\u00d6ZET
\n Desel\u00fclerize edilmi\u015f doku ve organlar \u00e7e\u015fitli doku m\u00fchendisli\u011fi ve rejeneratif t\u0131p uygulamalar\u0131nda ba\u015far\u0131yla kullan\u0131lmaktad\u0131r. Ekstrasel\u00fcler matristen elde edilen biyolojik bir iskele, i\u015flem g\u00f6recek dokudan h\u00fccreleri etkin bir \u015fekilde ortadan kald\u0131ran \u00e7e\u015fitli desel\u00fclerizasyon y\u00f6ntemleri ile \u00fcretilebilir. Kullan\u0131lan desel\u00fclerizasyon y\u00f6ntemleri dokular\u0131n ve organlar\u0131n yap\u0131s\u0131na g\u00f6re de\u011fi\u015fir. H\u00fccrelerin bir dokudan \u00e7\u0131kar\u0131lmas\u0131, dokunun kayna\u011f\u0131na ve kullan\u0131lan spesifik fiziksel, kimyasal ve enzimatik y\u00f6ntemlere ba\u011fl\u0131d\u0131r. Bu y\u00f6ntemlerin her biri geri kalan h\u00fccre d\u0131\u015f\u0131 matrisin (ECM) yap\u0131s\u0131ndaki biyokimyasal bile\u015fimini, dokunun ince yap\u0131s\u0131n\u0131 (ultrastr\u00fckt\u00fcr\u00fcn\u00fc) ve mekanik davran\u0131\u015f\u0131n\u0131 etkiler. Bu makalede en s\u0131k kullan\u0131lan desel\u00fclerizasyon y\u00f6ntemleri tan\u0131t\u0131larak ve bu y\u00f6ntemlerin biyolojik doku iskelesi materyali \u00fczerindeki etkileri ele al\u0131nmaktad\u0131r.
\n Anahtar Kelimeler; Desel\u00fclerizasyon, Ekstrasel\u00fcler Matris, Doku m\u00fchendisli\u011fi (TE), Desel\u00fclerizasyon y\u00f6ntemleri
\n DECELLULARIZATION METHODS AND USING OF THE TISSUES
\n Abstract
\n Decellularized tissues and organs have been successfully used in various tissue engineering and regenerative medicine applications. A biological scaffold obtained from the extracellular matrix can be produced by various decellularization methods which efficiently remove the cells from the tissue to be treated. Deceleration methods used vary according to the structure of the tissues and organs. The removal of cells from a tissue depends on the source of the tissue and the specific physical, chemical, and enzymatic methods used. Each of these methods influences the biochemical composition of the remaining extracellular matrix (ECM) structure, the ultrastructure of the tissue (ultrastructural product), and its mechanical behavior. The most commonly used deceleration methods are introduced in this article and the effects of these methods on biological tissue scaffold material are discussed.
\n Keywords; Decellularization Extracellular Matrix, tissue engineering (TE), Decellularization methods
\n 1. G\u0130R\u0130\u015e
\n Doku ve organ nakli \u00e7al\u0131\u015fmalar\u0131n\u0131n k\u0131s\u0131tl\u0131 olmas\u0131, doku m\u00fchendisli\u011fi (TE) yap\u0131lar\u0131 dahil olmak \u00fczere \u00e7e\u015fitli alternatiflerin geli\u015ftirilmesine yol a\u00e7m\u0131\u015ft\u0131r. Biyo iskelesi tasar\u0131m\u0131ndaki ilerlemelere ra\u011fmen, do\u011fal doku ultrastr\u00fckt\u00fcr\u00fcn\u00fc ve biyomekanik \u00f6zelliklerini etkili bir \u015fekilde taklit eden e\u015fde\u011fer yap\u0131lar bulunmam\u0131\u015ft\u0131r. Bununla birlikte doku m\u00fchendisli\u011finde biyo iskelesi \u00fcretim y\u00f6ntemlerinden biri olan desel\u00fclerizasyon y\u00f6ntemi s\u0131k kullan\u0131lan ve en uygun y\u00f6ntem olarak g\u00f6zlemlenmi\u015ftir (Mertsching vd., 2005; Wilson vd., 2013).
