Steril Field 1. Kurulum <ol><li> Kap ve maske de dahil olmak üzere, Don uygun kıyafetleri. Ne zaman işleme odasında göz koruması takın. </li><li> Işleme alanı hazırlayın. Bir işleme oda kullanırken, odada pozitif basınç oluşturmak için fanın sürekli çalışmasını sağlamak. </li><li> Tüm malzemeleri toplayın. Son kullanma tarihleri, ambalaj bütünlüğü ve sterilizasyon göstergesini kontrol edin. </li><li> Ellerinizi yıkayın ve non-steril eldiven yok. </li><li> Steril bir alan kurmak için steril kitleri açın. Bir işleme oda kullanırken, masaya kitleri yerleştirin ve aseptik tamamladı açın. Bu segment steril olmayan kabul edilir gibi steril alanın bir parçası olarak şal dış 1½ inç kullanmak etmeyin. </li><li> Açık aseptik teknik kullanılarak ve premade paketinin zaten bir parçası olmadıkça steril alan üzerine steril öğeleri bırakın. </li><li> Bu steril alana yukarıda olacak şekilde, mikroskop taşıyın mikroskop ışık kaynağı açmak ve gerektiğinde Okülerin ayarlayın. </li><li> Obtain kornea dokusu ve hazırlanan steril işleme alanının yanında yer (koruma medya) elimden. Görüntüleme odasının kapağını gevşetin ve meşgul hiçbir parçacıkları ile oda tabanını inceleyen üstüne dinlensin. Non-steril eldiven atın. </li><li> Steril eldiven ve havlu ile steril önlük açık bir çift. </li><li> Cerrahi fırçalayın gerçekleştirin. Daha sonra, steril önlük, steril eldiven koyun. </li><li> Açık alet kutuları, böylece tüm malzemeleri erişilebilir. Açık nem çalışma alanı oluşturmak için geçirimsiz drape. </li><li> 5 cc şırınga iğnesi takın. Bir hemostat kullanarak, eğim bir dirsek 1 mm yerleştirilerek 30 G iğne hazırlanması ve bazın başka bir dirsek 1 mm (&quot;S&quot; şeklini oluşturmak için). </li><li> Şırıngaya pistonu yerleştirilmesi ve parmak pistonu sıkarak, tripan mavi boyama için cam şırınga birleştirin. Kanülü takın. </li><li> Dengelenmiş tuz çözeltisi (BSS), şişeye kanülü takın. </li><li> Vakum bloktan trefin yumruk ayırınve bir kenara koyun. Mikroskop bakış alanı içinde vakum bloğunu. </li><li> Mikroskop ayarlarını odağa vakum blok getir. </li></ol> Donör Doku Hazırlama 2. Prosedür <ol><li> Bir bariyer olarak folyo kare kullanarak odasını inceleyen kapağı kaldırın. Sklera kornea kaldırmak için jant tarafından elinin tersiyle ve forseps kullanarak, hafifçe kornea kavramak. Odasını inceleyen kapağını değiştirin. </li><li> Tamamen emme şırınga bastırın, sonra aşağı vakum blok üzerinde kornea epitel tarafı yerleştirin ve merkezli olduğundan emin olun. (Şekil 1) </li><li> Kılavuz mesajları kullanarak vakum blok hafifçe daha düşük oturma halkası. Yavaş yavaş emme şırınga piston bırakılmasıyla vakum devreye sokun. Oturma halkasını çıkarın ve bir kenara koyun. (Şekil 2). </li><li> Descemet zarından skor kısmi trepanasyon gerçekleştirin. </li><li> Doku iyi görüntülenmiştir böylece mikroskop zoom ve odak ayarlayın. </li><li&gt; Vakum blok eğin ve aşırı orta kaldırmak için limbusta yerleştirilen steril çubukla mızrak kullanın. (Şekil 3) </li><li> Atılan Descemet zarının kenarına leke limbusta yeterince leke yerleştirin. Leke 60-90 saniye boyunca endotel üzerinde kalmasına izin verin. (Şekil 4) </li><li> Lekeyi: <ol><li> Kornea kapalı ve gazlı bezle üzerine lekeyi dökmek için vakum blok