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Mulpuri L, Ouano DP, Riaz KM, Warner EJ, Stone DU, Cheung AY, Gomez A, Rangu N, Sabater AL, Tonk RS. Intracameral Enoxaparin for Descemet Membrane Endothelial Keratoplasty: A Pilot Safety Study. Cornea 2025; 44:342-349. [PMID: 39902781 DOI: 10.1097/ico.0000000000003662] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/16/2024] [Accepted: 07/04/2024] [Indexed: 02/06/2025]
Abstract
PURPOSE The purpose of this study was to investigate the safety and outcomes of Descemet membrane endothelial keratoplasty (DMEK) performed with intracameral enoxaparin. METHODS Two arms were used: a clinical multicenter retrospective cohort arm (CA) and an ex vivo basic science arm (BSA). In CA, DMEKs were performed by 6 experienced corneal surgeons at multiple sites. Intracameral enoxaparin (40 mg/500 mL) was added to the irrigation fluid for all cases. Primary outcomes were measured at 6 and 12 months. In BSA, mated graft pairs were randomized to control or enoxaparin exposure (0.8 mg/mL × 1 hour) and assessed for endothelial cell death count at 0-, 1-, and 24-hour intervals and cellular stress by ELISA Annexin V protein quantification. RESULTS In the cohort arm, the mean age of 159 eyes of 134 patients was 69.3 years with Fuchs dystrophy as the primary diagnosis. Mean BCVA improved from 0.42 ± 0.3 logMAR preoperatively to 0.13 ± 0.1 logMAR postoperatively at 6 months (P < 0.001) and to 0.1 ± 0.1 logMAR at 12 months (P < 0.001). At 6 months, 58.4% of patients achieved a final BCVA of 20/25 or better and 91% improved to 20/40 or better. Rebubble rate was 13% (n = 21), with 6 of these 21 eyes requiring more than 1 rebubble. One total graft detachment was noted with no reports of intraoperative or postoperative hemorrhage. PGF occurred in 0 of 159 eyes. In BSA, enoxaparin had no significant effect on endothelial cell death count or cellular apoptosis compared with control. CONCLUSIONS Enoxaparin can safely be used in DMEK surgery without apparent increased risk of intraoperative hemorrhage, graft detachment/failure, or endothelial cell toxicity.
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Affiliation(s)
- Lakshman Mulpuri
- Department of Ophthalmology, Cook County Health System, Chicago, IL
| | | | - Kamran M Riaz
- Department of Ophthalmology, Dean McGee Eye Institute, University of Oklahoma, Oklahoma City, OK
| | - Evan J Warner
- Department of Ophthalmology, University of Wisconsin School of Medicine and Public Health, Madison, WI
| | | | | | - Angela Gomez
- Department of Ophthalmology, Cook County Health System, Chicago, IL
| | - Neal Rangu
- Department of Ophthalmology, Dean McGee Eye Institute, University of Oklahoma, Oklahoma City, OK
| | | | - Rahul S Tonk
- Department of Ophthalmology, Cook County Health System, Chicago, IL
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Walshe JA, Schmid KL, Toalster N, McGowan CC, Ekwe AP, McKirdy NC, Harkin DG. Current and emerging strategies for the manufacture, implantation, and clinical management of corneal tissue allografts. Clin Exp Optom 2024:1-12. [PMID: 39648366 DOI: 10.1080/08164622.2024.2434626] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/27/2024] [Revised: 11/07/2024] [Accepted: 11/20/2024] [Indexed: 12/10/2024] Open
Abstract
Approximately 40,000 Australians have received a donor corneal tissue transplant over the last 40 years, with the primary indications being keratoconus, Fuchs' endothelial dystrophy, bullous keratopathy, and failure of a prior corneal transplant. Although corneal cross-linking and rigid contact lenses have emerged as alternative strategies for the management of keratoconus, the demand for donor corneas is increasing in-line with the ageing population in Australia. Moreover, owing to the lack of tissue banking resources in less-developed countries, the global demand for donor corneas exceeds supply by 70-fold. These supply issues, combined with evolving tissue banking and surgical techniques, have led to the emergence of new strategies for the storage, processing and implantation of corneal cells and tissues. Organ culture techniques have been developed that support the storage of donor corneas for up to 30 days, facilitating improvements in tissue supply and surgery scheduling. Bespoke surgical methods have been developed that are tailored to the requirements of specific conditions, allowing reductions in both the volume of tissue required to be transplanted and the size of the necessary surgical incision. Further efficiencies and improvements in patient care may be achieved via exploitation of cell culture technologies as exemplified through use of cultured corneal epithelial cells for the treatment of limbal stem cell deficiency. Promising progress has also been made in developing a cultured corneal endothelial cell therapy for patients with corneal endothelial dysfunction. These evolving strategies are discussed with respect to their potential impact on the clinical presentation and management of patients who have received an implant of donor corneal tissue or cells.
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Affiliation(s)
- Jennifer A Walshe
- Centre for Vision and Eye Research, Queensland University of Technology, Brisbane, Queensland, Australia
| | - Katrina L Schmid
- Centre for Vision and Eye Research, Queensland University of Technology, Brisbane, Queensland, Australia
| | - Nicholas Toalster
- Ophthalmology, Royal Brisbane and Women's Hospital, Brisbane, Queensland, Australia
| | - Ceara C McGowan
- Centre for Vision and Eye Research, Queensland University of Technology, Brisbane, Queensland, Australia
| | - Adaeze P Ekwe
- Centre for Vision and Eye Research, Queensland University of Technology, Brisbane, Queensland, Australia
| | - Natalie C McKirdy
- Centre for Vision and Eye Research, Queensland University of Technology, Brisbane, Queensland, Australia
| | - Damien G Harkin
- Centre for Vision and Eye Research, Queensland University of Technology, Brisbane, Queensland, Australia
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Sneyers A, Daas L, Zemova E, Quintin A, Munteanu C, Seitz B. Impact of Donor, Host, and Surgical Parameters on High Endothelial Cell Density More Than 5 Years after Penetrating Keratoplasty. Klin Monbl Augenheilkd 2024; 241:1341-1348. [PMID: 39142340 DOI: 10.1055/a-2349-0770] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 08/16/2024]
Abstract
OBJECTIVES To investigate the correlation between postoperative endothelial cell loss (ECL) and donor, host, and surgical parameters, and to assess the clinical impact of maintaining a high endothelial cell density (ECD) of ≥ 1500 cells/mm2 5 years after penetrating keratoplasty (PKP). METHODS This retrospective cohort study included 216 eyes with 5 years of follow-up, of which 94 had annual visits, and who underwent normal-risk elective PKP for noninfectious indications by one corneal microsurgeon (B. S.) between 2009 and 2016. RESULTS Among the 216 eyes, ECL (39.1%) over 5 years postoperative exhibited weak positive correlations with storage solution time (p = 0.024) and postmortem time (p = 0.028), and moderately positively correlations with the preoperative ECD (p < 0.001). The 5-year postoperative ECL differed significantly between in domo-prepared (36.8%) and ex domo donor corneas (46.3%; p = 0.001). In the 94 eyes, no significant differences were found between the two groups for central pupil pachymetry (CCT) and BCVA (p > 0.074). However, CCT increased significantly between 1 and 4 years (p = 0.034) and 1 and 5 years postoperatively (p = 0.012), respectively. BCVA improved significantly at 1 year postoperatively and continued to improve until 2 years postoperatively (p < 0.001). CONCLUSION The Lions corneal bank Saar-Lor-Lux achieved a significantly reduced ECL (36.8%) over 5 years compared to ex domo donor corneas (46.3%). A weak positive correlation was found between ECL with the storage solution time and the postmortem time, as well as a moderate positive correlation with the preoperative ECD. Although CCT increased significantly over 5 years, BCVA improved significantly from the first to the second postoperative year and remained stable thereafter.
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Affiliation(s)
- Albéric Sneyers
- Department of Ophthalmology, Saarland University Medical Centre (UKS), Homburg/Saar, Germany
| | - Loay Daas
- Department of Ophthalmology, Saarland University Medical Centre (UKS), Homburg/Saar, Germany
| | - Elena Zemova
- Department of Ophthalmology, Saarland University Medical Centre (UKS), Homburg/Saar, Germany
| | - Adrien Quintin
- Department of Ophthalmology, Saarland University Medical Centre (UKS), Homburg/Saar, Germany
| | - Cristian Munteanu
- Department of Ophthalmology, Saarland University Medical Centre (UKS), Homburg/Saar, Germany
| | - Berthold Seitz
- Department of Ophthalmology, Saarland University Medical Centre (UKS), Homburg/Saar, Germany
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Hatzfeld AS, Germain N, Maboudou P, Dhayer M, Marchetti P. Medical and economic impacts of managing corneas from older donors at the tissue bank-a single-center retrospective study spanning over 12 years. Front Med (Lausanne) 2024; 11:1415515. [PMID: 39512619 PMCID: PMC11540627 DOI: 10.3389/fmed.2024.1415515] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/10/2024] [Accepted: 10/11/2024] [Indexed: 11/15/2024] Open
Abstract
Aim To evaluate the impact of corneas from donors over 80 years of age on the activity of the North of France Tissue Bank and to determine the potential cost implications for banks using corneas from older donors. Methods We analyzed data from a single-center retrospective cohort study of 6,023 corneas preserved at the Lille Tissue Bank between 2012 and 2023. Donors, unrestricted by age, were divided into two groups: younger (≤ 80 years) and older (> 80 years). Corneas were categorized based on endothelial cell density (ECD). Data were collected from patients who underwent corneal transplantation. A financial impact model was created to assess the effects of using corneas from different age groups on the overall benefits of corneal transplant procedures. Results The average donor age was 67.5 ± 14.5 years. The median age of donors gradually increased from 66 to 73 years over the 12-year study period, with donors over 80 years old representing more than 24% since 2021. Corneas from older donors had a higher discard rate (62.53% vs. 39.66%) due to poor endothelial quality and serological concerns (both p < 0.0001). Additionally, these corneas had lower ECD, with a larger proportion deemed unsuitable for grafting due to low ECD (30% vs. 8.2%). Corneas from younger donors were more often used for endothelial transplants, which require higher ECD. The mean economic benefit per cornea showed a moderate negative correlation with donor age. The net benefit of corneal transplants decreased as the proportion of donors aged over 80 years increased. It is predicted that a net benefit of zero would be attained when the proportion of donors over 80 years is 44.4%. Conclusion Using corneas from donors over 80 years of age can help alleviate the shortage of donor tissue and be effective if certain quality standards are met. However, additional costs incurred by eye banks must be factored into this equation.
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Affiliation(s)
| | - Nicolas Germain
- Tissue Bank of Lille, Biology Pathology Center, CHU Lille, Lille, France
- UMR 9020-U1277, CANTHER, University Lille, CNRS, Lille, France
| | - Patrice Maboudou
- Biochemistry Unit, Biology Pathology Center, CHU Lille, Lille, France
| | - Mélanie Dhayer
- UMR 9020-U1277, CANTHER, University Lille, CNRS, Lille, France
| | - Philippe Marchetti
- Tissue Bank of Lille, Biology Pathology Center, CHU Lille, Lille, France
- UMR 9020-U1277, CANTHER, University Lille, CNRS, Lille, France
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Alsalloum A, Gornostal E, Mingaleva N, Pavlov R, Kuznetsova E, Antonova E, Nadzhafova A, Kolotova D, Kadyshev V, Mityaeva O, Volchkov P. A Comparative Analysis of Models for AAV-Mediated Gene Therapy for Inherited Retinal Diseases. Cells 2024; 13:1706. [PMID: 39451224 PMCID: PMC11506034 DOI: 10.3390/cells13201706] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/12/2024] [Revised: 10/13/2024] [Accepted: 10/14/2024] [Indexed: 10/26/2024] Open
Abstract
Inherited retinal diseases (IRDs) represent a diverse group of genetic disorders leading to progressive degeneration of the retina due to mutations in over 280 genes. This review focuses on the various methodologies for the preclinical characterization and evaluation of adeno-associated virus (AAV)-mediated gene therapy as a potential treatment option for IRDs, particularly focusing on gene therapies targeting mutations, such as those in the RPE65 and FAM161A genes. AAV vectors, such as AAV2 and AAV5, have been utilized to deliver therapeutic genes, showing promise in preserving vision and enhancing photoreceptor function in animal models. Despite their advantages-including high production efficiency, low pathogenicity, and minimal immunogenicity-AAV-mediated therapies face limitations such as immune responses beyond the retina, vector size constraints, and challenges in large-scale manufacturing. This review systematically compares different experimental models used to investigate AAV-mediated therapies, such as mouse models, human retinal explants (HREs), and induced pluripotent stem cell (iPSC)-derived retinal organoids. Mouse models are advantageous for genetic manipulation and detailed investigations of disease mechanisms; however, anatomical differences between mice and humans may limit the translational applicability of results. HREs offer valuable insights into human retinal pathophysiology but face challenges such as tissue degradation and lack of systemic physiological effects. Retinal organoids, on the other hand, provide a robust platform that closely mimics human retinal development, thereby enabling more comprehensive studies on disease mechanisms and therapeutic strategies, including AAV-based interventions. Specific outcomes targeted in these studies include vision preservation and functional improvements of retinas damaged by genetic mutations. This review highlights the strengths and weaknesses of each experimental model and advocates for their combined use in developing targeted gene therapies for IRDs. As research advances, optimizing AAV vector design and delivery methods will be critical for enhancing therapeutic efficacy and improving clinical outcomes for patients with IRDs.
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Affiliation(s)
- Almaqdad Alsalloum
- Federal Research Center for Innovator and Emerging Biomedical and Pharmaceutical Technologies, 125315 Moscow, Russia (P.V.)
| | | | - Natalia Mingaleva
- Moscow Center for Advanced Studies, Kulakova Str. 20, 123592 Moscow, Russia
| | - Roman Pavlov
- Moscow Center for Advanced Studies, Kulakova Str. 20, 123592 Moscow, Russia
| | | | - Ekaterina Antonova
- Moscow Center for Advanced Studies, Kulakova Str. 20, 123592 Moscow, Russia
| | - Aygun Nadzhafova
- Moscow Center for Advanced Studies, Kulakova Str. 20, 123592 Moscow, Russia
| | - Daria Kolotova
- Institute of Higher Nervous Activity and Neurophysiology, Russian Academy of Sciences, 117485 Moscow, Russia
| | | | - Olga Mityaeva
- Federal Research Center for Innovator and Emerging Biomedical and Pharmaceutical Technologies, 125315 Moscow, Russia (P.V.)
- Department of Fundamental Medicine, Lomonosov Moscow State University, 119992 Moscow, Russia
| | - Pavel Volchkov
- Federal Research Center for Innovator and Emerging Biomedical and Pharmaceutical Technologies, 125315 Moscow, Russia (P.V.)
