Improving the success rate of human corneal endothelial cell cultures from single donor corneas with stabilization medium.

Main objective of this study was to improve the success rate of human corneal endothelial cell (hCEC) cultures from single donor corneas. We could show that the use of stabilization medium prior to cell isolation may have a positive effect on the success rate of hCEC cultures from single research-grade donor corneas by allowing growth of otherwise possibly not successful cultures and by improving their proliferative rate. hCEC were obtained from corneo-scleral rims of 7 discarded human research-grade cornea pairs. The Descemet membrane-endothelium (DM-EC) sheets of each pair were assigned to 2 experimental conditions: (1) immediate cell isolation after peeling, and (2) storage of the DM-EC sheet in a growth factor-depleted culture medium (i.e. stabilization medium) for up to 6 days prior to cell isolation. hCEC isolated by enzymatic digestion were then induced to proliferate on pre-coated culture plates. The success rate of primary cultures established from single donor corneas were higher for DM-EC sheets kept in stabilization medium before cell isolation. All cultures (7/7) initiated from stabilized DM-EC sheets were able to proliferate up to the third passage, while only 4 out of 7 cultures initiated from freshly peeled DM-EC sheets reached the third passage. In addition, for the 4 successful paired cultures we observed a faster growth rate if the DM-EC sheet was pre-stabilized prior to cell isolation (13.8 ± 1.8 vs 18.5 ± 1.5 days, P < 0.05). Expression of the phenotypical markers Na+/K+-ATPase and ZO-1 could be shown for the stabilized cultures that successfully proliferated up to the third passage.

La estandarización en el trasplante endotelial de membrana de Descemet: resultados de las primeras 450 cirugías / Standardization of the Descemet membrane endothelial keratoplasty technique: Outcomes of the first 450 consecutive cases

OBJETIVOS: Analizar los resultados de 450 casos con diferentes patologías endoteliales intervenidos mediante trasplante endotelial con la técnica de recambio de la membrana de Descemet (DMEK, por sus siglas en inglés: Descemet membrane endothelial keratoplasty) y evaluar las consecuencias de la estandarización de esta técnica. MATERIAL Y MÉTODOS: Se compararon 3 subgrupos consecutivos: el subgrupo I (casos 1-125) representaba la extensión de la curva de aprendizaje; el subgrupo II (casos 126-250) la transición a la estandarización de la técnica y el subgrupo III (casos 251-450) la estandarización propiamente dicha. Se registraron los resultados de agudeza visual mejor corregida pre- y postoperatorios, la densidad de células endoteliales, la paquimetría y las complicaciones. RESULTADOS: A los 6 meses de la cirugía, el 79% de los pacientes alcanzaron una agudeza visual mejor corregida ≥ 0,8 y el 43% ≥ 1. La densidad de células endoteliales media preoperatoria fue 2.530 ± 220 células/mm2, y 1.613 ± 495 células/mm2 al sexto mes poscirugía. La paquimetría era 668 ± 92 μm y 526 ± 46 μm pre- y postoperatoria a los 6 meses, respectivamente. No se encontraron diferencias en cuanto a la agudeza visual mejor corregida, la densidad de células endoteliales o la paquimetría entre los subgrupos (p > 0,05). El desprendimiento del injerto se observó en el 17,3% de los ojos. La tasa de desprendimientos disminuyó del 24 al 12%, y el número de reintervenciones, del 9,6 al 3,5%, del subgrupo I al III respectivamente. CONCLUSIONES: Los resultados visuales y la densidad de células endoteliales tras DMEK son independientes de la estandarización de la técnica quirúrgica. Sin embargo, la estandarización de la técnica podría haber contribuido con un descenso en el número de desprendimientos y con un relativamente bajo número de intervenciones secundarias. A la vista de estos resultados, DMEK podría convertirse en el tratamiento de elección para las enfermedades del endotelio corneal

