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Molecular mechanism of ocular surface damage: application to an in vitro dry eye model on human corneal epithelium.

Meloni M, De Servi B, Marasco D, Del Prete S - Mol. Vis. (2011)

Bottom Line: The effects of different commercially available tear substitutes on the induced dry eye condition were tested.The transcriptional study of selected biomarkers showed an increase in MUC4, MMP9, TNF-α, and hBD-2 (DEFB2) gene expression.By using a dynamic approach, we were able to define a biomarker gene signature of dry eye-induced effects that could be predictive of corneal damage in vivo and to discriminate the efficacy among different commercial artificial tears.

View Article: PubMed Central - PubMed

ABSTRACT

Purpose: The present study was concerned with the development of a new experimental model of dry eye using human reconstructed in vitro corneal epithelium (HCE). The model is based on the use of adapted culture conditions that induce relevant modifications at the cellular and molecular level thus mimicking dry eye.

Methods: The HCE model was maintained in a controlled environmental setting (relative humidity <40% and 40 °C temperature) for 24 h and up to 72 h to induce dry eye. The evolution of the dry eye condition was assessed by histology, immunohistochemistry staining, scanning electron microscopy, and gene expression by using TaqMan gene assay technology (mucin-4 [MUC4], matrix metallopeptidase-9 [MMP9], tumor necrosis factor-α [TNF-α], and defensin β-2 [DEFB2). The effects of different commercially available tear substitutes on the induced dry eye condition were tested.

Results: This in vitro dry eye HCE model, that was well established within 24 h, has the characteristic features of a dry eye epithelium and could be satisfactorily used for preliminary assessment of the protective activity of some artificial tears. The transcriptional study of selected biomarkers showed an increase in MUC4, MMP9, TNF-α, and hBD-2 (DEFB2) gene expression.

Conclusions: By using a dynamic approach, we were able to define a biomarker gene signature of dry eye-induced effects that could be predictive of corneal damage in vivo and to discriminate the efficacy among different commercial artificial tears.

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Related in: MedlinePlus

Reduction of AlamarBlue (%) resulting in % viability in EDEV-HCE compared to the CONTROL-HCE at 24 h, 48 h, and 72 h. The vertical lines represent the standard deviation (SD) between two different tissues. HCE: human corneal epithelium.
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f2: Reduction of AlamarBlue (%) resulting in % viability in EDEV-HCE compared to the CONTROL-HCE at 24 h, 48 h, and 72 h. The vertical lines represent the standard deviation (SD) between two different tissues. HCE: human corneal epithelium.

Mentions: The viability of HCE tissue was evaluated by AlamarBlue (AB) at 24, 48, and 72 h after dry eye induction and compared with control HCE. The test has been performed on duplicate cultures. As shown in Figure 2, after the maximum exposure time (72 h), the survival of HCE in EDEV conditions was still quite good (70% cellular viability) compared with the CONTROL-HCE (cellular viability 100%).


Molecular mechanism of ocular surface damage: application to an in vitro dry eye model on human corneal epithelium.

Meloni M, De Servi B, Marasco D, Del Prete S - Mol. Vis. (2011)

Reduction of AlamarBlue (%) resulting in % viability in EDEV-HCE compared to the CONTROL-HCE at 24 h, 48 h, and 72 h. The vertical lines represent the standard deviation (SD) between two different tissues. HCE: human corneal epithelium.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3021568&req=5

f2: Reduction of AlamarBlue (%) resulting in % viability in EDEV-HCE compared to the CONTROL-HCE at 24 h, 48 h, and 72 h. The vertical lines represent the standard deviation (SD) between two different tissues. HCE: human corneal epithelium.
Mentions: The viability of HCE tissue was evaluated by AlamarBlue (AB) at 24, 48, and 72 h after dry eye induction and compared with control HCE. The test has been performed on duplicate cultures. As shown in Figure 2, after the maximum exposure time (72 h), the survival of HCE in EDEV conditions was still quite good (70% cellular viability) compared with the CONTROL-HCE (cellular viability 100%).

Bottom Line: The effects of different commercially available tear substitutes on the induced dry eye condition were tested.The transcriptional study of selected biomarkers showed an increase in MUC4, MMP9, TNF-α, and hBD-2 (DEFB2) gene expression.By using a dynamic approach, we were able to define a biomarker gene signature of dry eye-induced effects that could be predictive of corneal damage in vivo and to discriminate the efficacy among different commercial artificial tears.

View Article: PubMed Central - PubMed

ABSTRACT

Purpose: The present study was concerned with the development of a new experimental model of dry eye using human reconstructed in vitro corneal epithelium (HCE). The model is based on the use of adapted culture conditions that induce relevant modifications at the cellular and molecular level thus mimicking dry eye.

Methods: The HCE model was maintained in a controlled environmental setting (relative humidity <40% and 40 °C temperature) for 24 h and up to 72 h to induce dry eye. The evolution of the dry eye condition was assessed by histology, immunohistochemistry staining, scanning electron microscopy, and gene expression by using TaqMan gene assay technology (mucin-4 [MUC4], matrix metallopeptidase-9 [MMP9], tumor necrosis factor-α [TNF-α], and defensin β-2 [DEFB2). The effects of different commercially available tear substitutes on the induced dry eye condition were tested.

Results: This in vitro dry eye HCE model, that was well established within 24 h, has the characteristic features of a dry eye epithelium and could be satisfactorily used for preliminary assessment of the protective activity of some artificial tears. The transcriptional study of selected biomarkers showed an increase in MUC4, MMP9, TNF-α, and hBD-2 (DEFB2) gene expression.

Conclusions: By using a dynamic approach, we were able to define a biomarker gene signature of dry eye-induced effects that could be predictive of corneal damage in vivo and to discriminate the efficacy among different commercial artificial tears.

Show MeSH
Related in: MedlinePlus