Potential contributors to endothelial cell loss encompass the donor's age and the interval between the donor's death and corneal culture. The data comparison included corneal transplants—PKPs, Corneae for DMEK, and pre-cut DMEK—evaluated from January 2017 to March 2021. A typical donor's age was 66 years, with a range from 22 to 88 years. Following death, enucleation occurred after a mean interval of 18 hours, with variations observed from 3 to 44 hours. The mean time required to cultivate the cornea to the point of pre-transplantation reevaluation was 15 days (7-29 days). The results remained unchanged when donors were classified into 10-year age groups. The cell count, initially assessed and subsequently re-evaluated, showed a persistent cell loss between 49% and 88%, exhibiting no increase in loss as donor age increased. The cultivation duration up to re-evaluation demonstrates identical characteristics. From the data comparison, it is concluded that donor age and cultivation time do not seem to be correlated with cell loss.

Corneas, intended for clinical use, have a maximum storage period of 28 days in organ culture medium after the donor's death. With the 2020 commencement of the COVID-19 pandemic, it became clear that an unusual situation was unfolding, one characterized by the cancellation of clinical procedures and the projection of a surplus in clinical-grade corneas. As a result, the corneas, having reached the end of their allotted storage time, were transferred to the Research Tissue Bank (RTB), provided the required consent was in place. However, the university's research activities were suspended due to the pandemic, creating a circumstance wherein the RTB possessed a high-grade tissue supply, unclaimed by any user. Cryopreservation of the tissue was the chosen method to store it for later use, avoiding its discard.
An adapted protocol now governs the cryopreservation of heart valves. Cryopreservation bags, fashioned from a Hemofreeze heart valve, each holding 100 ml of cryopreservation medium with 10% dimethyl sulfoxide, were then used to contain individual corneas previously embedded in wax histology cassettes. selleck At Planer, UK, they were kept at sub-zero temperatures inside a controlled-rate freezer, falling below -150°C, then stored in a vapor phase above liquid nitrogen at a temperature below -190°C. To evaluate corneal morphology, six corneas were bisected; one section was prepared for histological examination, while the other was cryopreserved, stored for seven days, and then subsequently processed for histology. The histological analysis employed Haematoxylin and Eosin (H&E) and Miller's with Elastic Van Gieson (EVG) stains.
No apparent, substantial, or detrimental alterations in morphology were identified in the cryopreserved samples during the comparative histological evaluation of the control group. Subsequently, an additional 144 corneas were cryopreserved, ensuring future availability. Ophthalmologists, in conjunction with eye bank technicians, examined the handling characteristics of the samples. The technicians at the eye bank identified the corneas as potentially appropriate for training exercises in procedures like DSAEK or DMEK. The ophthalmologists highlighted the equivalence of fresh and cryopreserved corneas, both being equally suitable for the purpose of training exercises.
An established cryopreservation protocol, adapted for storage containers and conditions, permits the successful preservation of organ-cultured corneas even after time expired. For training purposes, these corneas are appropriate, and this might avert the future discarding of corneas.
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 suitable for educational purposes, which may help prevent their future disposal.

A global tally of over 12 million people are awaiting corneal transplants, and the number of cornea donors has declined since the onset of the COVID-19 pandemic, leading to reduced availability for research purposes as well. Consequently, the application of ex vivo animal models proves extremely useful within this particular area.
Twelve fresh porcine eye bulbs were disinfected with orbital mixing in 10 mL of 5% povidone-iodine solution, for 5 minutes at room temperature. Corneoscleral rims were sectioned and placed in Tissue-C (Alchimia S.r.l., n=6) at 31°C and Eusol-C (Alchimia S.r.l., n=6) at 4°C for a maximum of 14 days. Endothelial cell density (ECD) and mortality were evaluated using Trypan Blue staining (TB-S, Alchimia S.r.l.). Digital 1X pictures of TB-stained corneal endothelium, their stained area percentages were determined using FIJI ImageJ software. Endothelial cell death (ECD) and endothelial mortality were measured on days 0, 3, 7, and 14 respectively.
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. The lamellar tissue enabled a superior, higher-magnification analysis of endothelium morphology, surpassing the limitations of the whole cornea.
To evaluate the performance and safety of storage conditions, the presented ex vivo porcine model is utilized. Further development of this method is expected to enable the preservation of porcine corneas for extended periods, reaching 28 days.
Evaluation of storage conditions' performance and safety is enabled by this presented ex vivo porcine model. Future investigations into this technique may involve extending the time porcine corneas can be stored to 28 days.

