Effects of ranibizumab on the extracellular matrix production by human Tenon's fibroblast
Anti-Vascular Endothelial Growth Factors (Anti-VEGF) agents have received recent interest as potential anti-fibrotic agents for their concurrent use with trabeculectomy. Preliminary cohort studies have revealed improved bleb morphology following trabeculectomy augmented with ranibizumab. The effects...
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| Veröffentlicht in: | Experimental eye research 2014-10, Vol.127, p.236-242 |
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| creator | Md Noh, Siti Munirah Sheikh Abdul Kadir, Siti H. Bannur, Zakaria M. Froemming, Gabriele Anisah Abdul Hamid Hasani, Narimah Mohd Nawawi, Hapizah Crowston, Jonathan G. Vasudevan, Sushil |
| description | Anti-Vascular Endothelial Growth Factors (Anti-VEGF) agents have received recent interest as potential anti-fibrotic agents for their concurrent use with trabeculectomy. Preliminary cohort studies have revealed improved bleb morphology following trabeculectomy augmented with ranibizumab. The effects of this humanized monoclonal antibody on human Tenon's fibroblast (HTF), the key player of post trabeculectomy scar formation, are not fully understood. This study was conducted to understand the effects of ranibizumab on extracellular matrix production by HTF. The effect of ranibizumab on HTF proliferation and cell viability was determined using MTT assay (3-(4,5-dimethylthiazone-2-yl)-2,5-diphenyl tetrazolium). Ranibizumab at concentrations ranging from 0.01 to 0.5 mg/mL were administered for 24, 48 and 72 h in serum and serum free conditions. Supernatants and cell lysates from samples were assessed for collagen type 1 alpha 1 and fibronectin mRNA and protein level using quantitative real time polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). After 48-h, ranibizumab at 0.5 mg/mL, significantly induced cell death under serum-free culture conditions (p |
| doi_str_mv | 10.1016/j.exer.2014.08.005 |
| format | Article |
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•This study aims to understand effects of ranibizumab on extracellular matrix component.•Analysis of gene and protein expression was done using PCR and ELISA techniques.•Ranibizumab inhibits collagen type 1 alpha 1 mRNA but increase its protein accumulation.•Ranibizumab caused insignificant impact on fibronectin mRNA and increase its protein accumulation.•We demonstrated that ranibizumab would be a potential anti-scarring agent in trabeculectomy.</description><identifier>ISSN: 0014-4835</identifier><identifier>EISSN: 1096-0007</identifier><identifier>DOI: 10.1016/j.exer.2014.08.005</identifier><identifier>PMID: 25139730</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>Angiogenesis Inhibitors - pharmacology ; Antibodies, Monoclonal, Humanized - pharmacology ; Cell Culture Techniques ; Cell Proliferation - drug effects ; Cell Survival - drug effects ; collagen type 1 alpha 1 ; Collagen Type I - genetics ; Collagen Type I - metabolism ; Enzyme-Linked Immunosorbent Assay ; Fibroblasts - drug effects ; Fibroblasts - metabolism ; fibronectin ; Fibronectins - genetics ; Fibronectins - metabolism ; Fluorescent Antibody Technique, Indirect ; Gene Expression Regulation - physiology ; Glaucoma, Open-Angle - surgery ; human Tenon's fibroblast ; Humans ; quantitative real time polymerase chain reaction ; Ranibizumab ; Real-Time Polymerase Chain Reaction ; RNA, Messenger - genetics ; Tenon Capsule - cytology ; Trabeculectomy ; Vascular Endothelial Growth Factor A - antagonists & inhibitors ; Vimentin - metabolism</subject><ispartof>Experimental eye research, 2014-10, Vol.127, p.236-242</ispartof><rights>2014 Elsevier Ltd</rights><rights>Copyright © 2014 Elsevier Ltd. