A new storage medium containing amphotericin B versus Optisol-GS for preservation of human donor corneas

2020 ◽  
pp. bjophthalmol-2020-317136
Author(s):  
Raffaela Mistò ◽  
Laura Giurgola ◽  
Francesca Pateri ◽  
Anna Limongelli ◽  
Eugenio Ragazzi ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Amphotericin B ◽  
Corneal Thickness ◽  
Storage Period ◽  
Blue Staining ◽  
Specular Microscopy ◽  
Storage Medium ◽  
Human Donor ◽  
Corneal Transparency ◽  
Donor Corneas

Background/AimWe 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.MethodsSixteen 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).ResultsAt 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.ConclusionsThe 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.

scholarly journals A novel human donor cornea preservation cocktail incorporating a thermo-reversible gelation polymer (TGP), enhancing the corneal endothelial cell density maintenance and explant culture of corneal limbal cells

2021 ◽  
Author(s):  
Bhuvaneshwari Namitha ◽  
Munusamy Rajendran Chitra ◽  
Mathevan Bhavya ◽  
Periasamy Parikumar ◽  
Shojiro Katoh ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Cell Density ◽  
Corneal Thickness ◽  
Explant Culture ◽  
Endothelial Cell Density ◽  
P Value ◽  
Specular Microscopy ◽  
Human Donor ◽  
Corneal Preservation

Abstract Purpose McCarey-Kaufman’s (MK) medium and Optisol-GS medium are the most commonly employed media for human donor corneal preservation. In this study, we evaluated the preservation efficacy of discarded human donor corneas using a Thermo-reversible gelation polymer (TGP) added to these two media. Methods Thirteen human corneal buttons collected from deceased donors, which were otherwise discarded due to low endothelial cell density (ECD) were used. They were stored in four groups: MK medium, MK medium with TGP, Optisol-GS and Optisol-GS with TGP at 4 °C for 96 h. Slit lamp examination and specular microscopy were performed. Corneal limbal tissues from these corneas were then cultured using explant methodology one with and the other without TGP scaffold, for 21 days. Results MK + TGP and Optisol-GS + TGP preserved corneas better than without TGP, which was observed by maintenance of ECD which was significantly higher in Optisol-GS + TGP than MK + TGP (p-value = 0.000478) and corneal thickness remaining the same for 96 h. Viable corneal epithelial cells could be grown from the corneas stored only in MK + TGP and Optisol-GS + TGP. During culture, the TGP scaffold helped maintain the native epithelial phenotype and progenitor/stem cell growth was confirmed by RT-PCR characterization. Conclusion TGP reconstituted with MK and Optisol—GS media yields better preservation of human corneal buttons in terms of relatively higher ECD maintenance and better in vitro culture outcome of corneal limbal tissue. This method has the potential to become a standard donor corneal transportation-preservation methodology and it can also be extended to other tissue or organ transportation upon further validation.

scholarly journals Evaluation of the HB&L System for the Microbiological Screening of Storage Medium for Organ-Cultured Corneas

2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
D. Camposampiero ◽  
S. Grandesso ◽  
E. Zanetti ◽  
S. Mazzucato ◽  
M. Solinas ◽  
...  
Keyword(s):  
Contamination Rate ◽  
Storage Medium ◽  
Human Donor ◽  
Predictive Values ◽  
Anaerobic Microorganisms ◽  
L System ◽  
Microbiological Control ◽  
Corneal Storage ◽  
Aerobic And Anaerobic ◽  
Donor Corneas

Aims. To compare HB&L and BACTEC systems for detecting the microorganisms contaminating the corneal storage liquid preserved at 31°C.Methods. Human donor corneas were stored at 4°C followed by preservation at 31°C. Samples of the storage medium were inoculated in BACTEC Peds Plus/F (aerobic microorganisms), BACTEC Plus Anaerobic/F (anaerobic microorganisms), and HB&L bottles. The tests were performed (a) after six days of storage, (b) end of storage, and (c) after 24 hours of preservation in deturgescent liquid sequentially. 10,655 storage and deturgescent media samples were subjected to microbiological control using BACTEC (6-day incubation) and HB&L (24-hour incubation) systems simultaneously. BACTEC positive/negative refers to both/either aerobic and anaerobic positives/negatives, whereas HB&L can only detect the aerobic microbes, and therefore the positives/negatives depend on the presence/absence of aerobic microorganisms.Results. 147 (1.38%) samples were identified positive with at least one of the two methods. 127 samples (134 identified microorganisms) were positive with both HB&L and BACTEC. 14 HB&L+/BACTEC− and 6 BACTEC+/HB&L− were identified. Sensitivity (95.5%), specificity (99.8%), and positive (90.1%) and negative predictive values (99.9%) were high with HB&L considering a 3.5% annual contamination rate.Conclusion. HB&L is a rapid system for detecting microorganisms in corneal storage medium in addition to the existing methods.

