4GMM0FabYHsddhUl1Zhw9W_CSfLr4D7UPZi_go9IEyD3wLPtYPepYXV_fXcxJ5LQUZrtpTH9SLX2Q3ApqP-_5Q0gqpS7</recordid><startdate>201702</startdate><enddate>201702</enddate><creator>Milne-Davies, Bailey A.</creator><creator>Antonelli, Patrick J.</creator><creator>Orobello, Nicklas C.</creator><creator>Dirain, Carolyn O.</creator><general>SAGE Publications</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>201702</creationdate><title>Collagen and α-Tubulin of Mouse Tympanic Membrane Fibroblasts Treated with Quinolones and Aminoglycosides</title><author>Milne-Davies, Bailey A. ; Antonelli, Patrick J. ; Orobello, Nicklas C. ; Dirain, Carolyn O.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3866-f74ca7852f2dd320d19932487842661c93ac27bea7fe84ac2bd5d88defb726393</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>aminoglycoside</topic><topic>Aminoglycosides - pharmacology</topic><topic>Animals</topic><topic>Blotting, Western</topic><topic>Cells, Cultured</topic><topic>collagen</topic><topic>Collagen - drug effects</topic><topic>eardrop</topic><topic>fibroblast</topic><topic>Fibroblasts - drug effects</topic><topic>Mice</topic><topic>Mice, Transgenic</topic><topic>Prospective Studies</topic><topic>quinolone</topic><topic>Quinolones - pharmacology</topic><topic>Tubulin - drug effects</topic><topic>Tympanic Membrane - cytology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Milne-Davies, Bailey A.</creatorcontrib><creatorcontrib>Antonelli, Patrick J.</creatorcontrib><creatorcontrib>Orobello, Nicklas C.</creatorcontrib><creatorcontrib>Dirain, Carolyn O.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Otolaryngology-head and neck surgery</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Milne-Davies, Bailey A.</au><au>Antonelli, Patrick J.</au><au>Orobello, Nicklas C.</au><au>Dirain, Carolyn O.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Collagen and α-Tubulin of Mouse Tympanic Membrane Fibroblasts Treated with Quinolones and Aminoglycosides</atitle><jtitle>Otolaryngology-head and neck surgery</jtitle><addtitle>Otolaryngol Head Neck Surg</addtitle><date>2017-02</date><risdate>2017</risdate><volume>156</volume><issue>2</issue><spage>341</spage><epage>349</epage><pages>341-349</pages><issn>0194-5998</issn><eissn>1097-6817</eissn><abstract>Objective
To assess collagen and α-tubulin levels of mouse tympanic membrane fibroblasts treated with quinolone and aminoglycoside antibiotics at concentrations found in eardrops.
Study Design
Prospective controlled cell culture study.
Setting
Academic tertiary medical center.
Subjects
Mouse tympanic membrane fibroblasts.
Methods
In experiment 1, fibroblasts were treated with the following for 24 or 48 hours: phosphate-buffered saline (negative control), dilute hydrochloric acid (positive control), 0.5% gatifloxacin, or commercially available 0.3% ciprofloxacin, 0.3% ciprofloxacin + 0.1% dexamethasone, 0.3% ofloxacin, 0.5% moxifloxacin, 0.3% gentamicin, or 3.5 mg/mL of neomycin + polymyxin B sulfate + hydrocortisone. In experiment 2, cells were treated with the pure form of gatifloxacin, gentamicin, ofloxacin, or ciprofloxacin. Cells were observed with phase-contrast microscope until harvested. Proteins were extracted for Western blotting with antibodies against collagen α1 type I (collagen 1A1) and α-tubulin, and for densitometry to quantify levels.
Results
Collagen and tubulin levels in fibroblasts treated with ofloxacin, moxifloxacin, gatifloxacin, or gentamicin for 24 hours were not different from the saline control. Fibroblasts treated with neomycin + polymyxin B + hydrocortisone, ciprofloxacin + dexamethasone, or ciprofloxacin for 24 hours had lower collagen 1A1 and α-tubulin levels (all P < .001) than the negative control. After 48 hours, fibroblasts treated with neomycin + polymyxin B sulfate + hydrocortisone, ciprofloxacin + dexamethasone, ciprofloxacin, or moxifloxacin had lower collagen 1A1 (P ≤ .007) and α-tubulin (P < .0001; except ciprofloxacin, P = .033) as compared with control. In experiment 2, only cells treated with ciprofloxacin had lower collagen 1A1 and α-tubulin levels and cell viability (all P < .0001) than control. Cytotoxicity assay and phase-contrast images mirrored the protein findings.
Conclusion
The adverse impact of topical antibiotic exposure on tympanic membrane collagen and tubulin protein levels is drug specific. This may be important for selection of ototopical therapy.</abstract><cop>Los Angeles, CA</cop><pub>SAGE Publications</pub><pmid>27677596</pmid><doi>10.1177/0194599816672627</doi><tpages>9</tpages></addata></record> |
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| ispartof | Otolaryngology-head and neck surgery, 2017-02, Vol.156 (2), p.341-349 |
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| language | eng |
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| source | MEDLINE; Wiley Online Library Journals Frontfile Complete; SAGE Complete |
| subjects | aminoglycoside Aminoglycosides - pharmacology Animals Blotting, Western Cells, Cultured collagen Collagen - drug effects eardrop fibroblast Fibroblasts - drug effects Mice Mice, Transgenic Prospective Studies quinolone Quinolones - pharmacology Tubulin - drug effects Tympanic Membrane - cytology |
| title | Collagen and α-Tubulin of Mouse Tympanic Membrane Fibroblasts Treated with Quinolones and Aminoglycosides |
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