From the start of the pandemic, there has been a steep decline in tissue donation across Catalonia in Spain. The enforced lockdown from March to May 2020 resulted in a decrease of approximately 70% in corneal donations and an approximately 90% decline in placental donations. Despite the rapid revisions to standard operating procedures, significant challenges persisted at various stages. Donor detection and evaluation by the transplant coordinator, adequate provision of personal protective equipment (PPE), and the quality control laboratory's screening resources are all crucial factors. The collapse of hospital resources, exacerbated by the daily influx of patients, caused donation levels to gradually recover, which was further compounded by this factor. Compared to 2019, a 60% decrease in corneal transplants marked the beginning of the confinement period. The Eye Bank tragically ran out of corneas by the end of March, impacting even emergency situations. This critical situation impelled the development of a new, innovative therapeutic method. A cryopreserved cornea, intended for tectonic procedures, is kept at a temperature of -196°C, a method that allows for up to five years of preservation. Consequently, this tissue enables us to address future similar crises effectively. This tissue necessitated an adjustment to our processing method, designed to serve two different functions. To ensure the inactivation of the SARS-CoV-2 virus, should it be present, was a priority. Unlike the current situation, a more expansive placental donation program is necessary. For this, alterations were made in the transport vehicle and the antibiotic mix. The final product now incorporates an irradiation stage. Consequently, the development of future contingency plans should address potential repeated donation stoppages.

To treat severe ocular surface diseases, patients can access a serum eyedrop (SE) service offered by NHS Blood and Transplant Tissue and Eye Services (TES). Serum collected during blood drives is used for SE preparation and diluted with 11 parts of physiological saline. Formerly, glass bottles in a Grade B cleanroom received 3 ml aliquots of the diluted serum. Since its inception, Meise Medizintechnik has crafted an automated, sealed filling system, utilizing interconnected chains of squeezable vials linked by tubing. Biolistic-mediated transformation Heat-sealing under sterile conditions is performed after filling the vials.
With the aim of improving SE production speed and efficiency, TES R&D undertook the task of validating the Meise system. To validate the closed system, a process simulation using bovine serum was conducted, encompassing every step from filling to freezing at -80°C, vial integrity testing, and packing into storage containers. Subsequently, they were placed in transport containers and dispatched on a journey, mimicking delivery to patients, that was round-trip. Upon their return, the vials were thawed, and the condition of each was inspected visually and by pressure testing with a plasma expander. Endocarditis (all infectious agents) Serum was dispensed into vials, flash-frozen using the previously described method, and stored for specific time points – 0, 1, 3, 6, and 12 months – within a household freezer set at a temperature between -15 and -20 degrees Celsius, to simulate the conditions of a patient's freezer. Ten random vial samples were removed at each data point. The outside containers were examined for damage or deterioration; the vials were tested for integrity; and the contents were tested for sterility and preservation. Stability was determined by examining serum albumin concentrations, and sterility was ascertained through the process of testing for microbial contamination.
Upon thawing, no structural damage or leakage was observed in any of the evaluated vials or tubing at any time point. Subsequently, all samples were free from any microbial contamination, and serum albumin levels consistently fell between 3 and 5 g/dL at each time point.
The successful dispensing of SE drops by Meise closed system vials, even after being stored frozen, is a testament to their maintained integrity, sterility, and stability, as demonstrated by these results.