Influence of subretinal fluid in advanced stage retinopathy of prematurity on proangiogenic response and cell proliferation
The clinical phenotype of advanced stage retinopathy of prematurity (ROP, stages 4 and 5) cannot be replicated in an animal model. To dissect the molecular events that can lead up to advanced ROP, we examined subretinal fluid (SRF) and surgically dissected retrolental membranes from patients with ad...
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| Veröffentlicht in: | Molecular vision 2014-06, Vol.20, p.881-893 |
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| creator | Ma, Jie Mehta, Manisha Lam, Godfrey Cyr, Desireé Ng, Tat Fong Hirose, Tatsuo Tawansy, Khaled A Taylor, Andrew W Lashkari, Kameran |
| description | The clinical phenotype of advanced stage retinopathy of prematurity (ROP, stages 4 and 5) cannot be replicated in an animal model. To dissect the molecular events that can lead up to advanced ROP, we examined subretinal fluid (SRF) and surgically dissected retrolental membranes from patients with advanced ROP to evaluate its influences on cell proliferation, angiogenic properties, and macrophage polarity.
We compared our findings to SRF collected from patients with uncomplicated rhegmatogenous retinal detachment (RD) without proliferative vitreoretinopathy and surgically dissected epiretinal membrane from eyes with macular pucker. All subretinal fluid samples were equalized for protein. The angiogenic potential of SRF from ROP eyes was measured using a combination of capillary cord formation in a fibrin clot assay, and its proliferative effect was tested with a DNA synthesis of human retinal microvascular endothelial cells. Findings were compared with SRF collected from participants with uncomplicated rhegmatogenous RD without proliferative vitreoretinopathy. The ability of SRF to induce nitric oxide production was measured in vitro using murine J774A.1 macrophages. Cytokine profiles of SRF from ROP and RD eyes were measured using a multienzyme-linked immunosorbent assay (ELISA). Fluorescent immunohistochemistry of retrolental membranes from ROP was performed to detect the presence of leukocytes and the composition of tissue macrophages using markers for M1 and M2 differentiation.
The cytokine composition in SRF revealed that in ROP, not only were several proangiogenic factors were preferentially elevated but also the profile of proinflammatory factors was also increased compared to the RD eyes. SRF from ROP eyes supported cell proliferation and endothelial cord formation while SRF from RD eyes had inhibitory effects. SRF from eyes with ROP but not RD robustly induced nitric oxide production in macrophages. Furthermore, fluorescent immunostaining revealed a preponderance of M1 over M2 macrophages in retrolental fibrous membranes from ROP eyes. The cytokine profile and biologic properties of SRF in ROP promote a proangiogenic environment, which supports the maintenance and proliferation of fibrous membranes associated with advanced stages of ROP. In contrast, SRF from RD eyes exhibits a suppressive environment for endothelial cell proliferation and angiogenesis.
Our investigation demonstrates that the microenvironment in advanced ROP eyes is proangiogenic a |
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We compared our findings to SRF collected from patients with uncomplicated rhegmatogenous retinal detachment (RD) without proliferative vitreoretinopathy and surgically dissected epiretinal membrane from eyes with macular pucker. All subretinal fluid samples were equalized for protein. The angiogenic potential of SRF from ROP eyes was measured using a combination of capillary cord formation in a fibrin clot assay, and its proliferative effect was tested with a DNA synthesis of human retinal microvascular endothelial cells. Findings were compared with SRF collected from participants with uncomplicated rhegmatogenous RD without proliferative vitreoretinopathy. The ability of SRF to induce nitric oxide production was measured in vitro using murine J774A.1 macrophages. Cytokine profiles of SRF from ROP and RD eyes were measured using a multienzyme-linked immunosorbent assay (ELISA). Fluorescent immunohistochemistry of retrolental membranes from ROP was performed to detect the presence of leukocytes and the composition of tissue macrophages using markers for M1 and M2 differentiation.
The cytokine composition in SRF revealed that in ROP, not only were several proangiogenic factors were preferentially elevated but also the profile of proinflammatory factors was also increased compared to the RD eyes. SRF from ROP eyes supported cell proliferation and endothelial cord formation while SRF from RD eyes had inhibitory effects. SRF from eyes with ROP but not RD robustly induced nitric oxide production in macrophages. Furthermore, fluorescent immunostaining revealed a preponderance of M1 over M2 macrophages in retrolental fibrous membranes from ROP eyes. The cytokine profile and biologic properties of SRF in ROP promote a proangiogenic environment, which supports the maintenance and proliferation of fibrous membranes associated with advanced stages of ROP. In contrast, SRF from RD eyes exhibits a suppressive environment for endothelial cell proliferation and angiogenesis.
