1. Configuración de campo estéril <ol><li> Don atuendo adecuado incluyendo gorro y mascarilla. Use protección para los ojos cuando en la sala de procesamiento. </li><li> Prepare el área de procesamiento. Cuando se utiliza una sala de procesamiento, mantener la operación continua del ventilador para crear presión positiva en la habitación. </li><li> Reúna todos los suministros. Compruebe las fechas de caducidad, la integridad del envase, y el indicador de esterilización. </li><li> Lavarse las manos y ponerse los guantes no estériles. </li><li> Abra los kits estériles para establecer un campo estéril. Al utilizar una sala de procesamiento, colocar los kits en la mesa y asépticamente abrir las envolturas. No utilizar los exteriores de 1 ½ pulgadas de la envoltura como parte del campo estéril ya que este segmento se considera no estéril. </li><li> Utilizando una técnica aséptica, abierta y soltar elementos estériles en campo estéril a menos que ya forman parte del paquete de preparado de antemano. </li><li> Mueva microscopio de modo que está por encima del campo estéril, encienda la fuente de luz de microscopio, y ajustar los oculares según sea necesario. </li><li> Obtain tejido corneal para ser despojado (en los medios de conservación) y el lugar al lado de la zona de procesamiento estéril preparada. Afloje la tapa de la cámara de observación y se deja reposar en la parte superior de la visualización de fondo de la cámara sin hilos dedicados. Deseche los guantes no estériles. </li><li> Abra un par de guantes estériles y bata estéril con una toalla. </li><li> Realizar lavado quirúrgico. Ponga en bata estéril, guantes estériles entonces. </li><li> Cajas de instrumentos abiertas para que todos los suministros son accesibles. Abrir la humedad cortina impermeable para crear el área de trabajo. </li><li> Coloque la aguja a una jeringa de 5 cc. Utilizando una pinza hemostática, preparar la aguja de 30 G mediante la colocación de una curva 1 mm del bisel, y otra curva 1 mm desde la base (para formar una forma de &quot;S&quot;). </li><li> Montar la jeringa de vidrio para la tinción de azul de tripano al colocar el émbolo en la jeringa y el dedo apretando el émbolo. Conecte la cánula. </li><li> Conecte la cánula a la botella de solución salina balanceada (BSS). </li><li> Separar el ponche trépano desde el bloque de vacíoy dejar de lado. Centre el bloque de vacío dentro del campo de visión del microscopio. </li><li> Traiga el bloque vacío en foco ajustando la configuración del microscopio. </li></ol> 2. Procedimiento para la Preparación del Tejido Donante <ol><li> Levante la tapa de la cámara de visión usando cuadrado de lámina como barrera. Con la mano opuesta y con unas pinzas, agarre suavemente córnea esclerótica llanta para eliminar córnea. Vuelva a colocar la tapa de la cámara de observación. </li><li> Completamente deprimir la jeringa de succión, a continuación, coloque el lado epitelial de la córnea hacia abajo el bloque de vacío y asegúrese de que está centrado. (Figura 1) </li><li> Baje suavemente el anillo de asientos en el bloque vacío utilizando los postes de guía. Enganche de vacío liberando lentamente el pistón de la jeringa de aspiración. Retire el anillo de asiento y reservar. (Figura 2). </li><li> Realizar una trepanación parcial para anotar a través de la membrana de Descemet. </li><li> Ajuste microscopio zoom y el enfoque de manera que el tejido se visualiza bien. </li><li&gt; Incline el bloque vacío y utilizar una lanza hisopo estéril colocado en el limbo para eliminar el exceso de medio. (Figura 3) </li><li> Coloque suficiente mancha en el limbo para teñir el borde de la membrana de Descemet marcado. Permita que la mancha de permanecer en el endotelio de 60-90 seg. (Figura 4) </li><li> Elimine las manchas: <ol><li> Incline el bloque de vacío para verter la mancha de la córnea y en la almohadilla de