- Department of Fundamental Medicine, Lomonosov Moscow State University, 119992 Moscow, Russia
- Moscow Clinical Scientific Center N.A. A.S. Loginov, 111123 Moscow, Russia
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Li Z, Böhringer D, Stachon T, Nastaranpour M, Fries FN, Seitz B, Ulrich M, Munteanu C, Langenbucher A, Szentmáry N. Culturing Limbal Epithelial Cells of Long-term Stored Corneal Donors (Organ Culture) In Vitro - A Stepwise Linear Regression Algorithm. Klin Monbl Augenheilkd 2024; 241:964-971. [PMID: 37130569 DOI: 10.1055/a-2084-7168] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 05/04/2023]
Abstract
PURPOSE To assess various potential factors on human limbal epithelial cell (LEC) outgrowth in vitro using corneal donor tissue following long-term storage (organ culture) and a stepwise linear regression algorithm. METHODS Of 215 donors, 304 corneoscleral rings were used for our experiments. For digestion of the limbal tissue and isolation of the limbal epithelial cells, the tissue pieces were incubated with 4.0 mg/mL collagenase A at 37 °C with 95% relative humidity and a 5% CO2 atmosphere overnight. Thereafter, limbal epithelial cells were separated from limbal keratocytes using a 20-µm CellTricks filter. The separated human LECs were cultured in keratinocyte serum-free medium medium, 1% penicillin/streptomycin (P/S), 0.02% epidermal growth factor (EGF), and 0.3% bovine pituitary extract (BPE). The potential effect of donor age (covariate), postmortem time (covariate), medium time (covariate), size of the used corneoscleral ring (360°, 270°180°, 120°, 90°, less than 90°) (covariate), endothelial cell density (ECD) (covariate), gender (factor), number of culture medium changes during organ culture (factor), and origin of the donor (donating institution and storing institution, factor) on the limbal epithelial cell outgrowth was analyzed with a stepwise linear regression algorithm. RESULTS The rate of successful human LEC outgrowth was 37.5%. From the stepwise linear regression algorithm, we found out that the relevant influencing parameters on the LEC growth were intercept (p < 0.001), donor age (p = 0.002), number of culture medium changes during organ culture (p < 0.001), total medium time (p = 0.181), and size of the used corneoscleral ring (p = 0.007), as well as medium time × size of the corneoscleral ring (p = 0.007). CONCLUSIONS The success of LEC outgrowth increases with lower donor age, lower number of organ culture medium changes during storage, shorter medium time in organ culture, and smaller corneoscleral ring size. Our stepwise linear regression algorithm may help us in optimizing LEC cultures in vitro.
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Affiliation(s)
- Zhen Li
- Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Congenital Aniridia Research, Saarland University, Homburg, Germany
| | - Daniel Böhringer
- Department of Ophthalmology, University of Freiburg, Freiburg im Breisgau, Germany
| | - Tanja Stachon
- Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Congenital Aniridia Research, Saarland University, Homburg, Germany
| | - Mahsa Nastaranpour
- Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Congenital Aniridia Research, Saarland University, Homburg, Germany
| | - Fabian Norbert Fries
- Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Congenital Aniridia Research, Saarland University, Homburg, Germany
- Department of Ophthalmology, Saarland University, Homburg, Germany
| | - Berthold Seitz
- Department of Ophthalmology, Saarland University, Homburg, Germany
| | - Myriam Ulrich
- Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Congenital Aniridia Research, Saarland University, Homburg, Germany
| | | | | | - Nóra Szentmáry
- Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Congenital Aniridia Research, Saarland University, Homburg, Germany
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Javadi F, Khorrami Z, Ashrafi S, Abolhosseini M, Kanavi MR, Safi S. Donor Risk Factors and Environmental Conditions Associated With Poor-Quality Corneas: An Analysis of the Central Eye Bank of Iran (2018-2021). Cornea 2024; 43:835-843. [PMID: 38016033 DOI: 10.1097/ico.0000000000003429] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/10/2023] [Accepted: 10/15/2023] [Indexed: 11/30/2023]
Abstract
PURPOSE The purpose of this study was to investigate the donor risk factors and environmental conditions associated with poor-quality corneas using the database of the Central Eye Bank of Iran over 4 years. METHODS This cohort study was conducted on the recorded data of all donated corneas at the Central Eye Bank of Iran database from March 2018 to March 2022. Donors' characteristics and tissue variables were extracted from the database. The final corneal quality was determined based on slitlamp biomicroscopic observations and the results of specular microscopy. Environmental variables were also obtained from reliable resources. Risk factors for poor-quality corneas were calculated using logistic mixed-effect regression analysis. All analyses were performed with STATA 17.0. The significance level of 0.05 was considered for all the analyses. RESULTS The data of 20,625 eyes of 10,601 donors were evaluated. We found that donor age had an inverse correlation with endothelial cell density (r = -0.28, P < 0.001). The trend of donated corneal poor quality decreased between 2018 and 2021. Several factors, including intoxication (odds ratio [OR] = 1.29), obesity (OR = 1.34), diabetes mellitus (OR = 1.63), hypertension (OR = 1.52), and pseudophakic eyes (OR = 1.56), were associated with the poor quality of donated corneal tissues. The outdoor temperature over 26°C was associated with higher odds of poor corneal quality (OR = 1.31), whereas high relative humidity decreased the odds of poor corneal quality (OR = 0.82). CONCLUSIONS This study revealed that the cause of donor death, obesity, donor comorbidities, pseudophakia, and environmental factors could affect the corneal quality and make the donated corneas less suitable for transplantation.
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Affiliation(s)
- Fatemeh Javadi
- Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Zahra Khorrami
- Ophthalmic Epidemiology Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Sadra Ashrafi
- Ophthalmic Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Mohammad Abolhosseini
- Ocular Tissue Engineering Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Mozhgan Rezaei Kanavi
- Ocular Tissue Engineering Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
- Central Eye Bank of Iran, Tehran, Iran; and
| | - Sare Safi
- Ophthalmic Epidemiology Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran
- Department of Optometry, School of Rehabilitation, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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Bu JB, Grabitz SD, Schön F, Apel M, Pusch T, Gericke A, Poplawski A, Pfeiffer N, Wasielica-Poslednik J. Utility of a Model to Predict Endothelial Cell Density of Donor Corneas to Determine Suitability for Transplantation. Transl Vis Sci Technol 2024; 13:21. [PMID: 39083373 PMCID: PMC11290561 DOI: 10.1167/tvst.13.7.21] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/07/2023] [Accepted: 06/02/2024] [Indexed: 08/02/2024] Open
Abstract
Purpose In Germany, approximately one-third of the harvested donor corneas are not suitable for transplantation, mostly due to insufficient endothelial cell density (ECD). The ECD can only be reliably determined after harvesting and processing of the cornea. Our group has previously developed a predictive model for corneal ECD: \( {Predicted\, ECD} = 2919-6^{\ast}\;{age}\; [{years}]-189\; [{if\, male}]\\ -7^{\ast}\;{death-to-explantation\, interval\,} [{hours}]\\ - 378\; [{if\, pseudophakic}] \;{cells/mm}^2 \). Methods A total of 2.999 consecutive donor corneas harvested between 2017 and 2021 from the Eye Bank of Rhineland-Palatinate in Mainz, Germany, were included. An actual ECD of >2000 cells/mm2 was defined as the cutoff value. To evaluate the clinical utility of the prognostic model as a screening instrument for transplant eligibility in an independent cohort, we performed a decision curve analysis. Results The median predicted ECD was 2061 cells/mm2 (interquartile range [IQR] = 1834 to 2221), whereas the median actual ECD was 2377 cells/mm2 (IQR = 1907 to 2624). There was a positive correlation between predicted and actual ECD (correlation coefficient = 0.411; P < 0.01). Our predictive model for ECD is a strong predictor for an actual ECD greater than 2000 (odds ratio = 1.374, 95% confidence interval [CI]) per 100 cells; P < 0.001, area under the curve [AUC] of 0.73). Decision curve analysis showed that the predictive model yielded a positive net benefit in clinical settings. Conclusions Decision curve analysis demonstrated a positive net benefit of the ECD predictive model in clinical settings with limited eye bank resources. Translational Relevance In possible scenarios where a choice between corneal grafts is required, or in countries with limited eye bank infrastructure and staff, the initial estimate of ECD from the formula may be beneficial.
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Affiliation(s)
- Julia B. Bu
- Department of Ophthalmology, University Medical Center of the Johannes Gutenberg-University Mainz, Germany
| | - Stephanie D. Grabitz
- Department of Ophthalmology, University Medical Center of the Johannes Gutenberg-University Mainz, Germany
| | - Franziska Schön
- Department of Ophthalmology, University Medical Center of the Johannes Gutenberg-University Mainz, Germany
| | - Melissa Apel
- Eye Bank of Rhineland-Palatinate, Koblenz, Germany
| | - Tobias Pusch
- Eye Bank of Rhineland-Palatinate, Koblenz, Germany
| | - Adrian Gericke
- Department of Ophthalmology, University Medical Center of the Johannes Gutenberg-University Mainz, Germany
| | - Alicia Poplawski
- Institute of Medical Biostatistics, Epidemiology and Informatics (IMBEI), University Medical Center of the Johannes Gutenberg-University Mainz, Mainz, Germany
| | - Norbert Pfeiffer
- Department of Ophthalmology, University Medical Center of the Johannes Gutenberg-University Mainz, Germany
| | - Joanna Wasielica-Poslednik
- Department of Ophthalmology, University Medical Center of the Johannes Gutenberg-University Mainz, Germany
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Romano V, Passaro ML, Ruzza A, Parekh M, Airaldi M, Levis HJ, Ferrari S, Costagliola C, Semeraro F, Ponzin D. Quality assurance in corneal transplants: Donor cornea assessment and oversight. Surv Ophthalmol 2024; 69:465-482. [PMID: 38199504 DOI: 10.1016/j.survophthal.2023.12.002] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/02/2023] [Revised: 12/12/2023] [Accepted: 12/29/2023] [Indexed: 01/12/2024]
Abstract
The cornea is the most frequently transplanted human tissue, and corneal transplantation represents the most successful allogeneic transplant worldwide. In order to obtain good surgical outcome and visual rehabilitation and to ensure the safety of the recipient, accurate screening of donors and donor tissues is necessary throughout the process. This mitigates the risks of transmission to the recipient, including infectious diseases and environmental contaminants, and ensures high optical and functional quality of the tissues. The process can be divided into 3 stages: (1) donor evaluation and selection before tissue harvest performed by the retrieval team, (2) tissue analysis during the storage phase conducted by the eye bank technicians after the retrieval, and, (3) tissue quality checks undertaken by the surgeons in the operating room before transplantation. Although process improvements over the years have greatly enhanced safety, quality, and outcome of the corneal transplants, a lack of standardization between centers during certain phases of the process still remains, and may impact on the quality and number of transplanted corneas. Here we detail the donor screening process for the retrieval teams, eye bank operators. and ophthalmic surgeons and examine the limitations associated with each of these stages.
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Affiliation(s)
- Vito Romano
- Eye Clinic, Department of Medical and Surgical Specialties, Radiological Sciences, and Public Health, University of Brescia, Brescia, Italy; Eye Clinic, ASST Spedali Civili di Brescia, Brescia, Italy; Department of Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy.
| | - Maria Laura Passaro
- Department of Neurosciences, Reproductive Sciences and Dentistry, University of Naples "Federico II", Naples, Italy; Department of Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy
| | - Alessandro Ruzza
- International Center for Ocular Physiopathology, Fondazione Banca Degli Occhi del Veneto Onlus, Venice, Italy; Department of Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy
| | - Mohit Parekh
- Schepens Eye Research Institute of Mass Eye and Ear, Dept. of Ophthalmology, Harvard Medical School, Boston, MA, USA; Department of Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy
| | - Matteo Airaldi
- Eye Clinic, Department of Medical and Surgical Specialties, Radiological Sciences, and Public Health, University of Brescia, Brescia, Italy; Eye Clinic, ASST Spedali Civili di Brescia, Brescia, Italy; Department of Neurosciences, Reproductive Sciences and Dentistry, University of Naples "Federico II", Naples, Italy; International Center for Ocular Physiopathology, Fondazione Banca Degli Occhi del Veneto Onlus, Venice, Italy; Schepens Eye Research Institute of Mass Eye and Ear, Dept. of Ophthalmology, Harvard Medical School, Boston, MA, USA; Department of Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy; Department of Eye and Vision Science, Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool, United Kingdom
| | - Hannah J Levis
- Department of Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy; Department of Eye and Vision Science, Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool, United Kingdom
| | - Stefano Ferrari
- International Center for Ocular Physiopathology, Fondazione Banca Degli Occhi del Veneto Onlus, Venice, Italy; Department of Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy
| | - Ciro Costagliola
- Department of Neurosciences, Reproductive Sciences and Dentistry, University of Naples "Federico II", Naples, Italy; Department of Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy
| | - Francesco Semeraro
- Eye Clinic, Department of Medical and Surgical Specialties, Radiological Sciences, and Public Health, University of Brescia, Brescia, Italy; Eye Clinic, ASST Spedali Civili di Brescia, Brescia, Italy; Department of Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy
| | - Diego Ponzin
- International Center for Ocular Physiopathology, Fondazione Banca Degli Occhi del Veneto Onlus, Venice, Italy; Department of Molecular and Translational Medicine, Università degli Studi di Brescia, Brescia, Italy
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10
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Polisetti N, Martin G, Ulrich E, Glegola M, Schlötzer-Schrehardt U, Schlunck G, Reinhard T. Influence of Organ Culture on the Characteristics of the Human Limbal Stem Cell Niche. Int J Mol Sci 2023; 24:16856. [PMID: 38069177 PMCID: PMC10706739 DOI: 10.3390/ijms242316856] [Citation(s) in RCA: 6] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/09/2023] [Revised: 11/23/2023] [Accepted: 11/24/2023] [Indexed: 12/18/2023] Open
Abstract
Organ culture storage techniques for corneoscleral limbal (CSL) tissue have improved the quality of corneas for transplantation and allow for longer storage times. Cultured limbal tissue has been used for stem cell transplantation to treat limbal stem cell deficiency (LSCD) as well as for research purposes to assess homeostasis mechanisms in the limbal stem cell niche. However, the effects of organ culture storage conditions on the quality of limbal niche components are less well described. Therefore, in this study, the morphological and immunohistochemical characteristics of organ-cultured limbal tissue are investigated and compared to fresh limbal tissues by means of light and electron microscopy. Organ-cultured limbal tissues showed signs of deterioration, such as edema, less pronounced basement membranes, and loss of the most superficial layers of the epithelium. In comparison to the fresh limbal epithelium, organ-cultured limbal epithelium showed signs of ongoing proliferative activity (more Ki-67+ cells) and exhibited an altered limbal epithelial phenotype with a loss of N-cadherin and desmoglein expression as well as a lack of precise staining patterns for cytokeratin ((CK)14, CK17/19, CK15). The analyzed extracellular matrix composition was mainly intact (collagen IV, fibronectin, laminin chains) except for Tenascin-C, whose expression was increased in organ-cultured limbal tissue. Nonetheless, the expression patterns of cell-matrix adhesion proteins varied in organ-cultured limbal tissue compared to fresh limbal tissue. A decrease in the number of melanocytes (Melan-A+ cells) and Langerhans cells (HLA-DR+, CD1a+, CD18+) was observed in the organ-cultured limbal tissue. The organ culture-induced alterations of the limbal epithelial stem cell niche might hamper its use in the treatment of LSCD as well as in research studies. In contrast, reduced numbers of donor-derived Langerhans cells seem associated with better clinical outcomes. However, there is a need to consider the preferential use of fresh CSL for limbal transplants and to look at ways of improving the limbal stem cell properties of stored CSL tissue.
