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Abstract
To develop a culture regime for the in vitro human lens capsular bag model that better reflects clinical events following cataract surgery and to use this refined model to evaluate the putative impact of IOLs on PCO formation. Capsulorhexis and lens extraction were performed on human donor eyes to generate capsular bags attached to the ciliary body by the zonules. Preparations were secured by pinning the ciliary body to a silicone ring and maintaining in 6 mL serum-free EMEM for 28 days or in a graded culture system (days 1-3, 5% human serum and 10 ng/mL TGFβ2; days 4-7, 2% human serum and 1 ng/mL TGFβ2; days 8-14, 1% human serum and 0.1 ng/mL TGFβ2; days 15-28, serum-free EMEM), which better mimics clinical changes. Preparations were monitored with phase-contrast and modified-dark-field microscopy. Cell coverage and light scatter were quantified using image analysis software. The transdifferentiation marker, α-SMA and matrix component, fibronectin were assessed by immunocytochemistry. To assess IOLs in the model, Alcon Acrysof or Hoya Vivinex IOLs were implanted in match-paired capsular bags. Match-paired experiments showed that graded culture enhanced growth, facilitated matrix contraction, increased transdifferentiation, and promoted matrix deposition relative to serum-free culture. The graded culture protocol was applied to match-paired bags implanted with a Hoya Vivinex or an Alcon Acrysof IOL. The Vivinex demonstrated a lag in growth across the posterior capsule. However, by day 28, coverage was similar, but light-scatter was greater with Acrysof implanted. Cell growth on the Acrysof IOL anterior surface was significantly greater than Vivinex. The graded culture human capsular bag model serves as an excellent system to evaluate and develop intraocular lenses. The Hoya Vivinex IOL showed an overall better level of performance against postsurgical wound healing and PCO than the Alcon Acrysof using this model.
Highlights
The graded culture human capsular bag model reflects the pattern of postsurgical changes in the ocular environment
enables IOLs to be assessed for PCO formation tendency
It therefore serves as an excellent system to evaluate
Summary
Capsulorhexis and lens extraction were performed on human donor eyes to generate capsular bags attached to the ciliary body by the zonules. The second matchpaired series was used to evaluate two IOLs using the gradedculture system, the match-paired experiments were carried out using two lenses from the same donor both maintained in the graded culture system, where one capsular bag was implanted with an Alcon Acrysof IOL and its partner was implanted with a Hoya Vivinex IOL. Low-power dark-field images of capsular bags, with IOLs removed, were captured at endpoint and analyzed, such that the central visual axis (outlined by the capsulorhexis) was defined and its area quantified. The areas within this region of interest covered by cells was manually defined and the area of all cell covered regions quantified. The area of cell covered regions was expressed as a percentage of the central visual axis area
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