1 Configuração de Estéril Campo <ol><li> Don traje adequado incluindo boné e máscara. Use óculos de proteção quando na sala de processamento. </li><li> Prepare a área de processamento. Quando se utiliza um local de tratamento, manter o funcionamento contínuo do ventilador para criar uma pressão positiva no quarto. </li><li> Reúna todos os suprimentos. Verifique as datas de validade, integridade da embalagem, e indicador de esterilização. </li><li> Lavar as mãos e vestir luvas não esterilizadas. </li><li> Abra os kits estéreis para estabelecer um campo estéril. Ao utilizar uma sala de processamento, colocar os kits em cima da mesa e assepticamente abrir os wraps. Não utilizar os exteriores 1 ½ polegadas do envoltório como parte do campo estéril como este segmento é considerado não esterilizado. </li><li> Utilizando uma técnica asséptica, aberto e soltar itens estéreis sobre campo estéril, a menos que já fazem parte do pacote premade. </li><li> Mover microscópio de modo que seja acima do campo estéril, ligar fonte de luz de microscópio, e ajustar oculares como necessário. </li><li> OBTtecido da córnea ain a ser desmontado (em meio de preservação) e coloque ao lado da área de processamento estéril preparado. Solte a tampa da câmara de visionamento e deixe descansar em cima de ver a base da câmara sem fios envolvidos. Descarte as luvas não esterilizadas. </li><li> Abra um par de luvas estéreis e bata esterilizada com uma toalha. </li><li> Realize paramentação. Revesti-vos de avental estéril, em seguida, luvas estéreis. </li><li> Caixas de instrumento aberto para que todos os suprimentos são acessíveis. Abrir humidade cortina impermeável para criar a área de trabalho. </li><li> Anexar agulha para uma seringa de 5 cc. Utilizando uma tesoura cirúrgica, preparar a agulha G 30, por colocação de uma curva a 1 mm do bisel, e outra curva a 1 mm da base (de modo a formar uma forma de &quot;S&quot;). </li><li> Montar a seringa de vidro para o corante azul de tripano, colocando o êmbolo para dentro da seringa e o dedo de aperto do êmbolo. Anexar cânula. </li><li> Fixe a cânula ao frasco de solução salina balanceada (BSS). </li><li> Separe o soco trephine do bloco de vácuoe reserve. O centro do bloco de vácuo no interior do campo de visão do microscópio. </li><li> Traga o bloco de vácuo em foco, ajustando as configurações de microscópio. </li></ol> 2 Processo de Preparação tecido do doador <ol><li> Levante a tampa da câmara de visualização utilizando quadrado folha como uma barreira. Com a mão oposta e usando uma pinça, segure suavemente córnea, esclera rim para remover córnea. Recoloque a tampa da câmara de visualização. </li><li> Completamente deprimir a seringa de sucção, em seguida, coloque o lado epitelial da córnea para baixo no bloco de vácuo e garantir que ele está centrado. (Figura 1) </li><li> Gentilmente menor Lugares para o bloco de vácuo com as mensagens de orientação. Envolver-se vácuo, liberando lentamente pistão da seringa de sucção. Remova o anel de estar e reserve. (Figura 2). </li><li> Realize uma trepanação parcial para marcar através da membrana de Descemet. </li><li> Ajuste o zoom e focagem do microscópio de modo que o tecido é bem visualizado. </li><li&gt; Inclinação do bloco de vácuo e usar uma lança colocada zaragatoa estéril no limbo para remover o excesso de meio. (Figura 3) </li><li> Coloque mancha suficiente no limbo para corar a borda da membrana de Descemet marcados. Permitir que a mancha de permanecer no endotélio de 60-90 seg. (Figura 4) </li><li> Remover manchas: <ol><li> Inclinação do bloco de vácuo para verter a mancha da córnea e na almofada de gaze. </li><li> Use as lanças cirúrgicas para remover o excesso de corante da área do limbo da córnea, tomando cuidado para