gasa. </li><li> Utilice las lanzas quirúrgicas para eliminar el exceso de colorante de la zona del limbo de la córnea, con cuidado de no tocar el endotelio. (Figura 5) </li></ol></li><li> Utilice varias gotas de BSS para enjuagar la córnea de solución de tinción, y lanzas de hisopos estériles para eliminar el exceso de BSS. (Figura 6) </li><li> Coloque 1-2 gotas de BSS en el borde esclerótica para evitar que el tejido de secado. Repita según sea necesario a lo largo del procedimiento. </li><li> Lleve a cabo la disección parcial de la membrana de Descemet: <ol><li> Comenzando en el que el tubo de conectars para el bloque de vacío, utilizando fórceps abierto, coloque una punta en el borde de la marca de puntuación y la membrana de Descemet separada suavemente y el endotelio lejos de la estroma. (Figura 7). </li><li> Membrana de Descemet separada y el endotelio no más de 1 mm del borde marcado y continuar la disección alrededor de 360 ​​°. (Figura 8). </li><li> Nota ubicación de cualquier lágrimas micro mediante la colocación de un punto en el bloque de vacío con el marcador de la piel. NOTA: Si el trépano no penetra completamente a través de la membrana de Descemet, utilizar el borde biselado de la aguja de 30 g preparado para completar la puntuación a través de la membrana de Descemet cuando sea necesario, seguido por una segunda aplicación de tinción según sea necesario. Si la adherencia de la membrana de Descemet está impidiendo la disección parcial, eliminar el endotelio fuera del área trephinated según sea necesario. </li></ol></li><li> El uso de fórceps para quitar cualquier etiqueta endoteliales que se superponen la marca de puntuación. </li> &lt;li&gt; Tenga en cuenta la ubicación de los previamente marcados micro desgarros y girar el bloque vacío para que el más grande es a las 6 en punto. Esto se convertirá en la bisagra del colgajo. </li><li> Usando las pinzas, agarre la membrana de Descemet a las 12 horas. Separar suavemente la membrana de Descemet del estroma por pelado hacia la bisagra. Crear la bisagra al detener la separación de 2 mm desde la marca de puntuación. NOTA: los bordes periféricos de aleta comenzarán a desplazarse durante la separación. (Figura 9). </li><li> Coloque con cuidado la tapa en su lugar en el estroma. Utilice BSS y de frotis lanzas a unscroll el endotelio y devolverlo a su posición original. (Figura 10A-C) </li><li> Extraer el líquido residual de entre la membrana y el estroma de Descemet con lanzas de hisopos estériles por lo que el colgajo se mantiene en su posición. (Figura 11) </li><li> Utilice lanzas hisopos estériles para absorber el exceso de humedad de la esclerótica cerca de la bisagra. Utilice el marcador de la piel para dibujar unn punta de flecha en el borde esclerótica apuntando al centro de la bisagra, eliminar cualquier exceso de tinta usando una lanza hisopo estéril. (Figura 12) </li><li> Liberación de vacío presionando el émbolo de la jeringa conectada al bloque de vacío. Si bien se desengancha de vacío, el uso de fórceps para quitar córnea desde el bloque de vacío. </li><li> Levante la tapa de la cámara de visión usando cuadrado de lámina como barrera. Coloque la córnea en la cámara de visión, lado endotelial, y coloque la tapa de la cámara de observación. </li><li> Repita los pasos anteriores procedimientos con compañero de córnea, en su caso. Utilice una gasa para limpiar el bloque de vacío antes de procesar el segundo tejido. </li><li> Apriete la tapa (s) cámara de visualización y extirpar tejido de la zona de elaboración. </li><li> Divida el campo estéril, por deshacerse de artículos según Biopeligroso Plan de Gestión de Residuos. Quítese los guantes contaminados, y vestido. Lavarse las manos y ponerse los guantes no estériles. </li><li> Realizar post-corte evaluación lámpara de hendidura y microscopía especular de la córnea (s)y registrar los resultados. </li></ol>