OBJECTIVES: To evaluate the clinical outcome of the first 450 consecutive cases after Descemet membrane endothelial keratoplasty (DMEK), as well as the effect of standardization of the technique. MATERIAL AND METHODS: Comparison between 3 groups: Group I: (cases 1-125), as the extended learning curve; Group II: (cases 126-250), transition to technique standardization; Group III: (cases 251-450), surgery with standardized technique. Best corrected visual acuity, endothelial cell density, pachymetry and intra- and postoperative complications were evaluated before, and 1, 3 and 6 months after DMEK. RESULTS: At 6 months after surgery, 79% of eyes reached a best corrected visual acuity of≥0.8 and 43%≥1.0. Mean preoperative endothelial cell density was 2,530±220 cells/mm2 and 1,613±495 at 6 months after surgery. Mean pachymetry measured 668±92 μm and 526±46 μm pre- and (6 months) postoperatively, respectively. There were no significant differences in best corrected visual acuity, endothelial cell density and pachymetry between the 3 groups (P > .05). Graft detachment presented in 17.3% of the eyes. The detachment rate declined from 24% to 12%, and the rate of secondary surgeries from 9.6% to 3.5%, from group I to III respectively. CONCLUSIONS: Visual outcomes and endothelial cell density after DMEK are independent of the technique standardization. However, technique standardization may have contributed to a lower graft detachment rate and a relatively low number of secondary interventions required. As such, DMEK may become the first choice of treatment in corneal endothelial disease

Standardization of the Descemet membrane endothelial keratoplasty technique: Outcomes of the first 450 consecutive cases.

OBJECTIVES: To evaluate the clinical outcome of the first 450 consecutive cases after Descemet membrane endothelial keratoplasty (DMEK), as well as the effect of standardization of the technique. MATERIAL AND METHODS: Comparison between 3 groups: Group I: (cases 1-125), as the extended learning curve; Group II: (cases 126-250), transition to technique standardization; Group III: (cases 251-450), surgery with standardized technique. Best corrected visual acuity, endothelial cell density, pachymetry and intra- and postoperative complications were evaluated before, and 1, 3 and 6 months after DMEK. RESULTS: At 6 months after surgery, 79% of eyes reached a best corrected visual acuity of≥0.8 and 43%≥1.0. Mean preoperative endothelial cell density was 2,530±220 cells/mm2 and 1,613±495 at 6 months after surgery. Mean pachymetry measured 668±92 µm and 526±46 µm pre- and (6 months) postoperatively, respectively. There were no significant differences in best corrected visual acuity, endothelial cell density and pachymetry between the 3 groups (P > .05). Graft detachment presented in 17.3% of the eyes. The detachment rate declined from 24% to 12%, and the rate of secondary surgeries from 9.6% to 3.5%, from group I to III respectively. CONCLUSIONS: Visual outcomes and endothelial cell density after DMEK are independent of the technique standardization. However, technique standardization may have contributed to a lower graft detachment rate and a relatively low number of secondary interventions required. As such, DMEK may become the first choice of treatment in corneal endothelial disease.

Agudeza visual y densidad de células endoteliales tras queratoplastia endotelial de membrana de Descemet (DMEK) / Visual acuity and endothelial cell density following Descemet membrane endothelial keratoplasty (dmek)

Objetivo: Evaluación de los resultados visuales y de la densidad de células endoteliales en los 120 primeros ojos sometidos a queratoplastia endotelial de membrana de Descemet (DMEK) como tratamiento para la distrofia endotelial de Fuchs. Material y métodos: Se evaluaron los primeros 120 ojos sometidos a DMEK. En todos ellos se midió la agudeza visual mejor corregida (AVMC) antes y al 1, 3 y 6 meses después de la cirugía, así como la densidad celular endotelial (DCE) antes y a los 6 meses. Resultados: En los ojos con trasplante exitoso y sin patologías concomitantes (n=96), se observo una AVMC en el 77% de los casos al mes de la cirugía, en el 92% a los 3 meses y en el 95% a los 6 meses. La AVMC fue mayor o igual 0,8 en el 50%, 63% y 73% de los casos y mayor o igual 1,0 en el 23%, 34% y 45% al 1, 3 y 6 meses después de la cirugía, respectivamente. La DCE preoperatoria fue 2.610 (más menos,185) células/mm2 y 1.770 (más menos, 520) células/mm2 a los 6 meses postcirugía. En quince ojos se realizó una queratoplastia endotelial con pelado de la membrana de Descemet (DSEK) secundaria. En este grupo, el 91% de los pacientes alcanzó una AVMC mayor o igual 0,5, alcanzando solo uno de ellos una AVMC de 0,8 (n=11) a los 6 meses. Además, la DCE media fue de 2.580 (más menos, 185) células/mm2 antes y de 1.310 (más menos, 740) células/mm2 a los 6 meses de la operación (n=13). Conclusiones: DMEK permite una rápida y casi completa rehabilitación visual. La DCE postoperatoria observada es comparable al obtenido con técnicas precedentes de queratoplastia endotelial(AU)

