The cellular basis for biocide-induced fluorescein hyperfluorescence in mammalian cell culture
Clinical examination of the ocular surface is commonly carried out after application of sodium fluorescein in both veterinary and medical practice by assessing the resulting 'staining'. Although localized intensely stained regions of the cornea frequently occur after exposure to 'adve...
Gespeichert in:
| Veröffentlicht in: | PloS one 2014-01, Vol.9 (1), p.e84427 |
|---|---|
| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | |
|---|---|
| container_issue | 1 |
| container_start_page | e84427 |
| container_title | PloS one |
| container_volume | 9 |
| creator | Bakkar, May M Hardaker, Luke March, Peter Morgan, Philip B Maldonado-Codina, Carole Dobson, Curtis B |
| description | Clinical examination of the ocular surface is commonly carried out after application of sodium fluorescein in both veterinary and medical practice by assessing the resulting 'staining'. Although localized intensely stained regions of the cornea frequently occur after exposure to 'adverse' clinical stimuli, the cell biology underlying this staining is unknown, including whether intense fluorescein staining indicates the presence of damaged cells. Ocular exposure to certain contact lens multipurpose solutions (MPS) gives rise to intense fluorescein staining referred to as solution induced corneal staining (SICS), and we have made use of this phenomenon with Vero and L929 cell culture models to investigate the fundamental biology of fluorescein interactions with cells. We found that all cells take up fluorescein, however a sub-population internalize much higher levels, giving rise to brightly staining 'hyperfluorescent' cells within the treated cultures, which contain fluorescein throughout the cell cytoplasm and nucleus. The numbers of these hyperfluorescent cells are significantly increased after exposure to MPS associated with SICS. Surprisingly, hyperfluorescent cells did not show higher levels of staining with propidium iodide, a marker of lysed cells. Consistently, treatment with the cytolytic toxin benzalkonium chloride resulted in almost all cells staining with propidium iodide, and the complete abolition of fluorescein hyperfluorescence. Finally we found that internalization of fluorescein and its loss from treated cells both require cellular activity, as both processes were halted after incubation at 4 °C. We conclude that fluorescein hyperfluorescence can be replicated in three diverse cell cultures, and is increased by MPS-treatment, as occurs clinically. The process involves the concentration of fluorescein by a sub-population of cells that are active, and does not occur in lysed cells. Our data suggest that corneal staining in the clinic reflects active living cells, and is not directly caused by dead cells being produced in response to adverse clinical stimuli. |
| doi_str_mv | 10.1371/journal.pone.0084427 |
| format | Article |
| fullrecord | <record><control><sourceid>gale_plos_</sourceid><recordid>TN_cdi_plos_journals_1492273842</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A478836049</galeid><doaj_id>oai_doaj_org_article_a5e17aa643194b22ae4fc37ad4b5bcd6</doaj_id><sourcerecordid>A478836049</sourcerecordid><originalsourceid>FETCH-LOGICAL-c692t-6a0ac5f752bad9ef695f0dbac17b863c03f2190c6a7eaa08c76b6af38f94b38a3</originalsourceid><addsrcrecordid>eNqNkl-L1DAUxYso7rr6DUQLguDDjGmSpumLsCz-GVhY0NVHw216M5MhbWaTVtxvb3anO0xBQfKQy83vnlwOJ8teFmRZsKp4v_Vj6MEtd77HJSGSc1o9yk6LmtGFoIQ9PqpPsmcxbgkpmRTiaXZCOZe1KMlp9vN6g7lG50YHIW8g2pgbnyrrtW1xYft21Njmxo0-YNRo-3xzu8NwaPQa89TsoOvAWejv1XI9umEM-Dx7YsBFfDHdZ9n3Tx-vL74sLq8-ry7OLxda1HRYCCCgS1OVtIG2RiPq0pC2AV1UjRRME2ZoURMtoEIAInUlGgGGSVPzhklgZ9nrve7O-agma6IqeE1pxSSniVjtidbDVu2C7SDcKg9W3Td8WCsIg9UOFZRYVACCsyLJUwrIjWYVtLwpG92KpPVh-m1sOmyTB0MANxOdv_R2o9b-l2I14ZKRJPBmEgj-ZsQ4_GPliVpD2sr2xicx3dmo1TmvpGSC8DpRy79Q6bTYWZ3CYWzqzwbezQYSM-DvYQ1jjGr17ev_s1c_5uzbI3aD4IZN9G4crO_jHOR7UAcfY0BzcK4g6i7bD26ou2yrKdtp7NWx64ehhzCzP6pq9rg</addsrcrecordid><sourcetype>Open