Area and depth of surfactant-induced corneal injury correlates with cell death

T. F. Li, J. K. Maurer, R. D. Parker, Walter M Petroll, Harrison D Cavanagh, J. V. Jester

Research output: Contribution to journalArticle

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Abstract

Purpose: Our previous in vivo confocal microscopic (CM) studies identified quantifiable differences in the corneal epitl elium and stroma for surfactants of different irritancy. This study characterized tbe relationship of area and depth of the initial corneal changes measured by in vivo CM to cell death as measured by ex vivo, Live/Dead Assay (L/D, Molecular Probes) Methods; For 4 groups of rabbits (8 animals each), 10 pi of a surfactant known to produce slight, mild, moderate, or severe irritation was placed on the central cornea of one eye; 2 untreated rabbits served as controls. In vivo CM measurement of groip total mean epithelial thickness and depth of keratocyte loss of 4 corneal regions weie made in 4 rabbits of each group and 2 controls at 3 hr and in the remaining rabbits at 3 hr and 1 day. Corneas were then removed, regions excised and placed in culture media containing 2 pM calcein AM and 4 uM ethidium homodimer. Using las>:r scanning CM, number of dead epithelial or stromal cells in a 300x300x170 (W> (x>z) volume of the cornea was determined. Results: (All p<0.05) By in vivo CM, the slight irritant resulted in epithelial thinning at 3 hr (41.0 ±3.7 pm vs. 45.6 ±2.5 urn fo control, n=4 and 2), while on day 1 mild, moderate and severe irritants caused complete epithelial loss and keratocyte disappearance to a depth of 26.4 ±15.5 u.m, 44.1 ±21.6 urn and 796.0 ±172.3 urn, respectively. At 3 hr, L/D assay detected more dead epithelial cells in slightly irritated eyes (59.8 ±19.5 vs. 14.9 11,6 cells for control); significantly correlating with the in vivo CM measurement of epithelial thinning (r=0.89). At day 1, mild and moderate irritants showed increasing stromal cell death from 9.8 ±16.2 to 36.4 ±31.2 cells which significantly correlated with in vivo CM keratocyte loss (r=0.78). No surviving keratocytes were detected in severely irritated eyes. Conclusions The data supports our hypothesis that differences in surfactant induced ocular irritation are directly related to area and depth of acute corneal injury.

Original languageEnglish (US)
JournalInvestigative Ophthalmology and Visual Science
Volume38
Issue number4
StatePublished - 1997

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Surface-Active Agents
Irritants
Cell Death
Cornea
Rabbits
Stromal Cells
Epithelial Cells
Molecular Probes
Culture Media
Control Groups
Corneal Injuries

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Area and depth of surfactant-induced corneal injury correlates with cell death. / Li, T. F.; Maurer, J. K.; Parker, R. D.; Petroll, Walter M; Cavanagh, Harrison D; Jester, J. V.

In: Investigative Ophthalmology and Visual Science, Vol. 38, No. 4, 1997.

Research output: Contribution to journalArticle

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abstract = "Purpose: Our previous in vivo confocal microscopic (CM) studies identified quantifiable differences in the corneal epitl elium and stroma for surfactants of different irritancy. This study characterized tbe relationship of area and depth of the initial corneal changes measured by in vivo CM to cell death as measured by ex vivo, Live/Dead Assay (L/D, Molecular Probes) Methods; For 4 groups of rabbits (8 animals each), 10 pi of a surfactant known to produce slight, mild, moderate, or severe irritation was placed on the central cornea of one eye; 2 untreated rabbits served as controls. In vivo CM measurement of groip total mean epithelial thickness and depth of keratocyte loss of 4 corneal regions weie made in 4 rabbits of each group and 2 controls at 3 hr and in the remaining rabbits at 3 hr and 1 day. Corneas were then removed, regions excised and placed in culture media containing 2 pM calcein AM and 4 uM ethidium homodimer. Using las>:r scanning CM, number of dead epithelial or stromal cells in a 300x300x170 (W> (x>z) volume of the cornea was determined. Results: (All p<0.05) By in vivo CM, the slight irritant resulted in epithelial thinning at 3 hr (41.0 ±3.7 pm vs. 45.6 ±2.5 urn fo control, n=4 and 2), while on day 1 mild, moderate and severe irritants caused complete epithelial loss and keratocyte disappearance to a depth of 26.4 ±15.5 u.m, 44.1 ±21.6 urn and 796.0 ±172.3 urn, respectively. At 3 hr, L/D assay detected more dead epithelial cells in slightly irritated eyes (59.8 ±19.5 vs. 14.9 11,6 cells for control); significantly correlating with the in vivo CM measurement of epithelial thinning (r=0.89). At day 1, mild and moderate irritants showed increasing stromal cell death from 9.8 ±16.2 to 36.4 ±31.2 cells which significantly correlated with in vivo CM keratocyte loss (r=0.78). No surviving keratocytes were detected in severely irritated eyes. Conclusions The data supports our hypothesis that differences in surfactant induced ocular irritation are directly related to area and depth of acute corneal injury.",
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T1 - Area and depth of surfactant-induced corneal injury correlates with cell death

AU - Li, T. F.