<ol>
	<li>Fernandez, M., Afshari, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+Keratoplasty:+From+DLEK+to+DMEK.">Endothelial Keratoplasty: From DLEK to DMEK.</a> Middle East African Journal of Ophthalmology. 17 (1), 5-8 (2010).</li><li>Kruse, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Stepwise+Approach+to+Donor+Preparation+and+Insertion+Increases+Safety+and+Outcome+of+Descemet+Membrane+Endothelial+Keratoplasty.">A Stepwise Approach to Donor Preparation and Insertion Increases Safety and Outcome of Descemet Membrane Endothelial Keratoplasty.</a> Cornea. 30 (5), 580-587 (2011).</li><li>Dapena, I., Moutsouris, K., Droutsas, K., Ham, L., van Dijk, K., Melles, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Standardized+'No-Touch'+Technique+for+Descemet+Membrane+Endothelial+Keratoplasty.">Standardized 'No-Touch' Technique for Descemet Membrane Endothelial Keratoplasty.</a> Archives of Ophthalmology. 129, 88-94 (2011).</li><li>Tourtas, T., Laaser, K., Bachmann, B., Cursiefen, C., Kruse, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Descemet+Membrane+Endothelial+Keratoplasty+versus+Descemet+Stripping+Automated+Endothelial+Keratoplasty.">Descemet Membrane Endothelial Keratoplasty versus Descemet Stripping Automated Endothelial Keratoplasty.</a> American Journal of Ophthalmology. 153 (6), 1082-1090 (2012).</li><li>Guerra, F., Anshu, A., Price, M., Price, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+Keratoplasty:+fellow+eyes+comparison+of+Descemet+stripping+automated+endothelial+keratoplasty+and+Descemet+membrane+endothelial+keratoplasty.">Endothelial Keratoplasty: fellow eyes comparison of Descemet stripping automated endothelial keratoplasty and Descemet membrane endothelial keratoplasty.</a> Cornea. 30 (12), 1382-1386 (2011).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> 2012 Eye Banking Statistical Report: Surgical Use and Indications for Corneal Transplant Statistical Report Analysis. , (2012).</li><li>Price, M., Price, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Descemet's+stripping+endothelial+keratoplasty.">Descemet's stripping endothelial keratoplasty.</a> Current Opinion Ophthalmology. 18 (4), 290-294 (2007).</li><li>Woodward, M., Titus, M., Mavin, K., Shtein, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Corneal+Donor+Tissue+Preparation+for+Endothelial+Keratoplasty.">Corneal Donor Tissue Preparation for Endothelial Keratoplasty.</a> Journal of Visualized Experiments. (64), e3847 (2012).</li><li>Price, M., Price, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Descemet's+membrane+endothelial+keratoplasty+surgery:+update+on+the+evidence+and+hurdles+to+acceptance.">Descemet's membrane endothelial keratoplasty surgery: update on the evidence and hurdles to acceptance.</a> Current Opinion Ophthalmology. 24 (4), 329-335 (2013).</li><li>Venzano, D., Pagani, P., Randazzo, N., Cabiddu, F., Traverrso, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Descemet+Membrane+air-bubble+separation+in+donor+cornea.">Descemet Membrane air-bubble separation in donor cornea.</a> Journal of Cataract and Refractive Surgery. 36 (12), 2022-2027 (2010).</li><li>Price, F., Price, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> DSEK: What you need to know about endothelial keratoplasty. , (2009).</li><li>Studeny, P., Sivekova, D., Lieheova, K., Vokrojova, M., Kuchynka, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hybrid+technique+of+lamellar+keratoplasty+(DMEK-S).">Hybrid technique of lamellar keratoplasty (DMEK-S).</a> Journal of Ophthalmology. 2013, 2013.254383 (2013).</li><li>Muraine, M., Gueudry, J., He, Z., Piselli, S., Lefevre, S., Toubeau, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+technique+for+the+preparation+of+corneal+grafts+for+Descemet+membrane+endothelial+keratoplasty.">Novel technique for the preparation of corneal grafts for Descemet membrane endothelial keratoplasty.</a> American Journal of Ophthalmology. 156 (5), 851-859 (2013).</li><li>Lie, J. T., Birbal, R., Ham, L., van der Wees, J., Melles, G. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Donor+tissue+preparation+for+Descemet+membrane+endothelial+keratoplasty.">Donor tissue preparation for Descemet membrane endothelial keratoplasty.</a> Journal of Cataract Refractive Surgery. 34 (9), 1578-1583 (2008).</li><li>Kruse, F. E., Laaser, K., Cursiefen, C., Heindl, L. M., Schlotzer-Schrehardt, U., Riss, S., Bachmann, B. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+stepwise+approach+to+donor+preparation+and+insertion+increases+safety+and+outcome+of+Descemet+membrane+endothelial+keratoplasty.">A stepwise approach to donor preparation and insertion increases safety and outcome of Descemet membrane endothelial keratoplasty.</a> Cornea. 30 (5), 580-587 (2011).</li><li>Rudolph, M., Kaaser, L., Bachmann, B., Cursiefen, C., Epstein, D., Kruse, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Corneal+higher-order+aberrations+after+Descemet&#8217;s+membrane+endothelial+keratoplasty.">Corneal higher-order aberrations after Descemet&#8217;s membrane endothelial keratoplasty.</a> Ophthalmology. 119 (3), 528-535 (2012).</li><li>Price, F., Price, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evolution+of+endothelial+keratoplasty.">Evolution of endothelial keratoplasty.</a> Cornea. 32, 28-32 (2013).</li><li>Dapena, I., Dapena, L., Dirisamer, M., Ham, L., Melles, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visual+acuity+and+endothelial+cell+density+following+Descemet+membrane+endothelial+keratoplasty+(DMEK).">Visual acuity and endothelial cell density following Descemet membrane endothelial keratoplasty (DMEK).</a> Arch Soc Esp Oftalmol. 86 (12), 395-401 (2011).</li></ol>