não tocar no endotélio. (Figura 5) </li></ol></li><li> Use algumas gotas de BSS para enxaguar a córnea de solução de coloração, e lanças swab estéril para remover o excesso de BSS. (Figura 6) </li><li> Colocar 1-2 gotas de BSS no aro esclerótica para impedir o tecido de secagem. Repita conforme necessário durante todo o procedimento. </li><li> Realize dissecação parcial da membrana de Descemet: <ol><li> Começando onde a tubulação se conectars para o bloco de vácuo, com a pinça aberta, colocar uma ponta na extremidade da marca de pontuação e membrana e endotélio suavemente separado de Descemet longe do estroma. (Figura 7). </li><li> Membrana separado de Descemet e endotélio não superior a 1 mm da margem marcou e continuar a dissecção de 360 ​​° ao redor. (Figura 8). </li><li> Nota localização de quaisquer rasgos micro colocando um ponto no bloco de vácuo, com o marcador de pele. NOTA: Se a trefina não penetrar completamente através da membrana de Descemet, utilizar a borda chanfrada da agulha G 30 preparado para completar o marcador através da membrana de Descemet, quando necessário, seguida de uma segunda aplicação de coloração conforme a necessidade. Se adesividade da membrana de Descemet está impedindo o esvaziamento parcial, remova o endotélio fora da área trepanação, conforme necessário. </li></ol></li><li> Use uma pinça para remover quaisquer etiquetas endoteliais que se sobrepõem a marca de pontuação. </li> &lt;li&gt; Considere o local de micro lágrimas marcadas anteriormente e girar o bloco de vácuo para que o maior é às 6 horas. Isso vai se tornar a dobradiça da tampa. </li><li> Usando a pinça, segure membrana de Descemet em 12 horas. Suavemente separar a membrana de Descemet do estroma por raspagem para a dobradiça. Criar a dobradiça parando a separação 2 mm da marca de pontuação. NOTA: bordas periféricas de retalho vai começar a rolar durante a separação. (Figura 9). </li><li> Cuidadosamente, coloque a tampa de volta no lugar no estroma. Use BSS e swab lanças para unscroll o endotélio e devolvê-lo à sua posição original. (Figura 10A-C) </li><li> Remover líquido residual entre a membrana de Descemet eo estroma com lanças swab estéril de modo a aba fica na posição. (Figura 11) </li><li> Use lanças swab estéril para absorver o excesso de umidade da esclera perto da dobradiça. Use o marcador pele para desenhar uman arrowhead na esclera aro que aponta para o centro da articulação, remover o excesso de tinta usando uma lança zaragatoa estéril. (Figura 12) </li><li> Soltar vácuo pressionando o êmbolo da seringa ligado ao bloco de vácuo. Enquanto o vácuo é desengatada, fórceps para remover a córnea a partir do bloco de vácuo. </li><li> Levante a tampa da câmara de visualização utilizando quadrado folha como uma barreira. Coloque a córnea na câmara de visualização, lado endotelial, e substituir a tampa da câmara de visualização. </li><li> Repita os passos acima de procedimento com o companheiro de córnea, se for o caso. Use gaze para limpar o bloco de vácuo, antes de processar o segundo tecido. </li><li> Apertar a tampa (s) da câmara de visualização e remover o tecido da área de processamento. </li><li> Quebrar o campo estéril pela eliminação de itens de acordo com o Plano de Gerenciamento de Resíduos de Risco Biológico. Retire as luvas contaminadas e vestido. Lavar as mãos e vestir luvas não esterilizadas. </li><li> Realizar pós-corte de avaliação com lâmpada de fenda e microscopia especular da córnea (s)e registrar os 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 e Dr. Navasuja Chandrasekaran tem nada a revelar. Drs. Mian, Shtein e Woodward são diretores médicos para o Michigan Banco de Olhos. Mr. Mavin, Sr. Tito, e Ms. Johnson são funcionários da Midwest Banco de Olhos.