Purpose: To assess the clinical outcomes of DMEK in the first series of 120 eyes operated for the treatment of Fuchs endothelial dystrophy in terms of visual acuity and endothelial cell density. Methods: The first 120 consecutive eyes that underwent DMEK (i.e. transplantation of an isolated donor Descemet membrane with its endothelium) were evaluated. In all eyes, the best corrected visual acuity (BCVA) before and at 1, 3 and 6 months after surgery, as well as the endothelial cell density (ECD) before and at 6 months were measured. Results: In eyes with a functional DMEK graft and good visual potential (n=96), the BCVA was major or equal 20/40 (major or equal 0.5) in 77% after 1 month, 92% after 3 months, and 95% after six months; major or equal 20/25 (major or equal 0.8) in 50%, 63%, and 73% of the cases, and major or equal 20/20 (major or equal 1.0) in 23%, 34%, and 45% of the cases at 1, 3, and 6 months respectively. In this group, ECD averaged 2610 (more less 185) cells/mm2 before, and 1770 (more less 520) cells/mm2 at six months after surgery (n=96). In 15 eyes, a secondary Descemet stripping endothelial keratoplasty (DSEK) was performed. In this group, 91% of patients reached a BCVA of major or equal 20/40 (major or equal 0.5) and only one patient achieved a BCVA of 0.8 at 6 months after surgery (n=11). Furthermore, ECD averaged 2580 (more less 185) cells/mm2 before and 1310 (more less 740) cells/mm2 at six months (n=13). Conclusion: DMEK provides a fast and high visual rehabilitation. Endothelial cell density loss may be similar to earlier types of endothelial keratoplasty(AU)

[Visual acuity and endothelial cell density following Descemet membrane endothelial keratoplasty (DMEK)]. / Agudeza visual y densidad de células endoteliales tras queratoplastia endotelial de membrana de Descemet (DMEK).

PURPOSE: To assess the clinical outcomes of DMEK in the first series of 120 eyes operated for the treatment of Fuchs endothelial dystrophy in terms of visual acuity and endothelial cell density. METHODS: The first 120 consecutive eyes that underwent DMEK (i.e. transplantation of an isolated donor Descemet membrane with its endothelium) were evaluated. In all eyes, the best corrected visual acuity (BCVA) before and at 1, 3 and 6 months after surgery, as well as the endothelial cell density (ECD) before and at 6 months were measured. RESULTS: In eyes with a functional DMEK graft and good visual potential (n=96), the BCVA was ≥ 20/40 (≥ 0.5) in 77% after 1 month, 92% after 3 months, and 95% after six months; ≥ 20/25 (≥ 0.8) in 50%, 63%, and 73% of the cases, and ≥ 20/20 (≥ 1.0) in 23%, 34%, and 45% of the cases at 1, 3, and 6 months respectively. In this group, ECD averaged 2610 (± 185) cells/mm(2) before, and 1770 (± 520) cells/mm(2) at six months after surgery (n=96). In 15 eyes, a secondary Descemet stripping endothelial keratoplasty (DSEK) was performed. In this group, 91% of patients reached a BCVA of ≥ 20/40 (≥ 0.5) and only one patient achieved a BCVA of 0.8 at 6 months after surgery (n=11). Furthermore, ECD averaged 2580 (± 185) cells/mm(2) before and 1310 (± 740) cells/mm(2) at six months (n=13). CONCLUSION: DMEK provides a fast and high visual rehabilitation. Endothelial cell density loss may be similar to earlier types of endothelial keratoplasty.

[Visual acuity following Descemet-membrane endothelial keratoplasty (DMEK): first 100 cases operated on for Fuchs endothelial dystrophy]. / Visus nach Descemet-Membran Endothelkeratoplastik (DMEK): Ergebnisse der ersten 100 Eingriffe bei Fuchs'scher Endotheldystrophie.