Hassan Tausif y Dr. Navasuja Chandrasekaran tienen nada que revelar. Dres. Mian, Shtein, y Woodward son directores médicos para el Banco de Ojos Michigan. Sr. Mavin, el Sr. Tito, y la señora Johnson son empleados de Midwest Bancos de Ojos.

La capa de células endoteliales de la córnea juega un papel crítico en el mantenimiento de la claridad de la córnea a través de la deshidratación. Enfermedades de las células endoteliales pueden resultar en discapacidad visual significativa. DMEK pretende reemplazar las células endoteliales enfermas con un injerto de donante que consiste en membrana y capa de células endoteliales de Descemet. La preparación del tejido de los donantes con un DM intacto y recuento de células endoteliales adecuada son fundamenta...

Queratoplastia endotelial (EK) ha reemplazado rápidamente queratoplastia penetrante (PKP) como el procedimiento de elección para el tratamiento de enfermedades endoteliales que representan más del 50% de todos los trasplantes realizados en los Estados Unidos en 2012 6. PKP está limitada por la rehabilitación visual prolongado con alto astigmatismo y la inestabilidad de refracción, la pérdida de la integridad de globo y las complicaciones relacionadas sutura. Procedimientos EK primera generación, incluyendo profunda lamelar queratoplastia endotelial (DLEK) involucrados disección manual estroma corneal con el trasplante de tejido de grosor parcial de los donantes. Estos procedimientos tuvieron éxito en el trasplante de tejido endotelial donante, manteniendo una superficie corneal intacto pero eran técnicamente desafiante y dio lugar a limitado la agudeza visual mejor corregida debido a una irregular corneal lamelar interfaz 1. El procedimiento EK actual de elección es de Descemet Stripping queratoplastia endotelial (DSEK) que ofrece clar cornealdad con una recuperación más rápida de la visión, minimiza el error de refracción postoperatoria y astigmatismo, y disminuye el riesgo de dehiscencia de la herida traumática mientras se mantiene la superficie corneal 7. Este procedimiento implica la eliminación de la membrana de Descemet y el endotelio enfermo y el reemplazo con donante de espesor parcial tejido corneal que comprende una nueva capa de células endoteliales. La córnea donante se diseca con un microqueratomo para preparar el tejido posterior de ser trasplantadas 8. Preparación del tejido del donante por los ojos-los bancos ha sido fundamental en el rápido crecimiento y, sin duda, el éxito de este procedimiento. El aumento de 16 veces cerca de procedimientos DSEK en los Estados Unidos entre 2005 y 2012 corresponde a casi la mitad del tejido que está preparando los ojos-los bancos 6. Aunque DSEK puede mejorar la agudeza visual de 20/40 o mejor, la mayoría de los pacientes no logran 20/20 visión debido a la neblina interfaz del estroma y el aumento de la luzdispersar desde la adición de donante de tejido estromal 7. La técnica quirúrgica EK ha progresado a la membrana de Descemet queratoplastia endotelial (DMEK) que implica el trasplante de membrana de Descemet único donante (DM) y el endotelio. Este procedimiento tiene una mayor proporción de pacientes que alcanzan 20/20 visión, la recuperación visual más rápida y un menor riesgo de rechazo endotelial 9. Sin embargo, la preparación de tejido del donante es más difícil debido a las dimensiones delgadas de injerto.

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			<td height="42" style="height:42px;width:216px;">Name of Material/ Equipment</td>
			<td style="width:268px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">VisionBlue&#160;</td>
			<td style="width:268px;">D.O.R.C., Zuiland, The Netherlands</td>
			<td style="width:119px;">NA</td>
			<td></td>
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			<td height="21" style="height:21px;width:216px;">Barron vacuum punch</td>
			<td style="width:268px;">Katena, Denville, New Jersey</td>
			<td style="width:119px;">K20-2112</td>
			<td></td>
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corneal donor tissue preparation for descemet's membrane endothelial keratoplasty

Queratoplastia endotelial de membrana de Descemet (DMEK) es una forma de trasplante de córnea en el que sólo una única capa de células, el endotelio corneal, junto con su membrana basal (membrana de Descemet) se introduce en el estroma posterior del receptor 3. A diferencia de Descemet Stripping Automated queratoplastia endotelial (DSAEK), donde se introduce estroma adicional de los donantes, no se crea una interfaz natural de estroma a estroma. Como resultado, la anatomía natural de la córnea se conserva tanto como sea posible que permita mejorar el tiempo de recuperación y la agudeza visual 4. Queratoplastia endotelial (EK) es el procedimiento de elección para el tratamiento de la disfunción endotelial. Las ventajas de EK incluyen la rápida recuperación de la visión, la preservación de la integridad ocular y el cambio de refracción mínimo debido al uso de una pequeña incisión, periférica 1. DSAEK utiliza tejido de donante preparado con espesor parcial estroma y el endotelio. El rápido success y la utilización de este procedimiento se puede atribuir a la disponibilidad de ojo-Banco preparó tejido precortado. Los beneficios de la preparación ojo-banco de tejido donante incluyen la eliminación de la necesidad de equipo especializado en la sala de operaciones y la disponibilidad de copia de seguridad de tejido del donante en caso de perforación de tejido durante la preparación. Además, la preparación de alto volumen de tejido del donante por técnicos ojo-banco puede proporcionar una mejor calidad de tejido del donante. DSAEK puede haber limitado la agudeza visual mejor corregida debido a la creación de una interfaz de estroma entre el donante y la córnea destinatario. La eliminación de esta interfaz con el trasplante de membrana y endotelio único donante de Descemet en DMEK puede mejorar los resultados visuales y reducir las complicaciones después de EK 5. Similar a DSAEK, el éxito a largo plazo y la aceptación de DMEK depende de la facilidad de disponibilidad de precorte, ojo-banco prepara tejido del donante. Aquí presentamos un enfoque paso a paso para la preparación del tejido donante que puede reducir barriERS oculares bancos enfrentan en la prestación de los injertos DMEK.