\n Desel\u00fclerizasyon dokunun veya organ\u0131n kendine \u00f6zg\u00fcn mimarisini koruyarak b\u00fct\u00fcn organlar\u0131n h\u00fccrelerinden ar\u0131nd\u0131r\u0131p, iskeleleri \u00fcreten bir y\u00f6ntemdir. Bu, organ yap\u0131s\u0131n\u0131 taklit de\u011fil, ayn\u0131 zamanda h\u00fccre d\u0131\u015f\u0131 matrise (ECM) yard\u0131m h\u00fccre \u00e7o\u011falmas\u0131 ve farkl\u0131la\u015fmas\u0131 i\u00e7inde kimyasal bile\u015fenleri i\u00e7erir (Wilson vd., 2013; Hrebikova vd., 2015).
\n Dokular\u0131n desel\u00fclerizasyonu i\u00e7in en yayg\u0131n kullan\u0131lan y\u00f6ntemler, fiziksel, kimyasal ve enzimatik i\u015flemleri bir arada i\u00e7erir. Fiziksel i\u015flemde ajitasyon veya sonikasyon, mekanik masaj veya bas\u0131n\u00e7 ya da donma ve \u00e7\u00f6z\u00fclme i\u00e7erebilir. Bu y\u00f6ntemler, ECM h\u00fccre i\u00e7eri\u011finin sonradan durulama ve \u00e7\u0131kar\u0131lmas\u0131n\u0131 kolayla\u015ft\u0131rmak i\u00e7in h\u00fccre zar\u0131n\u0131 bozar. Bu fiziksel i\u015flemler tam desel\u00fclerize elde etmek i\u00e7in genellikle yetersizdir ve bir kimyasal i\u015flem ile kombine edilmelidir. Enzimatik i\u015flemler, tripsin gibi kimyasal i\u015flemler veya iyonik \u00e7\u00f6zeltiler, h\u00fccre zar\u0131n\u0131n, h\u00fccreler aras\u0131 ve h\u00fccre d\u0131\u015f\u0131 ba\u011flant\u0131lar\u0131ndan sorumlu ba\u011flar\u0131n\u0131 bozmaktad\u0131rlar. Dokular hem h\u00fccresel malzemeden hem de ECM\u2019den olu\u015fur ve ECM\u2019nin doku kayna\u011f\u0131na ba\u011fl\u0131 kompaktl\u0131\u011f\u0131 de\u011fi\u015fken derecede d\u00fczenlenmi\u015ftir. ECM, desel\u00fclerizasyon i\u015flemi s\u0131ras\u0131nda kaotropik maddeler ile b\u00fct\u00fcn h\u00fccrelerin yeterli s\u00fcrede i\u015fleme maruz kalmas\u0131 malzemenin dokudan \u00e7\u0131kar\u0131lmas\u0131na bir yol sa\u011flamak i\u00e7in kesintiye u\u011framaktad\u0131r. \u00c7o\u011fu desel\u00fclerizasyon s\u00fcre\u00e7lerinin amac\u0131 kesintileri en aza indirmek ve standart fiziksel \u00f6zellikleri ve biyolojik \u00f6zellikleri muhafaza etmektir (Gilbert vd., 2006; Akbay ve Onur, 2016).
\n Yap\u0131lan bu \u00e7al\u0131\u015fmada desel\u00fclerizasyon y\u00f6ntemleri ve bu y\u00f6ntemlerin nas\u0131l kullan\u0131ld\u0131\u011f\u0131, ekstrasel\u00fcler matris karakterizasyonu ve desel\u00fclerizasyon y\u00f6ntemlerinin dokularda kullan\u0131m\u0131 tan\u0131t\u0131lm\u0131\u015ft\u0131r.
\n 2. ECM (EKSTRASEL\u00dcLER MATR\u0130S) KARAKTER\u0130ZASYONU
\n Bir h\u00fccre ile \u00e7evresi veya di\u011fer h\u00fccreler aras\u0131ndaki etkile\u015fimler, h\u00fccre y\u00fczey proteinleri taraf\u0131ndan y\u00f6netilir (Wong, 2009). H\u00fccreler biyolojik ortamlar\u0131nda nanofiber formdaki proteinlerden olu\u015fan bir ekstrasel\u00fcler matris (ECM) i\u00e7erisinde bulunmaktad\u0131r (Bayram, 2012). Ekstrasel\u00fcler matris, \u00e7ok h\u00fccreli bir organizmada baz\u0131 h\u00fccreler taraf\u0131ndan salg\u0131lanan, h\u00fccreler aras\u0131n\u0131 dolduran ve tan\u0131mlanm\u0131\u015f bir alanda h\u00fccreleri tutan ba\u011flay\u0131c\u0131 madde olarak i\u015flev g\u00f6ren \u00e7e\u015fitli proteinler ve polisakkaritler i\u00e7in genel bir terimdir (Wong, 2009; \u015een, 2012; Yi\u011fit vd., 2016).
