Трансплантация суспензии эндотелиальных клеток в эксперименте ex vivo

Abstract

Relevance. Corneal transplantation with entire cornea or anormal layers replacement remains the only treatment option for patients with corneal endothelium pathology. Selective endothelial replacement was introduced into practice two decades ago, namely posterior lamellar keratoplasty, which has advantages over penetrating keratoplasty. Improvements of this method led to technique creation where isolated Descemet's membrane with endothelium monolayer without stromal layer is transplanted. This technology is associated with risks of intra- and postoperative complications, and is difficult to perform. Nowadays, development is in progress of introducing corneal endothelial cells (CECs) transplantation without stromal layer and even without Descemet's membrane. CECs transplantation in suspension form has potential to change treatment approach of corneal endothelium pathologies giving possibility of rapid eyesight recovery and reducing need for graft material. Purpose. To evaluate effectiveness of corneal CECs suspension transplantation on cadaver eye in ex vivo experiment. Material and methods. CECs suspension was obtained by modified enzymatic method. CECs transplantation experiment consisted of 3 stages. Firstly, CECs loss was calculated depending on cell administration method. Secondly, corneoscleral button was used as recipient for the obtained suspension. The third stage was carried out on cadaver eyeball. Results. The viability of transplanted CECs and their effective adhesion to Descemet's membrane was confirmed in this experiment. The characteristic markers of CECs ZO-1, Na+/K+ -ATPase, Ki67 were determined in corneal samples by immunohistochemistry assay and single cells expressing Vimentin were detected after one-week cultivation. Conclusion. Based on transplantation experiment results, it can be noted this technique is quite promising in surgical rehabilitation of patients with corneal endothelial dysfunction. Thus, it seems relevant to continue research to achieve the main goal – to fully cover the defect of central zone of corneal posterior surface with isolated CECs given that their morphology and functional activity are preserved. Key words: cornea, endothelial corneal dystrophy, endothelial cell suspension, corneal endothelial cell transplantation