eğin. </li><li> Endoteli dokunmamaya özen, korneanın limbal alanından fazla lekeyi kaldırmak için cerrahi mızrak kullanın. (Şekil 5) </li></ol></li><li> Aşırı BSS kaldırmak için boyama çözüm kornea durulama BSS birkaç damla kullanın ve steril swab mızrak. (Şekil 6) </li><li> Kurutma doku önlemek için sklera jant üzerinde BSS 1-2 damla koyun. Işlem boyunca gerektiği kadar tekrarlayın. </li><li> Descemet zarının kısmi diseksiyon: <ol><li> Boru bağlamak nerede başlayanVakum blok s, açık forseps kullanılarak, skor işareti kenarı ile uzak stromadan hafifçe ayrı Descemet membran ve endotel az bir ucu ile. (Şekil 7). </li><li> Ayrı Descemet membranı ve endotel fazla 1 attı kenarından mm ve devam diseksiyon etrafında 360 °. (Şekil 8). </li><li> Cilt markeri ile vakum blok üzerinde bir nokta koyarak herhangi bir mikro gözyaşları yerini unutmayın. Not: trepan Descemet zar içinden tamamen nüfuz etmediği gerektiği gibi boyama bir ikinci uygulamasında, ardından gerektiğinde, Descemet zardan skoru tamamlamak için hazırlanan 30 G iğne eğimli kenar kullanın. Descemet zarının adhesivite kısmi diseksiyonu engelleyen ise gerektiği gibi, trephinated alanı dışında endotelyumu çıkarın. </li></ol></li><li> Skor işareti üst üste herhangi bir endotel etiketlerini çıkarmak için forseps kullanın. </li> &lt;li&gt; önceden işaretlenmiş mikro gözyaşları konumunu düşünün ve büyük 06:00 olacak şekilde vakum blok döndürmek. Bu, kapağın menteşe olacaktır. </li><li> Forseps kullanarak, saat 12 Descemet membran kavramak. Yavaşça menteşe doğru soyma tarafından stromadan Descemet zarı ayırmak. Puan işaretinden ayırma 2 mm durdurarak menteşe oluşturun. NOT: flep Periferik kenarları ayrılması sırasında kaydırma başlayacak. (Şekil 9). </li><li> Yavaşça yerine geri stroma üzerinde flep yatıyordu. Endotelyumu unscroll ve özgün konumuna geri dönmek için BSS ve sürüntü mızrak kullanın. (Şekil 10A-C) </li><li> Flep konumda kalır böylece steril çubukla mızraklarla Descemet membran ve stroma arasındaki kalıntı sıvıyı çıkarın. (Şekil 11) </li><li> Menteşe yakın skleradan fazla nemi emmek için steril çubukla mızrak kullanın. Çizmek için cilt işaretleyici kullanınn menteşenin merkezine işaret sklera jant üzerine ok ucu steril pamuklu çubuk mızrak ile fazla mürekkep çıkarın. (Şekil 12) </li><li> Vakum bloğuna bağlı, şırınganın piston basmak suretiyle vakum bırakın. Vakum devre dışı iken, vakum bloktan kornea kaldırmak için forseps kullanabilir. </li><li> Bir bariyer olarak folyo kare kullanarak odasını inceleyen kapağı kaldırın. Yukarı, görüntüleme odasında endotel tarafı kornea yerleştirin ve odasını görüntüleme kapağını değiştirin. </li><li> Varsa, dostum kornea ile prosedür yukarıdaki adımları tekrarlayın. İkinci doku işlemeden önce vakum blok temizlemek için gazlı bez kullanın. </li><li> Görüntüleme odası kapak (lar) sıkın ve işleme alanından doku kaldırmak. </li><li> Biyolojik zararlı Atık Yönetim Planına göre öğelerin atılması ile steril alanı kırın. Kirlenmiş eldiven ve önlük çıkarın. Ellerinizi yıkayın ve non-steril eldiven yok. </li><li> Kesme sonrası biomikroskop değerlendirme ve korneanın speküler mikroskopi (ler) gerçekleştirinve bulguları kaydetmeye. </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>

