Objective: This validation study investigates the treatment of cornea organ culture medium (Modified Eagle Medium, Biochrom GmbH, Berlin, Germany) with RESEP, a new medical device for antibiotics removal, before microbiological testing with BACTEC^TM blood culture bottles.

Methods and analysis: 10–100 colony forming units of Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, Bacillus subtillis, Aspergillus brasiliensis, Clostridium sporogenes, Enterobacter cloacae and Staphylococcus epidermidis were inoculated in 9mL of cornea organ culture medium.
In group A, the medium was withdrawn with RESEP and treated for 20 min at room temperature, and then inoculated in BACTEC Plus Aerobic/F/Anaerobic/F blood culture bottles.
In group B, the medium, spiked by the inoculation of microorganism, was injected directly. For each strain, a growth control was performed, by direct inoculation of the microorganisms in BACTEC^TM vials (positive control). All samples were incubated in the automated BACTEC^TM blood culture system at 36°C ±1°C for maximum of 14 days or until a positive reading. The elimination of antibiotics from the medium by RESEP was determined by high-performance liquid chromatography.

Results: After 20 min of RESEP treatment, 100% (n=9) of streptomycin, 100% (n=9) of amphotericin B and 99.7% (n=9) of penicillin G were eliminated.
In group A , all microorganisms were detected within 3 days of incubation with a sensitivity of 100% (n=99) and showed no significant delay compared with the positive controls.
In group B, the overall sensitivity was 67.9% (n=96) with a significant delay until detection of microbial growth for all tested microorganisms except for A. brasiliensis.

Conclusion: The use of RESEP to eliminate the antibiotics from cornea organ culture medium increases the sensitivity of the microbiological testing with BACTEC^TM Plus blood culture bottles significantly and fulfils the requirements of the European Pharmacopoeia method suitability test.

Purpose: To test the feasibility of a cell therapy approach to treat corneal endothelial (CE) disorders using an in vitro model of human corneal decompensation.

Methods: A CE decompensation model was established by removal of the Descemet membrane/endothelium complex from cadaveric human corneas in an air interface organ culture system (group 2) and compared with normal corneas (group 1). The posterior stroma of decompensated corneas was seeded with immortalized human corneal endothelial cells (HCEC-12) in group 3 and passage 0 primary human CE cells in group 4 corneas. Functional effects on stromal thickness were determined with histological analysis 3 to 10 days after cell therapy treatment.

Results: Removal of the Descemet membrane/endothelium complex in group 2 corneas resulted in a stromal thickness of 903 ± 86 μm at 12 hours compared with 557 ± 72 μm in group 1 corneas. Stromal thickness reduced from 1218 ± 153 μm to 458 ± 90 μm (63% ± 6%, P = 0.001) after cell transplantation in group 3 and from 1100 ± 86 μm to 489 ± 94 μm (55% ± 7%, P = 0.00004) in group 4. Posttransplantation histology demonstrated formation of a monolayer of corneal endothelium attached to the posterior stromal surface.

Conclusions: Direct transplantation of cultured human CE cells and immortalized HCEC-12 to bare posterior corneal stroma resulted in formation of anendothelial monolayer and restoration of stromal hydration to physiological thickness, demonstrating the feasibility of cell therapy in treatment of CE decompensation in a human in vitro model.

Purpose: To compare the protective properties and ease of manipulation during cataract surgery of corneal coating with a gel (eyeDRO; AL.CHI.MI.A. S.R.L, Italy) and corneal irrigation with balanced salt solution (BSS).

Methods: We analyzed the data of 51 patients receiving either eyeDRO or BSS during routine cataract surgery performed within a 20-day period in 2016. The selected parameters were intraoperative clarity and ease of manipulation; postoperative epithelial integrity; and patient discomfort.

Results: Compared with BSS irrigation, eyeDRO coating significantly increased intraoperative clarity and ease of manipulation (P < 0.01). Single application was required in eyeDRO-treated eyes, whereas BSS was applied 5.3 ± 0.4 times on average (P < 0.01). Two hours postoperatively, a normal epithelium was observed in 90.0% and 60.0% of eyeDRO-coated and BSS-irrigated eyes, respectively; punctate epithelial damage was observed in 9.7% and 40.0% (P < 0.05) of eyeDRO-coated and BSS-irrigated eyes, respectively; eye irritation and foreign body sensation were experienced by 13.0% and 37.0% of eyeDRO-treated patients and by 65.0% and 100% of BSS-treated patients, respectively (P < 0.01). Twenty-four hours postoperatively, 80.0% of BSS-treated patients versus 19.0% of eyeDRO-treated patients still experienced foreign body sensation (P < 0.01).

Conclusions: eyeDRO coating was shown to be a safer and more effective option than BSS irrigation in cataract surgery because single application provided optimal hydration and intraoperative clarity during the entire surgery, better preserved the corneal epithelium, and offered postoperative comfort to the patient.

Summary: A spanish study by electronical microscopy on corneas for DMEK, stored in Tissue-C at 31°C, shows that there are morphological and structural differences between the center and the periphery of the grafted tissue.

Purpose: The vitreous cortex, epiretinal membrane (ERM), and inner limiting membrane (ILM) are transparent tissues and are then difficult to visualize. Staining these structures can increase the efficiency of a nontraumatic removal.