AU - Maurer, J. K.

AU - Parker, R. D.

AU - Petroll, Walter M

AU - Cavanagh, Harrison D

AU - Jester, J. V.

PY - 1997

Y1 - 1997

N2 - Purpose: Our previous in vivo confocal microscopic (CM) studies identified quantifiable differences in the corneal epitl elium and stroma for surfactants of different irritancy. This study characterized tbe relationship of area and depth of the initial corneal changes measured by in vivo CM to cell death as measured by ex vivo, Live/Dead Assay (L/D, Molecular Probes) Methods; For 4 groups of rabbits (8 animals each), 10 pi of a surfactant known to produce slight, mild, moderate, or severe irritation was placed on the central cornea of one eye; 2 untreated rabbits served as controls. In vivo CM measurement of groip total mean epithelial thickness and depth of keratocyte loss of 4 corneal regions weie made in 4 rabbits of each group and 2 controls at 3 hr and in the remaining rabbits at 3 hr and 1 day. Corneas were then removed, regions excised and placed in culture media containing 2 pM calcein AM and 4 uM ethidium homodimer. Using las>:r scanning CM, number of dead epithelial or stromal cells in a 300x300x170 (W> (x>z) volume of the cornea was determined. Results: (All p<0.05) By in vivo CM, the slight irritant resulted in epithelial thinning at 3 hr (41.0 ±3.7 pm vs. 45.6 ±2.5 urn fo control, n=4 and 2), while on day 1 mild, moderate and severe irritants caused complete epithelial loss and keratocyte disappearance to a depth of 26.4 ±15.5 u.m, 44.1 ±21.6 urn and 796.0 ±172.3 urn, respectively. At 3 hr, L/D assay detected more dead epithelial cells in slightly irritated eyes (59.8 ±19.5 vs. 14.9 11,6 cells for control); significantly correlating with the in vivo CM measurement of epithelial thinning (r=0.89). At day 1, mild and moderate irritants showed increasing stromal cell death from 9.8 ±16.2 to 36.4 ±31.2 cells which significantly correlated with in vivo CM keratocyte loss (r=0.78). No surviving keratocytes were detected in severely irritated eyes. Conclusions The data supports our hypothesis that differences in surfactant induced ocular irritation are directly related to area and depth of acute corneal injury.

AB - Purpose: Our previous in vivo confocal microscopic (CM) studies identified quantifiable differences in the corneal epitl elium and stroma for surfactants of different irritancy. This study characterized tbe relationship of area and depth of the initial corneal changes measured by in vivo CM to cell death as measured by ex vivo, Live/Dead Assay (L/D, Molecular Probes) Methods; For 4 groups of rabbits (8 animals each), 10 pi of a surfactant known to produce slight, mild, moderate, or severe irritation was placed on the central cornea of one eye; 2 untreated rabbits served as controls. In vivo CM measurement of groip total mean epithelial thickness and depth of keratocyte loss of 4 corneal regions weie made in 4 rabbits of each group and 2 controls at 3 hr and in the remaining rabbits at 3 hr and 1 day. Corneas were then removed, regions excised and placed in culture media containing 2 pM calcein AM and 4 uM ethidium homodimer. Using las>:r scanning CM, number of dead epithelial or stromal cells in a 300x300x170 (W> (x>z) volume of the cornea was determined. Results: (All p<0.05) By in vivo CM, the slight irritant resulted in epithelial thinning at 3 hr (41.0 ±3.7 pm vs. 45.6 ±2.5 urn fo control, n=4 and 2), while on day 1 mild, moderate and severe irritants caused complete epithelial loss and keratocyte disappearance to a depth of 26.4 ±15.5 u.m, 44.1 ±21.6 urn and 796.0 ±172.3 urn, respectively. At 3 hr, L/D assay detected more dead epithelial cells in slightly irritated eyes (59.8 ±19.5 vs. 14.9 11,6 cells for control); significantly correlating with the in vivo CM measurement of epithelial thinning (r=0.89). At day 1, mild and moderate irritants showed increasing stromal cell death from 9.8 ±16.2 to 36.4 ±31.2 cells which significantly correlated with in vivo CM keratocyte loss (r=0.78). No surviving keratocytes were detected in severely irritated eyes. Conclusions The data supports our hypothesis that differences in surfactant induced ocular irritation are directly related to area and depth of acute corneal injury.

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