To improve procedure safety and simplicity, we evaluated dextran-based freezing media and a dry, no-medium approach at -80°C.
Three different donors yielded five samples of human amniotic membrane. For each donor, the preservation conditions included dimethyl sulfoxide at -160°C, dimethyl sulfoxide at -80°C, dextran-based medium at -160°C, dextran-based medium at -80°C, and dry freezing at -80°C without a medium. After four months in storage, the adhesive qualities and structural form were investigated.
The newer preservation protocols exhibited no variations in the adhesive or structural properties of the examined tissues. The preservation protocol had no effect on either the structure or the basement membrane, yet the stromal layer maintained its adhesiveness.
By opting for -80°C storage instead of liquid nitrogen cryopreservation, the manipulation steps would be reduced, the procedure simplified, and the cost lowered. The use of a dextran-based freezing solution, or the complete absence of any medium (a dry environment), serves to mitigate the potential toxicity that might stem from dimethyl sulfoxide-based freezing media.
A move to -80°C storage from liquid nitrogen cryopreservation would reduce the handling involved, simplify the protocol, and contribute to a decrease in financial costs. To circumvent the potential toxicity inherent in dimethyl sulfoxide-based cryopreservation media, dextran-based freezing media, or even no medium (dry freezing), can be employed.

This study sought to evaluate the effectiveness of Kerasave (AL.CHI.MI.A Srl), a corneal cold storage medium containing antimycotic tablets, in eliminating nine corneal infection-causing contaminants.
Incubation of Kerasave medium containing 10⁵ to 10⁶ CFUs of Candida albicans, Fusarium solani, Aspergillus brasiliensis, Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis spizizenii, Pseudomonas aeruginosa, Enterobacter cloacae, and Klebsiella pneumoniae at 4°C for 0, 3, and 14 days allowed for the determination of Kerasave's killing efficacy. By employing the serial dilution plating technique, log10 reductions at different time intervals were assessed.
Three days post-treatment, Kerasave produced the maximum log10 decrease in the concentrations of KP, PA, CA, and EC. For both SA and EF, a two-unit decrease in the log10 scale was observed. A minimal log10 decrease in concentration was noticed for BS, AB, and FS. By day 14, the microbial populations of CA, FS, SA, EF, PA, and EC were demonstrably lower.
Three days post-application, Kerasave yielded the highest log10 decrease in the measured concentrations of KP, PA, CA, and EC. A 2-log10 decrease was seen in both SA and EF measurements. BS, AB, and FS concentrations displayed the smallest reduction in log10 values. A 14-day period resulted in a further decrease in microbial counts across CA, FS, SA, EF, PA, and EC specimens.

Analysis of corneal guttae post-Descemet membrane endothelial keratoplasty (DMEK) in eyes treated for Fuchs endothelial corneal dystrophy (FECD).
Ten patients, all undergoing FECD surgery at a tertiary referral center between 2008 and 2019, contributed 10 eyes to this case series. The average age of the patients was 6112 years, with 3 females and 6 males among them. From the total patient population, five were phakic and the remaining four, pseudophakic. The average age of donors was 679 years old.
Specular microscopy images, obtained during a standard postoperative consultation, indicated a potential guttae recurrence in ten eyes subsequent to DMEK. In 9 instances, confocal microscopy subsequently established the presence of guttae; in one, histology confirmed the presence. Sixty percent of the patients (six out of ten) who underwent bilateral DMEK procedures, unfortunately, experienced guttae recurrence in only one eye. Primary DMEK resulted in guttae recurrence in nine eyes, while a single eye experienced recurrence after a re-DMEK procedure performed 56 months later, showing no signs of guttae after the primary DMEK. Specular microscopy, performed one month following DMEK, often displayed the presence of suspected guttae in the observed samples. Eight donors' preoperative endothelial cell density (ECD) count, initially registering 2,643,145 cells/mm2, saw a reduction to 1,047,458 cells/mm2 one year after the surgical procedure.
Guttae recurrence following DMEK is frequently attributed to undetectable guttae on the donor cornea, obscured from standard slit-lamp and light microscopy screening at the eye bank. AG 825 in vivo Improved diagnostic procedures for guttae, imperative for eye banks, are crucial to prevent the transplantation of tissue containing guttae or predisposed to guttae formation post-operatively.
The reappearance of guttae following DMEK surgery is frequently attributed to undetectable guttae present on the donor cornea, which eluded detection by routine slit-lamp and light microscopy at the eye bank. The release of guttae-containing or guttae-prone tissue for transplantation by eye banks should be circumvented through the development of more sensitive guttae detection methodologies.