\n Kalp kapak\u00e7\u0131klar\u0131, kan damarlar\u0131, deri, sinirler, iskelet kas\u0131, tendon, ba\u011f, ince ba\u011f\u0131rsak alt mukoza (SIS), idrar kesesi ve karaci\u011fer d\u00e2hil olmak \u00fczere, \u00e7e\u015fitli dokularda, ECM doku m\u00fchendisli\u011fi ve rejeneratif ila\u00e7 uygulamalar\u0131 i\u00e7in ele al\u0131nm\u0131\u015ft\u0131r. Bir desel\u00fclerizasyon protokol\u00fcn\u00fcn amac\u0131, bile\u015fimin biyolojik aktivitesi ve kalan ECM mekanik b\u00fct\u00fcnl\u00fc\u011f\u00fc \u00fczerinde herhangi bir olumsuz etkisini en aza indirirken, verimli bir \u015fekilde t\u00fcm h\u00fccre ve h\u00fccre membranlar\u0131n\u0131n \u00e7\u0131kar\u0131lmas\u0131d\u0131r (Gilbert vd., 2006; Fu vd., 2014; Akbay ve Onur, 2016).
\n 3. DESEL\u00dcLER\u0130ZASYON Y\u00d6NTEMLER\u0130N\u0130N A\u00c7IKLANMASI
\n En sa\u011flam ve etkili desel\u00fclerizasyon y\u00f6ntemleri fiziksel, kimyasal bile\u015fimi ve enzimatik yakla\u015f\u0131mlar\u0131 i\u00e7ermektedir. Desel\u00fclerizasyon y\u00f6ntemleri genellikle, h\u00fccre bile\u015fenlerinin ayr\u0131lmas\u0131 ve ard\u0131ndan fiziksel tedaviler veya iyonik \u00e7\u00f6zeltiler kullan\u0131larak h\u00fccre membran lizizi ile ba\u015flar, ECM kullan\u0131ld\u0131\u011f\u0131nda enzimatik i\u015flemler deterjan kullanarak sitoplazmik ve h\u00fccre membran bile\u015fenlerin \u00e7\u00f6z\u00fcn\u00fcrl\u00fc\u011f\u00fc ve doku h\u00fccrelerinin kal\u0131nt\u0131lar\u0131n\u0131n kald\u0131r\u0131lmas\u0131 ile sonland\u0131r\u0131l\u0131r. Bu ad\u0131mlar\u0131n etkinli\u011fini geli\u015ftirmek i\u00e7in mekanik \u00e7alkalama yap\u0131lmaktad\u0131r. Desel\u00fclerizasyondan sonra kalan t\u00fcm kimyasallar, kimyasal\u0131n olumsuz konak doku yan\u0131t\u0131n\u0131 \u00f6nlemek i\u00e7in \u00e7\u0131kart\u0131lmal\u0131d\u0131r. Desel\u00fclerizasyon sonucu ECM koruma etkinli\u011fi \u00e7e\u015fitli y\u00f6ntemlerle tespit edilebilir. Dokular\u0131n \u00e7e\u015fitli fiziksel, enzimatik ve kimyasal mekanizmalar\u0131 a\u015fa\u011f\u0131daki b\u00f6l\u00fcmlerde ve tablo 1\u2019de g\u00f6zden ge\u00e7irilmi\u015ftir.