Website</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1492273842</pqid></control><display><type>article</type><title>The cellular basis for biocide-induced fluorescein hyperfluorescence in mammalian cell culture</title><source>MEDLINE</source><source>TestCollectionTL3OpenAccess</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>PubMed Central</source><source>Free Full-Text Journals in Chemistry</source><source>Public Library of Science (PLoS)</source><creator>Bakkar, May M ; Hardaker, Luke ; March, Peter ; Morgan, Philip B ; Maldonado-Codina, Carole ; Dobson, Curtis B</creator><creatorcontrib>Bakkar, May M ; Hardaker, Luke ; March, Peter ; Morgan, Philip B ; Maldonado-Codina, Carole ; Dobson, Curtis B</creatorcontrib><description>Clinical examination of the ocular surface is commonly carried out after application of sodium fluorescein in both veterinary and medical practice by assessing the resulting 'staining'. Although localized intensely stained regions of the cornea frequently occur after exposure to 'adverse' clinical stimuli, the cell biology underlying this staining is unknown, including whether intense fluorescein staining indicates the presence of damaged cells. Ocular exposure to certain contact lens multipurpose solutions (MPS) gives rise to intense fluorescein staining referred to as solution induced corneal staining (SICS), and we have made use of this phenomenon with Vero and L929 cell culture models to investigate the fundamental biology of fluorescein interactions with cells. We found that all cells take up fluorescein, however a sub-population internalize much higher levels, giving rise to brightly staining 'hyperfluorescent' cells within the treated cultures, which contain fluorescein throughout the cell cytoplasm and nucleus. The numbers of these hyperfluorescent cells are significantly increased after exposure to MPS associated with SICS. Surprisingly, hyperfluorescent cells did not show higher levels of staining with propidium iodide, a marker of lysed cells. Consistently, treatment with the cytolytic toxin benzalkonium chloride resulted in almost all cells staining with propidium iodide, and the complete abolition of fluorescein hyperfluorescence. Finally we found that internalization of fluorescein and its loss from treated cells both require cellular activity, as both processes were halted after incubation at 4 °C. We conclude that fluorescein hyperfluorescence can be replicated in three diverse cell cultures, and is increased by MPS-treatment, as occurs clinically. The process involves the concentration of fluorescein by a sub-population of cells that are active, and does not occur in lysed cells. Our data suggest that corneal staining in the clinic reflects active living cells, and is not directly caused by dead cells being produced in response to adverse clinical stimuli.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0084427</identifier><identifier>PMID: 24489650</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Animals ; Anti-Infective Agents, Local - pharmacology ; Benzalkonium chloride ; Benzalkonium Compounds - pharmacology ; Biocides ; Biology ; Cell culture ; Cell cycle ; Cell Line ; Cells (Biology) ; Cercopithecus aethiops ; Chlorides ; Contact lenses ; Cornea ; Cytoplasm ; Data processing ; Drug Interactions ; Exposure ; Fibroblasts ; Fluorescein ; Fluorescein - pharmacology ; Hydrogels ; Internalization ; Iodides ; Life sciences ; Medical equipment ; Medicine ; Mice ; Microscopy ; Nuclei ; Nuclei (cytology) ; Penicillin ; Propidium ; Propidium iodide ; Sodium ; Solutions ; Staining ; Stimuli ; Surface active agents ; Toxins ; Vero Cells ; Veterinary medicine</subject><ispartof>PloS one, 2014-01, Vol.9 (1), p.e84427</ispartof><rights>COPYRIGHT 2014 Public Library of Science</rights><rights>2014 Bakkar et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2014 Bakkar et al 2014 Bakkar