Studies of recent clinical subjects indicate that replacing RPE cells could potentially maintain sight and rebuild retinal tissue in degenerative retinal ailments. Recent breakthroughs allowed the separation of RPE cells from induced pluripotent stem cells. The effectiveness of scaffold-based techniques in delivering these cells to the back of the eye is currently being investigated through ongoing clinical trials. Borrowed materials from donor tissues provide a cell support framework in subretinal transplantation procedures. These biological matrices closely resemble the microenvironment of the extracellular matrix in the native tissue. As an illustration of a basement membrane (BM), the Descemet's membrane (DM) contains an abundance of collagen. Further investigation is needed to determine the potential of this tissue for retinal repair.
A study examining the survival and characteristics of human embryonic stem cell-retinal pigment epithelium (hESC-RPE) cells on a decellularized matrix (DM), focusing on possible application in retinal transplantation.
The treatment of DMs, extracted from human donor corneas, involved thermolysin. Atomic force microscopy, coupled with histology, provided the means to evaluate the DM's surface topology and the effectiveness of the denudation technique. To ascertain the membrane's capacity to sustain hESC-RPE cell cultivation and preserve their vitality, hESC-RPE cells were seeded onto the acellular DM's endothelial surface. Transepithelial resistance was employed to determine the degree of integrity present in the hESC-RPE monolayer. Confirmation of cellular maturation and functionality on the novel substrate involved the assessment of RPE-specific gene expression, protein expression, and growth factor secretions.
A thermolysin treatment did not compromise the tissue integrity, therefore enabling a reliable method for standardizing decellularized DM preparations. The cell graft demonstrated a morphology that was indicative of RPE. The accurate RPE phenotype was further substantiated by the expression of typical RPE genes, the precise cellular location of proteins, and the secretion of essential growth factors. Maintaining the viability of the cells in culture was accomplished for up to four weeks.
Acellular DM's capacity to nurture the growth of hESC-RPE cells underscores its potential as an alternative to Bruch's membrane. Further in vivo studies are needed to determine if it is a viable tool for transporting RPE cells into the eye's posterior area.
Acellular dermal matrix (ADM) successfully fostered the expansion of human embryonic stem cell-derived retinal pigment epithelial (RPE) cells, effectively confirming its potential as an alternative to Bruch's membrane. Subsequent in vivo investigations will evaluate the feasibility of using this material to introduce RPE cells into the posterior segment of the eye. Our study signifies the opportunity to repurpose unsuitable corneal tissue, usually discarded by eye banks, for clinical purposes.

The current shortfall in ophthalmic tissue supply in the UK calls for the investigation and implementation of alternative supply routes. In response to this significant necessity, the NIHR funded the EDiPPPP project, a partnered initiative with NHSBT Tissue Services, now rebranded as Organ, Tissue Donation, and Transplantation.
This presentation, based on work package one of EDiPPPP, reports on a large-scale, multi-site, retrospective case notes review across England. The review sought to determine the size of the potential eye donation (ED) population, its clinical characteristics, and the obstacles faced by clinicians in using standard ED criteria to evaluate patient eligibility.
Reviewers, healthcare professionals stationed at research sites, retrospectively assessed 1200 deceased patient case notes (600 HPC; 600 HPCS). These assessments were subsequently evaluated by specialists at NHSBT-TS against current ED criteria. Analyzing the records of 1200 deceased patients, the study found that 46% (n=553) qualified for eye donation. In hospice care, the rate of suitability was 56% (n=337), and in palliative care, it was 36% (n=216). However, the referral rate to NHSBT-TS for actual eye donation was only 12% (4 hospice, 3 palliative), indicating a need for better protocols. natural biointerface Including cases (n=113) where assessments varied but NHSBT determined eligibility, the potential donor pool increases from 553 (46% of the total cases) to 666 (representing 56% of eligible cases).
Clinical sites in this study hold substantial potential for eye donations. Bioresearch Monitoring Program (BIMO) Currently, this potential is not being manifested. Bearing in mind the projected rise in the need for ophthalmic tissue, the outlined method for increasing the supply of this tissue, as observed in this retrospective case review, requires immediate attention. Finally, the presentation will offer suggestions for enhancing service provision.