Differentiation and proliferation potential of lens epithelial cells ex vivo - possible cause for late intraocular lens dislocation

Abstract

Purpose : Late spontaneous in-the-bag intraocular lens (IOL) dislocation presents a severe complication occurring three or more months after cataract surgery. Lens epithelial cells (LECs) can proliferate, migrate and modulate under stressful conditions. We aimed to test the proliferative and pluripotent properties of LECs during ex vivo cultivation of explanted dislocated IOLs with their lens capsule present in toto.

Methods : All tissue collection complied with the Guidelines of the Helsinki Declaration and was approved by the Local Ethical Committee, S. Fyodorov Eye Microsurgery Federal State Institution, Russian Federation (No.81.3). Explanted IOLs with lens capsules were plated into 6-well plates and cultured in DMEM:F12 supplemented with 10% FCS, antibiotics, and maintained in a humidified 5% CO₂, 95% air incubator at 37⁰C. Light microscopy was performed. After two weeks, the tissue was fixed in 4% paraformaldehyde for 24h and processed for immunohistochemistry (IHC). Fluorescent images were obtained by Zeiss Axio Observer Z1microscope.

Results : Direct microscopy imaging of the cultured lens capsules with the IOLs showed presence of fibroblastoid cells on the anterior side of the IOLs, which showed ability to proliferate during the cultivation period. Lens capsules were opacified and appeared to have fibrotic and contractile elements. IHC staining revealed the presence of proliferating cells, which were positive for proliferation (Ki-67) and stemness markers (Sox2, ABCG2). In addition, IHC confirmed the ability of LECs to migrate and undergo epithelial-to-mesenchymal transition towards myofibroblasts (alphaSMA positivity). The cells produced fibrotic and contractile elements in the extracellular matrix (ECM).

Conclusions : LECs found in dislocated IOLs with lens capsules present in toto can divide, migrate, proliferate, as well as transform into myofibroblasts under ex vivo culture conditions. In addition, LECs can produce contractile elements in the ECM. Altogether, this indicates that LECs on the lens capsule can provoke pathogenic mechanisms during IOL dislocation and contribute towards further complications in the affected human eye.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.