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Affiliation(s)
- Naresh Polisetti
- Eye Center, Medical Center—Faculty of Medicine, University of Freiburg, Killianstrasse 5, 79106 Freiburg, Germany
| | - Gottfried Martin
- Eye Center, Medical Center—Faculty of Medicine, University of Freiburg, Killianstrasse 5, 79106 Freiburg, Germany
| | - Eva Ulrich
- Eye Center, Medical Center—Faculty of Medicine, University of Freiburg, Killianstrasse 5, 79106 Freiburg, Germany
| | - Mateusz Glegola
- Eye Center, Medical Center—Faculty of Medicine, University of Freiburg, Killianstrasse 5, 79106 Freiburg, Germany
| | - Ursula Schlötzer-Schrehardt
- Department of Ophthalmology, University Hospital Erlangen, Friedrich-Alexander-University of Erlangen-Nürnberg, Schwabachanlage 6, 91054 Erlangen, Germany
| | - Günther Schlunck
- Eye Center, Medical Center—Faculty of Medicine, University of Freiburg, Killianstrasse 5, 79106 Freiburg, Germany
| | - Thomas Reinhard
- Eye Center, Medical Center—Faculty of Medicine, University of Freiburg, Killianstrasse 5, 79106 Freiburg, Germany
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11
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Hopkinson C, Curnow E, Larkin DFP, Prydal J, Tuft S. Graphical comparison of surgeon outcomes for the audit of a national corneal transplant registry (OTAG study 32). Eye (Lond) 2023; 37:1236-1241. [PMID: 35590105 PMCID: PMC10101946 DOI: 10.1038/s41433-022-02100-8] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/31/2022] [Revised: 04/09/2022] [Accepted: 05/09/2022] [Indexed: 11/08/2022] Open
Abstract
PURPOSE To compare Kaplan-Meier survival curves and funnel plots for the audit of surgeon-specific corneal transplantation outcomes. METHODS We obtained data on all patients with Fuchs endothelial dystrophy (FED) receiving a first corneal transplant in one eye between January 2012 and December 2017. We produced 2-year Kaplan-Meier graft survival curves to compare a simulated individual surgeon's graft survival rate to national pooled data. We used funnel plots to compare all surgeon outcomes to the national graft survival rate with superimposed 95 and 99.8% confidence limits. We defined an outlier as a surgeon who performed ≥10 transplants and had graft survival below the 99.8% national lower limit. To assess the effect of the surgeon case mix, we also compared unadjusted and risk-adjusted graft survival rates. RESULTS There were 3616 first corneal transplants for FED patients with complete data, performed or overseen by 196 surgeons. The 2-year national graft survival rate was 88%. The median change from the unadjusted to the risk-adjusted graft survival rate for individual surgeons was 0% (IQR: 0%- -2%). Of the 108 surgeons who had performed ≥10 transplants, we identified two outliers based on the unadjusted graft survival funnel plot, compared to four outliers based on the risk-adjusted graft survival funnel plot. CONCLUSION Funnel plots provide a visually accessible method for comparing individual graft survival rates to the national rate. Risk-adjustment accounts for clinical factors, and this has advantages for audit and clinical governance.
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Affiliation(s)
- Cathy Hopkinson
- NHS Blood and Transplant, Fox Den Road, Stoke Gifford, Bristol, BS34 8RR, UK.
| | - Elinor Curnow
- NHS Blood and Transplant, Fox Den Road, Stoke Gifford, Bristol, BS34 8RR, UK
| | | | - Jeremy Prydal
- Department of Ophthalmology, University Hospitals of Leicester NHS Trust, Infirmary Square, Leicester, LE1 5WW, UK
| | - Stephen Tuft
- Moorfields Eye Hospital, 162 City Road, London, EC1V 2PD, UK
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12
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Chu HS, Hu FR, Liu HY, Srikumaran D. Keratoplasty Registries: Lessons Learned. Cornea 2023; 42:1-11. [PMID: 36459579 DOI: 10.1097/ico.0000000000003088] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/04/2022] [Accepted: 05/19/2022] [Indexed: 11/03/2022]
Abstract
ABSTRACT Clinical registries have been developed for decades in the field of ophthalmology, and they are especially well-suited to the study of keratoplasty practices. A comprehensive donor/recipient registry system can provide insight into donor, recipient, and surgical factors associated with immediate and long-term outcomes and adverse reactions. Furthermore, linkage with demographic databases can elucidate relationships with social determinants of health and potentially shape public policy. The vast sample size and multicenter nature of registries enable researchers to conduct sophisticated multivariate or multilayered analyses. In this review, we aim to emphasize the importance of registry data for keratoplasty practice and 1) summarize the structure of current keratoplasty registries; 2) examine the features and scientific contributions of the registries from Australia, the United Kingdom, Singapore, the Netherlands, Sweden, Eye Bank Association of America, and European Cornea and Cell Transplant registries; 3) compare registry-based studies with large single-site clinical studies; 4) compare registry-based studies with randomized control studies; and 5) make recommendations for future development of keratoplasty registries. Keratoplasty registries have increased our knowledge of corneal transplant practices and their outcomes. Future keratoplasty registry-based studies may be further strengthened by record linkage, data sharing, and international collaboration.
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Affiliation(s)
- Hsiao-Sang Chu
- Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan
- National Eye Bank of Taiwan, Ministry of Health and Welfare, Taipei, Taiwan
- Graduate Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei, Taiwan; and
- Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD
| | - Fung-Rong Hu
- Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan
- National Eye Bank of Taiwan, Ministry of Health and Welfare, Taipei, Taiwan
| | - Hsin-Yu Liu
- Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan
- National Eye Bank of Taiwan, Ministry of Health and Welfare, Taipei, Taiwan
- Graduate Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei, Taiwan; and
| | - Divya Srikumaran
- Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD
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13
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Kounatidou NE, Kopsini D, Gibbons A, Crane AM, Palioura S, Alfonso EC. Semi-Autologous Corneal Transplantation with Simultaneous Bilateral Surgery: A Case Report. Case Rep Ophthalmol 2023; 14:439-447. [PMID: 37901627 PMCID: PMC10601773 DOI: 10.1159/000531990] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/10/2023] [Accepted: 06/29/2023] [Indexed: 10/31/2023] Open
Abstract
The present report describes a case of semi-autologous corneal transplantation with bilateral surgery using two operating microscopes simultaneously. An 86-year-old man with history of six prior failed penetrating keratoplasties in his right eye presented with decreased vision. His other eye was deeply amblyopic but had a clear 30-year-old Castroviejo-square graft with an endothelial cell count of 803 cells/mm2. A semi-autologous graft was performed from the left eye to the right. Surgery was performed simultaneously on both eyes by two different surgeons using a standard ophthalmic operating microscope as well as a second ENT microscope. Upon trephination of the right failed corneal graft, vitreous opacities were noted and sent for culture. The semi-autologous tissue was directly transferred from the left eye to the right without any storage in preservation media to avoid endothelial cell loss. The semi-autologous graft remained clear in the immediate postoperative period. However, the vitreous cultures grew coagulase-negative Staphylococcus. Despite all efforts, the patient eventually developed a retinal detachment and vision in the right eye decreased to light perception. Autologous penetrating keratoplasty is an option for patients with loss of corneal function in a potentially seeing eye and a clear cornea in a contralateral eye with poor visual potential due to non-corneal disease. This case is unique in that part of the autologous penetrating keratoplasty had an old square graft in the center and corneal transplant surgery was done simultaneously in both eyes. It also highlights chronic indolent endophthalmitis as a potential cause of multiple graft failures.
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Affiliation(s)
- Nefeli E. Kounatidou
- Department of Ophthalmology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
| | | | - Allister Gibbons
- Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami, Miller School of Medicine, Miami, FL, USA
| | - Ashley M. Crane
- Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami, Miller School of Medicine, Miami, FL, USA
| | - Sotiria Palioura
- Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami, Miller School of Medicine, Miami, FL, USA
| | - Eduardo C. Alfonso
- Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami, Miller School of Medicine, Miami, FL, USA
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14
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Camposampiero D, Fasolo A, Saccon G, Donisi PM, Zanetti E, Ponzin D. Gram stain and addition of amphotericin B to improve the microbial safety of human donor corneas. Cell Tissue Bank 2022; 23:707-715. [PMID: 34791554 PMCID: PMC9675658 DOI: 10.1007/s10561-021-09981-1] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/29/2021] [Accepted: 11/07/2021] [Indexed: 12/19/2022]
Abstract
To determine the effectiveness of two methods to improve the microbial safety of human corneas preserved in organ culture. We compared the number of positive preservation solutions of corneas in organ culture in which the initial short-term hypothermic corneal maintenance solution was supplemented with amphotericin B 2.5 µg/mL and the historical data of microbial test results (2015-2019). In addition, we appraised the efficacy of Gram stain to detect bacterial or fungal contamination in the organ culture solutions of corneas from at-risk donors compared to the culture tests of corneas from not-at-risk donors. Statistical analysis was performed using STATA and statistical significance set at p < 0.05. The number of positive culture tests after preservation was 15 (0.5%) in 2020 compared to a mean of 37 (1.2%) in the period 2015-2019 (p < 0.01), with 10 (1.0%) positive samples in the cohort of 998 corneas from at-risk donors and 5 (0.2%) in the 2046 corneas from not-at-risk donors (p < 0.01). All corneas from at-risk donors tested positive at Gram stain and the results were available 1-3 days before those of the conventional culture tests. Amphotericin B supplementation in the short-term maintenance solution markedly reduced the number of positive microbial tests after organ culture and the early detection of contaminants, including slow-growing microorganisms, by Gram stain before the standard culture results. This meant fewer corneas being discarded and a greater likelihood of preventing post-graft infections.
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Affiliation(s)
- Davide Camposampiero
- The Veneto Eye Bank Foundation, Padiglione. G. Rama, Via Paccagnella 11, Zelarino, 30174 Venice, Italy
| | - Adriano Fasolo
- The Veneto Eye Bank Foundation, Padiglione. G. Rama, Via Paccagnella 11, Zelarino, 30174 Venice, Italy
| | - Giuseppe Saccon
- The Veneto Eye Bank Foundation, Padiglione. G. Rama, Via Paccagnella 11, Zelarino, 30174 Venice, Italy
| | - Pietro M. Donisi
- Pathological Anatomy Unit, SS. Giovanni e Paolo Hospital, Aulss3 Serenissima, Venice, Italy
| | - Elisa Zanetti
- The Veneto Eye Bank Foundation, Padiglione. G. Rama, Via Paccagnella 11, Zelarino, 30174 Venice, Italy
| | - Diego Ponzin
- The Veneto Eye Bank Foundation, Padiglione. G. Rama, Via Paccagnella 11, Zelarino, 30174 Venice, Italy
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15
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Hjortdal J, Griffin MD, Cadoux M, Armitage WJ, Bylesjo M, Gabhann PM, Murphy CC, Pleyer U, Tole D, Vabres B, Walkinshaw MD, Gourraud P, Karakachoff M, Brouard S, Degauque N. Peripheral blood immune cell profiling of acute corneal transplant rejection. Am J Transplant 2022; 22:2337-2347. [PMID: 35704290 PMCID: PMC9796948 DOI: 10.1111/ajt.17119] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/15/2022] [Revised: 05/17/2022] [Accepted: 06/09/2022] [Indexed: 01/25/2023]
Abstract
Acute rejection (AR) of corneal transplants (CT) has a profound effect on subsequent graft survival but detailed immunological studies in human CT recipients are lacking. In this multi-site, cross-sectional study, clinical details and blood samples were collected from adults with clinically diagnosed AR of full-thickness (FT)-CT (n = 35) and posterior lamellar (PL)-CT (n = 21) along with Stable CT recipients (n = 177) and adults with non-transplanted corneal disease (n = 40). For those with AR, additional samples were collected 3 months later. Immune cell analysis was performed by whole-genome microarrays (whole blood) and high-dimensional multi-color flow cytometry (peripheral blood mononuclear cells). For both, no activation signature was identified within the B cell and T cell repertoire at the time of AR diagnosis. Nonetheless, in FT- but not PL-CT recipients, AR was associated with differences in B cell maturity and regulatory CD4+ T cell frequency compared to stable allografts. These data suggest that circulating B cell and T cell subpopulations may provide insights into the regulation of anti-donor immune response in human CT recipients with differing AR risk. Our results suggest that, in contrast to solid organ transplants, genetic or cellular assays of peripheral blood are unlikely to be clinically exploitable for prediction or diagnosis of AR.
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Affiliation(s)
- Jesper Hjortdal
- Department of OphthalmologyAarhus University HospitalAarhusDenmark,Department of Clinical MedicineAarhus UniversityAarhusDenmark
| | - Matthew D. Griffin
- Regenerative Medicine Institute (REMEDI) at CÚRAM SFI Centre for Research in Medical DevicesSchool of Medicine, National University of Ireland GalwayGalwayIreland
| | - Marion Cadoux
- Nantes Université, INSERM, Center for Research in Transplantation and Translational Immunology, UMR 1064NantesFrance,CHU Nantes, Institut De Transplantation Urologie Néphrologie (ITUN)NantesFrance
| | - W. John Armitage
- Translational Health SciencesUniversity of BristolBristolUK,Tissue and Eye ServicesNHS Blood and TransplantBristolUK
| | - Max Bylesjo
- Fios Genomics Ltd, Nine Edinburgh BioquarterEdinburghUK
| | | | - Conor C. Murphy
- Royal Victoria Eye and Ear HospitalDublinIreland,Royal College of Surgeons in Ireland University of Medicine and Health SciencesDublinIreland
| | - Uwe Pleyer
- Department of OphthalmologyCharité University HospitalBerlinGermany
| | - Derek Tole
- University Hospitals Bristol NHS Foundations TrustBristol Eye HospitalBristolUK
| | - Bertrand Vabres
- Nantes Université, CHU Nantes, Service OphtalmologieNantesFrance
| | - Malcolm D. Walkinshaw
- Wellcome Centre for Cell Biology, School of Biological SciencesUniversity of EdinburghEdinburghUK
| | - Pierre‐Antoine Gourraud
- Nantes Université, INSERM, Center for Research in Transplantation and Translational Immunology, UMR 1064NantesFrance,CHU Nantes, Institut De Transplantation Urologie Néphrologie (ITUN)NantesFrance,CHU de Nantes, INSERM, CIC 1413, Pôle Hospitalo‐Universitaire 11: Santé Publique, Clinique des donnéesNantesFrance
| | - Matilde Karakachoff
- CHU de Nantes, INSERM, CIC 1413, Pôle Hospitalo‐Universitaire 11: Santé Publique, Clinique des donnéesNantesFrance
| | - Sophie Brouard
- Nantes Université, INSERM, Center for Research in Transplantation and Translational Immunology, UMR 1064NantesFrance,CHU Nantes, Institut De Transplantation Urologie Néphrologie (ITUN)NantesFrance
| | - Nicolas Degauque
- Nantes Université, INSERM, Center for Research in Transplantation and Translational Immunology, UMR 1064NantesFrance,CHU Nantes, Institut De Transplantation Urologie Néphrologie (ITUN)NantesFrance
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16
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TUTAŞ GÜNAYDIN N, TANYILDIZ B. Donor and recipient characteristics associated with rebubbling rate, endothelial cell loss, and graft failure in primary descemet membrane endothelial keratoplasty. JOURNAL OF HEALTH SCIENCES AND MEDICINE 2022. [DOI: 10.32322/jhsm.1158938] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/05/2022] Open
Abstract
Aim: To determine whether donor and recipient characteristics are associated with rebubbling rate, endothelial cell loss (ECL), and graft failure 3 years after primary Descemet membrane endothelial keratoplasty (DMEK).
Material and Method: Records of 295 consecutive DMEK surgery and match with corresponding donor data were reviewed at a tertiary referral clinic. Recipients with intraoperative complications and coexisting ocular pathologies were excluded. Age, sex of donor and recipient, cause of donor death, death-to-preservation time (DtPT), storage time, donor endothelial cell density (ECD), and indications for surgery were analyzed for correlation with rebubbling rate, postoperative ECL, and graft failure. Further, subgroup analyses of the cause of death, donor sex, DtPT (median value, 3.5 h), and indications were performed. Multiple regression and receiver operating characteristics (ROC) analysis were used to determine the independent risk factors for graft failure.
Results: This study included 114 eyes that underwent DMEK for bullous keratopathy (BK; 64%) and for Fuchs’ endothelial corneal dystrophy (FECD; 36%). The graft failure percentage was the only parameter that was higher in patients with DtPT > 3.5 h (p=0.047) than those with shorter DtPT. The probability of graft failure was seven times higher in eyes with DtPT > 3.5 h than with shorter DtPT (odds ratio 7.36, 95% confidence interval CI 1.34‒40.53) and 10 times higher in eyes with BK than those with FECD (odds ratio 10.29, 95% CI 1.01‒104.54).
