Long-term preservation of donor corneas in glycerol for keratoplasty: exploring new protocols

Tripathi, Himi; Mehdi, Manzoor Ul; Gupta, Deepika; Sen, Seema; Kashyap, Seema; Nag, Tapas Chandra; Purwar, Manisha; Jassal, Manjeet; Agrawal, Ashwini K.; Mohanty, Sujata; Tandon, Radhika

doi:10.1136/bjophthalmol-2015-306944

Cited by 26 publications

(18 citation statements)

References 20 publications

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“…The use of different media to increase the shelf life of corneal buttons received at the eye bank includes the practice of recovering tissues in MK-medium with subsequent transfer to intermediate storage media. [3132] Tissue preservation in anhydrous glycerol at −80°C was found to be effective to maintain sterility of the tissue, corneal thickness, better optical transparency, tensile strength, and ultrastructural features. [32]…”

Section: Resultsmentioning

confidence: 99%

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Upgradation and modernization of eye banking services: Integrating tradition with innovative policies and current best practices

Tandon

Singh

Gupta

et al. 2017

Indian J Ophthalmol

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Purpose:The purpose of this study is to review the history and evolution of the National Eye Bank (NEB) and analyze the impact over the years and report the outcome of the invested resources.Methods:Review of archival material, records, project reports, policy and procedures' manuals, and publications was done. Descriptive and analytical processing of data obtained was undertaken. Parameters evaluated included total collection, transplantation, utilization rates of donor cornea, changing trends over time in terms of numbers and duration of recipients waiting, impactful research translated into changes in standard operating protocols, new facilities, and subsequent effects on numbers or quality assurance measures and overview of major achievements. Periodic situational analysis with contextual relevance and interpretation of outcomes was done pertaining to national goals and international standards.Results:The NEB and cornea services have played a key leadership role in furthering the development of eye banking and corneal transplantation services. The contribution extends beyond routine patient care to education, training, generation of resources, advocacy, and policymaking. In quantifiable terms, the overall performance has steadily increased over the years. Major contributions include training of doctors, eye bank staff and corneal surgeons, introduction of innovative techniques for corneal transplantation, setting of national standards for eye banking and provision of preservation media, customized corneal, and ocular surface cell replacement therapy in collaboration with other departments and institutes.Conclusion:The eye banking and corneal transplantation facilities have evolved with time providing quality services, modernized as appropriate with updated knowledge and incorporating technological advances supported by the systematic evidence-based approach.

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Section: Resultsmentioning

confidence: 99%

“…[3132] Tissue preservation in anhydrous glycerol at −80°C was found to be effective to maintain sterility of the tissue, corneal thickness, better optical transparency, tensile strength, and ultrastructural features. [32]…”

Section: Resultsmentioning

confidence: 99%

Upgradation and modernization of eye banking services: Integrating tradition with innovative policies and current best practices

Tandon

Singh

Gupta

et al. 2017

Indian J Ophthalmol

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“…10 Other studies have shown that freeze-dried tissues can also be used for conditions like epikeratophakia. 11,12 Tripathi et al 7 pro-posed preserving tissues at 280°C, which maintained the sterility, thickness, optical transparency, mechanical strength, and ultrastructural features. However, there is also contradic-tory evidence suggesting that viable donor keratocytes are necessary for a successful graft until the donor graft is completely attached to the recipient cornea.…”

Section: Discussionmentioning

confidence: 99%

“…Differ-ent dehydration storage conditions have been evaluated by several research groups that include organ culture medium supplemented with 5% PEG 35,000 for the dehydration of anterior donor grafts, 6 anhydrous glycerol with and without silica gel, and various temperatures for the preservation of corneal tissues. 7 To increase the availability of these tissues, we therefore investigated if human donor corneal stroma stored in a dehydrated condition and rehydrated after 2 weeks retained the same biological and biomechanical properties. We used silica gel at room temperature as a dehydrating agent, conditions easily repeatable even in developing countries.…”

Section: Introductionmentioning

confidence: 99%

Biobanking of Dehydrated Human Donor Corneal Stroma to Increase the Supply of Anterior Lamellar Grafts

