Semaphorin 7A promotes angiogenesis in an experimental corneal neovascularization model

Ramon C. Ghanem, Kyu Yeon Han, Juan Rojas, Okan Ozturk, David J. Kim, Sandeep Jain, Jin Hong Chang, Dimitri T. Azar

Research output: Contribution to journalArticle

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Abstract

Purpose: To characterize the involvement of Semaphorin 7A (Sema7a) in corneal neovascularization (NV). Methods: We generated anti-Sema7A antibodies to detect protein expression in corneal fibroblasts. Corneal fibroblast cells were cultured, stimulated with basic fibroblast growth factor (bFGF or FGF-2), immunostained with anti-Sema7A antibodies, and visualized by confocal microscopy. bFGF pellets were implanted in mouse corneal micropockets for 310 days, and corneal sections were immunostained with anti-Sema7A antibodies. Mouse corneas were injected with a Sema7A expression vector or a control vector for 3, 7, and 10 days. Mouse corneas were imaged by slit lamp microscopy, and areas of corneal NV were calculated using the ImageJ program. Mouse corneal sections were also immunostained with anti-macrophage marker (F4/80) and anti-vascular endothelial growth factor (VEGF)-A antibodies. Results: Our data showed enhanced Sema7A expression levels in bFGF-stimulated cultured corneal fibroblasts. bFGF corneal implantation also demonstrated enhanced Sema7A expression. Corneas injected with a Sema7A expression vector showed evidence of significant corneal NV compared to controls on day 10 (1.8mm 2 vs. 0.11mm 2; p<0.02). Additionally, immunolocalization of Sema7A expression vector-injected corneas (at day 7) revealed macrophage recruitment and enhanced VEGF-A levels. Conclusions: We demonstrated that Sema7A was expressed in vascularized corneas and showed pro-angiogenic properties in our corneal model. Understanding the mechanism of Sema7A in angiogenesis may provide a therapeutic target for the treatment of corneal angiogenesis-related disorders. © 2011 Informa Healthcare USA, Inc.
Original languageAmerican English
Pages (from-to)989-996
Number of pages889
JournalCurrent Eye Research
DOIs
StatePublished - 1 Nov 2011
Externally publishedYes

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Semaphorins
Corneal Neovascularization
Cornea
Anti-Idiotypic Antibodies
Fibroblasts
Fibroblast Growth Factor 2
Vascular Endothelial Growth Factor A
Macrophages
Confocal Microscopy
Cultured Cells
Delivery of Health Care
Antibodies
Proteins

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Ghanem, Ramon C. ; Han, Kyu Yeon ; Rojas, Juan ; Ozturk, Okan ; Kim, David J. ; Jain, Sandeep ; Chang, Jin Hong ; Azar, Dimitri T. / Semaphorin 7A promotes angiogenesis in an experimental corneal neovascularization model. In: Current Eye Research. 2011 ; pp. 989-996.
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abstract = "Purpose: To characterize the involvement of Semaphorin 7A (Sema7a) in corneal neovascularization (NV). Methods: We generated anti-Sema7A antibodies to detect protein expression in corneal fibroblasts. Corneal fibroblast cells were cultured, stimulated with basic fibroblast growth factor (bFGF or FGF-2), immunostained with anti-Sema7A antibodies, and visualized by confocal microscopy. bFGF pellets were implanted in mouse corneal micropockets for 310 days, and corneal sections were immunostained with anti-Sema7A antibodies. Mouse corneas were injected with a Sema7A expression vector or a control vector for 3, 7, and 10 days. Mouse corneas were imaged by slit lamp microscopy, and areas of corneal NV were calculated using the ImageJ program. Mouse corneal sections were also immunostained with anti-macrophage marker (F4/80) and anti-vascular endothelial growth factor (VEGF)-A antibodies. Results: Our data showed enhanced Sema7A expression levels in bFGF-stimulated cultured corneal fibroblasts. bFGF corneal implantation also demonstrated enhanced Sema7A expression. Corneas injected with a Sema7A expression vector showed evidence of significant corneal NV compared to controls on day 10 (1.8mm 2 vs. 0.11mm 2; p<0.02). Additionally, immunolocalization of Sema7A expression vector-injected corneas (at day 7) revealed macrophage recruitment and enhanced VEGF-A levels. Conclusions: We demonstrated that Sema7A was expressed in vascularized corneas and showed pro-angiogenic properties in our corneal model. Understanding the mechanism of Sema7A in angiogenesis may provide a therapeutic target for the treatment of corneal angiogenesis-related disorders. {\circledC} 2011 Informa Healthcare USA, Inc.",
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Semaphorin 7A promotes angiogenesis in an experimental corneal neovascularization model. / Ghanem, Ramon C.; Han, Kyu Yeon; Rojas, Juan; Ozturk, Okan; Kim, David J.; Jain, Sandeep; Chang, Jin Hong; Azar, Dimitri T.

