Effect of Trypan Blue on Descemet Membrane Elasticity

John, Thomas; Patel, Ayush; Vasavada, Abhay R.; Singh, Man; Nath, Vandana; Cheng, Anny M. S.; Sheha, Hosam

doi:10.1097/ico.0000000000000986

Cited by 7 publications

(3 citation statements)

References 16 publications

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“…When we compared the effect of exposure to either TB or MB on the elasticity of the 10–20 μm thick human DM, we observed a significant stiffening of up to 17.7% (median elasticity of MB1 compared to baseline). The above results are in line with recent reports, in which DMs were exposed to TB and assessed by AFM; a 60-second exposure to 0.06% Trypan blue led to increased rigidity of DMs [ 26 – 28 ]. In their study, the authors had highlighted the need to assess the time-dependent effects of vital dyes on human DMs, which we now present.…”

Section: Discussionsupporting

confidence: 92%

“…There is a constant drive to refine the surgical technique DMEK to minimise intraoperative complications and improve transplant attachment to the host posterior corneal stroma [ 1 – 2 ]. Adequate visibility of DM scrolls during surgery is a key factor to achieve unfolding and accurate placement of the transplant [ 25 – 26 ]. For this purpose, vital dyes, in particular, Trypan Blue (TB) and to a lesser extent Brilliant Blue (BB)[ 25 ], have been used.…”

Section: Discussionmentioning

confidence: 99%

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Effect of vital dyes on human corneal endothelium and elasticity of Descemet’s membrane

et al. 2017

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The purpose of this study was to evaluate the effects of vital dyes on human Descemet's membranes (DMs) and endothelia. DMs of 25 human cadaveric corneas with research consent were treated with dyes routinely used in Descemet membrane endothelial keratoplasty (DMEK), 0.05% Trypan blue (TB) or a combination of 0.15% Trypan blue, 0.025% Brilliant blue and 4% Polyethylene glycol (commercial name Membrane Blue Dual; MB). The effects of these two dyes on (i) endothelial cell viability, (ii) DM mechanical properties as assessed by atomic force microscopy, and iii) qualitative DM dye retention were tested for two varying exposure times (one or four minutes). No significant differences in cell toxicity were observed between treatments with TB and MB at the two different exposure times (P = 0.21). Further, both dyes led to a significant increase in DM stiffness: exposure to TB and MB for one minute increased the apparent elastic modulus of the DM by 11.2% (P = 8*10−3) and 17.7%, respectively (P = 4*10−6). A four-minute exposure led to an increase of 8.6% for TB (P = 0.004) and 13.6% for MB (P = 0.03). Finally, at 25 minutes, the dye retention of the DM was considerably better for MB compared to TB. Taken together, a one-minute exposure to MB was found to improve DM visibility compared to TB, with a significant increase in DM stiffness and without detrimental effects on endothelial cell viability. The use of MB could therefore improve (i) visibility of the DM scroll, and (ii) intraoperative unfolding, enhancing the probability of successful DMEK surgery.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Effect of vital dyes on human corneal endothelium and elasticity of Descemet’s membrane

et al. 2017

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“…Using AFM, the Young's modulus, a mechanical property that is indicative of tissue stiffness, can be determined quantitatively [30]. In the eye, AFM has been successfully used to evaluate stiffness across various ocular tissues and in a broad range of mammals, including corneal stiffness in mice [31] and humans [32,33]; trabecular meshwork stiffness in rats [34]; lens stiffness in mice [35] and non-human primates [36]; and the sclera and optic nerve head stiffness in pigs [37,38], mice [39], rats [40], and humans [41].…”

Section: Of 12mentioning

confidence: 99%

Collagen Mimetic Peptides Promote Repair of MMP-1-Damaged Collagen in the Rodent Sclera and Optic Nerve Head

Bou Ghanem,

Koktysh,

Baratta

et al. 2023

IJMS

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The structural and biomechanical properties of collagen-rich ocular tissues, such as the sclera, are integral to ocular function. The degradation of collagen in such tissues is associated with debilitating ophthalmic diseases such as glaucoma and myopia, which often lead to visual impairment. Collagen mimetic peptides (CMPs) have emerged as an effective treatment to repair damaged collagen in tissues of the optic projection, such as the retina and optic nerve. In this study, we used atomic force microscopy (AFM) to assess the potential of CMPs in restoring tissue stiffness in the optic nerve head (ONH), including the peripapillary sclera (PPS) and the glial lamina. Using rat ONH tissue sections, we induced collagen damage with MMP-1, followed by treatment with CMP-3 or vehicle. MMP-1 significantly reduced the Young’s modulus of both the PPS and the glial lamina, indicating tissue softening. Subsequent CMP-3 treatment partially restored tissue stiffness in both the PPS and the glial lamina. Immunohistochemical analyses revealed reduced collagen fragmentation after MMP-1 digestion in CMP-3-treated tissues compared to vehicle controls. In summary, these results demonstrate the potential of CMPs to restore collagen stiffness and structure in ONH tissues following enzymatic damage. CMPs may offer a promising therapeutic avenue for preserving vision in ocular disorders involving collagen remodeling and degradation.

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Trephine- and dye-free technique for eye bank preparation of pre-stripped Descemet membrane endothelial keratoplasty tissue

2019

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Effect of Trypan Blue on Descemet Membrane Elasticity

Abstract: Our results suggest that trypan blue may decrease DM elasticity and consequently increase its stiffness. This may influence the graft adherence when used for endothelial keratoplasty.

Cited by 7 publications

References 16 publications

Effect of vital dyes on human corneal endothelium and elasticity of Descemet’s membrane

Effect of vital dyes on human corneal endothelium and elasticity of Descemet’s membrane

Collagen Mimetic Peptides Promote Repair of MMP-1-Damaged Collagen in the Rodent Sclera and Optic Nerve Head

Trephine- and dye-free technique for eye bank preparation of pre-stripped Descemet membrane endothelial keratoplasty tissue

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