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c422t-9761b125fc192e49344df34774bd93b59fbfb13fcecc99324ce7e36cf4df82583</citedby><cites>FETCH-LOGICAL-c422t-9761b125fc192e49344df34774bd93b59fbfb13fcecc99324ce7e36cf4df82583</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0014483514002267$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25139730$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Md Noh, Siti Munirah</creatorcontrib><creatorcontrib>Sheikh Abdul Kadir, Siti H.</creatorcontrib><creatorcontrib>Bannur, Zakaria M.</creatorcontrib><creatorcontrib>Froemming, Gabriele Anisah</creatorcontrib><creatorcontrib>Abdul Hamid Hasani, Narimah</creatorcontrib><creatorcontrib>Mohd Nawawi, Hapizah</creatorcontrib><creatorcontrib>Crowston, Jonathan G.</creatorcontrib><creatorcontrib>Vasudevan, Sushil</creatorcontrib><title>Effects of ranibizumab on the extracellular matrix production by human Tenon's fibroblast</title><title>Experimental eye research</title><addtitle>Exp Eye Res</addtitle><description>Anti-Vascular Endothelial Growth Factors (Anti-VEGF) agents have received recent interest as potential anti-fibrotic agents for their concurrent use with trabeculectomy. Preliminary cohort studies have revealed improved bleb morphology following trabeculectomy augmented with ranibizumab. The effects of this humanized monoclonal antibody on human Tenon's fibroblast (HTF), the key player of post trabeculectomy scar formation, are not fully understood. This study was conducted to understand the effects of ranibizumab on extracellular matrix production by HTF. The effect of ranibizumab on HTF proliferation and cell viability was determined using MTT assay (3-(4,5-dimethylthiazone-2-yl)-2,5-diphenyl tetrazolium). Ranibizumab at concentrations ranging from 0.01 to 0.5 mg/mL were administered for 24, 48 and 72 h in serum and serum free conditions. Supernatants and cell lysates from samples were assessed for collagen type 1 alpha 1 and fibronectin mRNA and protein level using quantitative real time polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). After 48-h, ranibizumab at 0.5 mg/mL, significantly induced cell death under serum-free culture conditions (p < 0.05). Ranibizumab caused significant reduction of collagen type 1 alpha 1 (COL1A1) mRNA, but not for fibronectin (FN). Meanwhile, COL1A1 and FN protein levels were found upregulated in treated monolayers compared to control monolayers. Ranibizumab at 0.5 mg/mL significantly reduced cell viability in cultured HTF. From this study, we found that single application of ranibizumab is inadequate to induce the anti-fibrotic effects on HTF, suggesting the importance of adjunctive therapy. Further studies are underway to understand mechanism of actions of ranibizumab on HTF.
•This study aims to understand effects of ranibizumab on extracellular matrix component.•Analysis of gene and protein expression was done using PCR and ELISA techniques.•Ranibizumab inhibits collagen type 1 alpha 1 mRNA but increase its protein accumulation.•Ranibizumab caused insignificant impact on fibronectin mRNA and increase its protein accumulation.•We demonstrated that ranibizumab would be a potential anti-scarring agent in trabeculectomy.</description><subject>Angiogenesis Inhibitors - pharmacology</subject><subject>Antibodies, Monoclonal, Humanized - pharmacology</subject><subject>Cell Culture Techniques</subject><subject>Cell Proliferation - drug effects</subject><subject>Cell Survival - drug effects</subject><subject>collagen type 1 alpha 1</subject><subject>Collagen Type I - genetics</subject><subject>Collagen Type I - metabolism</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Fibroblasts - drug effects</subject><subject>Fibroblasts - metabolism</subject><subject>fibronectin</subject><subject>Fibronectins - genetics</subject><subject>Fibronectins - metabolism</subject><subject>Fluorescent Antibody Technique, Indirect</subject><subject>Gene Expression Regulation - physiology</subject><subject>Glaucoma, Open-Angle - surgery</subject><subject>human Tenon's fibroblast</subject><subject>Humans</subject><subject>quantitative real time polymerase chain reaction</subject><subject>Ranibizumab</subject><subject>Real-Time Polymerase Chain Reaction</subject><subject>RNA, Messenger - genetics</subject><subject>Tenon Capsule - cytology</subject><subject>Trabeculectomy</subject><subject>Vascular Endothelial Growth Factor A - antagonists & inhibitors</subject><subject>Vimentin - metabolism</subject><issn>0014-4835</issn><issn>1096-0007</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kDtLBDEYRYMouj7-gIWk02bGPGcmYCPiCwQbLaxCkvmCWeahyYys_nqzrFpaBZJzLzcHoWNKSkpodb4sYQWxZISKkjQlIXILLShRVUEIqbfRguSXQjRc7qH9lJb5lota7KI9JilXNScL9HLtPbgp4dHjaIZgw9fcG4vHAU-vgGE1ReOg6-bORNybKYYVfotjO7spZMZ-4tfMD_gJhnE4TdgHG0fbmTQdoh1vugRHP-cBer65frq6Kx4eb--vLh8KJxibClVX1FImvaOKgVBciNbnmbWwreJWKm-9pdw7cE4pzoSDGnjlfMYaJht-gM42vXnW-wxp0n1I68lmgHFOmspKSsJVRTLKNqiLY0oRvH6LoTfxU1Oi10r1Uq-V6rVSTRqdlebQyU__bHto_yK_DjNwsQEg__Ij5HhyAQYHbYhZrW7H8F__N7jwiMQ</recordid><startdate>20141001</startdate><enddate>20141001</enddate><creator>Md Noh, Siti Munirah</creator><creator>Sheikh Abdul Kadir, Siti H.</creator><creator>Bannur, Zakaria M.</creator><creator>Froemming, Gabriele Anisah</creator><creator>Abdul Hamid Hasani, Narimah</creator><creator>Mohd Nawawi, Hapizah</creator><creator>Crowston, Jonathan G.</creator><creator>Vasudevan, Sushil</creator><general>Elsevier Ltd</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>20141001</creationdate><title>Effects of ranibizumab on the extracellular matrix production by human Tenon's fibroblast</title><author>Md Noh, Siti Munirah ; Sheikh Abdul Kadir, Siti H. ; Bannur, Zakaria M. ; Froemming, Gabriele Anisah ; Abdul Hamid Hasani, Narimah ; Mohd Nawawi, Hapizah ; Crowston, Jonathan G. ; Vasudevan, Sushil</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c422t-9761b125fc192e49344df34774bd93b59fbfb13fcecc99324ce7e36cf4df82583</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Angiogenesis Inhibitors - pharmacology</topic><topic>Antibodies, Monoclonal, Humanized - pharmacology</topic><topic>Cell Culture Techniques</topic><topic>Cell Proliferation - drug effects</topic><topic>Cell Survival - drug effects</topic><topic>collagen type 1 alpha 1</topic><topic>Collagen Type I - genetics</topic><topic>Collagen Type I - metabolism</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>Fibroblasts - drug effects</topic><topic>Fibroblasts - metabolism</topic><topic>fibronectin</topic><topic>Fibronectins - genetics</topic><topic>Fibronectins - metabolism</topic><topic>Fluorescent Antibody Technique, Indirect</topic><topic>Gene Expression Regulation - physiology</topic><topic>Glaucoma, Open-Angle - surgery</topic><topic>human Tenon's fibroblast</topic><topic>Humans</topic><topic>quantitative real time polymerase chain reaction</topic><topic>Ranibizumab</topic><topic>Real-Time Polymerase Chain Reaction</topic><topic>RNA, Messenger - genetics</topic><topic>Tenon Capsule - cytology</topic><topic>Trabeculectomy</topic><topic>Vascular Endothelial Growth Factor A - antagonists & inhibitors</topic><topic>Vimentin - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Md Noh, Siti Munirah</creatorcontrib><creatorcontrib>Sheikh Abdul Kadir, Siti H.</creatorcontrib><creatorcontrib>Bannur, Zakaria M.</creatorcontrib><creatorcontrib>Froemming, Gabriele Anisah</creatorcontrib><creatorcontrib>Abdul Hamid Hasani, Narimah</creatorcontrib><creatorcontrib>Mohd Nawawi, Hapizah</creatorcontrib><creatorcontrib>Crowston, Jonathan G.</creatorcontrib><creatorcontrib>Vasudevan, Sushil</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>Experimental eye research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Md Noh, Siti Munirah</au><au>Sheikh Abdul Kadir, Siti H.</au><au>Bannur, Zakaria M.