Applied Microscopy

Korean Society of Microscopy (한국현미경학회)
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The Cytotoxicity and Effect on the Experimental Rabbit Cornea of Soft Contact Lens Multi-Purpose Solution (MPS)

콘택트렌즈용 다목적용액(MPS)의 세포증식저해 및 실험용 토끼 각막에 미치는 영향

  • Koh, Eun-Gyeong (Department of Biology, College of Natural Science, Chonnam National University) ;
  • Chae, Soo-Chul (Department of Biology, College of Natural Science, Chonnam National University) ;
  • Park, Su-Kyoung (Department of Biology, College of Natural Science, Chonnam National University) ;
  • Kim, Deok-Song (Department of Biology, College of Natural Science, Chonnam National University) ;
  • Lee, Jong-Bin (Department of Biology, College of Natural Science, Chonnam National University)
  • 고은경 (전남대학교 자연과학대학 생물학과) ;
  • 채수철 (전남대학교 자연과학대학 생물학과) ;
  • 박수경 (전남대학교 자연과학대학 생물학과) ;
  • 김덕송 (전남대학교 자연과학대학 생물학과) ;
  • 이종빈 (전남대학교 자연과학대학 생물학과)
  • Published : 2006.09.30
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Abstract

The purpose of this study is to find out how soft contact lens multi-purpose solution (MPS), often used for medical treatment, effects the inhibition on cell growth and research the result of using MPS, suspected to demage eye cells, on rabbit eye's corneal epithelium and endothelium tissue. In this treatise, $ReNu^{(R)}$ (Baush & Lomb, USA), Opti-free $express^{(R)}$ (Alcon, USA), Free-sol $plus^{(R)}$ (Hanamedicon, Korea) had been selected among the MPS. After culturing L929 roil line, cell growth inhibition rate was measured by MTT assay, and by making Hematoxylin and Eosin stain specimen. the morphology was observed by optical microscope. In the In vivo experiment,9 white rabbit eyes (18 eyes) were classified into 3 groups. The experimental group is left eyes (9 eyes) of rabbit, and MPS were dropped; however. the control group, the right eyes (9 eyes), were only used a saline solution without preservatives. After the dropping within the period, the cornea surface of rabbit eyes were stained by Rose bengal and observed. To figure out the changes of the corneal epithelium and endothelium tissue scanning electron microscopy (SEM) has been used. As the result, the rate of cell growth inhibition was 54%. 73% and 36%, respectively. Morphological changes represented that the shape has been changed into oval or round shape and those are not considered as a common formation of L929 cell line. When it comes to staining Rose bengal, each experiment group was stained red which is not shown in controls. The polygonal mosaic pattern of a corneal epithelium was disturbed in the picture taken by SEM; furthermore, the shape of the corneal endothelium was irregular. In conclusion, as we consider antimicrobial effect and the safety on living cells, it is necessary that we should improve concentration of preservatives and study continuously to develop a new preservatives without a toxic effect on the cornea surface.

본 연구는 시중에서 유통되고 있는 콘택트렌즈 관리용액인 다목적용액 (multi-purpose solution, MPS)이 세포에 미치는 증식저해정도와 실험용 토끼눈의 각막 상피 및 내피조직에 미치는 손상정도를 비교 관찰하고자 시행하였다. MPS는 $ReNu^{(R)}$ (Baush & Lomb, USA), Opti-free $express^{(R)}$ (Alcon, USA), Free-sol $plus^{(R)}$ (Hanamedicon, Korea)를 사용하였다. 세포증식 저해율은 L929세포주를 배양 후 MTT assay로 검정하였고, 형태학적으로는 광학 현미경과 Hematoxylin & Eosin staining 표본을 제작하여 관찰하였다. In vivo 실험은 백색 가토 9마리 (18안)를 3군으로 분류하여 실험군인 좌안(9안)에는 각 MPS제품을, 대조군인 우안(9안)에는 보존제가 포함되지 않은 멸균생리식염수를 점안하였다. 일정기간 점안 후, 실험용 토끼눈의 각막표면을 Rose bengal로 염색하여 관찰하였고 각막상피 및 내피조직의 변화는 주사전자현미경 (scanning electron microscopy, SEM)으로 관찰하였다. 세포증식 저해율은 각각 54, 73, 36%로 나타났으며, 형태학적 변화는 L929세포주의 일반적인 형태와 달리 타원형 또는 원형으로 위축되었다. Rose bengal 염색 결과, 각 실험군은 대조군과 달리 전반적으로 붉게 염색되었고 주사전자현미경 (SEM)관찰 결과, 각막상피는 다각형 모자이크(polygonal mosaic)형태의 균일도가 감소하였고, 각막내피 또한 형태가 불규칙하였으며 섬유아세포만 국소적으로 관찰되었다. 따라서 MPS의 항미생물의 효과와 더불어 생체세포의 안전성면에서 고려해 볼 때 합성보존제의 농도개선과 세포독성이 없는 새로운 보존제 개발의 지속적인 연구가 필요할 것으로 판단된다.

Keywords

References

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