Romano

Levis

Gallon

et al. 2019

Cornea

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Purpose: To investigate the effect of dehydration on human donor corneal stroma for biobanking. Methods: Epithelium and endothelium of research-grade human donor corneas (n = 12) were scraped off, leaving a bare stroma with attached sclera. The tissues were placed in a large Petri dish prefilled with silica gel in the periphery and stored at room temperature for 14 days. At the end of preservation, the tissues were rehydrated by being submerged in phosphate-buffered saline for 15 minutes. Transparency (using a custom-built device) and thickness (using optical coherence tomography) measurements were recorded before dehydration, after dehydration, and after rehydration of the tissues. Periodic acid-Schiff and alpha-smooth muscle actin (a-SMA) staining before dehydration and after rehydration were performed to determine the presence of keratocytes and expression of a-SMA. Tensile stress-strain before dehydration and after rehydration was performed to evaluate the biomechanical properties. Results: No difference in corneal transparency before dehydration (69.57 6 6.41%) and after rehydration (67.37 6 2.82%), P = 0.36, was observed. The corneas were more compact after dehydration. A significant change in thickness between before dehydration (625.8 6 75.58 mm) and after rehydration (563.6 6 15.77 mm) stage, P = 0.03, was noticed. The thickness was reduced to 147.6 6 3.71 mm when dehydrated. Periodic acid-Schiff staining showed presence of stromal keratocytes and a-SMA protein expressed in control, dehydrated, and rehydrated corneas. There was no significant difference in the stiffness between control (27.86 6 11.65 MPa) and rehydrated corneas (31.46 6 11.41 MPa). Conclusions: Human donor corneal stroma can be biobanked for up to 2 weeks in a dehydrated condition without losing their molecular or biomechanical properties after rehydration.

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“…Various reports have indicated that glycerol-dehydrated donor corneas can effectively substitute fresh corneas in patch grafts [ 21 ]. Storing corneal tissues in anhydrous glycerol at 4 °C has been shown to be as effective as storing these tissues at sub-zero temperatures [ 22 ]. Glycerol is a dehydrating agent with antimicrobial and antiprotease properties that also helps to maintain corneal structure [ 23 , 24 ].…”

Section: Discussionmentioning

confidence: 99%

Comparison of femtosecond laser-assisted corneal intrastromal xenotransplantation and the allotransplantation in rhesus monkeys

et al. 2017

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BackgroundIn our previous study, we showed that both allogeneic and autogeneic small-incision femtosecond laser-assisted corneal intrastromal transplantation are safe and effective surgeries. However, the results of small-incision femtosecond laser-assisted intrastromal xenotransplantation have not yet been explored. Additionally, we suggest that glycerol-dehydrated corneal lamellae might provide a possible alternative for this xenogenic implantation approach.MethodsCorneal inlay lamellae were produced from rabbits and humans using femtosecond laser-assisted surgeries and were dehydrated in glycerol for 1 week at 4 °C. These xenogeneic glycerol-dehydrated grafts and fresh allogeneic monkey lamellae were then implanted into rhesus monkeys using small-incision femtosecond laser assistance. Postoperatively, clinical examinations, AS-OCT measurements and tear inflammatory mediator assays were performed.ResultsThere were no significant changes in the transparency of the corneal lamellae after glycerol dehydration. Following implantation, no evidence of tissue rejection or severe inflammatory responses was observed in the monkeys, and the host corneas remained transparent throughout a 6-month observation period. The grafts were clearly visible via AS-OCT. Corneal thickness increased 1 week postoperatively but subsequently declined and remained unchanged 1 month after surgery. Significant changes were observed in all tear inflammatory mediators in the ‘Rabbit to Monkey’ group. The trends in changes of tear inflammatory mediators in the ‘Human to Monkey’ group were similar to those in the ‘Rabbit to Monkey’ group. At 1 month post-surgery, the levels of most tear inflammatory mediators had decreased, with the exception of IL-1β, TGF-β1 and IFN-γ in the allotransplantation group.ConclusionSmall-incision femtosecond laser-assisted intrastromal transplantation minimized invasiveness and improved surgical efficiency. In addition, the host cornea maintained a high level of biocompatibility. Glycerol-dehydrated corneal lamellae might be potentially useful as an alternative inlay xenogeneic material. In this study, we also describe a new treatment that can be used in keratoconus, corneal ectasia, presbyopia, hyperpresbyopia and other diseases.

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Long-term preservation of donor corneas in glycerol for keratoplasty: exploring new protocols

Abstract: Corneal tissue preserved at -80°C was the best method for preservation as it maintained the sterility, thickness, optical transparency, mechanical strength and ultrastructural features.

Cited by 26 publications

References 20 publications

Upgradation and modernization of eye banking services: Integrating tradition with innovative policies and current best practices

Upgradation and modernization of eye banking services: Integrating tradition with innovative policies and current best practices

Biobanking of Dehydrated Human Donor Corneal Stroma to Increase the Supply of Anterior Lamellar Grafts

Comparison of femtosecond laser-assisted corneal intrastromal xenotransplantation and the allotransplantation in rhesus monkeys

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