Effects of Mitomycin C on Scleral Collagen Fibrils According to Atomic Force Microscopy

Hui-Jae Lee; Samjin Choi; Youjin Cheong; Gyeong Bok Jung; Kyung-Hyun Jin; Hun-Kuk Park; Seung Jun Lee

doi:10.3341/jkos.2011.52.6.671

Journal List > J Korean Ophthalmol Soc > v.52(6) > 1009055

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Lee, Choi, Cheong, Jung, Jin, Park, and Lee: Effects of Mitomycin C on Scleral Collagen Fibrils According to Atomic Force Microscopy

Original Article

J Korean Ophthalmol Soc 2011;52(6):671-678.

Published online: 31 August 2011

DOI: https://doi.org/10.3341/jkos.2011.52.6.671

Effects of Mitomycin C on Scleral Collagen Fibrils According to Atomic Force Microscopy

Hui-Jae Lee, MD¹, Samjin Choi, PhD², Youjin Cheong, MD, PhD Candidate², Gyeong Bok Jung, PhD², Kyung-Hyun Jin, MD³, Hun-Kuk Park, MD, PhD^2,⁴, Seung Jun Lee, MD¹

¹Department of Ophthalmology, School of Medicine, Kangwon National University, Chuncheon, Korea

²Departments of Biomedical Engineering, Program of Medical Engineering, Kyung Hee University, Seoul, Korea

³Ophthalmology, College of Medicine, Kyung Hee University, Seoul, Korea

⁴Program of Medical Engineering, Kyung Hee University, Seoul, Korea

Address reprint requests to Seung Jun Lee, MD, Department of Ophthalmology, Kangwon National University Hospital #156 Baengryeong-ro, Chuncheon 200-722, Korea Tel: 82-33-258-2443, Fax: 82-33-258-2191, E-mail: opticus@kangwon.ac.kr

Received 15 September 2010 Revised 15 December 2010 Accepted 22 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 mitomycin C on the scleral collagen surfaces using atomic force microscopy (AFM).

Methods

Two non-contact mode AFM machines were used to observe changes in the morphological characteristics of human scleral surfaces before and after one, three, and five minutes of 0.02% mitomycin C application. Based on AFM topography and deflection images of the collagen fibril, the morphological characteristics of scleral fibrils including the fibril diameter and D-period were measured using the line profile.

Results

The sclera collagen fibril treated with 0.02% mitomycin C for one minute did not show any significant increases in mean fibril diameter (155.04 ± 17.46 nm) or mean D-periodicity (70.02 ± 3.33 nm), compared to those of the control group. However, the scleral collagen fibrils treated with 0.02% mitomycin C for three and five minutes showed significant increases in mean fibril diameter (182.33 ± 16.33 nm, 199.20 ± 12.40 nm, respectively) and mean D-periodicity (70.27 ± 13.66 nm, 72.75 ± 19.32 nm, respectively), compared to those of the control group.

Conclusions

The present study examined the structural changes in the scleral collagen fibrils before and after mitomycin C application according to atomic force microscopy. The results indirectly suggest that three or more minutes of 0.02% mitomycin C application affects the morphology of scleral collagen.

Keywords: Atomic force microscopy, Collagen fibrils, Diameter, D-banding, Human sclera, Mitomycin C

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Figure 1.

(A) Atomic force microscopy (AFM) block diagram, and (B) force vs. distance and relative zone modes.

Figure 2.

Representative atomic force microscopy topography images of the dehydrated human sclera in various scan sizes such as (A) 20 × 20 µm², (B) 5 × 5 µm², and (C) 1 × 1 µm².

Figure 3.

Representative three-dimensional images of the dehydrated human sclera in various scan sizes such as (A) 20 × 20 µm², (B) 5 × 5 µm², and (C) 1 × 1 µm².

Figure 4.

Representative examples of the diameter (C) and D-banding (D) measurements using line profiling plots on AFM topography (A) and deflection images (B) of the dehydrated scleral collagen fibrils in a scan size of 5 × 5 μ m².

Figure 5.

Representative AFM topography (A), deflection (B), and three-dimensional images (C) of the dehydrated scleral collagen fibrils in a scan size of 5 × 5 μ m².

Figure 6.

Representative AFM topography (A), deflection (B), and three-dimensional images (C) of the dehydrated scleral collagen fibrils with 0.02% mitomycin C application for 5 minutes in a scan size of 5 × 5 μ m².

Table 1.

Morphological changes in the sclera fibrils for control and MMC-treated groups

Parameter	Control	MMC 1 min	MMC 3 min	MMC 5 min
Diameter (mean ± SD, nm)	145.22 ± 17.78	155.04 ± 17.46	182.33 ± 16.33^*	199.20 ± 12.40^†
D-banding (mean ± SD, nm)	69.14 ± 14.15	70.02 ± 3.33	70.27 ± 13.66	72.75 ± 19.32

MMC = Mitomycin C.

^* p-value < 0.005 vs. Control group

^† p-value < 0.0001 vs. Control group.

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