Separación adecuada de DM y el endotelio de estroma es esencial en la preparación de tejido precortado para DMEK. Cincuenta córneas de donantes con una edad media de 65 ± 6 años (rango 55-75), muerte a tiempo de procesamiento de 5,9 ± 2,3 días (rango 1-10) y la densidad de células endoteliales (ECD) de 2.616 ± 321 células / mm 2 ( rango de 2,049-3,247) se utilizaron para evaluar la viabilidad y la reproducibilidad de la técnica de preparación quirúrgica por dos técnicos del banco de ojos. La tas...

Watch this Scientific Journal Video about Corneal Donor Tissue Preparation for Descemet's Membrane Endothelial Keratoplasty at JoVE.com

Corneal Donor Tissue Preparation for Descemet&#039;s Membrane Endothelial Keratoplasty

Queratoplastia endotelial es una técnica quirúrgica en continua evolución como resultado avances en el trasplante de injertos más delgadas con cada sucesión 1. Aquí presentamos una técnica para la disección del tejido manual de control para permitir la separación segura y repetible del endotelio y de las membranas de Descemet de botones corneoesclerales donantes para el trasplante 2.

Córnea Preparación de Tejidos de Donantes para la Membrana de Descemet queratoplastia endotelial

Descemet&#8217;s Membrane Endothelial Keratoplasty (DMEK) is a form of corneal transplantation in which only a single cell layer, the corneal endothelium, ...

corneal-donor-tissue-preparation-for-descemet-s-membrane-endothelial

Research

JoVE Journal

Medicine

Corneal Donor Tissue Preparation for Descemet's Membrane Endothelial Keratoplasty

15.2K vistas.  University of Michigan.  Queratoplastia endotelial es una técnica quirúrgica en continua evolución como resultado avances en el trasplante de injertos más delgadas con cada sucesión 1. Aquí presentamos una técnica para la disección del tejido manual de control para permitir la separación segura y repetible del endotelio y de las membranas de Descemet de botones corneoesclerales donantes para el trasplante 2. 

Video: Corneal Donor Tissue Preparation for Descemet's Membrane Endothelial Keratoplasty

Single Port Donor Nephrectomy

In 2007, Rane presented the first single port nephrectomy for a small non-functioning kidney at the World Congress of Endourology. Since that time, the use of single port surgery for nephrectomy has expanded to include donor nephrectomy. Over the next two years the technique was adopted for many others types of nephrectomies to include donor nephrectomy. We present our technique for single port donor nephrectomy using the Gelpoint device. We have successfully performed this surgery in over 100 patients and add this experience to our experience of over 1000 laparoscopic nephrectomies. With the proper equipment and technique, single port donor nephrectomy can be performed safely and effectively in the majority of live donors. We have found that our operative times and most importantly our transplant outcomes have not changed significantly with the adoption of the single port donor nephrectomy. We believe that single port donor nephrectomy represents a step forward in the care of living donors.

Single port laparoscopic surgery is changing the standard of care in surgical care like nothing since the laparoscopic technique was introduced 20 years ago. We present out technique of single port donor nephrectomy using the Gelpoint device. We have successfully performed this surgery in 100 patients.

De un puerto nefrectomía del donante

In 2007, Rane presented the first single port nephrectomy for a small non-functioning kidney at the World Congress of Endourology.  Since that time, the ...

Investigación

Medicina

Corneal Donor Tissue Preparation for Endothelial Keratoplasty

Over the past ten years, corneal transplantation surgical techniques have undergone revolutionary changes1,2. Since its inception, traditional full thickness corneal transplantation has been the treatment to restore sight in those limited by corneal disease. Some disadvantages to this approach include a high degree of post-operative astigmatism, lack of predictable refractive outcome, and disturbance to the ocular surface. The development of Descemet's stripping endothelial keratoplasty (DSEK), transplanting only the posterior corneal stroma, Descemet's membrane, and endothelium, has dramatically changed treatment of corneal endothelial disease. DSEK is performed through a smaller incision; this technique avoids 'open sky' surgery with its risk of hemorrhage or expulsion, decreases the incidence of postoperative wound dehiscence, reduces unpredictable refractive outcomes, and may decrease the rate of transplant rejection3-6.
Initially, cornea donor posterior lamellar dissection for DSEK was performed manually1 resulting in variable graft thickness and damage to the delicate corneal endothelial tissue during tissue processing. Automated lamellar dissection (Descemet's stripping automated endothelial keratoplasty, DSAEK) was developed to address these issues. Automated dissection utilizes the same technology as LASIK corneal flap creation with a mechanical microkeratome blade that helps to create uniform and thin tissue grafts for DSAEK surgery with minimal corneal endothelial cell loss in tissue processing.
Eye banks have been providing full thickness corneas for surgical transplantation for many years. In 2006, eye banks began to develop methodologies for supplying precut corneal tissue for endothelial keratoplasty. With the input of corneal surgeons, eye banks have developed thorough protocols to safely and effectively prepare posterior lamellar tissue for DSAEK surgery. This can be performed preoperatively at the eye bank. Research shows no significant difference in terms of the quality of the tissue7 or patient outcomes8,9 using eye bank precut tissue versus surgeon-prepared tissue for DSAEK surgery. For most corneal surgeons, the availability of precut DSAEK corneal tissue saves time and money10, and reduces the stress of performing the donor corneal dissection in the operating room. In part because of the ability of the eye banks to provide high quality posterior lamellar corneal in a timely manner, DSAEK has become the standard of care for surgical management of corneal endothelial disease.
The procedure that we are describing is the preparation of the posterior lamellar cornea at the eye bank for transplantation in DSAEK surgery (Figure 1).

