Abstract Title

Discovery of new agents to inhibit scarring and improve the success of glaucoma surgery

Presenter Name

Callie Hoelscher

RAD Assignment Number

901

Abstract

Purpose:

Glaucoma is a common ophthalmological condition related to elevated intraocular pressure (IOP). Glaucoma can sometimes be treated surgically with a procedure called a trabeculectomy that allows for increased drainage of the aqueous humor of the eye to reduce the IOP. After this surgery, many patients develop a scar at the surgical site that can reverse the benefits of the surgery. This study aims to compare multiple inhibitors of the TGFβ2-TLR4 signaling pathway against the current standard treatment for scar inhibition to determine if they could be more effective. Less scarring would lead to better maintainence of lower IOP and more successful glaucoma surgeries.

Methods:

Primary human conjunctiva/tenon fibroblast cell strains were treated with Mitomycin C (MMC), the standard treatment to prevent scarring, TAK-242 (a selective inhibitor of TLR4), or NBP2-29328 (a selective inhibitor of MyD88). NBP2-29328, TAK-242, and untreated cells were incubated with 0.5% FBS for 24 hours, and MMC cells were incubated for 5 minutes. Next, a wound scratch assay was performed. Each well was photographed with a Keyence imager at 0, 6, 24, 48, 72, and 96 hours post-scratch. Using ImageJ software, the 0-hour images were analyzed to get an area value and a ROI (Region of Interest) outline of the 0-hour scratch. The later images were analyzed to get values for the number of cells present and their total area covered. The four treatments were compared for efficacy of inhibition of cell migration and proliferation. This protocol has been performed in two studies.

Results:

There were significant differences between treatments at 72 and 96 hours. In the first study, inhibition from the MMC treatment was significant compared to control (p

Conclusions:

The data from the studies suggest that these treatments may be useful for scar inhibition after glaucoma trabeculectomy surgery.

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Research Area

Eye/Vision

Presentation Type

Poster

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Discovery of new agents to inhibit scarring and improve the success of glaucoma surgery

Purpose:

Glaucoma is a common ophthalmological condition related to elevated intraocular pressure (IOP). Glaucoma can sometimes be treated surgically with a procedure called a trabeculectomy that allows for increased drainage of the aqueous humor of the eye to reduce the IOP. After this surgery, many patients develop a scar at the surgical site that can reverse the benefits of the surgery. This study aims to compare multiple inhibitors of the TGFβ2-TLR4 signaling pathway against the current standard treatment for scar inhibition to determine if they could be more effective. Less scarring would lead to better maintainence of lower IOP and more successful glaucoma surgeries.

Methods:

Primary human conjunctiva/tenon fibroblast cell strains were treated with Mitomycin C (MMC), the standard treatment to prevent scarring, TAK-242 (a selective inhibitor of TLR4), or NBP2-29328 (a selective inhibitor of MyD88). NBP2-29328, TAK-242, and untreated cells were incubated with 0.5% FBS for 24 hours, and MMC cells were incubated for 5 minutes. Next, a wound scratch assay was performed. Each well was photographed with a Keyence imager at 0, 6, 24, 48, 72, and 96 hours post-scratch. Using ImageJ software, the 0-hour images were analyzed to get an area value and a ROI (Region of Interest) outline of the 0-hour scratch. The later images were analyzed to get values for the number of cells present and their total area covered. The four treatments were compared for efficacy of inhibition of cell migration and proliferation. This protocol has been performed in two studies.

Results:

There were significant differences between treatments at 72 and 96 hours. In the first study, inhibition from the MMC treatment was significant compared to control (p

Conclusions:

The data from the studies suggest that these treatments may be useful for scar inhibition after glaucoma trabeculectomy surgery.