Hasan TAUSIF ve Dr Navasuja Chandrasekaran ifşa hiçbir şey yok. Dr. Mian, Shtein ve Woodward Michigan Göz Bankası tıbbi yönetmenleri. Sayın Mavin, Sayın Titus ve Bayan Johnson Midwest Göz Bankaları çalışanlarıdır.

Kornea endotel hücre tabakası dehidratasyon ile kornea netlik sürdürülmesinde önemli bir rol oynar. Endotel hücrelerinin hastalıkları önemli görme kaybı ile sonuçlanabilir. DMEK Descemet membranı ve endotel hücre tabakasını kapsayan bir verici greft ile hastalıklı endotel hücrelerinin yerine amaçlamaktadır. Sağlam bir DM ve yeterli endotel hücre sayımı ile Donör doku hazırlanması greft dokusunun yaşam için önemlidir. Donör doku hazırlanması sırasında DM yırtılma doku kaybı ve don?...

Endotel Keratoplasti (EK) hızla 2012 6 Amerika Birleşik Devletleri&#39;nde yapılan tüm nakiller üzerinden% 50 için muhasebe endotel hastalıkların tedavisi için tercih prosedür olarak keratoplasti (PKP) delici yerini aldı. PKP yüksek astigmatizma ile uzun süreli görsel rehabilitasyon ile sınırlıdır ve refraktif istikrarsızlık, dünya bütünlüğü ve dikiş-ilişkili komplikasyonların kaybı. Derin Yapraklı endotelyal keratoplasti (dlek) dahil Erken nesil EK işlemleri, kısmi kalınlıkta donör doku nakli ile korneal stroma manuel diseksiyon çıkıyor. Bu prosedürler sağlam bir kornea yüzeyini korurken donör endotel doku nakli başarılı ama teknik olarak zor ve bağlı düzensiz kornea lamelli arabirimi 1 ile sınırlı iyi düzeltilmiş görme keskinliği sonuçlandı. Seçim mevcut EK cerrahisinin kornea Clar sunuyor Descemet sıyırma endotelyal keratoplasti (DSEK) olduğunuvizyon hızlı iyileşme Sığ, postoperatif refraksiyon kusuru ve astigmat en aza indirir ve kornea yüzeyini 7 korurken travmatik yara açılma riskini azaltır. Bu prosedür, yeni bir endotel hücre tabakası içeren kısmi kalınlıkta donör kornea dokusu ile hastalıklı Descemet zarının çıkarılması ve endotelyumu ve değiştirme içerir. Donör kornea 8 nakledildikten arka doku hazırlamak için bir mikrokeratomu ile disseke edilir. Göz bankalar tarafından donör doku hazırlanması, tartışmalı, bu prosedürün başarısı hızla büyümesinde etkili olmuştur ve. 2005 ve 2012 yılları arasında Amerika Birleşik Devletleri&#39;nde DSEK prosedürleri yakın 16 kat artış şu anda göz bankalar 6 tarafından hazırlanan doku neredeyse yarısı gelir. DSEK 20/40 veya daha iyi görme keskinliğini artırmak rağmen, hastaların çoğu nedeniyle stromal arayüzü bulanıklık ve artan ışığa 20/20 vizyona ulaşmak değilDonör stromal dokusunun 7 eklenmesinden dağıtır. EK cerrahi teknik, yalnızca verici Descemet zarı (DM) ve endotel nakli içerir Descemet Membran endotelyal keratoplasti (DMEK) kadar ilerlemiştir. Bu prosedür 20/20 vizyon, daha hızlı görsel iyileşme, ve endotelyal red 9 daha düşük bir risk ulaşan hasta daha yüksek bir paya sahiptir. Bununla birlikte, verici dokunun hazırlanması nedeniyle ince greft boyutlarına daha zordur.