PURPOSE: The aim of this study was to evaluate the visual outcome and endothelial cell density in the first series of 100 eyes after Descemet membrane endothelial keratoplasty (DMEK) for the treatment of Fuchs endothelial dystrophy. DESIGN: This was a non-randomised prospective clinical trial. METHODS: The first 100 consecutive eyes (87 patients) that underwent DMEK (i. e., transplantation of an isolated donor Descemet membrane carrying its endothelium) for Fuchs endothelial dystrophy were evaluated. In all eyes, the best corrected visual acuity (BCVA) as well as the endothelial cell density (ECD) were measured before and at one, three and six months after surgery. RESULTS: In eyes with a functional DMEK graft and good visual potential (n = 78), the BCVA was > or = 20 / 40 (> or = 0.5) in 79 % after 1 month, 94 % after 3 months, and 96 % after six months; > or = 20 / 25 (> or = 0.8) in respectively 54 %, 64 %, and 74 %, and > or = 20 / 20 (> or = 1.0) in respectively 26 %, 36 %, and 45 %. In this group, ECD averaged 2630 (+/- 190) cells/mm (2) before, and 1800 (+/- 540) cells/mm (2) at six months after surgery (n = 76). In 15 eyes, a secondary "Descemet stripping endothelial keratoplasty" (DSEK) was performed. When the outcomes of DMEK and secondary DSEK procedures were combined, 91 % reached a BCVA of > or = 20 / 40 (> or = 0.5), 65 % > or = 20 / 25 (> or = 0.8) and 38 % > or = 20 / 25 (> or = 1.0) at six months after surgery (n = 93). For the whole group, ECD averaged 2620 (+/- 190) cells/mm (2) before, and 1730 (+/- 590) cells/mm (2) at six months after surgery (n = 89). CONCLUSION: DMEK may provide relatively quick and nearly complete visual rehabilitation in a majority of patients operated on for Fuchs endothelial dystrophy. Endothelial cell survival may be similar to earlier types of endothelial keratoplasty. Early graft detachment (19 cases) was the main complication in this first series of DMEK surgeries.

[Descemet membrane endothelial keratoplasty (DMEK): two-year results]. / Queratoplastia endotelial de membrana de Descemet (DMEK): resultados a dos años.

PURPOSE: To report the two-year results of Descemet membrane endothelial keratoplasty (DMEK) for managing corneal endothelial disorders. METHODS: Non-randomized prospective clinical trial. A DMEK was performed in ten patients with Fuchs' endothelial dystrophy or bullous keratopathy. A 3.5 mm clear corneal incision was made and "under air" DM was stripped off from the posterior stroma. A 9.0 mm diameter, organ cultured donor DM roll was inserted into a recipient anterior chamber, positioned into the posterior stroma and secured by completely filling the anterior chamber with air for 30 minutes. RESULTS: Three eyes showed complete detachment of the tissue; this was managed by a secondary Descemet stripping endothelial keratoplasty procedure. The remaining seven eyes had a best corrected visual acuity of >or=0.7 in three eyes (43%) at one month, in five eyes (71%) at six months, and in six eyes (86%) at one and two years. At six months, the endothelial cell density averaged 2039 (+/-373) cells/mm2 (n=7), at one year 1925 (+/-267) cells/mm2 (n=7) and at two years 1730 (+/-400) cells/mm2 (n=6). CONCLUSIONS: DMEK may provide quick and nearly complete visual rehabilitation. Since the donor tissue can be stripped from donor corneo-scleral rims, the procedure may be readily accessible to most corneal surgeons.

Queratoplastia endotelial de membrana de Descemet (DMEK): resultados a dos años / Descemet membrane endothelial keratoplasty (DMEK): Two-year results

