Microplasma Induced Cell Morphological Changes and Apoptosis of Ex Vivo Cultured Human Anterior Lens Epithelial Cells - Relevance to Capsular Opacification.

Nina Recek,Saša Lazović,Marko Hawlina,Nataša Hojnik,Alenka Vesel,Miran Mozetič,Sofija Andjelić,Gregor Filipič,Gregor Primc,Uroš Cvelbar,Goran Petrovski,Mohammed Yousfi

doi:10.1371/journal.pone.0165883

Abstract

Inducing selective or targeted cell apoptosis without affecting large number of neighbouring cells remains a challenge. A plausible method for treatment of posterior capsular opacification (PCO) due to remaining lens epithelial cells (LECs) by reactive chemistry induced by localized single electrode microplasma discharge at top of a needle-like glass electrode with spot size ~3 μm is hereby presented. The focused and highly-localized atmospheric pressure microplasma jet with electrode discharge could induce a dose-dependent apoptosis in selected and targeted individual LECs, which could be confirmed by real-time monitoring of the morphological and structural changes at cellular level. Direct cell treatment with microplasma inside the medium appeared more effective in inducing apoptosis (caspase 8 positivity and DNA fragmentation) at a highly targeted cell level compared to treatment on top of the medium (indirect treatment). Our results show that single cell specific micropipette plasma can be used to selectively induce demise in LECs which remain in the capsular bag after cataract surgery and thus prevent their migration (CXCR4 positivity) to the posterior lens capsule and PCO formation.

Highlights

The applications of cold atmospheric pressure plasmas (CAP) in biomedicine has been growing enormously in the recent years.[1, 2] The CAPs have been applied for stem cell manipulation, cancer, skin treatments, wound healing and the like [3,4,5] To the best of our knowledge, this is the first to report highly selective use of CAP upon lens epithelial cells (LECs)
Weak membrane blebs appeared after 30 min, following the 30 s of plasma treatment applied on top of the medium for a twin cell system Fig 3
No mechanical effect of the gas flow on the cell morphology could be observed, which proved that the observed viability changes in the LECs were generated by the reactive oxygen species (ROS) species induced by the plasma

Summary

Introduction

The applications of cold atmospheric pressure plasmas (CAP) in biomedicine has been growing enormously in the recent years.[1, 2] The CAPs have been applied for stem cell manipulation, cancer, skin treatments, wound healing and the like [3,4,5] To the best of our knowledge, this is the first to report highly selective use of CAP upon lens epithelial cells (LECs). Microplasma Induced Cell Morphological Changes and Apoptosis of Single LECs cells are responsible for posterior capsular opacification (PCO), which is a major cause of postoperative or secondary visual loss that develops after cataract surgery in approximately 20% of cases within 5 years.[6] Cataract is still the leading cause of blindness worldwide, while PCO is caused by proliferation and migration of LECs remaining in the capsular bag after cataract surgery. The remaining cells can re-colonize the posterior lens capsule which was otherwise cellfree, and obstruct the visual axis contributing to light scattering and secondary visual loss

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Results

Discussion

Conclusion