Endothelial cell loss could be influenced by factors like the donor's age and the time between the donor's death and the cornea's preparation. The study period, spanning from January 2017 to March 2021, included an evaluation of corneal transplants in this data comparison; these included PKPs, Corneae for DMEK, and pre-cut DMEK. On average, donors were 66 years old, with ages ranging from 22 to 88 years. On average, enucleation occurred 18 hours subsequent to death, with a range between 3 and 44 hours. Cultivating the cornea until reevaluation before transplantation took an average of 15 days, fluctuating between 7 and 29 days. No notable disparities were found when donors were grouped by 10-year age intervals. Cell loss, assessed at the initial and follow-up evaluations, consistently demonstrated a loss between 49% and 88%, without a trend of increasing cell loss as donor age increased. Cultivation duration until reevaluation exhibits a comparable characteristic. From the data comparison, it is concluded that donor age and cultivation time do not seem to be correlated with cell loss.

A maximum of 28 days is the timeframe for storing corneas, intended for clinical application, in organ culture medium after the donor's demise. Early in the COVID-19 pandemic of 2020, a significant situation emerged: clinical operations were being halted, resulting in an expected surplus of corneas graded for clinical use. Accordingly, once the corneas reached the terminus of their storage timeframe, and with the grant of consent for the tissue, they were transferred to the Research Tissue Bank (RTB). Nevertheless, university research endeavors were halted by the pandemic, leading to a surplus of high-quality tissue samples at the RTB, devoid of any assigned users. Rather than immediate disposal, the tissue was decided to be stored for future use by cryopreservation.
A previously established protocol for cryopreservation of heart valves underwent modification. Individual corneas were first placed inside wax histology cassettes and then introduced into Hemofreeze heart valve cryopreservation bags, which were filled with 100 ml of cryopreservation medium containing 10% dimethyl sulfoxide. Diving medicine Using a controlled-rate freezer at Planer, UK, they were frozen to a temperature below -150°C, and subsequently stored in a vapor phase above liquid nitrogen at temperatures below -190°C. To examine corneal morphology, six corneas underwent bisection; one half was processed for histology, and the other half was cryopreserved for one week before histological analysis. Haematoxylin and Eosin (H&E) and Miller's with Elastic Van Gieson (EVG) were the stains selected for this study.
In the comparative histological evaluation of the cryopreserved group against the controls, there were no observable, significant, detrimental alterations in morphology. Following this, an additional 144 corneas underwent cryopreservation. Handling assessments of the samples were conducted by eye bank technicians and ophthalmologists in concert. Based on their assessment, the eye bank technicians considered the corneas a possible resource for training in procedures like DSAEK or DMEK. According to the ophthalmologists, both fresh and cryopreserved corneas were considered equally suitable resources for training applications.
The established cryopreservation protocol, tailored to utilize modified storage containers and conditions, permits the successful cryopreservation of organ-cultured corneas beyond the expiration time. The suitability of these corneas for training purposes suggests that future disposal of corneas may be avoided.
Despite the expiration of time, organ-cultured corneas are successfully cryopreserved by adjusting the storage protocol, specifically concerning the storage container and environmental conditions. These corneas are appropriate for training applications and may avert future discarding.

Across the globe, over 12 million individuals are anticipating corneal transplantation, and a decrease in corneal donors has been noted since the COVID-19 pandemic's onset, which has unfortunately also impacted the supply of human corneas for scientific research. Subsequently, the employment of ex vivo animal models within this field demonstrates substantial merit.
Under the influence of orbital mixing, twelve fresh porcine eye bulbs were disinfected by immersing them in 10 mL of 5% povidone-iodine solution for 5 minutes at room temperature. The corneoscleral rims were excised and preserved in Tissue-C (Alchimia S.r.l., n=6) at 31°C and in Eusol-C (Alchimia S.r.l., n=6) at 4°C for up to 14 days. The assessment of Endothelial Cell Density (ECD) and endothelial cell viability was carried out using the vital dye Trypan Blue staining (TB-S, Alchimia S.r.l.). To quantify the percentage of stained area, digital 1X images of TB-stained corneal endothelium were acquired and analyzed using FIJI ImageJ software. Endothelial cell death (ECD) and mortality were quantified at intervals of 0, 3, 7, and 14 days.
After 14 days of incubation in Tissue-C and Eusol-C, both whole corneas and separated lamellae displayed a comparable endothelial structure when stained with TB and AR. Higher magnification analysis of endothelium morphology was achieved with the lamellar tissue, thus exceeding the capability of analyzing the whole cornea.
The presented porcine ex vivo model is instrumental in evaluating the safety and performance of storage conditions. The prospective applications of this methodology include the storage of porcine corneas for up to 28 days, and this is a focus of future work.
The presented ex vivo porcine model provides a means for evaluating the performance and safety of storage conditions. Possible future developments with this method may include the extension of porcine cornea storage to a period of up to 28 days.