scholarly journals Corneal thickness and endothelial cell density in diabetic and non- diabetic patients: a hospital based comparative study

2018 ◽  
Vol 5 (3) ◽  
pp. 694 ◽  
Author(s):  
Nagaraj G. ◽  
Anasuya Sangaraj Desai ◽  
Nagesh Jayaram
Keyword(s):  
Endothelial Cell ◽  
Cell Density ◽  
Central Corneal Thickness ◽  
Corneal Thickness ◽  
Endothelial Cell Density ◽  
Diabetic Group ◽  
Control Group ◽  
Diabetic Patients ◽  
Specular Microscopy ◽  
The Mean

Background: Diabetes mellitus is associated with structural changes in corneal endothelial cells and their thickness. The present study was done to compare the endothelial cell density (ECD), central corneal thickness (CCT) and morphology in diabetic and non-diabetic patients.Methods: A cross-sectional study was conducted at Minto Ophthalmic hospital, BMC and RI Bangalore for a period of 20 months (October 2013 - May 2015). A total of 200 study subjects, 100 diabetics and 100 non-diabetic age matched controls were selected, and complete timed ophthalmic evaluation was performed. Specular microscopy was performed on all patients for endothelial cell count assessment and corneal thickness was measured by Pachymeter. The data was analyzed and represented using descriptive statistics. ‘t’ test was used for comparing the two groups.Results: The mean endothelial cell density in diabetic group was significantly lower (2438.73±250.23cells/mm2) compared to non-diabetic group (2599.88±168.16cells/mm2) (p<0.0001). The mean Central corneal thickness in diabetic group was significantly higher (518.40±28.13 μm) compared to control group (490.14±24.31 μm) (p<0.001). The Co-efficient of variation percentage of the diabetics was higher than the non-diabetics but this difference was not statistically significant (P>0.05). The hexagonality percentage was significantly lower in diabetic group compared to the controls suggesting less pleomorphism in the diabetic group.Conclusions: The study concludes that the endothelial cell density was lower and central corneal thickness was higher in diabetic patients compared with the non-diabetics. The altered endothelial morphology was significantly seen in the form of pleomorphism (hexagonality) but polymegathism was not significantly altered.

scholarly journals Corneal Thickness and Endothelial Morphology in Normal Thai Eyes

2020 ◽  
Author(s):  
Napaporn Tananuvat ◽  
Natawan Khumchoo
Keyword(s):  
Endothelial Cell ◽  
Cell Density ◽  
Central Corneal Thickness ◽  
Corneal Thickness ◽  
Age Groups ◽  
Inverse Correlation ◽  
Cell Loss ◽  
Endothelial Cell Density ◽  
Specular Microscopy ◽  
The Mean

Abstract Background: This study aimed to determine the influence of age on central corneal thickness and corneal endothelial morphology as well as to identify the relationship between them in normal Thai eyes.Methods: Non-contact specular microscopy was performed in volunteers stratified into seven age groups ranging from 11-88 years. The corneal endothelial parameters studied included central corneal thickness (CCT), endothelial cell density (ECD), coefficient of variation in cell size (CV), cell area (CA) and percentage of regular hexagonal cells.Results: In a total of 501 subjects (1002 eyes), the mean age was 43.12 ± 18.80 years and 347(69.3%) were females. The mean CCT, ECD, CV, CA, and hexagonality was 533.80 ± 33.00 μm, 2,732±258 cell/mm2, 37.61 ± 6.76%, 369.04 ± 37.90 mm, and 49.03% ± 7.53%, respectively. There was a statistically significant inverse correlation between age and CCT (r = -0.212, P <0.001), ECD (r = -0.484, P <0.001), and hexagonality (r = -0.193, P <0.001). The estimate rate of endothelial cell loss was 0.2% per year, whereas CV and CA had statistically significant correlation with age (r = 0.200, P <0.001 and r = 0.475, P <0.001). CCT correlated directly with ECD (r = 0.177, P <0.001).Conclusion: Normative data for corneal endothelial morphology in healthy Thai eyes showed that ECD, CCT, and hexagonality were significantly decreased, while CV and CA were increased with aging. The central corneal thickness was correlated with the endothelial cell density.