Conclusion:. DtPT and recipients with BK diagnosis were found to be independent risk factors for graft failure. Therefore, surgeons should consider DtPT for DMEK in eyes with BK.
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Affiliation(s)
- Nesrin TUTAŞ GÜNAYDIN
- UNIVERSITY OF HEALTH SCIENCES, İSTANBUL KARTAL DR. LÜTFİ KIRDAR HEALTH RESEARCH CENTER
| | - Burak TANYILDIZ
- UNIVERSITY OF HEALTH SCIENCES, İSTANBUL KARTAL DR. LÜTFİ KIRDAR HEALTH RESEARCH CENTER
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17
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Downward L, Ahmed M, Hopkinson C, Romano V, Curnow E, Kaye SB. Endothelial failure and rejection in recipients of corneas from the same donor. BMJ Open Ophthalmol 2022; 7:bmjophth-2021-000965. [PMID: 36161852 PMCID: PMC9389126 DOI: 10.1136/bmjophth-2021-000965] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/30/2021] [Accepted: 04/08/2022] [Indexed: 11/12/2022] Open
Abstract
Objective To determine whether patients who receive corneas from the same donor have similar risks of endothelial failure and rejection. Methods and Analysis Patients with Fuchs endothelial dystrophy (FED) and pseudophakic bullous keratopathy (PBK) who received their first corneal transplant between 1999 and 2016 were analysed. Patients receiving corneas from donors who donated both corneas for the same indication were defined as ‘paired’. Gray’s test was used to compare the cumulative incidence of endothelial failure and rejection within 5 years post-transplant for ‘paired’ and ‘unpaired’ groups. Cox regression models were fitted to determine whether there was an association between recorded donor characteristics (endothelial cell density (ECD), age and sex and endothelial graft failure and rejection. Results 10 838 patients were analysed of whom 1536 (14%) were paired. The unpaired group comprised 1837 (69%) recipients of single corneal donors and 7465 (69%) donors who donated both corneas for another indication. ECD was lower for unpaired single cornea donors (p<0.01). There was no significant difference in endothelial graft failure or rejection between paired and unpaired groups for FED (p=0.37, p=0.99) or PBK (p=0.88, p=0.28) nor for donor ECD, age, sex and paired donation after adjusting for transplant factors (across all models p>0.16 for ECD, p>0.32 for donor age, p>0.14 for sex match and p>0.17 for the donor effect). Conclusion The absence of a significant difference in graft outcome for corneal transplants for FED and PBK between paired and unpaired donors may reflect a homogeneous donor pool in the UK.
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Affiliation(s)
- Lewis Downward
- Statistics, NHS Blood and Transplant Organ Donation and Transplantation Directorate, Bristol, UK
| | - Mahmoud Ahmed
- Ophthalmology, Royal Liverpool University Hospital, Liverpool, UK
| | - Cathy Hopkinson
- Statistics, NHS Blood and Transplant Organ Donation and Transplantation Directorate, Bristol, UK
| | - Vito Romano
- Department of Eye and Vision Science, University of Liverpool, Liverpool, UK
| | - Elinor Curnow
- Statistics, NHS Blood and Transplant Organ Donation and Transplantation Directorate, Bristol, UK
| | - Stephen B Kaye
- Ophthalmology, Royal Liverpool University Hospital, Liverpool, UK
- Department of Eye and Vision Science, University of Liverpool, Liverpool, UK
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18
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Steger B, Kaye SB, Romano V. Corneal transplantation: the fine line between donor shortage and tissue quality. BMJ Open Ophthalmol 2022; 7:e001046. [PMID: 36161834 PMCID: PMC9389094 DOI: 10.1136/bmjophth-2022-001046] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/24/2022] Open
Affiliation(s)
- Bernhard Steger
- Department of Ophthalmology, Medizinische Universitat Innsbruck, Innsbruck, Austria
| | - Stephen B Kaye
- Department of Eye and Vision Science, University of Liverpool, Liverpool, UK
| | - Vito Romano
- Department of Ophthalmology, University of Brescia, Brescia, Italy
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19
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P-Cadherin Is Expressed by Epithelial Progenitor Cells and Melanocytes in the Human Corneal Limbus. Cells 2022; 11:cells11121975. [PMID: 35741104 PMCID: PMC9221557 DOI: 10.3390/cells11121975] [Citation(s) in RCA: 10] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/12/2022] [Revised: 06/09/2022] [Accepted: 06/13/2022] [Indexed: 12/13/2022] Open
Abstract
Interactions between limbal epithelial progenitor cells (LEPC) and surrounding niche cells, which include limbal mesenchymal stromal cells (LMSC) and melanocytes (LM), are essential for the maintenance of the limbal stem cell niche required for a transparent corneal surface. P-cadherin (P-cad) is a critical stem cell niche adhesion molecule at various epithelial stem cell niches; however, conflicting observations were reported on the presence of P-cad in the limbal region. To explore this issue, we assessed the location and phenotype of P-cad+ cells by confocal microscopy of human corneoscleral tissue. In subsequent fluorescence-activated cell sorting (FACS) experiments, we used antibodies against P-cad along with CD90 and CD117 for the enrichment of LEPC, LMSC and LM, respectively. The sorted cells were characterized by immunophenotyping and the repopulation of decellularized limbal scaffolds was evaluated. Our findings demonstrate that P-cad is expressed by epithelial progenitor cells as well as melanocytes in the human limbal epithelial stem cell niche. The modified flow sorting addressing P-cad as well as CD90 and CD117 yielded enriched LEPC (CD90−CD117−P-cad+) and pure populations of LMSC (CD90+CD117−P-cad−) and LM (CD90−CD117+P-cad+). The enriched LEPC showed the expression of epithelial progenitor markers and better colony-forming ability than their P-cad− counterparts. The cultured LEPC and LM exhibited P-cad expression at intercellular junctions and successfully repopulated decellularized limbal scaffolds. These data suggest that P-cad is a critical cell–cell adhesion molecule, connecting LEPC and LM, which may play an important role in the long-term maintenance of LEPC at the limbal stem cell niche; moreover, these findings led to further improvement of cell enrichment protocols to enhance the yield of LEPC.
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20
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Gram N, Shehab A, Ivarsen A, Hjortdal J. Influence of time to procurement, incubation and release of organ cultured donor corneas on graft failure after Descemet Stripping Automated Endothelial Keratoplasty. Acta Ophthalmol 2022; 100:414-421. [PMID: 34318589 DOI: 10.1111/aos.14994] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/20/2021] [Accepted: 07/16/2021] [Indexed: 11/28/2022]
Abstract
PURPOSE The aim of the present study was to investigate whether the time from death to procurement, to preservation or the storage time of donor corneas preserved in organ culture influenced the clinical outcome of patients undergoing Descemet stripping automated endothelial keratoplasty (DSAEK) for Fuchs endothelial keratoplasty. METHODS We conducted a registry-based study on 776 patients undergoing DSAEK. Data on time from donor death to cornea retrieval (DRT), time from death to preservation (DPT), the preservation time and donor cornea characteristics: age, sex and endothelial cell density (ECD) at the time of release for surgery, were extracted from The Danish Cornea Bank Registry. Data on recipient follow-up were collected from a corneal graft registry. The primary outcome was presence of graft failure within a period from 2 months to 2 years after surgery. Secondary outcomes were DRT, DPT, ECD ≤2300 and gender mismatch between donor and recipient. RESULTS Graft failure occurred in 26 patients. The mean preservation time for failed grafts was 34.1 ± 10.0 days (mean ± SD) and 27.3 ± 10.6 days (mean ± SD) for the clear, functional grafts at the 2-year follow-up. A preservation time of >29 days compared with ≤29 days was associated with a lower survival (HR 2.33, 95% CI on 1.06-5.14, p = 0.036) and an increased risk of graft failure (RR 1.53, 95% CI on 1.11-2.10, p = 0.009). For the secondary outcome variables, no difference in the risk of graft failure was observed and did not appear to impact the survival rate of DSAEK patients. CONCLUSION Preservation time of donor cornea was associated with graft survival and a prolonged preservation time of more than 4 weeks seemed to lower the 2-year survival.
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Affiliation(s)
- Niels Gram
- Department of Ophthalmology Aarhus University Hospital Aarhus Denmark
- Department of Clinical Medicine Aarhus University Aarhus Denmark
| | - Anders Shehab
- Department of Ophthalmology Aarhus University Hospital Aarhus Denmark
- Department of Clinical Medicine Aarhus University Aarhus Denmark
| | - Anders Ivarsen
- Department of Ophthalmology Aarhus University Hospital Aarhus Denmark
- Department of Clinical Medicine Aarhus University Aarhus Denmark
| | - Jesper Hjortdal
- Department of Ophthalmology Aarhus University Hospital Aarhus Denmark
- Department of Clinical Medicine Aarhus University Aarhus Denmark
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Shehab A, Gram N, Ivarsen A, Hjortdal J. The importance of donor characteristics, post-mortem time and preservation time for use and efficacy of donated corneas for posterior lamellar keratoplasty. Acta Ophthalmol 2022; 100:269-276. [PMID: 34173345 DOI: 10.1111/aos.14943] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/20/2021] [Accepted: 05/20/2021] [Indexed: 11/30/2022]
Abstract
PURPOSE The aim of this study is to examine whether donor age, death-to-retrieval time (DRT) and death-to-preservation time (DPT) as well as total preservation time affect donor cornea suitability for endothelial keratoplasty (EK) or penetrating keratoplasty (PK). METHODS A registry-based study was performed identifying 3248 corneas donated between 2011 and 2017. Data regarding donated corneas were extracted from The Danish Cornea Bank and donor medical records and evaluated for missing information. The primary outcome was whether ECD at preservation (ECD-P) or at release (ECD-R) was >2000 cells/mm2 . RESULTS Logistic regression for ECD-P showed a significant negative effect of increasing age (OR: 1.07, 95%CI: 1.05;1.08, p < 0.001) on donor suitability. Higher ECD-P had a significant positive effect on graft eligibility (OR: 1.007 95%CI: 1.003;1.010, p < 0.001). No significant effect of donor sex (p = 0.547), DRT (p = 0.289) or DPT (p = 0.102) on donor suitability for EK or PK (Chi-squared test). CONCLUSION High donor age and low ECD-P negatively affect the suitability of donor corneas for EK/PK whereas DRT and DPT did not affect graft suitability.
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Affiliation(s)
- Anders Shehab
- Department of Ophthalmology Aarhus University Hospital Aarhus Denmark
- Department of Clinical Medicine Aarhus University Aarhus Denmark
| | - Niels Gram
- Department of Ophthalmology Aarhus University Hospital Aarhus Denmark
- Department of Clinical Medicine Aarhus University Aarhus Denmark
| | - Anders Ivarsen
- Department of Ophthalmology Aarhus University Hospital Aarhus Denmark
- Department of Clinical Medicine Aarhus University Aarhus Denmark
| | - Jesper Hjortdal
- Department of Ophthalmology Aarhus University Hospital Aarhus Denmark
- Department of Clinical Medicine Aarhus University Aarhus Denmark
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22
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Xu TT, Cao R, Dong YL, Xie LX, Cheng J. Analysis of risk factors of rapid attenuation of graft endothelium in the early stage after penetrating keratoplasty. PLoS One 2022; 17:e0266072. [PMID: 35381040 PMCID: PMC8982837 DOI: 10.1371/journal.pone.0266072] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/19/2021] [Accepted: 12/25/2021] [Indexed: 01/27/2023] Open
Abstract
This study aimed to analyze the factors of rapid attenuation of graft endothelium in the early stage after penetrating keratoplasty (PKP), with a view to guiding patients with PKP to better long-term outcomes. This study included 226 patients who underwent PKP with follow-up time >1 year at the Qingdao Eye Hospital of Shandong First Medical University from January 2018 to June 2020. Medical records were retrospectively studied, and donor factors, patient factors, and surgical factors were comparatively analyzed to clarify those affecting the rapid decay of graft endothelium after PKP. The median time between excision and death >60 min and patient age >60 years were risk factors for endothelial cell loss (ECL) rate >30% at 1 month postoperatively. However, a higher percentage of patients with donor age ≤60 years and Optisol preservation solution had endothelial cell density (ECD) >2000 cells/mm2 in the graft at postoperative 1 year. A year after the surgery, patients with corneal endothelial decompensation and immune rejection were at risk for ECD < 1000 cells/mm2. The combined operations had a significant effect on the ECL in the early postoperative period. Patients who underwent combined extracapsular cataract extraction or intraocular lens implantation had a significantly higher rate of ECL at postoperative 1 month than other patients, and no significant effect at postoperative 1 year. However, patients without combined operations have a higher probability of maintaining a high level of graft ECD. The graft diameter also affected postoperative ECL. In patients with a larger graft diameter, attenuation of ECD was slower. The ultimate goal of PKP is to maintain graft transparency for extended periods. The use of younger donors, minimizing unnecessary operation in the anterior chamber, and minimizing immune rejection may maintain a greater donor corneal endothelium in the long term.
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Affiliation(s)
- Ting-ting Xu
- Weifang Medical University, Institute of Clinical Medicine, Weifang, China
- State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Qingdao, China
| | - Rui Cao
- State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Qingdao, China
- Eye Institute of Shandong First Medical University, Qingdao Eye Hospital of Shandong First Medical University, Qingdao, China
- School of Ophthalmology, Shandong First Medical University, Jinan, China
| | - Yan-ling Dong
- State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Qingdao, China
- Eye Institute of Shandong First Medical University, Qingdao Eye Hospital of Shandong First Medical University, Qingdao, China
- School of Ophthalmology, Shandong First Medical University, Jinan, China
| | - Li-xin Xie
- State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Qingdao, China
- Eye Institute of Shandong First Medical University, Qingdao Eye Hospital of Shandong First Medical University, Qingdao, China
- School of Ophthalmology, Shandong First Medical University, Jinan, China
| | - Jun Cheng
- State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Qingdao, China
- Eye Institute of Shandong First Medical University, Qingdao Eye Hospital of Shandong First Medical University, Qingdao, China
- School of Ophthalmology, Shandong First Medical University, Jinan, China
- * E-mail:
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Romano V, Parekh M, Virgili G, Coco G, Leon P, Islein K, Ponzin D, Ferrari S, Fasolo A, Yu AC, Lucenteforte E, Busin M, Kaye SB. Gender Matching Did Not Affect 2-year Rejection or Failure Rates Following DSAEK for Fuchs Endothelial Corneal Dystrophy. Am J Ophthalmol 2022; 235:204-210. [PMID: 34626575 DOI: 10.1016/j.ajo.2021.09.029] [Citation(s) in RCA: 6] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/29/2021] [Revised: 09/29/2021] [Accepted: 09/29/2021] [Indexed: 11/01/2022]
Abstract
PURPOSE To investigate whether donor to recipient gender or H-Y mismatching was associated with graft rejection or failure following Descemet stripping automated endothelial keratoplasty (DSAEK) in patients with Fuchs endothelial corneal dystrophy (FECD). DESIGN Clinical cohort study. METHODS This study used a multi-center registry including patients aged >18 years who had undergone their first DSAEK for FECD between January 2008 and March 2018. The impact of donor and recipient gender incompatibility (including H-Y mismatches) on corneal graft rejection and failure was evaluated using Kaplan-Meier curves and univariable and multivariable Cox models. RESULTS Outcome data from 4341 eyes (3915 from the UK and 426 from Italy) were analyzed. Graft failure at 2-year follow-up occurred in 477 (11.0%) cases. Graft rejection at 2-year follow-up occurred in 175 cases (4.0%), 58 (1.3%) of whom developed graft failure. There was no significant effect of gender or H-Y mismatching on either rejection (P = .12, P = .06) or failure (P = .28, P = .14), respectively. CONCLUSIONS In patients with FECD undergoing DSAEK, no significant influence of gender and or H-Y mismatch on graft rejection or failure was found.