Our investigation demonstrates that the microenvironment in advanced ROP eyes is proangiogenic and proinflammatory. These findings suggest that management of advanced ROP should not be limited to the surgical removal of the fibrovascular membranes and antiangiogenic therapy but also directed to anti-inflammatory therapy and to promote M2 activation over M1 activity.</description><identifier>ISSN: 1090-0535</identifier><identifier>EISSN: 1090-0535</identifier><identifier>PMID: 24966660</identifier><language>eng</language><publisher>United States: Molecular Vision</publisher><subject>Animals ; Capillaries - metabolism ; Capillaries - pathology ; Capillaries - physiopathology ; Cell Polarity ; Cell Proliferation ; Cells, Cultured ; Cytokines - metabolism ; Humans ; Immunohistochemistry ; Infant ; Inflammation Mediators - metabolism ; Macrophages - metabolism ; Macrophages - pathology ; Mice ; Neovascularization, Physiologic ; Nitric Oxide - biosynthesis ; Nitrites - metabolism ; Retinal Detachment - metabolism ; Retinopathy of Prematurity - metabolism ; Retinopathy of Prematurity - pathology ; Retinopathy of Prematurity - physiopathology ; Subretinal Fluid - metabolism</subject><ispartof>Molecular vision, 2014-06, Vol.20, p.881-893</ispartof><rights>Copyright © 2014 Molecular Vision. 2014 Molecular Vision</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4067231/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4067231/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,53790,53792</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24966660$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ma, Jie</creatorcontrib><creatorcontrib>Mehta, Manisha</creatorcontrib><creatorcontrib>Lam, Godfrey</creatorcontrib><creatorcontrib>Cyr, Desireé</creatorcontrib><creatorcontrib>Ng, Tat Fong</creatorcontrib><creatorcontrib>Hirose, Tatsuo</creatorcontrib><creatorcontrib>Tawansy, Khaled A</creatorcontrib><creatorcontrib>Taylor, Andrew W</creatorcontrib><creatorcontrib>Lashkari, Kameran</creatorcontrib><title>Influence of subretinal fluid in advanced stage retinopathy of prematurity on proangiogenic response and cell proliferation</title><title>Molecular vision</title><addtitle>Mol Vis</addtitle><description>The clinical phenotype of advanced stage retinopathy of prematurity (ROP, stages 4 and 5) cannot be replicated in an animal model. To dissect the molecular events that can lead up to advanced ROP, we examined subretinal fluid (SRF) and surgically dissected retrolental membranes from patients with advanced ROP to evaluate its influences on cell proliferation, angiogenic properties, and macrophage polarity.
We compared our findings to SRF collected from patients with uncomplicated rhegmatogenous retinal detachment (RD) without proliferative vitreoretinopathy and surgically dissected epiretinal membrane from eyes with macular pucker. All subretinal fluid samples were equalized for protein. The angiogenic potential of SRF from ROP eyes was measured using a combination of capillary cord formation in a fibrin clot assay, and its proliferative effect was tested with a DNA synthesis of human retinal microvascular endothelial cells. Findings were compared with SRF collected from participants with uncomplicated rhegmatogenous RD without proliferative vitreoretinopathy. The ability of SRF to induce nitric oxide production was measured in vitro using murine J774A.1 macrophages. Cytokine profiles of SRF from ROP and RD eyes were measured using a multienzyme-linked immunosorbent assay (ELISA). Fluorescent immunohistochemistry of retrolental membranes from ROP was performed to detect the presence of leukocytes and the composition of tissue macrophages using markers for M1 and M2 differentiation.
The cytokine composition in SRF revealed that in ROP, not only were several proangiogenic factors were preferentially elevated but also the profile of proinflammatory factors was also increased compared to the RD eyes. SRF from ROP eyes supported cell proliferation and endothelial cord formation while SRF from RD eyes had inhibitory effects. SRF from eyes with ROP but not RD robustly induced nitric oxide production in macrophages. Furthermore, fluorescent immunostaining revealed a preponderance of M1 over M2 macrophages in retrolental fibrous membranes from ROP eyes. The cytokine profile and biologic properties of SRF in ROP promote a proangiogenic environment, which supports the maintenance and proliferation of fibrous membranes associated with advanced stages of ROP. In contrast, SRF from RD eyes exhibits a suppressive environment for endothelial cell proliferation and angiogenesis.
