Development optimal conditions for cryopreservation of tissue-engineered corneal constructs

Abstract

Relevance. In recent years, due to the shortage of donor corneas, the need to creating conditions for storing stromal lenticules in eye banks for their clinical use in ophthalmosurgery have been actively discussed. Currently, scientists are looking for optimal conditions for storage native lenticules. There were no reports of storage of decellularized lenticules in the literature. Purpose. To develop optimal conditions for cryopreservation of stromal tissue-engineered constructs for the subsequent creation of a cryobank. Material and methods. The optical properties of native lenticules and tissue–engineered corneal constructs (TCs) were assessed using spectrophotometry. We used a decellularization protocol with 1.5 M NaCl with DNase 5 U/ml and RNase 5 U/ml to create TC. Dispersed viscoelastic agent approved for clinical use in ophthalmology was used for dehydration of TC. Three comparison groups were formed: 1st – control group (native lenticules), 2nd – group without dehydration of TC, 3rd – group with dehydration of TC. The spectrophotometer data was evaluated in 2 stages. The transparency of the control group was measured at 1 stage. At the second stage, the transparency of two experimental groups after storage in DMSO was investigated (a group without dehydration of TC and a group with dehydration of TC). Results. When compared between groups without dehydration of TC; with dehydration of TC and the control group (p≥0.05), no statistical difference was revealed, and when comparing groups without dehydration of TC and groups with dehydration of TC (p≥0.05), no statistical difference was revealed. Conclusion. Groups with dehydration of TC and without dehydration of TC after storage in DMSO did not differ in transparency. In this regard, these groups should be considered as interchangeable in terms of optical properties. Key words: storage, cryopreservation, lenticule, cornea, decellularization, tissue engineering