<table><tbody><tr><td>VisionBlue</td><td>D.O.R.C., Zuiland, The Netherlands</td><td>NA</td><td></td></tr><tr><td>Barron vacuum punch</td><td>Katena, Denville, New Jersey</td><td>K20-2112</td><td></td></tr></tbody></table>

corneal donor tissue preparation for descemet's membrane endothelial keratoplasty

Descemet Membran Endotel Keratoplasi (DMEK) olan bir temel zar (Descemet membranı) ile birlikte tek bir hücre tabakası, kornea endotelyum, alıcının arka stromada 3 üzerine zerk edilmekte olduğu kornea şeklidir. Ek bağış stroma tanıtıldı Descemet sıyırma Otomatik endotelyal keratoplasti (DSAEK), aksine, hiçbir doğal olmayan stroma-to-stroma arayüzü oluşturulur. Bunun bir sonucu olarak, korneanın doğal anatomisi geliştirilmiş geri kazanım süresi ve görme keskinliği 4 izin mümkün olduğu kadar korunur. Endotel Keratoplasti (EK) endotel disfonksiyonu tedavisi için tercih işlemdir. EK avantajları nedeniyle küçük, periferik kesi 1 kullanımı hızlı görme kurtarma, gözün bütünlüğü ve minimal refraktif değişim korunması bulunmaktadır. DSAEK kısmi kalınlıkta stroma ve endotel ile hazırlanan donör doku kullanmaktadır. Hızlı succesini ve bu prosedürün kullanımı göz bankanın durumuna isnat edilebilir precut doku hazırlanmıştır. Donör doku göz banka hazırlık faydaları hazırlanması sırasında doku perforasyon durumunda geri donör doku çalışma odası ve kullanılabilirlik özel ekipman ihtiyacı ortadan kaldırılmasını içerir. Buna ek olarak, göz-banka teknisyenler tarafından donör doku yüksek hacimli hazırlanması donör doku kalitesi iyileştirilmiş sağlayabilir. DSAEK nedeniyle donör ve alıcı kornea arasında stromal arayüz oluşturulması için en iyi düzeltilmiş görme keskinliği sınırlı olabilir. Görsel sonuçlarını iyileştirmek ve EK 5 sonra komplikasyonları azaltabilir DMEK sadece donör Descemet membran ve endotel nakli ile bu arayüzü ortadan kaldırılması. DSAEK, uzun süreli başarı ve DMEK kabul benzer hazır kesilmiş kullanılabilirlik kolaylığı bağlıdır, göz banka donör doku hazırladı. Burada bazı BARRI azaltabilir donör doku hazırlanması için aşamalı bir yaklaşım sunmakers göz bankalar DMEK greft sağlayan karşıya.

Stromadan DM ve endotel Uygun ayırma DMEK için hazır kesilmiş doku hazırlanması esastır. 65 ± 6 yıl (aralık 55-75), 2.616 ± 321 hücre / mm 2, 5.9 ± 2.3 gün (dağılım 1-10) ve endotel hücre yoğunluğu işlem süresi ölüm (AKD) (yaş ortalaması elli donör kornealar aralığı 2,049-3,247) iki göz bankası teknisyenleri tarafından cerrahi hazırlama tekniği fizibilite ve tekrarlanabilirliği değerlendirmek için kullanıldı. Başarı oranı ilk 25 olguda% 72 ve% 76 genel başarı oran?...

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's Membrane Endothelial Keratoplasty

Endotel keratoplasti gelişmeler her arka arkaya 1 ile ince greft nakli neden olarak sürekli gelişen bir cerrahi tekniktir. Burada endotel ve transplantasyon 2 donör korneoskleral düğmeleri Descemet zarının güvenli ve tekrarlanabilir ayrılmasına izin kontrollü manuel doku diseksiyonu için bir teknik sunuyoruz.

Descemet Membran endotelyal keratoplasti için Kornea Donör doku hazırlanması

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

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JoVE Journal

Medicine

15.2K Görüntüleme.  University of Michigan.  Endotel keratoplasti gelişmeler her arka arkaya 1 ile ince greft nakli neden olarak sürekli gelişen bir cerrahi tekniktir. Burada endotel ve transplantasyon 2 donör korneoskleral düğmeleri Descemet zarının güvenli ve tekrarlanabilir ayrılmasına izin kontrollü manuel doku diseksiyonu için bir teknik sunuyoruz. 