The pandemic's impact has been profoundly felt on tissue donation figures in Catalonia, Spain. A noteworthy drop of approximately 70% in corneal donations and a significant decrease of roughly 90% in placental donations occurred during the lockdown period from March to May 2020. Despite the accelerated updating of standard operating procedures, considerable difficulties were encountered across multiple points. In terms of the transplant coordinator's availability for donor detection and evaluation, the procurement of sufficient personal protective equipment (PPE), and the resources available in quality control laboratories for screening, several factors are critical. The sheer volume of patients needing hospitalization, and the subsequent strain on hospital systems, caused a gradual increase in donation levels The initial confinement period witnessed a significant 60% drop in cornea transplants compared to 2019. This resulted in an alarming shortage of corneal donations by the end of March, impacting even emergency cases. Our Eye Bank responded by developing a new therapeutic approach to this problem. The tissue of a cryopreserved cornea, earmarked for tectonic surgery, is kept at -196°C, allowing a lifespan of up to five years. Subsequently, this is a tissue that enables us to proactively handle future similar emergencies. An adaptation of our processing protocol was implemented for this particular tissue, for the achievement of two distinct purposes. To guarantee the SARS-CoV-2 virus could be rendered inactive, if it existed, was a crucial goal. In contrast, a greater number of placentas should be donated. In order to accomplish this, the transport medium and the antibiotic cocktail were changed. In the final processing stage, an irradiation step was included. However, it is imperative to prepare for future possibilities and develop contingency strategies if donations are halted again.

NHS Blood and Transplant Tissue and Eye Services (TES) provides a serum eyedrop (SE) service for those experiencing severe ocular surface diseases. The serum collected from blood donation drives is further processed for SE preparation, where it is diluted eleven times with a physiological saline solution. Previously, 3 milliliter portions of diluted serum were dispensed into glass bottles within a Grade B cleanroom. With the initiation of this service, Meise Medizintechnik has implemented a system of automated, closed filling, characterized by squeezable vials arranged in tubing chains. genetic evolution The filling of vials is followed by their heat-sealing in a sterile environment.
For the purpose of enhancing SE production's speed and efficiency, TES R&D was tasked with validating the Meise system. A simulation of the closed system's validation process involved assessing bovine serum's performance during each stage of the filling process, followed by freezing to -80°C, vial integrity checks, and storage container packing. The items were then transported in containers on a round-trip journey to simulate the delivery process for patients. Upon their return, the vials were thawed, and the condition of each was inspected visually and by pressure testing with a plasma expander. MitoSOXRed Serum was delivered into pre-labelled vials, frozen in accordance with the protocol outlined earlier, and preserved for specific time periods of 0, 1, 3, 6, and 12 months within a standard household freezer that was temperature-controlled to -15 to -20 degrees Celsius, simulating a patient's home freezer. Every time point witnessed the extraction of ten random vial samples, with subsequent assessment of the outer packaging for damage or deterioration, and the vials for structural integrity, and their contents for sterility and stability. Stability was established via serum albumin concentration measurements, with sterility determined by testing for microbial contamination.
An assessment of the vials and tubing, performed after thawing at various intervals, indicated no instances of structural damage or leakage. All tested samples lacked microbial contamination, and serum albumin levels remained consistently within the anticipated range of 3-5 grams per deciliter at each respective time point.
The frozen storage of Meise closed system vials did not compromise the integrity, sterility, or stability of the dispensed SE drops, as demonstrated by these results.