Endothelial corneal transplantation is a surgical technique for treatment of posterior corneal diseases. Mechanical microkeratome dissection to prepare tissue results in thinner, more symmetric grafts with less endothelial cell loss and improved outcomes. Dissections can be performed at the eye bank prior to corneal transplantation surgery.

Preparación del tejido corneal donante para la queratoplastia endotelial

Over the past ten years, corneal transplantation surgical techniques have undergone revolutionary changes1,2. Since its inception, traditional full ...

Preparation and In Vitro Characterization of Magnetized miR-modified Endothelial Cells

To date, the available surgical and pharmacological treatments for cardiovascular diseases (CVD) are limited and often palliative. At the same time, gene and cell therapies are highly promising alternative approaches for CVD treatment. However, the broad clinical application of gene therapy is greatly limited by the lack of suitable gene delivery systems. The development of appropriate gene delivery vectors can provide a solution to current challenges in cell therapy. In particular, existing drawbacks, such as limited efficiency and low cell retention in the injured organ, could be overcome by appropriate cell engineering (i.e., genetic) prior to transplantation. The presented protocol describes the efficient and safe transient modification of endothelial cells using a polyethyleneimine superparamagnetic magnetic nanoparticle (PEI/MNP)-based delivery vector. Also, the algorithm and methods for cell characterization are defined. The successful intracellular delivery of microRNA (miR) into human umbilical vein endothelial cells (HUVECs) has been achieved without affecting cell viability, functionality, or intercellular communication. Moreover, this approach was proven to cause a strong functional effect in introduced exogenous miR. Importantly, the application of this MNP-based vector ensures cell magnetization, with accompanying possibilities of magnetic targeting and non-invasive MRI tracing. This may provide a basis for magnetically guided, genetically engineered cell therapeutics that can be monitored non-invasively with MRI.

Preparation and In Vitro Characterization of Magnetized miR-modified Endothelial Cells

This manuscript describes the efficient, non-viral delivery of miR to endothelial cells by a PEI/MNP vector and their magnetization. Thus, in addition to genetic modification, this approach allows for magnetic cell guidance and MRI detectability. The technique can be used to improve the characteristics of therapeutic cell products.

Preparación y<em&gt; In Vitro</em&gt; Caracterización de Magnetized miR-modificado células endoteliales

To date, the available surgical and pharmacological treatments for cardiovascular diseases (CVD) are limited and often palliative. At the same time, gene ...

Ex Vivo Corneal Organ Culture Model for Wound Healing Studies

The cornea has been used extensively as a model system to study wound healing. The ability to generate and utilize primary mammalian cells in two dimensional (2D) and three dimensional (3D) culture has generated a wealth of information not only about corneal biology but also about wound healing, myofibroblast biology, and scarring in general. The goal of the protocol is an assay system for quantifying myofibroblast development, which characterizes scarring. We demonstrate a corneal organ culture ex vivo model using pig eyes. In this anterior keratectomy wound, corneas still in the globe are wounded with a circular blade called a trephine. A plug of approximately 1/3 of the anterior cornea is removed including the epithelium, the basement membrane, and the anterior part of the stroma. After wounding, corneas are cut from the globe, mounted on a collagen/agar base, and cultured for two weeks in supplemented-serum free medium with stabilized vitamin C to augment cell proliferation and extracellular matrix secretion by resident fibroblasts. Activation of myofibroblasts in the anterior stroma is evident in the healed cornea. This model can be used to assay wound closure, the development of myofibroblasts and fibrotic markers, and for toxicology studies. In addition, the effects of small molecule inhibitors as well as lipid-mediated siRNA transfection for gene knockdown can be tested in this system.

A protocol for an ex vivo corneal organ culture model useful for wound healing studies is described. This model system can be used to assess the effects of agents to promote regenerative healing or drug toxicity in an organized 3D multicellular environment.

Ex Vivo modelo de cultura órgano córneo para la curación de heridas estudios

The cornea has been used extensively as a model system to study wound healing. The ability to generate and utilize primary mammalian cells in two ...