Objetivo: Describir los resultados, dos años despuésde realizar una queratoplastia endotelial demembrana de Descemet (DMEK: Descemet membraneendothelial keratoplasty), para el tratamientode alteraciones del endotelio corneal.Métodos: Estudio clínico prospectivo no randomizado.En 10 pacientes con distrofia endotelial deFuchs o queratopatía bullosa, se practicó unaDMEK. A través de una incisión de 3,5 mm en córneaclara, la membrana de Descemet (MD) delreceptor fue desprendida del estroma posterior enpresencia de aire. Un disco de 9 mm de diámetroenrollado de MD donante preservada, fue insertadoen la cámara anterior del receptor, posicionado encontacto con el estroma posterior corneal y aseguradoen su posición mediante el llenado completode la cámara anterior con aire durante 30 minutos.Resultados: Tres ojos mostraron un desprendimientocompleto del tejido donante, por lo que fueronsometidos posteriormente a una queratoplastiaendotelial con «pelado» de la MD (DSEK: Descemet stripping endothelial keratoplasty). En los sieteojos restantes, se observó una agudeza visual mejorcorregida (AVMC) ≥ a 0,7 en 3 ojos (43%) en el primermes, en 5 ojos (71%) a los seis meses, y en seisojos (86%) al primer y segundo años. A los seismeses, la densidad celular endotelial media fue de2039 (DS: 373) cél/mm2 (n=7), al año de 1925 (DS:267 cél/mm2 (n=7) y a los 2 años de 1730 (DS: 400)cél/mm2 (n=6).Conclusión: DMEK podría proporcionar una recuperaciónrápida y casi completa de la visión. Debidoa que el tejido donante puede ser obtenido a partirde anillos córneo-esclerales donantes, el procedimientopodría ser fácilmente accesible para lamayoría de los cirujanos corneales(AU)

Purpose: To report the two-year results of Descemetmembrane endothelial keratoplasty (DMEK)for managing corneal endothelial disorders.Methods: Non-randomized prospective clinicaltrial. A DMEK was performed in ten patients withFuchs’ endothelial dystrophy or bullous keratopathy.A 3.5 mm clear corneal incision was madeand «under air» DM was stripped off from the posteriorstroma. A 9.0 mm diameter, organ cultureddonor DM roll was inserted into a recipient anteriorchamber, positioned into the posterior stroma andsecured by completely filling the anterior chamberwith air for 30 minutes.Results: Three eyes showed complete detachmentof the tissue; this was managed by a secondary Descemetstripping endothelial keratoplasty procedure.The remaining seven eyes had a best correctedvisual acuity of ≥ 0.7 in three eyes (43%) at onemonth, in five eyes (71%) at six months, and in sixeyes (86%) at one and two years. At six months, theendothelial cell density averaged 2039 (±373) cells/mm2 (n=7), at one year 1925 (±267) cells/mm2(n=7) and at two years 1730 (±400) cells/mm2(n=6).Conclusions: DMEK may provide quick andnearly complete visual rehabilitation. Since thedonor tissue can be stripped from donor corneoscleralrims, the procedure may be readily accessibleto most corneal surgeons(AU)

Descemet membrane endothelial keratoplasty (DMEK) for Fuchs endothelial dystrophy: review of the first 50 consecutive cases.

PURPOSE: To evaluate the clinical outcome and complications of Descemet membrane endothelial keratoplasty (DMEK), using Descemet-stripping endothelial keratoplasty (DSEK) as a back-up procedure, in the management of Fuchs endothelial dystrophy. DESIGN: Non-randomised prospective clinical study. METHODS: The first fifty consecutive eyes that underwent DMEK, that is, transplantation of an isolated donor Descemet membrane carrying its endothelium, for Fuchs endothelial dystrophy were evaluated. In all eyes, the best-corrected visual acuity (BCVA) as well as the endothelial cell density (ECD) was measured before and at 6 months after surgery, as clinical outcome parameters. RESULTS: Ten patients required a secondary DSEK for failed DMEK. In the remaining 40 DMEK eyes, 95% had a BCVA of > or = 20/40 (> or = 0.5) and 75% > or = 20/25 (> or = 0.8) at 6 months after surgery. ECD averaged 2618 (+ or - 201) cells/mm(2) before, and 1876 (+ or - 522) cells/mm(2) at 6 months after surgery (n = 35). When the outcomes of DMEK and secondary DSEK procedures were combined, 94% reached a BCVA of > or = 20/40 (> or = 0.5) and 66% > or = 20/25 (> or = 0.8) (n = 47), and ECD averaged 2623 (+ or - 193) cells/mm(2) before, and 1815 (+ or - 578) cells/mm(2) at 6 months after surgery (n = 43). CONCLUSION: With DSEK as a back-up procedure, DMEK may provide relatively quick and complete visual rehabilitation in a majority of patients operated on for Fuchs endothelial dystrophy. Endothelial cell survival may be similar to earlier types of (lamellar) keratoplasty. Early graft detachment was the main complication in this first series of DMEK surgeries for Fuchs endothelial dystrophy.