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

Adenoviral Gene Therapy for Diabetic Keratopathy: Effects on Wound Healing and Stem Cell Marker Expression in Human Organ-cultured Corneas and Limbal Epithelial Cells

The goal of this protocol is to describe molecular alterations in human diabetic corneas and demonstrate how they can be alleviated by adenoviral gene therapy in organ-cultured corneas. The diabetic corneal disease is a complication of diabetes with frequent abnormalities of corneal nerves and epithelial wound healing. We have also documented significantly altered expression of several putative epithelial stem cell markers in human diabetic corneas. To alleviate these changes, adenoviral gene therapy was successfully implemented using the upregulation of c-met proto-oncogene expression and/or the downregulation of proteinases matrix metalloproteinase-10 (MMP-10) and cathepsin F. This therapy accelerated wound healing in diabetic corneas even when only the limbal stem cell compartment was transduced. The best results were obtained with combined treatment. For possible patient transplantation of normalized stem cells, an example is also presented of the optimization of gene transduction in stem cell-enriched cultures using polycationic enhancers. This approach may be useful not only for the selected genes but also for the other mediators of corneal epithelial wound healing and stem cell function.

An example of adenoviral gene therapy in the human diabetic organ-cultured corneas is presented towards the normalization of delayed wound healing and markedly reduced epithelial stem cell marker expression in these corneas. It also describes the optimization of this process in stem cell-enriched limbal epithelial cultures.

Diyabetik Keratopatide için adenoviral Gen Tedavisi: İnsan Organ-kültürlü kornealar ve Limbal Epitel Hücreleri Yara İyileşmesi Üzerine Etkileri ve Hücre Marker İfade Kök

The goal of this protocol is to describe molecular alterations in human diabetic corneas and demonstrate how they can be alleviated by adenoviral gene ...

Gelişim Biyolojisi

A Simplified Technique for In situ Excision of Cornea and Evisceration of Retinal Tissue from Human Ocular Globe

Enucleation is the process of retrieving the ocular globe from a cadaveric donor leaving the rest of the globe undisturbed. Excision refers to the retrieval of ocular tissues, especially cornea, by cutting it separate from the ocular globe. Evisceration is the process of removing the internal organs referred here as retina. The ocular globe consists of the cornea, the sclera, the vitreous body, the lens, the iris, the retina, the choroid, muscles etc (Suppl. Figure 1). When a patient is suffering from corneal damage, the cornea needs to be removed and a healthy one must be transplanted by keratoplastic surgeries. Genetic disorders or defects in retinal function can compromise vision. Human ocular globes can be used for various surgical procedures such as eye banking, transplantation of human cornea or sclera and research on ocular tissues. However, there is little information available on human corneal and retinal excision, probably due to the limited accessibility to human tissues. Most of the studies describing similar procedures are performed on animal models. Research scientists rely on the availability of properly dissected and well-conserved ocular tissues in order to extend the knowledge on human eye development, homeostasis and function. As we receive high amount of ocular globes out of which approximately 40% (Table 1) of them are used for research purposes, we are able to perform huge amount of experiments on these tissues, defining techniques to excise and preserve them regularly.
The cornea is an avascular tissue which enables the transmission of light onto the retina and for this purpose should always maintain a good degree of transparency. Within the cornea, the limbus region, which is a reservoir of the stem cells, helps the reconstruction of epithelial cells and restricts the overgrowth of the conjunctiva maintaining corneal transparency and clarity. The size and thickness of the cornea are critical for clear vision, as changes in either of them could lead to distracted, unclear vision. The cornea comprises of 5 layers; a) epithelium, b) Bowman's layer, c) stroma, d) Descemet's membrane and e) endothelium. All layers should function properly to ensure clear vision4,5,6. The choroid is the intermediate tunic between the sclera and retina, bounded on the interior by the Bruch's membrane and is responsible for blood flow in the eye. The choroid also helps to regulate the temperature and supplies nourishment to the outer layers of the retina5,6. The retina is a layer of nervous tissue that covers the back of the ocular globe (Suppl. Figure 1) and consists of two parts: a photoreceptive part and a non-receptive part. The retina helps to receive the light from the cornea and lens and converts it into the chemical energy eventually transmitted to the brain with help of the optic nerve5,6.
The aim of this paper is to provide a protocol for the dissection of corneal and retinal tissues from human ocular globes. Avoiding cross-contamination with adjacent tissues and preserving RNA integrity is of fundamental importance as such tissues are indispensable for research purposes aimed at (i) characterizing the transcriptome of the ocular tissues, (ii) isolating stem cells for regenerative medicine projects, and (iii) evaluating histological differences between tissues from normal/affected subjects. In this paper we describe the technique we currently use to remove the cornea, the choroid and retinal tissues from an ocular globe. Here we provide a detailed protocol for the dissection of the human ocular globe and the excision of corneal and retinal tissues. The accompanying video will help researchers to learn an appropriate technique for the retrieval of precious human tissues which are difficult to find regularly.