et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-6a0ac5f752bad9ef695f0dbac17b863c03f2190c6a7eaa08c76b6af38f94b38a3</citedby><cites>FETCH-LOGICAL-c692t-6a0ac5f752bad9ef695f0dbac17b863c03f2190c6a7eaa08c76b6af38f94b38a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3904830/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3904830/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79342,79343</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24489650$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bakkar, May M</creatorcontrib><creatorcontrib>Hardaker, Luke</creatorcontrib><creatorcontrib>March, Peter</creatorcontrib><creatorcontrib>Morgan, Philip B</creatorcontrib><creatorcontrib>Maldonado-Codina, Carole</creatorcontrib><creatorcontrib>Dobson, Curtis B</creatorcontrib><title>The cellular basis for biocide-induced fluorescein hyperfluorescence in mammalian cell culture</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Clinical examination of the ocular surface is commonly carried out after application of sodium fluorescein in both veterinary and medical practice by assessing the resulting 'staining'. Although localized intensely stained regions of the cornea frequently occur after exposure to 'adverse' clinical stimuli, the cell biology underlying this staining is unknown, including whether intense fluorescein staining indicates the presence of damaged cells. Ocular exposure to certain contact lens multipurpose solutions (MPS) gives rise to intense fluorescein staining referred to as solution induced corneal staining (SICS), and we have made use of this phenomenon with Vero and L929 cell culture models to investigate the fundamental biology of fluorescein interactions with cells. We found that all cells take up fluorescein, however a sub-population internalize much higher levels, giving rise to brightly staining 'hyperfluorescent' cells within the treated cultures, which contain fluorescein throughout the cell cytoplasm and nucleus. The numbers of these hyperfluorescent cells are significantly increased after exposure to MPS associated with SICS. Surprisingly, hyperfluorescent cells did not show higher levels of staining with propidium iodide, a marker of lysed cells. Consistently, treatment with the cytolytic toxin benzalkonium chloride resulted in almost all cells staining with propidium iodide, and the complete abolition of fluorescein hyperfluorescence. Finally we found that internalization of fluorescein and its loss from treated cells both require cellular activity, as both processes were halted after incubation at 4 °C. We conclude that fluorescein hyperfluorescence can be replicated in three diverse cell cultures, and is increased by MPS-treatment, as occurs clinically. The process involves the concentration of fluorescein by a sub-population of cells that are active, and does not occur in lysed cells. Our data suggest that corneal staining in the clinic reflects active living cells, and is not directly caused by dead cells being produced in response to adverse clinical stimuli.</description><subject>Animals</subject><subject>Anti-Infective Agents, Local - pharmacology</subject><subject>Benzalkonium chloride</subject><subject>Benzalkonium Compounds - pharmacology</subject><subject>Biocides</subject><subject>Biology</subject><subject>Cell culture</subject><subject>Cell cycle</subject><subject>Cell Line</subject><subject>Cells (Biology)</subject><subject>Cercopithecus aethiops</subject><subject>Chlorides</subject><subject>Contact lenses</subject><subject>Cornea</subject><subject>Cytoplasm</subject><subject>Data processing</subject><subject>Drug Interactions</subject><subject>Exposure</subject><subject>Fibroblasts</subject><subject>Fluorescein</subject><subject>Fluorescein - pharmacology</subject><subject>Hydrogels</subject><subject>Internalization</subject><subject>Iodides</subject><subject>Life sciences</subject><subject>Medical equipment</subject><subject>Medicine</subject><subject>Mice</subject><subject>Microscopy</subject><subject>Nuclei</subject><subject>Nuclei (cytology)</subject><subject>Penicillin</subject><subject>Propidium</subject><subject>Propidium iodide</subject><subject>Sodium</subject><subject>Solutions</subject><subject>Staining</subject><subject>Stimuli</subject><subject>Surface