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Shah R, Hirwe S, Ashar J, Sengupta S. Utilization rate of corneal tissue obtained from donors over 75 years of age in Western India for keratoplasty. Indian J Ophthalmol 2022; 70:511-515. [PMID: 35086227 PMCID: PMC9023935 DOI: 10.4103/ijo.ijo_1329_21] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/18/2022] Open
Abstract
Purpose: To examine the utilization patterns of cornea procured from diseased individuals ≥75 years of age at an eye bank in western India. Methods: In this retrospective study, data from 1,217 eyes of 653 donors with age ≥75 years were reviewed from October 2008 to December 2019. Donor age, lens status, endothelial cell count (ECD), utilization of the tissue for transplantation or non-clinical purposes (e.g., research, training/discarded), and causes of non-utilization were noted. Results: The mean age of the donors was 80.9 ± 4.6 years and the tissue utilization rate was 36.5% (445 out of 1,217 eyes). The eyes used for keratoplasty procedures had a lower donor age (79.6 ± 5.7 vs. 81.5 ± 5.1; P < 0.001), a higher endothelial cell count (2493 ± 531 vs. 2034 ± 581; P < 0.001), and were more often phakic (61% vs. 36.6%) compared to the unused group. A multivariable logistic regression analysis showed that the likelihood of tissue utilization for keratoplasty was 13% higher with every 100-cell increment in donor ECD (odds ratio [OR] = 1.13, 95% CI = 1.10–1.16, P < 0.001) and 33% lower with having a pseudophakic status in the donor eye (OR = 0.67, 95% CI = 0.52–0.87, P = 0.03). Age was not a significant determinant of tissue utilization when used in the same multivariable model. Conclusion: More than one-third of the eyes (36.5%) can be utilized even when the donors are above 75 years of age. Eyes that were more likely to be utilized for keratoplasty were phakic and had a significantly higher ECD; age was not a determinant in tissue utilization.
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Affiliation(s)
- Rakesh Shah
- Swaraashi Netralaya, Mumbai, Maharashtra, India
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25
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Chen Y, Dong L, Kong B, Huang Y, Zhong S, Connon C, Tan J, Yang S, Sun W, Mi S. Effects of Gelatin Methacrylate Hydrogel on Corneal Repair and Regeneration in Rats. Transl Vis Sci Technol 2021; 10:25. [PMID: 34935910 PMCID: PMC8711000 DOI: 10.1167/tvst.10.14.25] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/03/2022] Open
Abstract
Purpose This study investigates the repairing process of rat cornea after surgery of lamellar keratoplasty (LKP) and evaluates the effects of gelatin methacrylate (GelMA) hydrogel. Methods In the LKP group, the lamellar stroma matrixes of Sprague-Dawley rats were transplanted to enhanced green fluorescent protein rats, whereas those in the GelMA group were also embedded with a GelMA hydrogel during the corneal transplantation. Grafted eyes were harvested on days seven, 30, and 90. Hematoxylin and eosin staining, immunofluorescence staining, scanning electron microscopy, optical coherence tomography, and a slit-lamp microscope were used to study the process of corneal restoration and regeneration. Results A total of 42 rats were analyzed, including 18 rats in each of the experimental group and six rats in the control group. After three months, the infiltration degree of inflammatory cells differed between the LKP group and the GelMA group (P < 0.001). Moreover, in multiple comparisons in corneal thickness, significant difference was observed between the LKP group and the GelMA group. There was also divergence in the results between the LKP group and the control group (P < 0.001, P < 0.001). At the same time, the expression of α-smooth muscle actin (α-SMA) and transforming growth factor (TGF)-β1 varied distinctly between the LKP group and the GelMA group (P < 0.05, P < 0.001). Conclusions Significant differences were demonstrated between the LKP group and the GelMA group in inflammatory cell infiltration, corneal thickness, as well as the expression of α-SMA and TGF-β1. Those differences indicate the ability of GelMA hydrogel to support alleviation in corneal stroma fibrosis and show the influences of fibrosis in the dysfunction of corneal refractive power. Translational Relevance Our research provides new ideas for the future development of LKP and tissue-engineered corneas.
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Affiliation(s)
- Yun Chen
- Macromolecular Platforms for Translational Medicine and Bio-Manufacturing Laboratory, Tsinghua-Berkeley Shenzhen Institute, Shenzhen, P.R. China.,Open FIESTA Center, International Graduate School at Shenzhen, Tsinghua University, Shenzhen, P.R. China
| | - Lina Dong
- Macromolecular Platforms for Translational Medicine and Bio-Manufacturing Laboratory, Tsinghua-Berkeley Shenzhen Institute, Shenzhen, P.R. China
| | - Bin Kong
- Macromolecular Platforms for Translational Medicine and Bio-Manufacturing Laboratory, Tsinghua-Berkeley Shenzhen Institute, Shenzhen, P.R. China
| | - Yu Huang
- Biomanufacturing Engineering Laboratory, International Graduate School at Shenzhen, Tsinghua University, Shenzhen, P.R. China
| | - Suyi Zhong
- Institute of Optical Imaging and Sensing, Shenzhen Key Laboratory for Minimal Invasive Medical Technologies, Graduate School at Shenzhen, Tsinghua University, Shenzhen, P.R. China
| | - Che Connon
- Biosciences Institute, Newcastle University
| | - Jiaqi Tan
- Open FIESTA Center, International Graduate School at Shenzhen, Tsinghua University, Shenzhen, P.R. China
| | - Siming Yang
- Key Laboratory of Wound Repair and Regeneration of PLA, Chinese PLA General Hospital, Medical College of PLA, Beijing, P.R. China
| | - Wei Sun
- Macromolecular Platforms for Translational Medicine and Bio-Manufacturing Laboratory, Tsinghua-Berkeley Shenzhen Institute, Shenzhen, P.R. China.,Department of Mechanical Engineering, Biomanufacturing Center, Tsinghua University, Beijing, P.R. China.,Department of Mechanical Engineering, Drexel University, Philadelphia, PA, USA
| | - Shengli Mi
- Macromolecular Platforms for Translational Medicine and Bio-Manufacturing Laboratory, Tsinghua-Berkeley Shenzhen Institute, Shenzhen, P.R. China.,Open FIESTA Center, International Graduate School at Shenzhen, Tsinghua University, Shenzhen, P.R. China.,Department of Mechanical Engineering, Biomanufacturing Center, Tsinghua University, Beijing, P.R. China
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Olszewski C, Maassen J, Guenther R, Skazik-Voogt C, Gutermuth A. Mechanotransductive Differentiation of Hair Follicle Stem Cells Derived from Aged Eyelid Skin into Corneal Endothelial-Like Cells. Stem Cell Rev Rep 2021; 18:1668-1685. [PMID: 34515937 PMCID: PMC9209348 DOI: 10.1007/s12015-021-10249-0] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 08/23/2021] [Indexed: 11/25/2022]
Abstract
Corneal endothelial insufficiency is one of the leading causes of blindness. The main contemporary treatment for corneal blindness is endothelial keratoplasty, which, however, is unsatisfactory as a medical therapy due to the lack of donor corneas and graft rejection. Therefore, autologous stem cell-based corneal endothelial tissue substitutes may be a promising alternative to conventional grafts in the future. To address the age of most patients suffering from corneal endothelial deficiencies, we investigated the presence and potential of hair-derived stem cells from older tissue donors. Our studies revealed the presence of pluripotency- and neural crest-associated markers in tissue sections from blepharoplasty patients aged 50 to 80 years. In vitro outgrowths from eyelid hair follicles on collagen-coated tissue culture plates revealed a weak decrease in stem-cell potency. In contrast, cells within the spheres that spontaneously formed from the adherent cell layer retained full stem-cell potency and could be differentiated into cells of the ecto- meso and endodermal lineages. Although these highly potent hair follicle derived stem cells (HFSC) were only very slightly expandable, they were able to recognize the biomimicry of the Descemet’s-like topography and differentiate into corneal endothelial-like cells. In conclusion, HFSCs derived from epidermal skin of eyelid biopsies are a promising cell source to provide autologous corneal endothelial replacement for any age group of patients.
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Affiliation(s)
- Christian Olszewski
- Fraunhofer Institute for Production Technology, Steinbachstraße 17, 52074, Aachen, Germany
| | - Jessika Maassen
- Fraunhofer Institute for Production Technology, Steinbachstraße 17, 52074, Aachen, Germany
| | - Rebecca Guenther
- Fraunhofer Institute for Production Technology, Steinbachstraße 17, 52074, Aachen, Germany
| | - Claudia Skazik-Voogt
- Fraunhofer Institute for Production Technology, Steinbachstraße 17, 52074, Aachen, Germany
| | - Angela Gutermuth
- Fraunhofer Institute for Production Technology, Steinbachstraße 17, 52074, Aachen, Germany.
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Romo-Valera C, Pérez-Garrastachu M, Hernáez-Moya R, Rodriguez-Astigarraga M, Romano-Ruiz P, Etxebarria J, Arluzea J, Andollo N. Characterisation of corneas following different time and storage methods for their use as a source of stem-like limbal epithelial cells. Exp Eye Res 2021; 211:108720. [PMID: 34389315 DOI: 10.1016/j.exer.2021.108720] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/29/2021] [Revised: 07/21/2021] [Accepted: 08/05/2021] [Indexed: 12/13/2022]
Abstract
The transplantation of expansions of limbal epithelial stem cells (LESC) remains one of the most efficient therapies for the treatment of limbal stem cell deficiency (LSCD) to date. However, the available donor corneas are scarce, and the corneas conserved for long time, under hypothermic conditions (after 7 days) or in culture (more than 28 days), are usually discarded due to poor viability of the endothelial cells. To establish an objective criterion for the utilisation or discarding of corneas as a source of LESC, we characterized, by immunohistochemistry analysis, donor corneas conserved in different conditions and for different periods of time. We also studied the potency of LESCs isolated from these corneas and maintained in culture up to 3 cell passages. We hoped that the study of markers of LESCs present in both the corneoscleral histological sections and the cell cultures would show the adequacy of the methods used for cell isolation and how fit the LESC enrichment of the obtained cell populations to be expanded was. Thus, the expressions of markers of the cells residing in the human limbal and corneal epithelium (cytokeratin CK15 and CK12, vimentin, Collagen VII, p63α, ABCG2, Ki67, Integrin β4, ZO1, and melan A) were analysed in sections of corneoscleral tissues conserved in hypothermic conditions for 2-9 days with post-mortem time (pmt) < 8 h or for 1 day with pmt > 16 h, and in sclerocorneal rims maintained in an organ culture medium for 29 days. Cell populations isolated from donor corneoscleral tissues were also assessed based on these markers to verify the adequacy of isolation methods and the potential of expanding LESCs from these tissues. Positivity for several putative stem cell markers such as CK15 and p63α was detected in all corneoscleral tissues, although a decrease was recorded in the ones conserved for longer times. The barrier function and the ability to adhere to the extracellular matrix were maintained in all the analysed tissues. In limbal epithelial cell cultures, a simultaneous decrease in the melan A melanocyte marker and the putative stem cell markers was detected, suggesting a close relationship between the melanocytes and the limbal stem cells of the niche. Holoclones stained with putative stem cell markers were obtained from long-term, hypothermic, stored sclerocorneal rims. The results showed that the remaining sclerocorneal rims after corneal transplantation, which were conserved under hypothermic conditions for up to 7 days and would have been discarded at a first glance, still maintained their potential as a source of LESC cultures.
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Affiliation(s)
- Cristina Romo-Valera
- Department of Cell Biology and Histology, School of Medicine and Nursing, University of the Basque Country, Sarriena, S/N, 48940, Leioa, Spain
| | - Miguel Pérez-Garrastachu
- Department of Cell Biology and Histology, School of Medicine and Nursing, University of the Basque Country, Sarriena, S/N, 48940, Leioa, Spain
| | - Raquel Hernáez-Moya
- Department of Cell Biology and Histology, School of Medicine and Nursing, University of the Basque Country, Sarriena, S/N, 48940, Leioa, Spain
| | - Maddalen Rodriguez-Astigarraga
- Department of Cell Biology and Histology, School of Medicine and Nursing, University of the Basque Country, Sarriena, S/N, 48940, Leioa, Spain
| | - Paula Romano-Ruiz
- Department of Cell Biology and Histology, School of Medicine and Nursing, University of the Basque Country, Sarriena, S/N, 48940, Leioa, Spain
| | - Jaime Etxebarria
- Department of Cell Biology and Histology, School of Medicine and Nursing, University of the Basque Country, Sarriena, S/N, 48940, Leioa, Spain; Department of Ophthalmology, University Hospital of Cruces, Cruces Plaza S/N, 48903, Barakaldo, Spain; BioCruces Bizkaia Health Research Institute, Begiker, Cruces Plaza S/N, 48903, Barakaldo, Spain
| | - Jon Arluzea
- Department of Cell Biology and Histology, School of Medicine and Nursing, University of the Basque Country, Sarriena, S/N, 48940, Leioa, Spain
| | - Noelia Andollo
- Department of Cell Biology and Histology, School of Medicine and Nursing, University of the Basque Country, Sarriena, S/N, 48940, Leioa, Spain; BioCruces Bizkaia Health Research Institute, Begiker, Cruces Plaza S/N, 48903, Barakaldo, Spain.
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How to Predict the Suitability for Corneal Donorship? J Clin Med 2021; 10:jcm10153426. [PMID: 34362207 PMCID: PMC8347780 DOI: 10.3390/jcm10153426] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/28/2021] [Revised: 07/26/2021] [Accepted: 07/30/2021] [Indexed: 01/24/2023] Open
Abstract
Background: In Germany, more than one-third of donor corneas harvested are not suitable for transplantation. We evaluated the factors associated with the usability of donor corneas. Method: Data from 2032 consecutive donor corneas harvested at the Rhineland-Palatinate Eye Bank in Mainz, Germany, were retrospectively analyzed. Factors of interest were age, sex, lens status, cause of death, cardiopulmonary resuscitation (CPR), death-to-explantation-interval (DEI), and the influence of these factors on the proportion of discarded donor corneas. Factors associated with endothelial cell density (ECD) were analyzed in a linear regression mixed model. Results: Higher donor age, male gender, pseudophakic lens status, and longer DEI were associated with significantly reduced ECD. With respect to DEI, the estimated cell loss was 7 ± 2 cells/mm2/hour (p < 0.001). Age was associated with a lower ECD of 6 ± 2 cells/mm2 per year (p = 0.001). Female ECD was 189 ± 44 cells/mm2 higher than male ECD (p < 0.001). Pseudophakic eyes had 378 ± 42 cells/mm2 less compared with phakic eyes (p < 0.001). Cause of death did not affect the ECD. Of note, 55% and 38% of corneas harvested on the second and third postmortem day, respectively, and 45% of corneas from donors older than 80 years were still suitable for transplantation. Conclusions: In the context of a growing need for donor corneas, we do not recommend limiting donor age and collection time to 24 h or excluding oncology donors, as is the practice in many countries. Therefore, we propose a mathematical model for better donor preselection.
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Outcomes of corneal transplantation in Europe: report by the European Cornea and Cell Transplantation Registry. J Cataract Refract Surg 2021; 47:780-785. [PMID: 33278237 DOI: 10.1097/j.jcrs.0000000000000520] [Citation(s) in RCA: 18] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/07/2020] [Accepted: 11/05/2020] [Indexed: 11/25/2022]
Abstract
PURPOSE To analyze real-world graft survival and visual acuity outcomes of corneal transplantation in Europe. SETTING Corneal clinics in 10 European Union member states, the United Kingdom, and Switzerland. DESIGN Multinational registry study. METHODS All corneal transplant procedures registered in the European Cornea and Cell Transplantation Registry (ECCTR) were identified. Graft survival of primary corneal transplants were analyzed using Kaplan-Meier survival curves with log-rank test and Cox regression. Corrected distance visual acuities (CDVAs) are reported at baseline and 2 years postoperatively using the Lundström distribution matrix. RESULTS A total of 12 913 corneal transplants were identified. Overall, 32-year graft survival of corneal transplants was high (89%) but differed between indications, ranging from 98% in keratoconus and 80% for trauma. Overall, CDVA improved postoperatively, but the risk for losing vision ranged from 7% (baseline vision ≤0.1 Snellen) to 58% (baseline vision ≥1.0 Snellen). CONCLUSIONS This report provides a comprehensive overview of graft survival and visual outcomes of corneal transplantation in Europe. In addition, it provides real-world estimates of outcomes for a variety of indications and surgical techniques to support benchmarking and demonstrates the relationship between baseline and postoperative vision.
