In vivo corneal endothelial cell findings after the infusion of cultivated HCECs into the anterior chamber with a slit-scanning wide-field contact specular microscope

Abstract

Purpose : We are currently in the process of developing a new therapy for bullous keratopathy that involves the infusion of cultivated human corneal endothelial cells (HCECs) into the anterior chamber. It is still unclear whether or not these cells adhere to the Descemet’s membrane, as well as how they are sustained post injection. It has been reported that the slit-scanning wide-field contact specular microscope allows for in vivo CECs to be observed more clearly and over a wider area compared with a non-contact specular microscope. In this present study, we report the CEC findings in patients who underwent cell infusion therapy via observation with a slit-scanning wide-field contact specular microscope.

Methods : A slit-scanning wide-field specular microscope (Konan Medical) was used to observe the CEC layer in 18 patients (9 male, 9 female, mean age 66.6 ± 10.8 years) with bullous keratopathy, who underwent cell infusion therapy from Dec 2013 to Sep 2014 in Kyoto Prefectural University of Medicine, before and at 1-, 3-, 6-months after surgery. In each patient, oxybuprocaine 0.4% eye drops were instilled to anaesthetize the operated eye and a contact cone lens was then attached. Next, images of the CEC layer were recorded via the slit-scan technique from the center to peripheral region. Focused still images were then extracted from the created MPEG-2 file and the CEC density in a 0.3 x 0.3mm central area of the cornea was then manually calculated.

Results : Pre surgery, in vivo CECs could not be observed due to the corneal edema, however, they could be observed post surgery via the use of wide-field specular microscopy. The injected CECs were observed attached to the cornea in all cases, and at a density of over 2000 cells/mm² in the early postoperative period in 13 of the 18 cases. Hexagonal-shape CECs were observed in the peripheral region as well as in the center.

Conclusions : The use of wide-field specular microscopy allowed us to obtain images of cultivated HCECs adhesion post injection into the anterior chamber and for the time-dependent change of those cells to be observed in vivo.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.