June 2021
Volume 62, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2021
Human Trabecular Meshwork Cells Appear Healthier & More Viable in Contact with Titanium Stents Than in Contact with Nickel-Titanium Stents
Author Affiliations & Notes
  • Hal Heitzmann
    Glaukos Corporation, San Clemente, California, United States
  • Barbara M Wirostko
    The University of Utah School of Medicine, Salt Lake City, Utah, United States
  • Feryan Ahmed
    Glauconix Biosciences, Albany, New York, United States
  • Andrea Unser
    Glauconix Biosciences, Albany, New York, United States
  • Vandhana Chari
    Glauconix Biosciences, Albany, New York, United States
  • Karen Yud Torrejon
    Glauconix Biosciences, Albany, New York, United States
  • Footnotes
    Commercial Relationships   Hal Heitzmann, Glaukos (C); Barbara Wirostko, Glauconix Biosciences (S); Feryan Ahmed, Glauconix Biosciences (E), Glaukos (F); Andrea Unser, Glauconix Biosciences (E), Glaukos (F); Vandhana Chari, Glauconix Biosciences (E), Glaukos (F); Karen Torrejon, Glauconix Biosciences (E), Glaukos (F)
  • Footnotes
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Investigative Ophthalmology & Visual Science June 2021, Vol.62, 3421. doi:
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    • Get Citation

      Hal Heitzmann, Barbara M Wirostko, Feryan Ahmed, Andrea Unser, Vandhana Chari, Karen Yud Torrejon; Human Trabecular Meshwork Cells Appear Healthier & More Viable in Contact with Titanium Stents Than in Contact with Nickel-Titanium Stents. Invest. Ophthalmol. Vis. Sci. 2021;62(8):3421.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Trabecular bypass stents (TBS) are implanted through the human trabecular meshwork (HTM) to improve physiologic outflow and lower intraocular pressure in open-angle glaucoma. Given the widespread use of TBS, improved understanding of their biocompatibility is imperative. Of the two commercially available TBS, one is comprised of titanium & the other of nickel-titanium (nitinol). This study tested the hypothesis that there would be differences in the viability of primary HTM cells in culture when contacted with one versus the other of these devices.

Methods : HTM cells were grown on glass or gelatin-coated glass substrates & then placed into contact with sterile stents. Cell morphology in the vicinity of the stents was monitored every day for 4 days via optical microscopy, and live-dead fluorescence staining was performed to assess cell viability. The corrected total fluorescence of micrographs imaged under identical conditions (N≥21 per timepoint) was quantitated w/ ImageJ software. Data were analyzed using two-way ANOVA & compared to stent-free samples w/ Tukey’s multiple comparison test.

Results : Cells cultured in contact with nitinol stents for 48, 72 & 96 hours demonstrated progressive cell necrosis, clumping, and in some cases, complete cell layer detachment. However, cells in contact with titanium stents remained attached to the substrate & showed little or no necrosis. Dead-cell staining intensity was greater after nitinol vs. titanium stent contact (p<0.0001, N=21/stent) across all timepoints examined.

Conclusions : HTM cells remained viable & morphologically intact when in contact with titanium stents, whereas contact with nitinol stents led to necrosis & morphological degradation. These differences were statistically significant & visually apparent.

This is a 2021 ARVO Annual Meeting abstract.

 

Live/Dead Assay: Fluorescence micrographs of HTM cells (left panel) in contact with nitinol-based stent (middle panel) or titanium stent (right panel) for 96 hours. Green(calcein AM)=alive, red(ethidium homodimer)=dead, blue (DAPI)=nuclei. The tip-end of the nitinol-based stent & the complete titanium stent are outlined in white. All scale bars=200μM (20x).

Live/Dead Assay: Fluorescence micrographs of HTM cells (left panel) in contact with nitinol-based stent (middle panel) or titanium stent (right panel) for 96 hours. Green(calcein AM)=alive, red(ethidium homodimer)=dead, blue (DAPI)=nuclei. The tip-end of the nitinol-based stent & the complete titanium stent are outlined in white. All scale bars=200μM (20x).

 

Fluorescence Intensity of Dead Cells. The difference between conditions was analyzed using one-way ANOVA followed by Bonferroni post-tests. **p<0.005, ***<0.001, ****p<0.0001. N=21.

Fluorescence Intensity of Dead Cells. The difference between conditions was analyzed using one-way ANOVA followed by Bonferroni post-tests. **p<0.005, ***<0.001, ****p<0.0001. N=21.

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