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Dunker SL, Armitage WJ, Armitage M, Brocato L, Figueiredo FC, Heemskerk MBA, Hjortdal J, Jones GLA, Konijn C, Nuijts RMMA, Lundström M, Dickman MM. Practice patterns of corneal transplantation in Europe: first report by the European Cornea and Cell Transplantation Registry. J Cataract Refract Surg 2021; 47:865-869. [PMID: 33577274 DOI: 10.1097/j.jcrs.0000000000000574] [Citation(s) in RCA: 23] [Impact Index Per Article: 5.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/07/2020] [Accepted: 11/21/2020] [Indexed: 11/25/2022]
Abstract
PURPOSE To report practice patterns of corneal transplantation in Europe. SETTING Corneal clinics in 10 European member states (MS), the United Kingdom, and Switzerland. DESIGN Multinational registry study. METHODS Corneal transplant procedures registered in the European Cornea and Cell Transplantation Registry were identified. Preoperative donor and recipient characteristics, indication and reason for transplantation, and surgical techniques were analyzed. RESULTS A total of 12 913 corneal transplants were identified from 10 European Union MS, the United Kingdom, and Switzerland. Most countries were self-sufficient with regard to donor tissue. Fuchs endothelial corneal dystrophy was the most common indication (41%, n = 5325), followed by regraft (16%, n = 2108), pseudophakic bullous keratopathy (12%, n = 1594), and keratoconus (12%, n = 1506). Descemet stripping automated endothelial keratoplasty (DSAEK, 46%, n = 5918) was the most commonly performed technique, followed by penetrating keratoplasty (30%, n = 3886) and Descemet membrane endothelial keratoplasty (9%, n = 1838). Vision improvement was the main reason for corneal transplantation (90%, n = 11 591). Surgical technique and reason for transplantation differed between indications. CONCLUSIONS This report provides the most comprehensive overview of corneal transplantation practice patterns in Europe to date. Fuchs endothelial dystrophy is the most common indication, vision improvement the leading reason, and DSAEK the predominant technique for corneal transplantation.
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Affiliation(s)
- Suryan L Dunker
- From the University Eye Clinic, Maastricht University Medical Center+, The Netherlands (Dunker, Nuijts, Dickman); Translational Health Sciences, University of Bristol, United Kingdom (Armitage); Tissue and Eye Services, NHS Blood and Transplant, Bristol, United Kingdom (Armitage); European Eye Bank Association, Venice, Italy (Armitage, Jones); Department of Ophthalmology, Sahlgrenska University Hospital, Gothenburg, Sweden (Armitage); European Society of Cataract and Refractive Surgeons, Dublin, Ireland (Brocato, Nuijts, Lundström); Department of Ophthalmology, Royal Victoria Infirmary and Newcastle University, Newcastle upon Tyne, United Kingdom (Figueiredo); Dutch Transplant Foundation, Leiden, The Netherlands (Heemskerk, Konijn); Department of Ophthalmology, Aarhus University Hospital, Denmark (Hjortdal); European Society of Cornea and Ocular Surface Disease Specialists, Dublin, Ireland (Hjortdal); The Veneto Eye Bank Foundation, Venice, Italy (Jones); Department of Clinical Sciences, Ophthalmology, Faculty of Medicine, Lund University, Lund, Sweden (Lundström)
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Kwon HY, Hyon JY, Jeon HS. Effect of Donor Age on Graft Survival in Primary Penetrating Keratoplasty with Imported Donor Corneas. KOREAN JOURNAL OF OPHTHALMOLOGY 2021; 34:35-45. [PMID: 32037748 PMCID: PMC7010478 DOI: 10.3341/kjo.2019.0086] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/25/2019] [Revised: 09/23/2019] [Accepted: 10/04/2019] [Indexed: 11/25/2022] Open
Abstract
Purpose To investigate the influence of donor age on corneal graft survival following primary penetrating keratoplasty (PK) with imported donor corneas. Methods The eyes of patients who underwent primary PK with imported donor corneas were classified retrospectively into two groups according to a donor-age cutoff of 65 years. Primary outcome measures were rejection-free graft survival and graft survival. Cox proportional hazard regression analysis was used to assess the factors affecting graft survival. Survival analysis was performed using the Kaplan-Meier method, while differences between groups were examined using a log-rank test. A subgroup analysis of low- and high-risk eyes according to preoperative diagnosis was also performed. Results A total of 140 eyes from 138 patients (age, 58 ± 18 years) were enrolled. Cox regression analysis revealed that the donor age of 65 years or older group presented an increased risk of both graft rejection and failure. Survival analysis revealed that rejection-free graft survival and graft survival rates were higher in eyes in the donor age of less than 65 years group. Finally, in the subgroup analysis, both rejection-free graft survival and graft survival rates were significantly higher in the donor age of less than 65 years group than in the donor age of 65 years or older group, but only in the low-risk subgroup. Conclusions Donor age may correlate with graft survival in primary PK performed with imported donor corneas. Donor age could be a considerable factor in primary PK with imported donor corneas, especially in preoperatively low-risk patients.
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Affiliation(s)
- Hyeon Yoon Kwon
- Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Korea
| | - Joon Young Hyon
- Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Korea.,Department of Ophthalmology, Seoul National University Bundang Hospital, Seongnam, Korea
| | - Hyun Sun Jeon
- Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Korea.,Department of Ophthalmology, Seoul National University Bundang Hospital, Seongnam, Korea.
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Usefulness of Prestorage Corneal Swab Culture in the Prevention of Contaminated Corneal Tissue in Corneal Transplantation. Cornea 2021; 39:827-833. [PMID: 31990848 DOI: 10.1097/ico.0000000000002267] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/11/2023]
Abstract
PURPOSE To assess the efficacy of the prestorage corneal swab (PCS) culture to screen for corneal graft contamination after storage in Optisol-GS. METHODS A retrospective analysis of all PCS cultures was performed at the Eye Bank of Québec in Hôpital Maisonneuve-Rosemont from September 2013 to June 2016. Whole corneal culture was performed on rejected grafts because of a positive PCS, and a contamination rate was calculated. In addition, contamination rates of corneoscleral rims were compared between corneas tested with PCS and those of imported corneas which did not have PCS. RESULTS Among the 1966 PCS cultures performed, 814 (41.4%) were positive for growth. Pathogenic bacteria were present in 144 (7.3%) corneas, including Staphylococcus aureus (n = 96, 11.8% of all positive cultures), Enterobacteriaceae (n = 14, 1.7%), and Pseudomonas aeruginosa (n = 6, 0.7%). After preservation in Optisol-GS, only 7 (6.9%) corneas remained contaminated (95% confidence interval 5.1-9.3). The sensitivity of the PCS culture was 87.5% (95% confidence interval 47.4-99.7). There was no significant difference in corneoscleral rim contamination between corneas tested with PCS (1/388; 0.2%) compared with imported, nonswabbed corneas (3/214; 1.4%) (P = 0.131). Therefore, the cost to recover the loss of tissue rejected because of false-positive PCS by purchasing corneal tissue was calculated to be $142,884 (CAD) per year. CONCLUSIONS Despite the high sensitivity of PCS cultures, there was no significant reduction of infection after corneal transplantation using this technique. In consequence, 93% of the corneas possibly suitable for transplantation were rejected. This suggests that the PCS culture alone is a poor test for detecting clinically relevant corneal contamination.
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Vasiliauskaitė I, de Jong M, Quilendrino R, van der Wees J, Oellerich S, Melles GRJ. Use of Corneas From Septic Donors for Descemet Membrane Endothelial Keratoplasty. Cornea 2021; 40:33-38. [PMID: 32769680 DOI: 10.1097/ico.0000000000002443] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
Abstract
PURPOSE To evaluate the suitability of corneas from septic donors for transplantation by analyzing the discard rate in the eye bank and the clinical outcome of Descemet membrane endothelial keratoplasty (DMEK) using organ-cultured corneal grafts from septic versus nonseptic donors. METHODS This retrospective study included 1554 corneas of which 456 corneas (29%) were from septic and 1072 corneas (69%) from nonseptic donors [for 26 corneas (2%) sepsis status was unknown]. The clinical outcome at 6 months after DMEK was evaluated for 82 grafts (26 from septic and 56 from nonseptic donors). Outcome measures were endothelial cell density, central corneal thickness, and postoperative complications. RESULTS Primary discard rates were higher for corneas from septic than from nonseptic donors (32.9% vs. 24.5%, P = 0.001). The main discard reason was poor endothelial cell quality for both septic (13.8%) and nonseptic (11.8%) donor corneas. Eye bank contamination rates for septic and nonseptic donor corneas were 1.1% and 1.7%, respectively (P = 0.102). After DMEK, donor endothelial cell density at 6m postoperatively was comparable between grafts from septic and nonseptic donors (1410 ± 422 cells/mm vs. 1590 ± 519 cells/mm, P = 0.140). No differences in 6m central corneal thickness and in the rebubbling rate were observed between the 2 groups (P = 0.780 and P = 0.396, respectively). None of the cases had graft rejection nor endophthalmitis in both groups. CONCLUSIONS Provided strict adherence to donor screening and evaluation protocols, the use of organ-cultured corneas from septic donors for DMEK does not seem to increase the risk for recipients and allows for expansion of the donor pool for corneal tissue.
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Affiliation(s)
- Indrė Vasiliauskaitė
- Netherlands Institute for Innovative Ocular Surgery (NIIOS), Rotterdam, the Netherlands
- Melles Cornea Clinic Rotterdam, the Netherlands; and
| | - Maloeke de Jong
- Netherlands Institute for Innovative Ocular Surgery (NIIOS), Rotterdam, the Netherlands
- Amnitrans Eye Bank Rotterdam, the Netherlands
| | - Ruth Quilendrino
- Netherlands Institute for Innovative Ocular Surgery (NIIOS), Rotterdam, the Netherlands
- Melles Cornea Clinic Rotterdam, the Netherlands; and
- Amnitrans Eye Bank Rotterdam, the Netherlands
| | - Jacqueline van der Wees
- Netherlands Institute for Innovative Ocular Surgery (NIIOS), Rotterdam, the Netherlands
- Amnitrans Eye Bank Rotterdam, the Netherlands
| | - Silke Oellerich
- Netherlands Institute for Innovative Ocular Surgery (NIIOS), Rotterdam, the Netherlands
| | - Gerrit R J Melles
- Netherlands Institute for Innovative Ocular Surgery (NIIOS), Rotterdam, the Netherlands
- Melles Cornea Clinic Rotterdam, the Netherlands; and
- Amnitrans Eye Bank Rotterdam, the Netherlands
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Armitage WJ, Winton HL, Jones MNA, Downward L, Crewe JM, Rogers CA, Tole DM, Dick AD. Corneal Transplant Follow-up Study II: a randomised trial to determine whether HLA class II matching reduces the risk of allograft rejection in penetrating keratoplasty. Br J Ophthalmol 2020; 106:42-46. [PMID: 33268345 DOI: 10.1136/bjophthalmol-2020-317543] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/20/2020] [Revised: 11/02/2020] [Accepted: 11/14/2020] [Indexed: 12/16/2022]
Abstract
PURPOSE A randomised trial to test the hypothesis that human leucocyte antigen (HLA) class II matching reduces the risk of allograft rejection in high-risk penetrating keratoplasty (PK). METHODS All transplants were matched for HLA class I antigens (≤2 mismatches at the A and B loci) and corneas were allocated to patients by cohort minimisation to achieve 0, 1 or 2 HLA class II antigen mismatches. The corneal transplants (n=1133) were followed for 5 years. The primary outcome measure was time to first rejection episode. RESULTS Cox regression analysis found no influence of HLA class II mismatching on risk of immunological rejection (HR 1.13; 95% CI 0.79 to 1.63; p=0.51). The risk of rejection in recipients older than 60 years was halved compared with recipients ≤40 years (HR 0.51; 95% CI 0.36 to 0.73; p=0.0003). Rejection was also more likely where cataract surgery had been performed after PK (HR 3.68; 95% CI 1.95 to 6.93; p<0.0001). In univariate analyses, preoperative factors including chronic glaucoma (p=0.02), vascularisation (p=0.01), inflammation (p=0.03), ocular surface disease (p=0.0007) and regrafts (p<0.001) all increased the risk of rejection. In the Cox model, however, none of these factors was individually significant but rejection was more likely where≥2 preoperative risk factors were present (HR 2.11; 95% CI 1.26 to 3.47; p<0.003). CONCLUSIONS HLA class II matching, against a background of HLA class I matching, did not reduce the risk of allograft rejection. Younger recipient age, the presence of ≥2 preoperative risk factors and cataract surgery after PK all markedly increased the risk of allograft rejection. TRIAL REGISTRATION NUMBER ISRCTN25094892.
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Affiliation(s)
- W John Armitage
- Translational Health Sciences, Bristol Medical School, University of Bristol, Bristol, United Kingdom
| | - Helen L Winton
- Translational Health Sciences, Bristol Medical School, University of Bristol, Bristol, United Kingdom
| | | | | | - Julie M Crewe
- Translational Health Sciences, Bristol Medical School, University of Bristol, Bristol, United Kingdom
| | - Chris A Rogers
- Bristol Trials Centre, Bristol Medical School, University of Bristol, Bristol, United Kingdom
| | | | - Andrew D Dick
- Translational Health Sciences, Bristol Medical School, University of Bristol, Bristol, United Kingdom
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Fabre L, Puyraveau M, Jeanvoine A, Thibaud G, Pizzuto J, Pouthier F, Delbosc B, Gauthier AS. Changes of Contamination Rate and Microorganism Evaluation in Organ-Cultured Human Corneas: A 14-Year Review From a French Regional Eye Bank. Cornea 2020; 40:696-703. [PMID: 33290322 DOI: 10.1097/ico.0000000000002618] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/08/2020] [Accepted: 10/07/2020] [Indexed: 11/25/2022]
Abstract
PURPOSE This study aimed to assess how the contamination rate of organ-cultured corneas has evolved and to analyze the evolution of microorganisms involved. METHODS Data from the Besançon eye bank were reviewed over 14 years (2005-2018). The changes in the contamination rate and the contaminant species found during the organ culture storage were analyzed. Microbiological tests were performed twice on the storage media-at day 5 and before the deswelling phase. RESULTS Among the 17,979 donor corneas collected, 1240 corneas were microbiological-test positive. The average annual contamination rate was 6.8% (range: 5.2%-8.9%). Seventy-five percent of contaminations were bacterial. The most frequently found bacterium was Staphylococcus spp. (31.3%), followed by non-Enterobacteriaceae Gram-negative Bacilli (GNB) (27.3%), with most Sphingomonas spp. and Pseudomonas spp. Fungal contamination (21.9%) was dominated by Candida (82.7%). Seventy-seven types of microorganisms were identified. The Staphylococcus rate tended to decrease, whereas non-Enterobacteriaceae GNB rate has increased in the past few years to reach 46% of bacteria. Most of the contaminations were detected in the early phase of organ culture at day 5 (89.2%). The second microbiological test found 44.8% of fungal contaminations (predominantly Candida spp.). CONCLUSIONS The annual contamination rate was stable and remains low, but the types of contaminating microorganisms varied from year to year. Staphylococcus spp. and non-Enterobacteriaceae GNB accounted for a significant proportion of the contaminations. We found a significant proportion of contamination, especially fungal, at the late phase of storage. Reassessing the antibiotics and antifungals in the storage medium may be useful to limit corneal disposal.
