Effect of Amniotic Membrane on Epithelial Thickness and Formation of Hemidesmosomes after Corneal Stromal Wound

Yong Jun Yun; Joo Young Kwag; Si Hwan Choi

doi:10.3341/jkos.2011.52.5.589

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Yun, Kwag, and Choi: Effect of Amniotic Membrane on Epithelial Thickness and Formation of Hemidesmosomes after Corneal Stromal Wound

Original Article

J Korean Ophthalmol Soc 2011;52(5):589-596.

Published online: 3 August 2011

DOI: https://doi.org/10.3341/jkos.2011.52.5.589

Effect of Amniotic Membrane on Epithelial Thickness and Formation of Hemidesmosomes after Corneal Stromal Wound

Yong Jun Yun, MD, Joo Young Kwag, MD, Si Hwan Choi, MD, PhD

Department of Ophthalmology, Chungnam National University College of Medicine, Daejeon, Korea

Address reprint requests to Si Hwan Choi, MD, PhD Department of Ophthalmology, Chungnam National University Hospital #640 Daesa-dong, Jung-gu, Daejeon 301-747, Korea Tel: 82-42-280-7609, Fax: 82-42-255-3745, E-mail: shchoi@cnu.ac.kr

Received 24 August 20096 December 2010 Accepted 8 March 2011

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Purpose

To investigate the effects of an amniotic membrane patch on corneal epithelial thickness and formation of hemidesmosomes during corneal stromal wound healing.

Methods

A stromal wound 9 mm in diameter and 130 μm in depth was created on rabbit cornea using a microkeratome. The changes in corneal epithelial thickness and hemidesmosome formations were compared between the amniotic membrane, contact lens, and control groups. Changes in the corneal epithelium were examined using H&E staining and hemidesmosome formation was examined using an electron microscope at 2 and 4 weeks after flap removal.

Results

Two weeks after treatment, the corneal epithelial thickness was 95.3 ± 6.3 μm in the amniotic membrane group being significantly thicker than 76.4 ± 5.1 μm in the contact lens group and 68.3 ± 6.1 μm in the control group. Furthermore, more hemidesmosome formations were observed in the amniotic membrane group compared to the other 2 groups. However, there were no significant differences in corneal epithelial thickness or hemidesmosome formation among the 3 groups at week 4.

Conclusions

The amniotic membrane group showed a thicker corneal epithelium and more hemidesmosome formation than the other 2 groups 2 weeks after flap removal. Thus, the use of an amniotic membrane patch appears to be effective in the early stages of corneal stromal wound healing.

Keywords: Amniotic membrane, Contact lens, Corneal epithelial thickness, Hemidesmosome

References

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Figure 1.

Photographs showing the making and removal of the corneal wound (diameter 9 mm, depth 130 μm). (A) Creating the corneal wound flap with Microkeratome (BDK3000, Becton & Dickinson, Franklin Lakes, NJ, USA). (B) Corneal flap. (C) Lifting of the corneal wound flap and cutting the hinge of the corneal flap using a blade. (D) Fluorescein dye staining after corneal flap removal.

Figure 2.

Photograph of the rabbit cornea after amniotic membrane patch graft. Arrowheads indicate the original amniotic membrane patch margin (diameter 14 mm) and arrows indicate 10-0 nylon sutures.

Figure 3.

Light microscopy of the central cornea at 2 weeks. (A) Normal cornea. (B) Amniotic membrane group. (C) Contact lens group. (D) Control group. Corneal epithelial thickness in the amniotic membrane group is thicker than the other groups (Hematoxylin-Eosin stain, original magnification ×100).

Figure 4.

Light microscopy of central cornea at 4 weeks. (A) Normal cornea. (B) Amniotic membrane group. (C) Contact lens group. (D) Control group. Corneal epithelial thickness is similar in all groups (Hematoxylin-Eosin stain, original magnification ×100).

Figure 5.

Comparison of central corneal epithelial thickness among the 3 groups at 2 weeks and 4 weeks. In all groups, the central corneal epithelial thickness was significantly thicker at 2 weeks than at 4 weeks. ^* Corneal epithelial thickness in the AM group is significantly thicker than the CL and control group at 2 weeks. ^† Corneal epithelial thickness in the CL group is significantly thicker than the control group. AM = amniotic membrane; CL = contact lens.

Figure 6.

Transmission electron photomicrographs (original magnification ×45,000) of the central cornea at 2 weeks. (A) Morphologically established hemidesmosome is found in the amniotic membrane group. (B) Morphologically identifiable hemidesmosomes appear in the contact lens group. (C) Hyperactive formation of adhesion complex in the control group.

Figure 7.

Transmission electron photomicrographs (original magnification ×45,000) of the central cornea at 4 weeks. (A) Amniotic membrane group. (B) Contact lens group. (C) Control group. Morphologically established hemidesmosomes (arrowheads) are detected in all groups. There are no significant differences in adhesion complex formation among the 3 groups.

Table 1.

The comparison of corneal epithelial thickness among the 3 groups 2 mm from the limbus (Corneal flap did not involve this area)

Groups	n	Epithelial thickness (mean ± SD, μm)	p-value^*
Control group	9	51.4 ± 4.3
AM group^†	9	50.3 ± 3.3	0.73
CL group^‡	8	52.6 ± 3.3

^* Kruskal-Wallis test;

^† AM = amniotic membrane;

^‡ CL = contact lens.

Table 2.

The comparison of the central corneal epithelial thickness among the 3 groups

Time	Control group (mean ± SD, n = 8)	AM group^*(mean ± SD, n = 9)	CL group^†(mean ± SD, n = 9)	p-value^‡
2 weeks	68.3 ± 6.1	95.3 ± 6.3	76.4 ± 5.1	0.012
4 weeks	51.4 ± 4.7	50.0 ± 6.5	51.8 ± 3.9	0.612

^* AM = amniotic membrane;

^† CL = contact lens;

^‡ Kruskal-Wallis test.

Table 3.

The comparison of hemidesmosome numbers in the central cornea among the 3 groups at 2 weeks

Groups	n	Hemidesmosomes (number/100 μm, mean ± SD)	p-value^*
Control group	9	0.80 ± 0.2
AM group^†	9	1.95 ± 0.3	0.002
CL group^‡	8	1.45 ± 0.2

^* Kruskal-Wallis test;

^† AM = amniotic membrane;

^‡ CL = contact lens.

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