A Simplified Technique for In situ Excision of Cornea and Evisceration of Retinal Tissue from Human Ocular Globe

The paper describes a simplified technique to excise corneal and to eviscerate retinal tissues from the ocular globe of human cadaveric donors. The technique described here will help to excise good quality tissues to be used for transplantation, surgical or research purposes without damaging other tissues of the ocular globe.

Için Basitleştirilmiş Tekniği<em&gt; Yerinde</emİnsan Oküler Globe gelen Kornea ve Retina Doku Eviserasyon of&gt; eksizyonu

Enucleation is the process of retrieving the ocular globe from a cadaveric donor leaving the rest of the globe undisturbed. Excision refers to the ...

Tıp

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.

Endotel Keratoplasti için Kornea Donör doku hazırlanması

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

Corneal Tissue Engineering: An In Vitro Model of the Stromal-nerve Interactions of the Human Cornea

Tissue engineering has gained substantial recognition due to the high demand for human cornea replacements with an estimated 10 million people worldwide suffering from corneal vision loss1. To address the demand for viable human corneas, significant progress in three-dimensional (3D) tissue engineering has been made2,3,4. These cornea models range from simple monolayer systems to multilayered models, leading to 3D full-thickness corneal equivalents2.
However, the use of a 3D tissue-engineered cornea in the context of in vitro disease models studied to date lacks resemblance to the multilayered 3D corneal tissue structure, function, and the networking of different cell types (i.e., nerve, epithelium, stroma, and endothelium)2,3. In addition, the demand for in vitro cornea tissue models has increased in an attempt to reduce animal testing for pharmaceutical products. Thus, more sophisticated models are required to better match systems to human physiological requirements, and the development of a model that is more relevant to the patient population is absolutely necessary. Given that multiple cell types in the cornea are affected by diseases and dystrophies, such as Keratoconus, Diabetic Keratopathy, and Fuchs, this model includes a 3D co-culture model of primary human corneal fibroblasts (HCFs) from healthy donors and neurons from the SH-SY5Y cell line. This allows us for the first time to investigate the interactions between the two cell types within the human corneal tissue. We believe that this model could potentially dissect the underlying mechanisms associated with the stromal-nerve interactions of corneal diseases that exhibit nerve damages. This 3D model mirrors the basic anatomical and physiological nature of the corneal tissue in vivo and can be used in the future as a tool for investigating corneal defects as well as screening the efficacy of various agents before animal testing.

Corneal Tissue Engineering: An In Vitro Model of the Stromal-nerve Interactions of the Human Cornea

This protocol describes a novel three-dimensional in vitro model, where corneal stromal cells and differentiated neuronal cells are cultured together to assist in the examination and understanding of the interactions of the two cell types.

Kornea doku Mühendisliği: İnsan kornea sinir Stromal etkileşimlerin bir Vitro modeli

Tissue engineering has gained substantial recognition due to the high demand for human cornea replacements with an estimated 10 million people worldwide ...