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Affiliation(s)
- Laura Fabre
- Department of Ophthalmology, J.Minjoz University Hospital, Besançon, France
| | - Marc Puyraveau
- Clinical Methodology Center, University Hospital, Besançon, France
| | | | - Garcin Thibaud
- Department of Ophthalmology, University Hospital, Saint Etienne, France
| | - Joëlle Pizzuto
- French Blood Establishment Bourgogne/Franche-Comté, Besançon, France; and
| | - Fabienne Pouthier
- French Blood Establishment Bourgogne/Franche-Comté, Besançon, France; and
| | - Bernard Delbosc
- Department of Ophthalmology, University Hospital, Besançon, France
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Mistò R, Giurgola L, Pateri F, Limongelli A, Ragazzi E, D'Amato Tóthová J. A new storage medium containing amphotericin B versus Optisol-GS for preservation of human donor corneas. Br J Ophthalmol 2020; 106:184-189. [PMID: 33172862 DOI: 10.1136/bjophthalmol-2020-317136] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/08/2020] [Revised: 09/21/2020] [Accepted: 10/17/2020] [Indexed: 11/03/2022]
Abstract
BACKGROUND/AIM We compared the quality of human donor corneas stored in a cold storage medium containing 2.5 μg/ml of amphotericin B (Kerasave, AL.CHI.MI.A. S.R.L., Ponte San Nicolò, Italy) and Optisol-GS (Bausch & Lomb Inc., Bridgewater, NJ, USA) for 14 days. METHODS Sixteen pairs of human donor corneas were collected in Eusol-C (AL.CHI.MI.A. S.R.L., Ponte San Nicolò, Italy). Next, all tissues underwent the first evaluation that included the assessments of central corneal thickness (CCT), endothelial cell density (ECD) measured using both trypan blue staining and specular microscopy, endothelial cell (EC) mortality and morphology, and corneal transparency within 24 hours from recovery (Day 1). Afterwards, one cornea of each pair was transferred into Kerasave or Optisol-GS. ECD and CCT were also assessed at Day 7, and all the metrics were evaluated again at the end of the storage period (Day 14). RESULTS At all tested time points, no differences were found in the qualitative (corneal transparency, EC morphology) and quantitative metrics (ECD, CCT, EC mortality) between the Kerasave and the Optisol-GS storage groups. At Day 14, the corneas stored in Kerasave and Optisol-GS showed ECD of 2312±98 and 2335±128 cells/mm2 (p=0.886), CCT of 717±17 and 697±19 μm (p=0.454) and central EC mortality of 0.54%±0.40% and 0.14%±0.14% (p=0.719), respectively. CONCLUSIONS The new amphotericin B-containing medium Kerasave was comparable to Optisol-GS in terms of preservation of corneal characteristics at 2-8°C for 14 days.
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Affiliation(s)
- Raffaela Mistò
- Eye Bank of Monza, Azienda Ospedaliera San Gerardo, Monza, Italy
| | | | - Francesca Pateri
- Eye Bank of Monza, Azienda Ospedaliera San Gerardo, Monza, Italy
| | - Anna Limongelli
- Eye Bank of Monza, Azienda Ospedaliera San Gerardo, Monza, Italy
| | - Eugenio Ragazzi
- Department of Pharmaceutical and Pharmacological Sciences, Università Degli Studi Di Padova, Padova, Italy
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Factors influencing endothelial cell density of corneas for transplantation. Cell Tissue Bank 2020; 22:263-275. [PMID: 33165826 DOI: 10.1007/s10561-020-09875-8] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/12/2019] [Accepted: 10/10/2020] [Indexed: 10/23/2022]
Abstract
To evaluate factors affecting corneal endothelial cell density (ECD) under enucleation and preservation time studies at Eye Bank of the Federal District of Brazil. We conducted a case-control study collecting data from 1128 corneas where death-to-enucleation time and enucleation-to-preservation time were within 24 h. Low cell count were those corneas with an ECD less than 2000 cells/mm2 and high cell count was defined as those with ECD greater than 2000 cells/mm2. We calculated the independent risk factors related to: cause of death, donor age, death-to-enucleation time, enucleation-to-preservation time and primary graft failure. Correlation analysis was used to assess which parameters influence ECD: death-to-enucleation time, enucleation-to-preservation time, average cell area (AVE), coefficient of variation and percentage of hexagonal cells. Of the total number of corneas, 1004 had ECD data and were selected for the study. 87.4% (n = 877) had high cell counts with 2699 ± 412 cells/mm2. The mean donor age was 38.8 ± 16 years. The most common causes of death were external causes (48.6%, n = 488). Longer times from death-to-enucleation, up to 24 h were not associated with a decrease in ECD (OR 0.58; P = 0.44) or risk of graft survival (P = 0.74). Enucleation-to-preservation intervals greater than 12 h showed increased risk of graft survival (P = 0.04). AVE was the main parameter for ECD (R2 = 0.96, P < 0.001). The overall graft survival rate was 98.2% (n = 761). Donors with 40 years of age and above did not present a higher risk of graft survival (P = 0.09). We suggest that the maximum time from death-to-enucleation should be 24 h, assuming the body has been refrigerated after 6 h; and from enucleation-to-preservation time of 12 h, followed by proper processing and cornea morphology examination.
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Wojcik G, Ferrari S, Romano V, Ponzin D, Ahmad S, Parekh M. Corneal storage methods: considerations and impact on surgical outcomes. EXPERT REVIEW OF OPHTHALMOLOGY 2020. [DOI: 10.1080/17469899.2021.1829476] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 10/23/2022]
Affiliation(s)
- Gabriela Wojcik
- International Center for Ocular Physiopathology, Fondazione Banca degli Occhi del Veneto, Venice, Italy
| | - Stefano Ferrari
- International Center for Ocular Physiopathology, Fondazione Banca degli Occhi del Veneto, Venice, Italy
| | - Vito Romano
- St. Paul’s Eye Unit, Royal Liverpool University Hospital, Liverpool, UK
| | - Diego Ponzin
- International Center for Ocular Physiopathology, Fondazione Banca degli Occhi del Veneto, Venice, Italy
| | - Sajjad Ahmad
- Institute of Ophthalmology, University College London, London, UK
- Cornea and external eye disease, Moorfields Eye Hospital NHS Trust Foundation, London, UK
| | - Mohit Parekh
- International Center for Ocular Physiopathology, Fondazione Banca degli Occhi del Veneto, Venice, Italy
- Institute of Ophthalmology, University College London, London, UK
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Hofmann N, Wittmershaus I, Salz AK, Börgel M. Cornea Procurement and Processing up to 72 Hours: No Risk for Cornea Transplant Quality. Transfus Med Hemother 2020; 48:3-11. [PMID: 33708047 DOI: 10.1159/000510588] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/26/2020] [Accepted: 07/22/2020] [Indexed: 11/19/2022] Open
Abstract
Background The realization of tissue donations is bound to a tight timeframe. Depending on the type of tissue, time limits are specified within which the donation must be procured and processed. Otherwise, there is a risk of tissue quality loss with increasing time intervals from cardiovascular arrest. According to the European Directorate for the Quality of Medicines and HealthCare (EDQM) guide, cornea must be procured and processed within 72 h after death. The question arises whether this time interval has an influence on the quality of transplanted tissues and how it affects the accomplishment of tissue donations. Methods In order to obtain information on this, the numbers of tissue donations in the network of the German Society for Tissue Transplantation (DGFG) were evaluated as a function of the death to retrieval time (DRT) as well as the death to preservation time (DPT). For this purpose, 21,454 database entries of cornea donations made in the period from 2014 to 2018 were included. Results The results show that nearly 50% of donations realized in the DGFG network could be processed only 48 h or later after cardiovascular death due to the opt-in regulation in Germany. For these donations, there seems to be a higher discard rate compared to donations taken earlier. Nevertheless, there is a transplantation rate for these grafts of more than 65%, which is comparable to average transplantation rates stated in the literature. Conclusion All corneas finally selected for transplantation must meet the specified quality parameters. Since this naturally also applies to transplants that could only be procured at later time points, it can be concluded that DPT up to 72 h for corneal tissue is adequate and has no influence on the quality of corneas that are ultimately transplanted.
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Affiliation(s)
- Nicola Hofmann
- German Society for Tissue Transplantation (DGFG) gGmbH, Hannover, Germany
| | - Ilka Wittmershaus
- German Society for Tissue Transplantation (DGFG) gGmbH, Hannover, Germany
| | | | - Martin Börgel
- German Society for Tissue Transplantation (DGFG) gGmbH, Hannover, Germany
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Medina IFB, Oguido APMT, Urbano MR, Casella AMB. Intensive care unit time and prolonged enucleation to processing interval are associated with donor cornea contamination. Graefes Arch Clin Exp Ophthalmol 2020; 258:2241-2249. [PMID: 32613575 DOI: 10.1007/s00417-020-04758-w] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/18/2019] [Revised: 03/30/2020] [Accepted: 05/19/2020] [Indexed: 11/25/2022] Open
Abstract
PURPOSE To determine donor cornea contamination rate and to determine factors associated with cornea contamination. To assess the effect of hospitalization, intensive care unit (ICU) time, and antibiotic use on corneal contamination rate. To determine the spectrum of the contaminating microorganisms. METHODS The contamination rate of 212 corneas, obtained by enucleation from April 2014 to January 2015 in a single eye bank, was assessed retrospectively according to age, sex, cause of death, systemic antibiotic use, hospitalization time, ICU time, mechanical ventilation (MV), death to enucleation interval (DEI), enucleation to processing interval (EPI), and corneal epithelial exposure grading. The relative risk (RR) and adjusted RR with a 95% confidence interval were calculated using IBM-SPSS 20.0. RESULTS The contamination rate was 35.6% (n = 75). On multivariate analysis, ICU stay of 4 days or longer and enucleation to processing interval (EPI) greater than 7.4 h (RR 1.58, CI 0.96-2.60, P = 0.06) were associated with donor cornea contamination. Corneal contamination risk was highest from 4 to 6 days at the ICU (RR 3.40, CI 1.54-7.51, P < 0.01) and decreased after 7 days (RR 2.22, CI 1.00-4.93, P = 0.05). Coagulase-negative Staphylococcus was the most common isolated bacteria (69.6%). The frequency of gentamicin-resistant bacteria was higher among patients who stayed 4 days or longer at the ICU. CONCLUSION Patients staying at the intensive care unit 4 days or longer showed increased risk of corneal contamination. This is an important result to consider further indication for cornea donation.
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da Mata Martins TM, da Silva Cunha P, Rodrigues MA, de Carvalho JL, de Souza JE, de Carvalho Oliveira JA, Gomes DA, de Goes AM. Epithelial basement membrane of human decellularized cornea as a suitable substrate for differentiation of embryonic stem cells into corneal epithelial-like cells. MATERIALS SCIENCE & ENGINEERING. C, MATERIALS FOR BIOLOGICAL APPLICATIONS 2020; 116:111215. [PMID: 32806330 DOI: 10.1016/j.msec.2020.111215] [Citation(s) in RCA: 15] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 12/20/2019] [Revised: 06/13/2020] [Accepted: 06/18/2020] [Indexed: 12/11/2022]
Abstract
The ability to decellularize and recellularize the corneas deemed unsuitable for transplantation may increase the number of available grafts. Decellularized corneas (DCs) may provide a natural microenvironment for cell adhesion and differentiation. Despite this, no study to date has evaluated their efficacy as a substrate for the induction of stem cell differentiation into corneal cells. The present study aimed to compare the efficiency of NaCl and NaCl plus nucleases methods to decellularize whole human corneas, and to investigate the effect of epithelial basement membrane (EBM) of whole DCs on the ability of human embryonic stem cells (hESCs) to differentiate into corneal epithelial-like cells when cultured in animal serum-free differentiation medium. As laminin is the major component of EBM, we also investigated its effect on hESCs differentiation. The decellularization efficiency and integrity of the extracellular matrix (ECM) obtained were investigated by histology, electron microscopy, DNA quantification, immunofluorescence, and nuclear staining. The ability of hESCs to differentiate into corneal epithelial-like cells when seeded on the EBM of DCs or laminin-coated wells was evaluated by immunofluorescence and RT-qPCR analyses. NaCl treatment alone, without nucleases, was insufficient to remove cellular components, while NaCl plus nucleases treatment resulted in efficient decellularization and preservation of the ECM. Unlike cells induced to differentiate on laminin, hESCs differentiated on DCs expressed high levels of corneal epithelial-specific markers, keratin 3 and keratin 12. It was demonstrated for the first time that the decellularized matrices had a positive effect on the differentiation of hESCs towards corneal epithelial-like cells. Such a strategy supports the potential applications of human DCs and hESCs in corneal epithelium tissue engineering.
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Affiliation(s)
- Thaís Maria da Mata Martins
- Department of Morphology, Institute of Biological Sciences, Federal University of Minas Gerais, Avenida Presidente Antônio Carlos, 6627, Belo Horizonte 31270-901, Minas Gerais, Brazil.
| | - Pricila da Silva Cunha
- Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais, Avenida Presidente Antônio Carlos, 6627, Belo Horizonte 31270-901, Minas Gerais, Brazil
| | - Michele Angela Rodrigues
- Department of Pathology, Institute of Biological Sciences, Federal University of Minas Gerais, Avenida Presidente Antônio Carlos, 6627, Belo Horizonte 31270-901, Minas Gerais, Brazil
| | - Juliana Lott de Carvalho
- Department of Genomic Sciences and Biotechnology, Catholic University of Brasilia, QS 07 - Lote 01, EPCT - Taguatinga, Brasília, Distrito Federal 71966-700, Brazil; Faculty of Medicine, University of Brasilia, Campus Universitário Darcy Ribeiro, Brasília, Distrito Federal 70910-900, Brazil
| | - Joyce Esposito de Souza
- Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais, Avenida Presidente Antônio Carlos, 6627, Belo Horizonte 31270-901, Minas Gerais, Brazil
| | - Junnia Alvarenga de Carvalho Oliveira
- Department of Microbiology, Federal University of Minas Gerais, Avenida Presidente Antônio Carlos, 6627, Belo Horizonte 31270-901, Minas Gerais, Brazil
| | - Dawidson Assis Gomes
- Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais, Avenida Presidente Antônio Carlos, 6627, Belo Horizonte 31270-901, Minas Gerais, Brazil
| | - Alfredo Miranda de Goes
- Department of Pathology, Institute of Biological Sciences, Federal University of Minas Gerais, Avenida Presidente Antônio Carlos, 6627, Belo Horizonte 31270-901, Minas Gerais, Brazil
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Ong HS, Peh G, Neo DJH, Ang HP, Adnan K, Nyein CL, Morales-Wong F, Bhogal M, Kocaba V, Mehta JS. A Novel Approach of Harvesting Viable Single Cells from Donor Corneal Endothelium for Cell-Injection Therapy. Cells 2020; 9:cells9061428. [PMID: 32526886 PMCID: PMC7349718 DOI: 10.3390/cells9061428] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/31/2020] [Revised: 05/25/2020] [Accepted: 06/02/2020] [Indexed: 12/13/2022] Open
Abstract
Donor corneas with low endothelial cell densities (ECD) are deemed unsuitable for corneal endothelial transplantation. This study evaluated a two-step incubation and dissociation harvesting approach to isolate single corneal endothelial cells (CECs) from donor corneas for corneal endothelial cell-injection (CE-CI) therapy. To isolate CECs directly from donor corneas, optimization studies were performed where donor Descemet’s membrane/corneal endothelium (DM/CE) were peeled and incubated in either M4-F99 or M5-Endo media before enzymatic digestion. Morphometric analyses were performed on the isolated single cells. The functional capacities of these cells, isolated using the optimized simple non-cultured endothelial cells (SNEC) harvesting technique, for CE-CI therapy were investigated using a rabbit bullous keratopathy model. The two control groups were the positive controls, where rabbits received cultured CECs, and the negative controls, where rabbits received no CECs. Whilst it took longer for CECs to dislodge as single cells following donor DM/CE incubation in M5-Endo medium, CECs harvested were morphologically more homogenous and smaller compared to CECs obtained from DM/CE incubated in M4-F99 medium (p < 0.05). M5-Endo medium was hence selected as the DM/CE incubation medium prior to enzymatic digestion to harvest CECs for the in vivo cell-injection studies. Following SNEC injection, mean central corneal thickness (CCT) of rabbits increased to 802.9 ± 147.8 μm on day 1, gradually thinned, and remained clear with a CCT of 385.5 ± 38.6 μm at week 3. Recovery of corneas was comparable to rabbits receiving cultured CE-CI (p = 0.40, p = 0.17, and p = 0.08 at weeks 1, 2, and 3, respectively). Corneas that did not receive any cells remained significantly thicker compared to both SNEC injection and cultured CE-CI groups (p < 0.05). This study concluded that direct harvesting of single CECs from donor corneas for SNEC injection allows the utilization of donor corneas unsuitable for conventional endothelial transplantation.