Our investigation demonstrates that the microenvironment in advanced ROP eyes is proangiogenic and proinflammatory. These findings suggest that management of advanced ROP should not be limited to the surgical removal of the fibrovascular membranes and antiangiogenic therapy but also directed to anti-inflammatory therapy and to promote M2 activation over M1 activity.</description><subject>Animals</subject><subject>Capillaries - metabolism</subject><subject>Capillaries - pathology</subject><subject>Capillaries - physiopathology</subject><subject>Cell Polarity</subject><subject>Cell Proliferation</subject><subject>Cells, Cultured</subject><subject>Cytokines - metabolism</subject><subject>Humans</subject><subject>Immunohistochemistry</subject><subject>Infant</subject><subject>Inflammation Mediators - metabolism</subject><subject>Macrophages - metabolism</subject><subject>Macrophages - pathology</subject><subject>Mice</subject><subject>Neovascularization, Physiologic</subject><subject>Nitric Oxide - biosynthesis</subject><subject>Nitrites - metabolism</subject><subject>Retinal Detachment - metabolism</subject><subject>Retinopathy of Prematurity - metabolism</subject><subject>Retinopathy of Prematurity - pathology</subject><subject>Retinopathy of Prematurity - physiopathology</subject><subject>Subretinal Fluid - metabolism</subject><issn>1090-0535</issn><issn>1090-0535</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFUU1PwzAMrRCIjcFfQDlyqZQmbdNckNDEx6RJXOBcua3TBXVJadJJE3-eFAYaJ3yxn5_99CyfRPOEShrTjGenR_UsunDujVKWZKk4j2YslXkIOo8-VkZ1I5oaiVXEjdWAXhvoSOjqhmhDoNlBoBviPLRIvnjbg9_sp41-wC34cdA-QBOgBdNq26LRdZh1vTUOCZiG1Nh1E99phQN4bc1ldKagc3h1yIvo9eH-ZfkUr58fV8u7ddwzKX1cC2wKmUtWJCIYL5TMoICsQiaB85wLldQMk1QUMi1y0QhGlaqQV2mmWCVyvohuv3X7sdpiU6PxA3RlP-gtDPvSgi7_MkZvytbuypTmgvEkCNwcBAb7PqLz5Va76R4waEdXJnlB0xBS_D-apQnPJc0m1etjW79-fp7DPwGuB497</recordid><startdate>20140621</startdate><enddate>20140621</enddate><creator>Ma, Jie</creator><creator>Mehta, Manisha</creator><creator>Lam, Godfrey</creator><creator>Cyr, Desireé</creator><creator>Ng, Tat Fong</creator><creator>Hirose, Tatsuo</creator><creator>Tawansy, Khaled A</creator><creator>Taylor, Andrew W</creator><creator>Lashkari, Kameran</creator><general>Molecular Vision</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope><scope>7TK</scope><scope>5PM</scope></search><sort><creationdate>20140621</creationdate><title>Influence of subretinal fluid in advanced stage retinopathy of prematurity on proangiogenic response and cell proliferation</title><author>Ma, Jie ; Mehta, Manisha ; Lam, Godfrey ; Cyr, Desireé ; Ng, Tat Fong ; Hirose, Tatsuo ; Tawansy, Khaled A ; Taylor, Andrew W ; Lashkari, Kameran</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p299t-c7ed896928172498f95a8a5be29a33637f1c2e147894867d720ffbe3b45f2b763</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Animals</topic><topic>Capillaries - metabolism</topic><topic>Capillaries - pathology</topic><topic>Capillaries - physiopathology</topic><topic>Cell Polarity</topic><topic>Cell Proliferation</topic><topic>Cells, Cultured</topic><topic>Cytokines - metabolism</topic><topic>Humans</topic><topic>Immunohistochemistry</topic><topic>Infant</topic><topic>Inflammation Mediators - metabolism</topic><topic>Macrophages - metabolism</topic><topic>Macrophages - pathology</topic><topic>Mice</topic><topic>Neovascularization, Physiologic</topic><topic>Nitric Oxide - biosynthesis</topic><topic>Nitrites - metabolism</topic><topic>Retinal Detachment - metabolism</topic><topic>Retinopathy of Prematurity - metabolism</topic><topic>Retinopathy of Prematurity - pathology</topic><topic>Retinopathy of Prematurity - physiopathology</topic><topic>Subretinal Fluid - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ma, Jie</creatorcontrib><creatorcontrib>Mehta, Manisha</creatorcontrib><creatorcontrib>Lam, Godfrey</creatorcontrib><creatorcontrib>Cyr, Desireé</creatorcontrib><creatorcontrib>Ng, Tat Fong</creatorcontrib><creatorcontrib>Hirose, Tatsuo</creatorcontrib><creatorcontrib>Tawansy, Khaled A</creatorcontrib><creatorcontrib>Taylor, Andrew W</creatorcontrib><creatorcontrib>Lashkari, Kameran</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><collection>Neurosciences Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Molecular