Combination of Microstereolithography and Electrospinning to Produce Membranes Equipped with Niches for Corneal Regeneration

Corneal problems affect millions of people worldwide reducing their quality of life significantly. Corneal disease can be caused by illnesses such as Aniridia or Steven Johnson Syndrome as well as by external factors such as chemical burns or radiation. Current treatments are (i) the use of corneal grafts and (ii) the use of stem cell expanded in the laboratory and delivered on carriers (e.g., amniotic membrane); these treatments are relatively successful but unfortunately they can fail after 3-5 years. There is a need to design and manufacture new corneal biomaterial devices able to mimic in detail the physiological environment where stem cells reside in the cornea. Limbal stem cells are located in the limbus (circular area between cornea and sclera) in specific niches known as the Palisades of Vogt. In this work we have developed a new platform technology which combines two cutting-edge manufacturing techniques (microstereolithography and electrospinning) for the fabrication of corneal membranes that mimic to a certain extent the limbus. Our membranes contain artificial micropockets which aim to provide cells with protection as the Palisades of Vogt do in the eye.

We report a technique for the fabrication of micropockets within electrospun membranes in which to study cell behavior. Specifically, we describe a combination of microstereolithography and electrospinning for the production of PLGA (Poly(lactide-co-glycolide)) corneal biomaterial devices equipped with microfeatures.

Microstereolithography ve ELEKTROÜRETİM kombinasyonu Kornea Yenilenme Nişli ile donatılmış Membranlarını Üretiliyor

Corneal problems affect millions of people worldwide reducing their quality of life significantly. Corneal disease can be caused by illnesses such as ...

Biyomühendislik

Murine Corneal Transplantation: A Model to Study the Most Common Form of Solid Organ Transplantation

Corneal transplantation is the most common form of organ transplantation in the United States with between 45,000 and 55,000 procedures performed each year. While several animal models exist for this procedure and mice are the species that is most commonly used. The reasons for using mice are the relative cost of using this species, the existence of many genetically defined strains that allow for the study of immune responses, and the existence of an extensive array of reagents that can be used to further define responses in this species. This model has been used to define factors in the cornea that are responsible for the relative immune privilege status of this tissue that enables corneal allografts to survive acute rejection in the absence of immunosuppressive therapy. It has also been used to define those factors that are most important in rejection of such allografts. Consequently, much of what we know concerning mechanisms of both corneal allograft acceptance and rejection are due to studies using a murine model of corneal transplantation. In addition to describing a model for acute corneal allograft rejection, we also present for the first time a model of late-term corneal allograft rejection.

Mice have been used as a model for studying many forms of transplantation, including corneal transplantation. We describe in this report a murine model for both acute and late-term corneal transplantation.

Sıçangiller Kornea Nakli: Katı Organ Nakli En Yaygın Formu Eğitim için Bir Model

Corneal transplantation is the most common form of organ transplantation in the United States with between 45,000 and 55,000 procedures performed each ...

An &#8220;All-laser&#8221; Endothelial Transplant

The &#8220;all laser&#8221; assisted endothelial keratoplasty is a procedure that is performed with a femtosecond laser used to cut the donor tissue at an intended depth, and a near infrared diode laser to weld the corneal tissue. The proposed technique enables to reach the three main goals in endothelial keratoplasty: a precise control in the thickness of the donor tissue; its easy insertion in the recipient bed and a reduced risk of donor lenticule dislocation. The donor cornea thickness is measured in the surgery room with optical coherence tomography (OCT), in order to correctly design the donor tissue dimensions. A femtosecond laser is used to cut the donor cornea. The recipient eye is prepared by manual stripping of the descemetic membrane. The donor endothelium is inserted into a Busin-injector, the peripheral inner side is stained with a proper chromophore (a water solution of Indocyanine Green) and then it is pulled in the anterior chamber. The transplanted tissue is placed in the final and correct location and then diode laser welding is induced from outside the eyeball. The procedure has been performed on more than 15 patients evidencing an improvement in surgery performances, with a good recovery of visual acuity and a reduced donor lenticule dislocation event.