In: Current Eye Research, 01.11.2011, p. 989-996.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Semaphorin 7A promotes angiogenesis in an experimental corneal neovascularization model

AU - Ghanem, Ramon C.

AU - Han, Kyu Yeon

AU - Rojas, Juan

AU - Ozturk, Okan

AU - Kim, David J.

AU - Jain, Sandeep

AU - Chang, Jin Hong

AU - Azar, Dimitri T.

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N2 - Purpose: To characterize the involvement of Semaphorin 7A (Sema7a) in corneal neovascularization (NV). Methods: We generated anti-Sema7A antibodies to detect protein expression in corneal fibroblasts. Corneal fibroblast cells were cultured, stimulated with basic fibroblast growth factor (bFGF or FGF-2), immunostained with anti-Sema7A antibodies, and visualized by confocal microscopy. bFGF pellets were implanted in mouse corneal micropockets for 310 days, and corneal sections were immunostained with anti-Sema7A antibodies. Mouse corneas were injected with a Sema7A expression vector or a control vector for 3, 7, and 10 days. Mouse corneas were imaged by slit lamp microscopy, and areas of corneal NV were calculated using the ImageJ program. Mouse corneal sections were also immunostained with anti-macrophage marker (F4/80) and anti-vascular endothelial growth factor (VEGF)-A antibodies. Results: Our data showed enhanced Sema7A expression levels in bFGF-stimulated cultured corneal fibroblasts. bFGF corneal implantation also demonstrated enhanced Sema7A expression. Corneas injected with a Sema7A expression vector showed evidence of significant corneal NV compared to controls on day 10 (1.8mm 2 vs. 0.11mm 2; p<0.02). Additionally, immunolocalization of Sema7A expression vector-injected corneas (at day 7) revealed macrophage recruitment and enhanced VEGF-A levels. Conclusions: We demonstrated that Sema7A was expressed in vascularized corneas and showed pro-angiogenic properties in our corneal model. Understanding the mechanism of Sema7A in angiogenesis may provide a therapeutic target for the treatment of corneal angiogenesis-related disorders. © 2011 Informa Healthcare USA, Inc.

AB - Purpose: To characterize the involvement of Semaphorin 7A (Sema7a) in corneal neovascularization (NV). Methods: We generated anti-Sema7A antibodies to detect protein expression in corneal fibroblasts. Corneal fibroblast cells were cultured, stimulated with basic fibroblast growth factor (bFGF or FGF-2), immunostained with anti-Sema7A antibodies, and visualized by confocal microscopy. bFGF pellets were implanted in mouse corneal micropockets for 310 days, and corneal sections were immunostained with anti-Sema7A antibodies. Mouse corneas were injected with a Sema7A expression vector or a control vector for 3, 7, and 10 days. Mouse corneas were imaged by slit lamp microscopy, and areas of corneal NV were calculated using the ImageJ program. Mouse corneal sections were also immunostained with anti-macrophage marker (F4/80) and anti-vascular endothelial growth factor (VEGF)-A antibodies. Results: Our data showed enhanced Sema7A expression levels in bFGF-stimulated cultured corneal fibroblasts. bFGF corneal implantation also demonstrated enhanced Sema7A expression. Corneas injected with a Sema7A expression vector showed evidence of significant corneal NV compared to controls on day 10 (1.8mm 2 vs. 0.11mm 2; p<0.02). Additionally, immunolocalization of Sema7A expression vector-injected corneas (at day 7) revealed macrophage recruitment and enhanced VEGF-A levels. Conclusions: We demonstrated that Sema7A was expressed in vascularized corneas and showed pro-angiogenic properties in our corneal model. Understanding the mechanism of Sema7A in angiogenesis may provide a therapeutic target for the treatment of corneal angiogenesis-related disorders. © 2011 Informa Healthcare USA, Inc.

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