vision</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ma, Jie</au><au>Mehta, Manisha</au><au>Lam, Godfrey</au><au>Cyr, Desireé</au><au>Ng, Tat Fong</au><au>Hirose, Tatsuo</au><au>Tawansy, Khaled A</au><au>Taylor, Andrew W</au><au>Lashkari, Kameran</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Influence of subretinal fluid in advanced stage retinopathy of prematurity on proangiogenic response and cell proliferation</atitle><jtitle>Molecular vision</jtitle><addtitle>Mol Vis</addtitle><date>2014-06-21</date><risdate>2014</risdate><volume>20</volume><spage>881</spage><epage>893</epage><pages>881-893</pages><issn>1090-0535</issn><eissn>1090-0535</eissn><abstract>The clinical phenotype of advanced stage retinopathy of prematurity (ROP, stages 4 and 5) cannot be replicated in an animal model. To dissect the molecular events that can lead up to advanced ROP, we examined subretinal fluid (SRF) and surgically dissected retrolental membranes from patients with advanced ROP to evaluate its influences on cell proliferation, angiogenic properties, and macrophage polarity.
We compared our findings to SRF collected from patients with uncomplicated rhegmatogenous retinal detachment (RD) without proliferative vitreoretinopathy and surgically dissected epiretinal membrane from eyes with macular pucker. All subretinal fluid samples were equalized for protein. The angiogenic potential of SRF from ROP eyes was measured using a combination of capillary cord formation in a fibrin clot assay, and its proliferative effect was tested with a DNA synthesis of human retinal microvascular endothelial cells. Findings were compared with SRF collected from participants with uncomplicated rhegmatogenous RD without proliferative vitreoretinopathy. The ability of SRF to induce nitric oxide production was measured in vitro using murine J774A.1 macrophages. Cytokine profiles of SRF from ROP and RD eyes were measured using a multienzyme-linked immunosorbent assay (ELISA). Fluorescent immunohistochemistry of retrolental membranes from ROP was performed to detect the presence of leukocytes and the composition of tissue macrophages using markers for M1 and M2 differentiation.
The cytokine composition in SRF revealed that in ROP, not only were several proangiogenic factors were preferentially elevated but also the profile of proinflammatory factors was also increased compared to the RD eyes. SRF from ROP eyes supported cell proliferation and endothelial cord formation while SRF from RD eyes had inhibitory effects. SRF from eyes with ROP but not RD robustly induced nitric oxide production in macrophages. Furthermore, fluorescent immunostaining revealed a preponderance of M1 over M2 macrophages in retrolental fibrous membranes from ROP eyes. The cytokine profile and biologic properties of SRF in ROP promote a proangiogenic environment, which supports the maintenance and proliferation of fibrous membranes associated with advanced stages of ROP. In contrast, SRF from RD eyes exhibits a suppressive environment for endothelial cell proliferation and angiogenesis.
Our investigation demonstrates that the microenvironment in advanced ROP eyes is proangiogenic and proinflammatory. These findings suggest that management of advanced ROP should not be limited to the surgical removal of the fibrovascular membranes and antiangiogenic therapy but also directed to anti-inflammatory therapy and to promote M2 activation over M1 activity.</abstract><cop>United States</cop><pub>Molecular Vision</pub><pmid>24966660</pmid><tpages>13</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Capillaries - metabolism Capillaries - pathology Capillaries - physiopathology Cell Polarity Cell Proliferation Cells, Cultured Cytokines - metabolism Humans Immunohistochemistry Infant Inflammation Mediators - metabolism Macrophages - metabolism Macrophages - pathology Mice Neovascularization, Physiologic Nitric Oxide - biosynthesis Nitrites - metabolism Retinal Detachment - metabolism Retinopathy of Prematurity - metabolism Retinopathy of Prematurity - pathology Retinopathy of Prematurity - physiopathology Subretinal Fluid - metabolism |
| title | Influence of subretinal fluid in advanced stage retinopathy of prematurity on proangiogenic response and cell proliferation |
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