scholarly journals Ophthalmic parameters in adult Shih Tzu dogs

2015 ◽  
Vol 45 (7) ◽  
pp. 1280-1285 ◽  
Author(s):  
Karina Kamachi Kobashigawa ◽  
Tiago Barbalho Lima ◽  
Ivan Ricardo Martinez Padua ◽  
Alexandre Augusto Franchi de Barros Sobrinho ◽  
Fabio de Andrade Marinho ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Standard Error ◽  
Corneal Thickness ◽  
Physical Restraint ◽  
Corneal Opacity ◽  
Applanation Tonometry ◽  
Specular Microscopy ◽  
Schirmer Tear Test ◽  
Time Test ◽  
Breakup Time

Ocular surface diseases are often diagnosed in brachycephalic dogs. The ophthalmic parameters of the Shih Tzu dogs are evaluated in this study since this breed is among the most commonly affected by these diseases. In this study, ophthalmic parameters of this breed were partly studied under physical restraint. Schirmer tear test, breakup time test, aesthesiometry, biomicroscopy, non-contact specular microscopy, laser flaremetry, applanation tonometry, ultrasonography and ophthalmoscopy were carried in 48 eyes of 24 male and female dogs, aged from two to four years, weighing between 5 and 10kg. All dogs were from a breeding kennel. Descriptive statistics were applied to the data. Mean and standard error for Schirmer tear test was 26.145±0.803mm min-1; breakup time test, 13.668±0.538s; and aesthesiometry, 2.395±0.071cm. The biomicroscopy evaluation showed that 70.83% of the eyes had medial entropion; 42% caruncular trichiasis; 33% distichiasis, 27% mild paracentral corneal opacity; and 13% corneal melanosis. Mean and standard error for endothelial cell density was 2221.591±20.161cells mm-2; endothelial cell hexagonality, 63.770±1.805%; endothelial cell area, 451.895±4.179mm2; central corneal thickness, 0.490±0.007 mm; laser flaremetry, 1.720±0.216PC ms-1; applanation tonometry, 16.118±0.460mmHg; axial length, 20.255±0.134mm; lens thickness, 6.624±0.031mm; anterior chamber, 4.064±0.109mm; and vitreous chamber, 9.565±0.054mm. Ophthalmoscopy findings were not different from previous reports for other breeds. The results showed that the ophthalmic Shih Tzu parameters did not differ from other breeds according to data in the literature, except for the breakup time and Cochet-Bonnet aesthesiometry.

scholarly journals A Comparative Study of Corneal Endothelial Cell Count and Central Corneal Thickness in Emmetropia and Axial Myopia

2020 ◽  
Vol 9 (44) ◽  
pp. 3300-3304
Author(s):  
Vandana Panjwani ◽  
Sachin Daigavane
Keyword(s):  
Endothelial Cell ◽  
Comparative Study ◽  
Cell Count ◽  
Axial Length ◽  
Central Corneal Thickness ◽  
Corneal Thickness ◽  
Corneal Endothelial Cell ◽  
Specular Microscopy ◽  
Endothelial Cell Count ◽  
Axial Myopia

BACKGROUND The main attributes of cornea which makes it optically important is its ability to maintain its dehydrated state. This corneal dehydration can be achieved and maintained by various factors among which corneal endothelial cell count and morphology play a significant role. Corneal attributes considered in this study i.e. the corneal endothelial cell count and central corneal thickness are extremely variable, and changes are seen even with smaller changes in anterior segment or overall structural changes in eyes. We wanted to compare central corneal thickness in emmetropia and axial myopia and compare corneal endothelial cell count in emmetropia and axial myopia. METHODS The sample size was decided taking into consideration various prevalence studies. The patients were consecutively recruited for the study considering the inclusion and exclusion criteria. The subjects were divided into two groups emmetropia and axial myopia and were differentiated on the basis of the axial lengths. Specular microscopy was performed for the subjects and corneal endothelial cell count, morphology and central corneal thickness were measured, and comparative study was performed. RESULTS This study of 80 eyes shows us that the difference between mean the corneal endothelial cell count between emmetrope (2812.80 cells / mm2) and axial myopes (2653 cells / mm2) is statistically significant where p was < 0.05 while the mean central corneal thickness measurements didn’t show statistically significant change between emmetrope (490.05 microns) and axial myope. (489.37 microns). CONCLUSIONS Axial length has an indirect correlation with the corneal endothelial cell count which is statistically significant. While significant correlation between the central corneal thickness and axial length cannot be established in this study. KEY WORDS Emmetropia, Axial Myopia, Corneal Endothelial Cell Count, Central Corneal Thickness, Comparison

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