A camada de células endoteliais desempenham um papel crítico na manutenção da transparência córnea através de desidratação. Doenças das células endoteliais pode resultar em deficiência visual significativo. DMEK visa substituir as células endoteliais doentes com um enxerto de doador constituído por membrana e camada de células endoteliais de Descemet. Preparação de tecido do dador com um DM intacta e contagem de células endoteliais adequada são críticos para a sobrevivência do tecido do enxerto. Ras...

Endotelial Keratoplasty (EK) substituiu rapidamente ceratoplastia penetrante (PKP), como o procedimento de escolha para o tratamento de doenças endoteliais representam mais de 50% de todos os transplantes realizados nos Estados Unidos em 2012 6. PKP é limitada pela reabilitação visual prolongado com alto astigmatismo e instabilidade de refração, perda da integridade globo e complicações relacionadas à sutura. Procedimentos EK primeira geração, incluindo profunda lamelar endotelial Keratoplasty (dlek) envolvidos dissecção Manual do estroma corneal com transplante de tecido doador espessura parcial. Estes procedimentos foram bem sucedidos em transplante de tecido endotelial doador, mantendo uma superfície da córnea intacta, mas eram tecnicamente desafiador e resultou na limitado melhor acuidade visual corrigida, devido a uma córnea irregular de interface lamelar 1. O procedimento EK atual de escolha é de Descemet Stripping endotelial Keratoplasty (DSEK), que oferece clar córneadade com recuperação mais rápida da visão, minimiza erros de refração pós-operatória e astigmatismo, e diminui o risco de deiscência da ferida traumática, mantendo a superfície da córnea 7. Este procedimento envolve a remoção da membrana de Descemet e endotélio doente e substituição por tecido doador de espessura parcial que compreende uma nova camada de células endoteliais. A córnea do doador é dissecado com um microcerátomo para preparar o tecido posterior serem transplantadas 8. Preparação do tecido do doador pelo olho bancos tem sido fundamental para o crescimento rápido e, sem dúvida, o sucesso deste procedimento. O aumento de 16 vezes perto em procedimentos DSEK nos Estados Unidos entre 2005 e 2012 corresponde a quase metade do tecido a ser preparada pelo olho bancos 6. Embora DSEK pode melhorar a acuidade visual de 20/40, ou melhor, a maioria dos pacientes não alcançam 20/20 visão devido a neblina interface de estroma e aumento de luzespalham a partir da adição de doador de tecido estromal 7. A técnica cirúrgica EK progrediu a membrana de Descemet ceratoplastia endotelial (DMEK) que envolve o transplante de membrana só doador de Descemet (DM) e endotélio. Este procedimento tem uma maior proporção de pacientes que alcançam 20/20 de visão, a recuperação visual mais rápida, e um menor risco de rejeição endotelial 9. No entanto, a preparação do tecido doador é mais difícil devido às dimensões do enxerto mais finas.

<table border="0" cellpadding="0" cellspacing="0" width="770">
	<tbody>
		<tr height="42">
			<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>
		</tr>
		<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>
		</tr>
		<tr height="21">
			<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>
		</tr>
	</tbody>
</table>

corneal donor tissue preparation for descemet's membrane endothelial keratoplasty

