A Human Three-Dimensional In Vitro Model of Lens Epithelial Cells as a Model to Study Mechanisms of Drug-Induced Posterior Subcapsular Cataracts

Abstract

Purpose: Cataract is a pathological opacification of the lens, which is still one of the leading causes of blindness in the world. Several etiologies are described, among them drug-induced cataract, for example, posterior subcapsular cataract (PSC) after steroid treatment. To investigate different mechanisms of drug-induced cataract a human three-dimensional (3D) lens in vitro model was developed, consisting of immortalized human lens epithelial cells. Methods: These cells were cultivated on 96-well, ultralow attachment plates, where they rapidly form spheroids. By gene expression analysis different markers were observed, which are important to maintain lens transparency, such as ephrin type-A receptor 2 (EphA2) or a-smooth muscle actin (a-SMA). Results: The lens epithelial cells form a spheroid within a few days and show stable expression of important lens marker, and size and viability remain stable up to 26 days in culture. The gene expression of the glucocorticoid-treated spheroids revealed a clear shift in the expression of EphA2, a-SMA, aB-crystallin (CRYAB), and heat shock protein beta-1 (HSPB1). Furthermore, the glucocorticoid treatment did not improve cell survival. Conclusions: This study proposes a useful 3D in vitro model, which expresses important lens markers and is capable of demonstrating features found in drug-induced cataracts. As the viability remains stable over long time, this model can also be used for long-term treatment. The main characteristics are the increased expression of a-SMA, CRYAB, and HSPB1 and the decreased expression of EphA2. The present data provide some first evidence on novel mechanisms involved in glucocorticoid-induced cataracts.

Factors Affecting Posterior Capsule Opacification in the Development of Intraocular Lens Materials

“…In addition to primary cell lines, the human lens epithelial cell line HLEB3 is one of the most established cell lines for research into the development and treatment of PCO [1,[105][106][107]. HLE-B3 cells possess receptors for growth factors and when incubated with these factors, elicit fibrotic pathways shown during PCO development.…”

“…The development of three-dimensional in vitro models of PCO provides greater functional and structural applicability as shown by Plüss [106] who created a three-dimensional in vitro model of HLE-B3 cells by seeding the cells within 96-well plates and centrifuging to aggregate the cells. The spheroid structure was established by day 1 and was maintained up to day 26 (Figure 3).…”

“…• Three-dimensional modelling [106,107,123,129] • Role of cytokines and inflammation [5,16,102] • Investigation of intraocular lenses [105,108,109,130] • Inhibition of molecular pathways [92,103,121,131,132] • Surgical technique evaluation [15] In vivo Porcine, murine, human, rabbit…”

“…The spheroids were maintained for 26 days. Stable expression of key markers N-cadherin (Hs00983056_mL) and αβcrystallin (Hs00157107_mL) was seen up to day 26 as determined by reverse transcription-quantitative polymerase chain reaction[106].…”

Zaćma posteroidowa – polekowe uszkodzenie narządu wzroku, na które warto zwrócić uwagę w terapii nieswoistych chorób zapalnych jelit

“…Szansa upatrywana jest w kwasie askorbinowym mającym właściwości przeciwutleniające, który nie hamując działania przeciwzapalnego GKS, pozwoliłby ograniczyć wpływ wolnych rodników [22]. Modulacja aktywności czynnika wzrostu fibroblastów [23] czy ekspresji receptora EphA2 [24] może być upatrywana jako szansa na hamowanie rozwoju zaćmy powstającej wskutek długotrwałej sterydoterapii w NZJ.…”