Methods: The surgeon performs a partial core vitrectomy and induces a posterior vitreous detachment. The vital dye is then injected into the retrohyaloid space in balanced salt solution (BSS). The dyes used are TWIN (AL.CHI.MI.A. Srl, Padova, Italy), MembraneBlue-Dual (DORC International, Zuidland, The Netherlands), and Doubledyne (Alfa Intes, Casoria, Italy). The surgeon can complete the vitrectomy and gradually aspirate the dye with a probe. Once the vitrectomy is complete, the surgeon can perform the peeling of the ERM without the need to reinject the vital dye over the macula.

Results: The presence of the dye over the macula facilitates the visualization of the vitreous cortex by blocking the red reflex and increasing the contrast power of the coaxial light probe during the vitrectomy. This allows a negative staining of the vitreous because the dye acts by increasing the visibility of the surrounding BSS and not the vitreous itself.

Conclusions: We describe a new chromovitrectomy technique using the same dye to increase the visualization of the vitreous, posterior hyaloid, ERM, and ILM.

Objective: This study aimed at validating the method for sterility testing of the corneal culture medium, TISSUE-C, and the transport/deswelling medium, CARRY-C, according to the method suitability test, as defined by the European Pharmacopoeia (EP), using RESEP, which is a new medical device for removal of antimicrobial agents and an automated culture system.

Methods and analysis: The six EP reference strains were inoculated in TISSUE-C and CARRY-C. Half of the samples were treated with RESEP (RESEP+ group) prior to the sterility testing, whereas the remaining samples were untreated (RESEP− group). Growth controls were obtained by direct inoculation of the micro-organisms in the culture broths. Microbial growth was read by an automated light scattering culture system within 48 hours.

Results: The use of RESEP allowed detection of microbial growth in 100% of the tested samples, with a mean time to detection (TTD) comparable with that of the growth control group. Significantly lower sensitivity (38.83%±20.03% for both media, P<0.05) and TTD variability, depending on the tested micro-organism, were observed in the RESEP− group. The method specificity was 100% for both groups.

Conclusion: The use of RESEP increased the sensitivity of the sterility testing method to 100% and, for the first time, allowed validation of the method for sterility testing of corneal storage media according to the EP method suitability test. This further increases the safety of the corneas intended for transplantation.

Objectives: To evaluate the efficacy of Eusol-C as a corneal storage medium on the survival of donor endothelium.

Materials and Methods: Twenty-seven corneas not suitable for transplant were included in this study. All donor corneas were stored in Eusol-C at 4°C. Daily donor corneal endothelial cell counting was performed with an eye bank specular microscope. All corneas were discarded after the study process.

Results: Mean donor age was 51.3 ± 18.8 years (range, 25-94 y). The mean duration between death and corneal excision was 9.5 ± 6.7 hours (range, 3-23 h). Mean endothelial cell density was 2195 ± 383 cells/mm² at the beginning of the preservation (range, 1361-2899 cells/mm²). Donor endothelial cell density was between 1500 to 2000 cells/mm² in 9 corneas, 2000 to 2500 in 11 corneas, 2500 to 3000 in 5, and higher than 3000 in 2 corneas at baseline. Mean endothelial cell density was found 1658 cells/mm² on the eighth day of storage, with a mean endothelial cell loss rate of 24.5%. Corneas stored 9 to 24 days in Eusol-C had a rate of endothelial cell damage of 3.1% per day.

Conclusions: Although our results revealed a higher endothelial cell loss than previous reports, overall performance of Eusol-C in preserving the donor endothelium may be satisfactory for clinical use.

Purpose: To report the surgical results of primary pseudophakic  rhegmatogenous retinal detachment (RRD) with only inferior retinal breaks (IRBs) repaired by pars plana vitrectomy (PPV) alone and complete drainage of subretinal fluid.

Design: Prospective, interventional cohort study.

Participants: A total of 147 consecutive pseudophakic eyes of 147 patients with primary RRD with causative inferior breaks.

Methods: All eyes underwent PPV alone and complete drainage of subretinal fluid, with air, 20% sulfur hexafluoride (SF6), or 12% perfluoropropane (C3F8) as tamponade and with no face-down position in the postoperative period.

Main Outcome Measures: Postoperative primary and final anatomic outcome, visual acuity, and complications.

Results: The patient population consisted of 44 women (30%) and 103 men (70%) with a mean age of 60.8
12.9 years. The mean follow-up period was 24.9
19.4 months. The mean number of quadrants affected was 2.4 (range, 1e4). A single break was present in 90 cases (61.2%), and 2 to 4 breaks were present in 57 cases (38.8%). The macula was found to be detached in 118 cases (80.3%) and attached in 29 cases (19.7%) intraoperatively.
Of 152 breaks located between 5 and 7 clock-hours, 124 breaks (81.6%) were located outside the limits of the gas bubble on the first or third day postoperatively. Initial reattachment was achieved in 139 cases (94.5%; 95% confidence interval, 89.5e97.6). Final reattachment was achieved in 147 cases (100%). Two cases (1.3%) redetached because of new or missed retinal breaks. Six cases (4.1%) redetached because of incomplete retinal adhesion of the treated break(s). Mean preoperative best-corrected visual acuity (BCVA) was 1.11
0.59 logarithm of the minimum angle of resolution (logMAR). The mean final postoperative BCVA was 0.42
0.33 logMAR.

Conclusions: Pars plana vitrectomy alone with complete drainage of subretinal fluid achieves a high reattachment rate in the management of primary pseudophakic RRD due to IRBs.