Membrana de Descemet ceratoplastia endotelial (DMEK) é uma forma de transplante de córnea, em que apenas uma camada única de células, o endotélio da córnea, juntamente com a sua membrana basal (membrana de Descemet) é introduzido no estroma posterior do recipiente 3. Ao contrário de Descemet Stripping Automated endotelial Keratoplasty (DSAEK), onde estroma adicional dos doadores é introduzida, sem interface antinatural estroma-to-estroma é criado. Como resultado, a anatomia natural da córnea é preservada, tanto quanto possível, permitindo o tempo de recuperação para melhorar a acuidade visual e 4. Endotelial Keratoplasty (EK) é o procedimento de escolha para o tratamento de disfunção endotelial. As vantagens de EK incluem a rápida recuperação da visão, a preservação da integridade do bulbo ocular e alteração refrativa mínimo devido ao uso de uma incisão pequena, periférica 1. DSAEK utiliza tecido do dador preparada com estroma de espessura parcial e endotélio. A rápida success e utilização deste procedimento pode ser atribuído à disponibilidade de olho banco preparado tecidos pré-cortados. Os benefícios da preparação olho banco de tecido do doador incluem a eliminação de necessidade de equipamento especializado na sala de operação e disponibilidade de back up tecido do doador em caso de perfuração do tecido durante a preparação. Além disso, a preparação de alto volume de tecido do doador por técnicos olho bancárias pode proporcionar a melhoria da qualidade do tecido do doador. DSAEK pode ter limitado melhor acuidade visual corrigida, devido à criação de uma interface do estroma entre o doador eo receptor da córnea. Eliminação desta interface com o transplante de membrana e endotélio único doador de Descemet em DMEK pode melhorar os resultados visuais e reduzir as complicações após EK 5. Semelhante ao DSAEK, o sucesso a longo prazo e aceitação de DMEK depende facilidade de disponibilidade de pré-cortados, olho-bank preparado tecido do doador. Aqui nós apresentamos uma abordagem passo a passo para a preparação do tecido doador, que pode reduzir alguns barriers oculares bancos enfrentam no fornecimento de enxertos DMEK.

Separação adequada do DM e endotélio de estroma é essencial na preparação de tecidos pré-cortados para DMEK. Cinquenta córneas de doadores com idade média de 65 ± 6 anos (variação 55-75), a morte para o tempo de processamento de 5,9 ± 2,3 dias (variação 1-10) e densidade de células endoteliais (ECD) de 2616 ± 321 células / mm 2 ( intervalo 2,049-3,247) foram usadas para avaliar a viabilidade e reprodutibilidade da técnica de preparação cirúrgica por dois técnicos banco de olhos. A taxa ...

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

Ceratoplastia endotelial é uma técnica cirúrgica em constante evolução como avanços resultam em transplante de enxertos mais finas com cada sucessão 1. Aqui apresentamos uma técnica de dissecção dos tecidos controlada manual para permitir uma separação segura e repetível do endotélio e membrana de Descemet do doador botões córneo para transplante 2.

Preparação tecido doador de córnea para Membrana de Descemet endotelial Keratoplasty

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 visualizações.  University of Michigan.  Ceratoplastia endotelial é uma técnica cirúrgica em constante evolução como avanços resultam em transplante de enxertos mais finas com cada sucessão 1. Aqui apresentamos uma técnica de dissecção dos tecidos controlada manual para permitir uma separação segura e repetível do endotélio e membrana de Descemet do doador botões córneo para transplante 2. 

Vídeo: 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.

Nefrectomia Single Port

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 ...

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.

Preparação do tecido da córnea Doadores de ceratoplastia 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.

preparação e<em&gt; In Vitro</em&gt; Caracterização de magnetizado miR-modificado Células Endoteliais

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 órgão da córnea para cicatrização de feridas estudos

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.

Um modelo de cultura de órgão endotelial corneano porcina usando Split córnea botões

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).

Desenvolvimento de um modelo experimental não invasivo, assistido a laser de perda de células endoteliais córneas

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.

Um modelo de abrasão epitelial para estudar a cicatrização da ferida córnea

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.

Um Protocolo de Transplante de Coração Heterotópico Porcino para Entrega de Terapêutica a um Aoenxerto 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).

Um modelo de cultura de órgãos córneas humanas de descascamento de descemet apenas com cura acelerada estimulada pelo fator de crescimento do fibroblasto projetado 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.

Preparação do tecido miocárdio humano para cultivo a longo prazo

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