Options in endothelial transplantation

Purpose To reduce DSAEK surgery time using pre-cut tissues.In addition real time valuation of specific parameters like cell density,endothelial mortality rate and finally thickeness of the posterior corneal lamella. Methods Corneo scleral discs were fixed in an artificial anterior chamber AC and Moria One microkeratome (Moria S.A., Antony, France) with a 350-micron Carriazo-Barraquer-type head was used to the prepare the lamellar corneal tissues.The artificial AC was cushioned with EUSOL-C (AL.CHI.MI.A.S.r.l,Ponte S.Nicolo,Italy).The sectioned tissues were stored at 4°C;post-cut and 24 hours pachymetry was repeated.Before the cutting,post-cut,at 4 and 24 hours cell viability and the same time cell morphology was measured with an inverted microscope.Cell density was measured at beginning and at the end of time course.Following this preliminary study,40 tissues were assigned for transplantation. Results Endothelial cell mortality was observed,which was characterized by trypan blue-positive cells diffused in the whole endothelial surface and corneal folds.The pattern of damage was typically due to mechanical factors associated to the passage of the microkeratome head.Cell morphology was well preserved at the end of the storage period at 4°C in EUSOL-C. We find a regular endothelial mosaic and absence of significant pleomorphism and polymegathism. An increase (approx.20%) in corneal thickness was observed at the end of time course. Finally no cases of graft dislocation or primary graft failure were observed. Conclusion The results have demonstrated the reproducibility of the procedure.In this way is possible to provide tissues ready to use,within 24h, and at the same time to certify tissue quality to prevent the primary graft failure and graft dislocation.