active agents</subject><subject>Toxins</subject><subject>Vero Cells</subject><subject>Veterinary medicine</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNqNkl-L1DAUxYso7rr6DUQLguDDjGmSpumLsCz-GVhY0NVHw216M5MhbWaTVtxvb3anO0xBQfKQy83vnlwOJ8teFmRZsKp4v_Vj6MEtd77HJSGSc1o9yk6LmtGFoIQ9PqpPsmcxbgkpmRTiaXZCOZe1KMlp9vN6g7lG50YHIW8g2pgbnyrrtW1xYft21Njmxo0-YNRo-3xzu8NwaPQa89TsoOvAWejv1XI9umEM-Dx7YsBFfDHdZ9n3Tx-vL74sLq8-ry7OLxda1HRYCCCgS1OVtIG2RiPq0pC2AV1UjRRME2ZoURMtoEIAInUlGgGGSVPzhklgZ9nrve7O-agma6IqeE1pxSSniVjtidbDVu2C7SDcKg9W3Td8WCsIg9UOFZRYVACCsyLJUwrIjWYVtLwpG92KpPVh-m1sOmyTB0MANxOdv_R2o9b-l2I14ZKRJPBmEgj-ZsQ4_GPliVpD2sr2xicx3dmo1TmvpGSC8DpRy79Q6bTYWZ3CYWzqzwbezQYSM-DvYQ1jjGr17ev_s1c_5uzbI3aD4IZN9G4crO_jHOR7UAcfY0BzcK4g6i7bD26ou2yrKdtp7NWx64ehhzCzP6pq9rg</recordid><startdate>20140128</startdate><enddate>20140128</enddate><creator>Bakkar, May M</creator><creator>Hardaker, Luke</creator><creator>March, Peter</creator><creator>Morgan, Philip B</creator><creator>Maldonado-Codina, Carole</creator><creator>Dobson, Curtis B</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</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>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20140128</creationdate><title>The cellular basis for biocide-induced fluorescein hyperfluorescence in mammalian cell culture</title><author>Bakkar, May M ; Hardaker, Luke ; March, Peter ; Morgan, Philip B ; Maldonado-Codina, Carole ; Dobson, Curtis B</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-6a0ac5f752bad9ef695f0dbac17b863c03f2190c6a7eaa08c76b6af38f94b38a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Animals</topic><topic>Anti-Infective Agents, Local - pharmacology</topic><topic>Benzalkonium chloride</topic><topic>Benzalkonium Compounds - pharmacology</topic><topic>Biocides</topic><topic>Biology</topic><topic>Cell culture</topic><topic>Cell cycle</topic><topic>Cell Line</topic><topic>Cells (Biology)</topic><topic>Cercopithecus aethiops</topic><topic>Chlorides</topic><topic>Contact lenses</topic><topic>Cornea</topic><topic>Cytoplasm</topic><topic>Data processing</topic><topic>Drug Interactions</topic><topic>Exposure</topic><topic>Fibroblasts</topic><topic>Fluorescein</topic><topic>Fluorescein - pharmacology</topic><topic>Hydrogels</topic><topic>Internalization</topic><topic>Iodides</topic><topic>Life sciences</topic><topic>Medical equipment</topic><topic>Medicine</topic><topic>Mice</topic><topic>Microscopy</topic><topic>Nuclei</topic><topic>Nuclei (cytology)</topic><topic>Penicillin</topic><topic>Propidium</topic><topic>Propidium iodide</topic><topic>Sodium</topic><topic>Solutions</topic><topic>Staining</topic><topic>Stimuli</topic><topic>Surface active agents</topic><topic>Toxins</topic><topic>Vero Cells</topic><topic>Veterinary medicine</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bakkar, May M</creatorcontrib><creatorcontrib>Hardaker, Luke</creatorcontrib><creatorcontrib>March, Peter</creatorcontrib><creatorcontrib>Morgan, Philip B</creatorcontrib><creatorcontrib>Maldonado-Codina, Carole</creatorcontrib><creatorcontrib>Dobson, Curtis B</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - Academic</collection><collection>ProQuest Engineering Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Engineering Database</collection><collection>Nursing & Allied Health Premium</collection><collection>Advanced Technologies & Aerospace Database</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>Materials Science Collection</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>Engineering Collection</collection><collection>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><collection>TestCollectionTL3OpenAccess</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bakkar, May M</au><au>Hardaker, Luke</au><au>March, Peter</au><au>Morgan, Philip B</au><au>Maldonado-Codina, Carole</au><au>Dobson, Curtis B</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The cellular basis for biocide-induced fluorescein hyperfluorescence in mammalian cell