An all laser procedure is proposed in the endothelial transplant. The surgical technique is based on the use of a femtosecond laser to prepare the donor tissue. Laser welding technique is then used to secure the donor endothelium in the correct position.

Bir &quot;All-lazer&quot; Endotelyal Nakli

The &#8220;all laser&#8221; assisted endothelial keratoplasty is a procedure that is performed with a femtosecond laser used to cut the donor tissue at an ...

Growth of Human and Sheep Corneal Endothelial Cell Layers on Biomaterial Membranes

Corneal endothelial cell cultures have a tendency to undergo epithelial-to-mesenchymal transition (EMT) after loss of cell-to-cell contact. EMT is deleterious for the cells as it reduces their ability to form a mature and functional layer. Here, we present a method for establishing and subculturing human and sheep corneal endothelial cell cultures that minimizes the loss of cell-to-cell contact. Explants of corneal endothelium/Descemet&#39;s membrane are taken from donor corneas and placed into tissue culture under conditions that allow the cells to collectively migrate onto the culture surface. Once a culture has been established, the explants are transferred to fresh plates to initiate new cultures. Dispase II is used to gently lift clumps of cells off tissue culture plates for subculturing. Corneal endothelial cell cultures that have been established using this protocol are suitable for transferring to biomaterial membranes to produce tissue-engineered cell layers for transplantation in animal trials. A custom-made device for supporting biomaterial membranes during tissue culture is described and an example of a tissue-engineered graft composed of a layer of corneal endothelial cells and a layer of corneal stromal cells on either side of a collagen type I membrane is presented.

This protocol describes the critical steps required to establish and grow corneal endothelial cell cultures from explants of human or sheep tissue. A method for subculturing corneal endothelial cells on membranous biomaterials is also presented.

Biyomalzeme Membranlar Üzerinde İnsan ve Koyun Korneal Endotel Hücre Tabakalarının Büyümesi

Corneal endothelial cell cultures have a tendency to undergo epithelial-to-mesenchymal transition (EMT) after loss of cell-to-cell contact. EMT is ...

Full-Field Optical Coherence Microscopy for Histology-Like Analysis of Stromal Features in Corneal Grafts

The quality of donor corneal stroma, which makes up about 90% of total corneal thickness, is likely to be one of the main, if not the major, limiting factor(s) for success of deep anterior lamellar and penetrating keratoplasty. These are surgical procedures that involve replacing part or all of the diseased corneal layers, respectively, by donated tissue, the graft, taken from a recently deceased individual. However, means to evaluate stromal quality of corneal grafts in eye banks are limited and lack the capability of high-resolution quantitative assessment of disease indicators. Full-field optical coherence microscopy (FF-OCM), permitting high-resolution 3D imaging of fresh or fixed ex vivo biological tissue samples, is a non-invasive technique well suited for donor cornea assessment. Here we describe a method for the qualitative and quantitative analysis of corneal stroma using FF-OCM. The protocol has been successfully applied to normal donor corneas and pathological corneal buttons, and can be used to identify healthy and pathologic features on both the macroscopic and microscopic level, thereby facilitating the detection of stromal disorders that could compromise the outcome of keratoplasty. By improving the graft quality control, this protocol has the potential to result in better selection (and rejection) of donor tissues and hence decreased graft failure.

We describe use of full-field optical coherence microscopy as a method for high quality assessment of corneal donor stroma. This protocol can be used to identify features indicative of health or disease, and is aimed at improving the screening and selection of donor tissues, and hence the results of keratoplasty.

Kornea Greftlerinde Stromal Özelliklerin Histoloji Benzeri Analizi için Tam Alan Optik Koherens Mikroskopisi

The quality of donor corneal stroma, which makes up about 90% of total corneal thickness, is likely to be one of the main, if not the major, limiting ...

İmmünoloji ve Enfeksiyon

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).

Kornea Endotel Hücre Kaybının Noninvaziv, Lazer Destekli Deneysel Modelinin Geliştirilmesi

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

Descemet Membran endotelyal keratoplasti için Kornea Donör doku hazırlanması

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