A Porcine Corneal Endothelial Organ Culture Model Using Split Corneal Buttons

Experimental research on corneal endothelial cells is associated with several difficulties. Human donor corneas are scarce and rarely available for experimental investigations as they are normally needed for transplantation. Endothelial cell cultures often do not translate well to in vivo situations. Due to the biostructural characteristics of non-human corneas, stromal swelling during cultivation induces substantial corneal endothelial cell loss, which makes it difficult to perform cultivation for an extended period of time. Deswelling agents such as dextran are used to counteract this response. However, they also cause significant endothelial cell loss. Therefore, an ex vivo organ culture model not requiring deswelling agents was established. Pig eyes from a local slaughterhouse were used to prepare split corneal buttons. After partial corneal trephination, the outer layers of the cornea (epithelium, bowman layer, parts of the stroma) were removed. This significantly reduces corneal endothelial cell loss induced by massive stromal swelling and Descemet&#39;s membrane folding throughout longer cultivation periods and improves general preservation of the endothelial cell layer. Subsequent complete corneal trephination was followed by the removal of the split corneal button from the remaining eye bulb and cultivation. Endothelial cell density was assessed at follow-up times of up to 15 days after preparation (i.e., days 1, 8, 15) using light microscopy. The preparation technique used allows a better preservation of the endothelial cell layer enabled by less stromal tissue swelling, which results in slow and linear decline rates in split corneal buttons comparable to human donor corneas. As this standardized organo-typically cultivated research model for the first time allows a stable cultivation for at least two weeks, it is a valuable alternative to human donor corneas for future investigations of various external factors with regards to their effects on the corneal endothelium.

Here, a step-by-step protocol for the preparation and cultivation of porcine split corneal buttons is presented. As this organo-typically cultivated organ culture model shows cell death rates within 15 days, comparable to human donor corneas, it represents the first model allowing long-term cultivation of non-human corneas without adding toxic dextran.

Un modelo de cultivo de órgano endotelial Corneal porcino usando botones de córnea divididos

Experimental research on corneal endothelial cells is associated with several difficulties. Human donor corneas are scarce and rarely available for ...

Development of a Noninvasive, Laser-Assisted Experimental Model of Corneal Endothelial Cell Loss

Nd:YAG lasers have been used to perform noninvasive intraocular surgery, such as capsulotomy for several decades now. The incisive effect relies on the optical breakdown at the laser focus. Acoustic shock waves and cavitation bubbles are generated, causing tissue rupture. Bubble sizes and pressure amplitudes vary with pulse energy and position of the focal point. In this study, enucleated porcine eyes were positioned in front of a commercially available Nd:YAG laser. Variable pulse energies as well as different positions of the focal spots posterior to the cornea were tested. Resulting lesions were evaluated by two-photon microscopy and histology to determine the best parameters for an exclusive detachment of corneal endothelial cells (CEC) with minimum collateral damage. The advantages of this method are the precise ablation of CEC, reduced collateral damage, and above all, the non-contact treatment.

Here, we present a protocol to detach corneal endothelial cells (CEC) from Descemet&#8217;s membrane (DM) using a neodymium:YAG (Nd:YAG) laser as an ex vivo disease model for bullous keratopathy (BK).

Desarrollo de un Modelo Experimental No Invasivo, Asistido por Láser de Pérdida Celular Endotelial Corneal

Nd:YAG lasers have been used to perform noninvasive intraocular surgery, such as capsulotomy for several decades now. The incisive effect relies on the ...

An Epithelial Abrasion Model for Studying Corneal Wound Healing

The cornea is critical for vision, accounting for about two-thirds of the refractive power of the eye. Crucial to the role of the cornea in vision is its transparency. However, due to its external position, the cornea is highly susceptible to a wide variety of injuries that can lead to the loss of corneal transparency and eventual blindness. Efficient corneal wound healing in response to these injuries is pivotal for maintaining corneal homeostasis and preservation of corneal transparency and refractive capabilities. In events of compromised corneal wound healing, the cornea becomes vulnerable to infections, ulcerations, and scarring. Given the fundamental importance of corneal wound healing to the preservation of corneal transparency and vision, a better understanding of the normal corneal wound healing process is a prerequisite to understanding impaired corneal wound healing associated with infection and disease. Toward this goal, murine models of corneal wounding have proven useful in furthering our understanding of the corneal wound healing mechanisms operating under normal physiological conditions. Here, a protocol for creating a central corneal epithelial abrasion in mouse using a trephine and a blunt golf club spud is described. In this model, a 2 mm diameter circular trephine, centered over the cornea, is used to demarcate the wound area. The golf club spud is used with care to debride the epithelium and create a circular wound without damaging the corneal epithelial basement membrane. The resulting inflammatory response proceeds as a well-characterized cascade of cellular and molecular events that are critical for efficient wound healing. This simple corneal wound healing model is highly reproducible and well-published and is now being used to evaluate compromised corneal wound healing in the context of disease.

Here, a protocol for creating a central corneal epithelial abrasion wound in the mouse using a trephine and a blunt golf club spud is described. This corneal wound healing model is highly reproducible and is now being used to evaluate compromised corneal wound healing in the context of diseases.

Un modelo de abrasión epitelial para estudiar la cicatrización de heridas corneales

The cornea is critical for vision, accounting for about two-thirds of the refractive power of the eye. Crucial to the role of the cornea in vision is its ...