culture</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2014-01-28</date><risdate>2014</risdate><volume>9</volume><issue>1</issue><spage>e84427</spage><pages>e84427-</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Clinical examination of the ocular surface is commonly carried out after application of sodium fluorescein in both veterinary and medical practice by assessing the resulting 'staining'. Although localized intensely stained regions of the cornea frequently occur after exposure to 'adverse' clinical stimuli, the cell biology underlying this staining is unknown, including whether intense fluorescein staining indicates the presence of damaged cells. Ocular exposure to certain contact lens multipurpose solutions (MPS) gives rise to intense fluorescein staining referred to as solution induced corneal staining (SICS), and we have made use of this phenomenon with Vero and L929 cell culture models to investigate the fundamental biology of fluorescein interactions with cells. We found that all cells take up fluorescein, however a sub-population internalize much higher levels, giving rise to brightly staining 'hyperfluorescent' cells within the treated cultures, which contain fluorescein throughout the cell cytoplasm and nucleus. The numbers of these hyperfluorescent cells are significantly increased after exposure to MPS associated with SICS. Surprisingly, hyperfluorescent cells did not show higher levels of staining with propidium iodide, a marker of lysed cells. Consistently, treatment with the cytolytic toxin benzalkonium chloride resulted in almost all cells staining with propidium iodide, and the complete abolition of fluorescein hyperfluorescence. Finally we found that internalization of fluorescein and its loss from treated cells both require cellular activity, as both processes were halted after incubation at 4 °C. We conclude that fluorescein hyperfluorescence can be replicated in three diverse cell cultures, and is increased by MPS-treatment, as occurs clinically. The process involves the concentration of fluorescein by a sub-population of cells that are active, and does not occur in lysed cells. Our data suggest that corneal staining in the clinic reflects active living cells, and is not directly caused by dead cells being produced in response to adverse clinical stimuli.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>24489650</pmid><doi>10.1371/journal.pone.0084427</doi><tpages>e84427</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2014-01, Vol.9 (1), p.e84427 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1492273842 |
| source | MEDLINE; TestCollectionTL3OpenAccess; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Free Full-Text Journals in Chemistry; Public Library of Science (PLoS) |
| subjects | Animals Anti-Infective Agents, Local - pharmacology Benzalkonium chloride Benzalkonium Compounds - pharmacology Biocides Biology Cell culture Cell cycle Cell Line Cells (Biology) Cercopithecus aethiops Chlorides Contact lenses Cornea Cytoplasm Data processing Drug Interactions Exposure Fibroblasts Fluorescein Fluorescein - pharmacology Hydrogels Internalization Iodides Life sciences Medical equipment Medicine Mice Microscopy Nuclei Nuclei (cytology) Penicillin Propidium Propidium iodide Sodium Solutions Staining Stimuli Surface active agents Toxins Vero Cells Veterinary medicine |
| title | The cellular basis for biocide-induced fluorescein hyperfluorescence in mammalian cell culture |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-12T19%3A20%3A27IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=The%20cellular%20basis%20for%20biocide-induced%20fluorescein%20hyperfluorescence%20in%20mammalian%20cell%20culture&rft.jtitle=PloS%20one&rft.au=Bakkar,%20May%20M&rft.date=2014-01-28&rft.volume=9&rft.issue=1&rft.spage=e84427&rft.pages=e84427-&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0084427&rft_dat=%3Cgale_plos_%3EA478836049%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1492273842&rft_id=info:pmid/24489650&rft_galeid=A478836049&rft_doaj_id=oai_doaj_org_article_a5e17aa643194b22ae4fc37ad4b5bcd6&rfr_iscdi=true |