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Affiliation(s)
- Hon Shing Ong
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore 169856, Singapore; (G.P.); (D.J.H.N.); (H.-P.A.); (K.A.); (C.L.N.); (F.M.-W.); (M.B.); (V.K.)
- Eye-Academic Clinical Program (ACP), Duke-National University of Singapore (NUS), Graduate Medical School, Singapore 169857, Singapore
- Corneal and External Diseases Department, Singapore National Eye Centre, Singapore 168751, Singapore
- Correspondence: (H.S.O.); (J.S.M.); Tel.: +65-6227-7255 (H.S.O. & J.S.M.); Fax: +65-6227-7290 (H.S.O. & J.S.M.)
| | - Gary Peh
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore 169856, Singapore; (G.P.); (D.J.H.N.); (H.-P.A.); (K.A.); (C.L.N.); (F.M.-W.); (M.B.); (V.K.)
- Eye-Academic Clinical Program (ACP), Duke-National University of Singapore (NUS), Graduate Medical School, Singapore 169857, Singapore
| | - Dawn Jin Hui Neo
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore 169856, Singapore; (G.P.); (D.J.H.N.); (H.-P.A.); (K.A.); (C.L.N.); (F.M.-W.); (M.B.); (V.K.)
| | - Heng-Pei Ang
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore 169856, Singapore; (G.P.); (D.J.H.N.); (H.-P.A.); (K.A.); (C.L.N.); (F.M.-W.); (M.B.); (V.K.)
| | - Khadijah Adnan
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore 169856, Singapore; (G.P.); (D.J.H.N.); (H.-P.A.); (K.A.); (C.L.N.); (F.M.-W.); (M.B.); (V.K.)
| | - Chan Lwin Nyein
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore 169856, Singapore; (G.P.); (D.J.H.N.); (H.-P.A.); (K.A.); (C.L.N.); (F.M.-W.); (M.B.); (V.K.)
| | - Fernando Morales-Wong
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore 169856, Singapore; (G.P.); (D.J.H.N.); (H.-P.A.); (K.A.); (C.L.N.); (F.M.-W.); (M.B.); (V.K.)
| | - Maninder Bhogal
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore 169856, Singapore; (G.P.); (D.J.H.N.); (H.-P.A.); (K.A.); (C.L.N.); (F.M.-W.); (M.B.); (V.K.)
- Cornea Unit, Guy’s & St Thomas’ Hospital, London SE1 7EH, UK
| | - Viridiana Kocaba
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore 169856, Singapore; (G.P.); (D.J.H.N.); (H.-P.A.); (K.A.); (C.L.N.); (F.M.-W.); (M.B.); (V.K.)
- Netherlands Institute for Innovative Ocular Surgery, Melles Cornea Clinic, Amnitrans EyeBank Rotterdam, 3071 AA Rotterdam, The Netherlands
| | - Jodhbir S. Mehta
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore 169856, Singapore; (G.P.); (D.J.H.N.); (H.-P.A.); (K.A.); (C.L.N.); (F.M.-W.); (M.B.); (V.K.)
- Eye-Academic Clinical Program (ACP), Duke-National University of Singapore (NUS), Graduate Medical School, Singapore 169857, Singapore
- Corneal and External Diseases Department, Singapore National Eye Centre, Singapore 168751, Singapore
- School of Material Science and Engineering, Nanyang Technological University, Singapore 639798, Singapore
- Correspondence: (H.S.O.); (J.S.M.); Tel.: +65-6227-7255 (H.S.O. & J.S.M.); Fax: +65-6227-7290 (H.S.O. & J.S.M.)
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Wong SW, Carley F, Jones NP. Corneal Decompensation in Uveitis Patients: Incidence, Etiology, and Outcome. Ocul Immunol Inflamm 2020; 29:771-775. [DOI: 10.1080/09273948.2019.1698747] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/17/2022]
Affiliation(s)
- Shiao W. Wong
- Manchester Royal Eye Hospital, Manchester University NHS Foundation Trust, Manchester, UK
| | - Fiona Carley
- Manchester Royal Eye Hospital, Manchester University NHS Foundation Trust, Manchester, UK
| | - Nicholas P. Jones
- Manchester Royal Eye Hospital, Manchester University NHS Foundation Trust, Manchester, UK
- Medical Academic Health Science Centre, University of Manchester, Manchester, UK
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Thareja T, Kowalski R, Kamyar R, Dhaliwal D, Jeng BH, Tu E, Jhanji V. Fungal infection after keratoplasty and the role of antifungal supplementation to storage solution: a review. Br J Ophthalmol 2019; 104:1036. [PMID: 31796428 DOI: 10.1136/bjophthalmol-2019-314664] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/28/2019] [Revised: 11/13/2019] [Accepted: 11/16/2019] [Indexed: 11/04/2022]
Abstract
Fungal infection after corneal transplantation is a rare, yet potentially devastating, postoperative complication and has become a growing concern for the transplant surgeon and eye banking community. The Eye Bank Association of America (EBAA) has reported an increasing trend in the rate of postkeratoplasty fungal infections and a reversal in the previously documented predominance of bacterial over fungal infections. Additionally, several studies have confirmed a high correlation between positive corneoscleral donor rim fungal cultures and postoperative infections. Optisol GS (Bausch & Lomb, Irvine, California, USA), the most extensively used corneal storage solution in US eye banks, does not currently contain any antifungal supplementation. Although large randomised control trials evaluating the efficacy and safety of routine antifungal supplementation to corneal storage solution are lacking, several investigative studies have assessed the role of antifungal agents in reducing fungal contamination of donor corneas without causing undue corneal toxicity. This review will present the current epidemiology of postkeratoplasty fungal infections and evidence for obtaining routine fungal rim cultures and antifungal supplementation of storage solution.
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Affiliation(s)
| | - Regis Kowalski
- Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.,Campbell Laboratory, UPMC Eye and Ear Institute, Pittsburgh, Pennsylvania, USA
| | - Roheena Kamyar
- UPMC Eye Center, Pittsburgh, Pennsylvania, USA.,Campbell Laboratory, UPMC Eye and Ear Institute, Pittsburgh, Pennsylvania, USA
| | - Deepinder Dhaliwal
- UPMC Eye Center, Pittsburgh, Pennsylvania, USA.,Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.,Campbell Laboratory, UPMC Eye and Ear Institute, Pittsburgh, Pennsylvania, USA
| | - Bennie H Jeng
- Department of Ophthalmology and Visual Sciences, University of Maryland School of Medicine, Baltimore, Maryland, USA
| | - Elmer Tu
- Department of Ophthalmology & Visual Sciences, University of Illinois-Chicago, Chicago, Illinois, USA
| | - Vishal Jhanji
- UPMC Eye Center, Pittsburgh, Pennsylvania, USA .,Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.,Campbell Laboratory, UPMC Eye and Ear Institute, Pittsburgh, Pennsylvania, USA
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A new approach to extend the storage of donor corneas after 28 days of corneal culture in an Italian eye bank. Cell Tissue Bank 2019; 21:47-55. [PMID: 31782073 DOI: 10.1007/s10561-019-09795-2] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/26/2019] [Accepted: 11/23/2019] [Indexed: 10/25/2022]
Abstract
This study aimed to evaluate the possibility to extend the storage of unused organ-cultured donor corneas. After 28 days of corneal culture in TISSUE-C (AL.CHI.MI.A. S.R.L., Italy) and 5-day storage in transport/deswelling medium CARRY-C (AL.CHI.MI.A. S.R.L., Italy), 25 corneas that were deemed suitable for transplantation were transferred in fresh TISSUE-C at 31 °C for additional 7 days and then in fresh CARRY-C at room temperature for 24 h. Tissues were assessed for endothelial cell density (ECD), endothelial mortality and morphology after the standard and the extended corneal storage. In addition, the effect of donor age < 85 years and ≥ 85 years on corneal characteristics was assessed. After the extended storage, 6 out of 25 tested corneas (24%) showed ECD values below the acceptance limit (< 2000 cells/mm2). 19 corneas (76%) were still suitable for transplantation and showed a 5.9% loss in ECD, which was not statistically significant (P = 0.0949) compared to standard storage period. The two donor age groups did not show statistically significant differences in any tested parameter, although a trend for lower ECD and higher mortality in Descemet's folds after standard storage was observed in the ≥ 85 age donor group. Thus, the attempt of the current study to provide new sight-restorative options for unused tissues and increasing the availability of corneas in case of shortage gave encouraging results. Although a higher vulnerability of corneas from very old donors could not be statistically demonstrated in the present study, higher sample size could be required for prolonging the shelf life of these tissues.
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Pessoa JLE, Schirmer J, Freitas DD, Knihs NDS, Roza BDA. Ocular tissue distribution in the State of São Paulo: analysis on corneal discarding reasons. Rev Lat Am Enfermagem 2019; 27:e3196. [PMID: 31618389 PMCID: PMC6792342 DOI: 10.1590/1518-8345.3041.3196] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/29/2018] [Accepted: 06/18/2019] [Indexed: 11/22/2022] Open
Abstract
Objective to identify the reasons for refusal of corneas. Method this was a cross-sectional, retrospective, descriptive and correlational
study composed of 5,560 optical corneas. The information was taken from the
notification, organ procurement and distribution centers database as well as
donor records. Descriptive statistics were used for the analysis of
categorical variables and specific tests with a significance level of 5% for
assessing the associations between variables. This study met the ethical
aspects of scientific research. Results 60% of the donors were male and 40% died by circulatory problems. The main
reason for refusal as informed by transplant teams is the donor’s age and
the endothelial cell count. For each year added to the donor’s age, there is
a 1% decrease in the chance that this cornea will be used for
transplantation, and the increase of 100 cells per mm2 increases the chances
that this cornea will be used by 9%. Conclusion the main cause of refusal in the acceptance of corneal tissue is related to
the age and the endothelial cell count.
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Affiliation(s)
- João Luis Erbs Pessoa
- Secretaria de Saúde do Estado de São Paulo, Central de Transplantes, São Paulo, SP, Brasil
| | - Janine Schirmer
- Universidade Federal de São Paulo, Escola Paulista de Enfermagem, São Paulo, SP, Brasil
| | - Denise de Freitas
- Universidade Federal de São Paulo, Escola Paulista de Medicina, São Paulo, SP, Brasil
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Antimycotic Efficacy and Safety of a New Cold Corneal Storage Medium by Time–Kill and Toxicity Studies. Cornea 2019; 38:1314-1321. [DOI: 10.1097/ico.0000000000002068] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
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Schnichels S, Kiebler T, Hurst J, Maliha AM, Löscher M, Dick HB, Bartz-Schmidt KU, Joachim SC. Retinal Organ Cultures as Alternative Research Models. Altern Lab Anim 2019; 47:19-29. [DOI: 10.1177/0261192919840092] [Citation(s) in RCA: 19] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/18/2023]
Abstract
Ex vivo organ cultures represent unique research models, as they combine the advantages of cell cultures with those of animal models. Being able to mimic in vivo situations through the use of organ cultures provides an excellent opportunity to investigate cellular processes, molecular pathways and cell–cell interactions, as well as structural and synaptic organisation. Human and animal organ cultures are now well established and comprise sensitive, easy-to-manipulate experimental systems that raise minimal ethical concerns. The eye, in particular, is a very complex organ that is not easy to reproduce in vitro. However, a lot of research has been dedicated to the development of suitable ocular organ cultures. This review covers the various ex vivo retinal organ culture systems available for use in ophthalmology research and compares them with commonly used animal models. In particular, bovine and porcine retinal organ culture systems are described, because the size, anatomy, physiology and vessel morphology of bovine and porcine eyes are similar to the human eye in an undisputed way, thus making them good models. In addition, these animals are widely used by the food industry and the eyes are considered surplus material. A short overview of murine, rat, rabbit, cat, canine and simian retinal organ cultures is also provided.
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Affiliation(s)
- Sven Schnichels
- Centre for Ophthalmology, University Eye Hospital Tübingen, Tübingen, Germany
| | - Tobias Kiebler
- Centre for Ophthalmology, University Eye Hospital Tübingen, Tübingen, Germany
| | - José Hurst
- Centre for Ophthalmology, University Eye Hospital Tübingen, Tübingen, Germany
| | - Ana M. Maliha
- Experimental Eye Research, University Eye Hospital, Ruhr-University Bochum, Bochum, Germany
| | - Marina Löscher
- Centre for Ophthalmology, University Eye Hospital Tübingen, Tübingen, Germany
| | - H. Burkhard Dick
- Experimental Eye Research, University Eye Hospital, Ruhr-University Bochum, Bochum, Germany
| | | | - Stephanie C. Joachim
- Experimental Eye Research, University Eye Hospital, Ruhr-University Bochum, Bochum, Germany
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Factors Affecting Corneal Organ Culture Contamination: A 6-year Study at the New South Wales Tissue Bank. Cornea 2019; 38:829-835. [PMID: 31170101 DOI: 10.1097/ico.0000000000001936] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
Abstract
PURPOSE To report the rate of microbial contamination and analyze possible risk factors for contamination of banked corneas stored using the organ culture method. METHODS Data from the New South Wales Tissue Banks incorporating the Lions NSW Eye Bank, between September 1, 2011, and November 30, 2017, were reviewed retrospectively. All corneas collected during this period and stored in organ culture storage media were tested for microbial contamination. The influence of potential factors on the rate of contamination was analyzed using the χ test and logistic regression using generalized estimating equations. RESULTS A total of 4410 corneas were included in this study, of which 110 were medium culture positive, representing a microbial contamination rate of 2.5%. The main contaminants were Candida species followed by Staphylococcus species. Corneal tissue collected in summer and autumn had a significantly higher contamination rate (P = 0.006). All other factors studied were not shown to have a statistically significant association with contamination after accounting for within-pair correlation and confounders. CONCLUSIONS A relatively low contamination rate of 2.5% observed in our study reflects the stringent laboratory protocols, strict donor selection criteria, and high level of experience among staff at the Lions NSW Eye Bank. Our study demonstrated that the season of collection had a strong association with the rate of organ culture contamination. Because Candida species contributed the largest percentage of contamination, specific measures to reduce and eliminate fungal proliferation should be considered by eye banks particularly in warm seasons.
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Efficacy and Safety of Various Amphotericin B Concentrations on Candida albicans in Cold Storage Conditions. Cornea 2019; 39:110-117. [DOI: 10.1097/ico.0000000000002019] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/25/2022]
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