Comparison of the Effects of Temperature and Dehydration Mode on Glycerin-Based Approaches to SMILE-Derived Lenticule Preservation

Abstract

The aim of this study was to explore the optimal method of small-incision lenticule extraction (SMILE)-derived lenticules, subjected to long-term preservation using glycerol, under a range of temperatures, and using an array of dehydration agents. Methods:In total, 108 myopic lenticules were collected from patients undergoing the SMILE procedure. Fresh lenticules served as a control group for this study, whereas all other lenticules were separated into 8 groups, which were preserved at 4 different temperatures (room temperature [RT], 4, 220, and 280°C) with or without silica gel in anhydrous glycerol. Evaluated parameters included thickness, transmittance, hematoxylin and eosin staining, transmission electron microscopy, and immunohistochemistry analyses.Results: After a 3-month preservation period, lenticular thickness in these different groups was significantly increased, particularly for samples stored at RT. The mean percentage transmittance of lenticules stored at 280°C with or without silica gel was closest to that of fresh lenticules. Hematoxylin and eosin staining revealed sparsely arranged collagen fibers that were more scattered in preserved lenticules relative to fresh lenticules, particularly in RT samples. Transmission electron microscopy revealed that the fibril bundles densities in lenticules stored at RT were significantly less than those stored at other temperatures. Immunohistochemistry analyses revealed reductions in or loss of CD45 and human leukocyte antigens in all preserved lenticules relative to control samples.Conclusions: Of the tested approaches, the preservation of SMILE-derived lenticules over a 3-month period was optimal at 280°C with or without silica gel in anhydrous glycerol.

Cutting Edge: Corneal Stromal Lenticule Implantation (Corneal Stromal Augmentation) for Ectatic Disorders

“…Animal models and initial clinical results have confirmed the efficacy of this preservation method. 11 Dehydration is mainly used to store anterior keratoplasty grafts. In this method, the organ culture medium is supplemented with a dehydrating agent, such as polyethylene glycol or silica gel, to preserve glycerol-preserved tissues at different temperatures.…”

“…Before use, the tissue should be rehydrated by submerging it in sterile phosphate-buffered saline for 15 minutes. 11 Decellularization has the advantage of making the cornea free from immunogenic cellular components while maintaining the integrity of the extracellular matrix. Various methods have been described using detergents and enzymes to efficiently remove cellular and nuclear materials.…”

“…Sodium dodecyl sulfate has been investigated as a way to decellularize thin human stromal lenticules, and the biocompatibility of the decellularized lenticules has been confirmed in patients with advanced KCN. 11…”

“…Animal models and initial clinical results have confirmed the efficacy of this preservation method. 11…”

“…Sodium dodecyl sulfate has been investigated as a way to decellularize thin human stromal lenticules, and the biocompatibility of the decellularized lenticules has been confirmed in patients with advanced KCN. 11 In a recent study on 108 myopic lenticules collected from patients who underwent the SMILE procedure, fresh lenticules were compared with those preserved at 4 different temperatures (room temperature, 4°C, 220°C, and 280°C) with or without silica gel in anhydrous glycerol. The 3-month follow-up results showed reductions in or loss of CD45 and human leukocyte antigens, as well as increased lenticular thickness, particularly for samples stored at room temperature with sparsely arranged collagen fibers and low fibril bundle densities.…”