</au><au>Froemming, Gabriele Anisah</au><au>Abdul Hamid Hasani, Narimah</au><au>Mohd Nawawi, Hapizah</au><au>Crowston, Jonathan G.</au><au>Vasudevan, Sushil</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Effects of ranibizumab on the extracellular matrix production by human Tenon's fibroblast</atitle><jtitle>Experimental eye research</jtitle><addtitle>Exp Eye Res</addtitle><date>2014-10-01</date><risdate>2014</risdate><volume>127</volume><spage>236</spage><epage>242</epage><pages>236-242</pages><issn>0014-4835</issn><eissn>1096-0007</eissn><abstract>Anti-Vascular Endothelial Growth Factors (Anti-VEGF) agents have received recent interest as potential anti-fibrotic agents for their concurrent use with trabeculectomy. Preliminary cohort studies have revealed improved bleb morphology following trabeculectomy augmented with ranibizumab. The effects of this humanized monoclonal antibody on human Tenon's fibroblast (HTF), the key player of post trabeculectomy scar formation, are not fully understood. This study was conducted to understand the effects of ranibizumab on extracellular matrix production by HTF. The effect of ranibizumab on HTF proliferation and cell viability was determined using MTT assay (3-(4,5-dimethylthiazone-2-yl)-2,5-diphenyl tetrazolium). Ranibizumab at concentrations ranging from 0.01 to 0.5 mg/mL were administered for 24, 48 and 72 h in serum and serum free conditions. Supernatants and cell lysates from samples were assessed for collagen type 1 alpha 1 and fibronectin mRNA and protein level using quantitative real time polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). After 48-h, ranibizumab at 0.5 mg/mL, significantly induced cell death under serum-free culture conditions (p < 0.05). Ranibizumab caused significant reduction of collagen type 1 alpha 1 (COL1A1) mRNA, but not for fibronectin (FN). Meanwhile, COL1A1 and FN protein levels were found upregulated in treated monolayers compared to control monolayers. Ranibizumab at 0.5 mg/mL significantly reduced cell viability in cultured HTF. From this study, we found that single application of ranibizumab is inadequate to induce the anti-fibrotic effects on HTF, suggesting the importance of adjunctive therapy. Further studies are underway to understand mechanism of actions of ranibizumab on HTF.
•This study aims to understand effects of ranibizumab on extracellular matrix component.•Analysis of gene and protein expression was done using PCR and ELISA techniques.•Ranibizumab inhibits collagen type 1 alpha 1 mRNA but increase its protein accumulation.•Ranibizumab caused insignificant impact on fibronectin mRNA and increase its protein accumulation.•We demonstrated that ranibizumab would be a potential anti-scarring agent in trabeculectomy.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>25139730</pmid><doi>10.1016/j.exer.2014.08.005</doi><tpages>7</tpages></addata></record> |
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| subjects | Angiogenesis Inhibitors - pharmacology Antibodies, Monoclonal, Humanized - pharmacology Cell Culture Techniques Cell Proliferation - drug effects Cell Survival - drug effects collagen type 1 alpha 1 Collagen Type I - genetics Collagen Type I - metabolism Enzyme-Linked Immunosorbent Assay Fibroblasts - drug effects Fibroblasts - metabolism fibronectin Fibronectins - genetics Fibronectins - metabolism Fluorescent Antibody Technique, Indirect Gene Expression Regulation - physiology Glaucoma, Open-Angle - surgery human Tenon's fibroblast Humans quantitative real time polymerase chain reaction Ranibizumab Real-Time Polymerase Chain Reaction RNA, Messenger - genetics Tenon Capsule - cytology Trabeculectomy Vascular Endothelial Growth Factor A - antagonists & inhibitors Vimentin - metabolism |
| title | Effects of ranibizumab on the extracellular matrix production by human Tenon's fibroblast |
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