A Porcine Heterotopic Heart Transplantation Protocol for Delivery of Therapeutics to a Cardiac Allograft

Cardiac transplantation is the gold standard treatment for end-stage heart failure. However, it remains limited by the number of available donor hearts and complications such as primary graft dysfunction and graft rejection. The recent clinical use of an ex vivo perfusion device in cardiac transplantation introduces a unique opportunity for treating cardiac allografts with therapeutic interventions to improve function and avoid deleterious recipient responses. Establishing a translational, large-animal model for therapeutic delivery to the entire allograft is essential for testing novel therapeutic approaches in cardiac transplantation. The porcine, heterotopic heart transplantation model in the intraabdominal position serves as an excellent model for assessing the effects of novel interventions and the immunopathology of graft rejection. This model additionally offers long-term survival for the pig, given that the graft is not required to maintain the recipient's circulation. The aim of this protocol is to provide a reproducible and robust approach for achieving ex vivo delivery of a therapeutic to the entire cardiac allograft prior to transplantation and provide technical details to perform a survival heterotopic transplant of the ex vivo perfused heart.

We present a protocol for utilizing a normothermic ex vivo sanguinous perfusion system for the delivery of therapeutics to an entire cardiac allograft in a porcine heterotopic heart transplant model.

Un protocolo de trasplante de corazón heterotópico porcino para la administración de terapias a un aloinjerto cardíaco

Cardiac transplantation is the gold standard treatment for end-stage heart failure. However, it remains limited by the number of available donor hearts ...

A Human Corneal Organ Culture Model of Descemet's Stripping Only with Accelerated Healing Stimulated by Engineered Fibroblast Growth Factor 1

Fuchs Endothelial Corneal Dystrophy (FECD) results from dysfunctional corneal endothelial cells (CECs) and is currently treated by transplantation of the whole cornea or Descemet's membrane. Recent developments in ocular surgery have established Descemet's Stripping Only (DSO), a surgical technique in which a central circle of guttae-dense Descemet's membrane is removed to allow for the migration of CECs onto the smooth stroma, restoring function and vision to the cornea. While this potential treatment option is of high interest in the field of ophthalmic research, no successful ex vivo models of DSO have been established and clinical data is limited. This work presents a novel wound-healing model simulating DSO in human donor corneas. Using this approach to evaluate the efficacy of the human engineered FGF1 (NM141), we found that treatment accelerated healing via stimulation of migration and proliferation of CECs. This finding was confirmed in 11 pairs of human corneas with signs of dystrophy reported by the eye banks in order to verify that these results can be replicated in patients with Fuchs' Dystrophy, as the target population of the DSO procedure.

A Human Corneal Organ Culture Model of Descemet&#039;s Stripping Only with Accelerated Healing Stimulated by Engineered Fibroblast Growth Factor 1

Descemet's Stripping Only is an experimental procedure wherein patients with central corneal guttae resulting from Fuchs Endothelial Corneal Dystrophy have Descemet's membrane stripped for peripheral cells to regenerate the endothelial layer. We present novel methodology simulating DSO in dystrophic human corneas ex vivo with accelerated healing stimulated by eFGF1 (NM141).

Un modelo de cultivo de órganos corneales humanos de la extracción de Descemet solo con curación acelerada estimulada por el factor de crecimiento de fibroblastos diseñado 1

Fuchs Endothelial Corneal Dystrophy (FECD) results from dysfunctional corneal endothelial cells (CECs) and is currently treated by transplantation of the ...

Preparation of Human Myocardial Tissue for Long-Term Cultivation

Cardiomyocyte cultivation has seen a vast number of developments, ranging from two-dimensional (2D) cell cultivation to iPSC derived organoids. In 2019, an ex vivo way to cultivate myocardial slices obtained from human heart samples was demonstrated, while approaching in vivo condition of myocardial contraction. These samples originate mostly from heart transplantations or left-ventricular assist device placements. Using a vibratome and a specially developed cultivation system, 300 &#181;m thick slices are placed between a fixed and a spring wire, allowing for stable and reproducible cultivation for several weeks. During cultivation, the slices are continuously stimulated according to individual settings. Contractions can be displayed and recorded in real-time, and pharmacological agents can be readily applied. User-defined stimulation protocols can be scheduled and performed to assess vital contraction parameters like post-pause-potentiation, stimulation threshold, force-frequency relation, and refractory period. Furthermore, the system enables a variable pre- and afterload setting for a more physiological cultivation.
Here, we present a step-by-step guide on how to generate a successful long-term cultivation of human left ventricular myocardial slices, using a commercial biomimetic cultivation solution.

We present a protocol for ex vivo cultivation of human ventricular myocardial tissue. It allows for detailed analysis of contraction force and kinetics, as well as the application of pre- and afterload to mimic the in vivo physiological environment more closely.

Preparación del tejido miocárdico humano para el cultivo a largo plazo

Cardiomyocyte cultivation has seen a vast number of developments, ranging from two-dimensional (2D) cell cultivation to iPSC derived organoids. In 2019, ...