\n Tablo 1. Yayg\u0131n Olarak Kullan\u0131lan Desel\u00fclerizasyon Y\u00f6ntemleri ve Kaotropik Maddeler
\n Y\u00f6ntem Etki \u015fekli ECM \u00fczerindeki etkileri Kaynaklar
\n Fiziksel
\n Ek bile\u015fenini dondurma H\u00fccre i\u00e7i buz kristalleri, h\u00fccre zar\u0131n\u0131 bozabilir ECM, bozulmu\u015f veya h\u0131zl\u0131 donma s\u0131ras\u0131nda hasar g\u00f6rm\u00fc\u015f olabilir. Jackson vd., 1987
\n Gulati, 1988
\n Mekanik kuvvet Bas\u0131n\u00e7la h\u00fccreleri patlama ve doku al\u0131nmas\u0131 h\u00fccrelerini ortadan kald\u0131r\u0131r Mekanik kuvvet ECM zarar verebilir Freytes vd., 2004
\n Lin vd., 2004
\n Mekanik \u00e7alkalama H\u00fccre par\u00e7alama neden olabilir, ancak daha genel olarak kimyasal maddeye maruz kalmas\u0131n\u0131 ve h\u00fccresel malzemenin \u00e7\u0131kart\u0131lmas\u0131n\u0131 kolayla\u015ft\u0131rmak i\u00e7in kullan\u0131l\u0131r H\u00fccresel malzeme bertaraf edildi\u011fi zaman agresif \u00e7alkalama ya da sonikasyon ECM\u2019yi bozabilir Schenke-Layland vd., 2003
\n Dahl vd., 2003
\n Freytes vd., 2004
\n Lin vd., 2004
\n Kimyasal
\n Alkali; asit H\u00fccrelerin sitoplazmik eritmesi; n\u00fckleik asitleri bozar Glikoz amino glikan\u2019lar\u0131 (GAG) kald\u0131rma olas\u0131l\u0131\u011f\u0131 var\u0131dr. Yoo vd., 1998
\n De Filippo vd., 2002
\n Freytes vd., 2004
\n \u0130yonik olmayan deterjanlar
\n Triton X-100 Sa\u011flam protein-protein etkile\u015fimlerini b\u0131rak\u0131rken, lipid-lipid ve lipid-protein etkile\u015fimlerini bozar Kar\u0131\u015f\u0131k sonu\u00e7lar; dokulara g\u00f6re verim sa\u011flar, GAG\u2019lar\u0131 kald\u0131rma olas\u0131l\u0131\u011f\u0131 var\u0131dr. Chen vd., 1999
\n Cartmell ve Dunn, 2000
\n De Filippo vd., 2002
\n Dahl vd., 2003
\n Grauss vd., 2003
\n Lin vd., 2004
\n Woods ve Gratzer, 2005
\n \u0130yonik deterjanlar
\n Sodyum dodesil s\u00fclfat (SDS) Sitoplazmik ve h\u00fccresel membranlar \u00e7\u00f6z\u00fcn\u00fcr; proteinleri denat\u00fcre olma e\u011filimindedir H\u00fccre membranlar\u0131nda ki kal\u0131nt\u0131lar\u0131 ve sitoplazmik proteinleri yok eder; yerli doku yap\u0131s\u0131n\u0131 bozabilir GAGlar ve hasarl\u0131 kollajenleri giderme e\u011filimindedir.
\n Chen vd., 2004
\n Hudson vd., 2004
\n Hudson vd., 2004
\n Lin vd., 2004
\n Rieder vd., 2004
\n Woods ve Gratzer, 2005
\n Ketchedjian vd., 2005
\n Sodyum deoksikolat Doku yap\u0131s\u0131 SDS\u2019den daha y\u0131k\u0131c\u0131d\u0131r. Chen vd., 2004
\n Hudson vd., 2004
\n Hudson vd., 2004
\n Lin vd., 2004
\n Rieder vd., 2004
\n Woods ve Gratzer, 2005
\n Ketchedjian vd., 2005
\n Triton X-200 Zwitteriyonik deterjanlar ile kullan\u0131ld\u0131\u011f\u0131nda etkili desel\u00fclerizasyon uygulan\u0131r. Chen vd., 2004
\n Hudson vd., 2004
\n Hudson vd., 2004
\n Lin vd., 2004
\n Rieder vd., 2004
\n Woods ve Gratzer, 2005
\n Ketchedjian vd., 2005
\n Zwitteriyonik deterjanlar
\n CHAPS
\n (3 \u2013 [(3-Kolamidopropil) -dimetilamonyo] -propan s\u00fclfonat) \u0130yonik olmayan ve iyonik deterjanlar \u00f6zelliklerini g\u00f6sterir. Triton X-100 ile verilene benzer bir ECM bozulma ile verimli desel\u00fclerizasyon uygulan\u0131r. Dahl vd., 2003
\n S\u00fclfobetain-10 ve -16 (SB-10, SB16) Triton X-200 ile sa\u011flanan h\u00fccre \u00e7\u0131karma ve hafif ECM bozulmas\u0131 Chen vd., 2004
\n Hudson vd., 2004
\n Hudson vd., 2004
\n Lin vd., 2004
\n Rieder vd., 2004
\n Woods ve Gratzer, 2005
\n Ketchedjian vd., 2005
\n Tri (n-butil) fosfat Protein-protein etkile\u015fimlerini bozan organik \u00e7\u00f6z\u00fcc\u00fcd\u00fcr. De\u011fi\u015fken desel\u00fclerizasyon; kolajen i\u00e7eri\u011fi kayb\u0131 fazla iken mekanik \u00f6zellikler \u00fczerindeki etki \u00e7ok azd\u0131r Cartmell ve Dunn, 2000
\n Woods ve Gratzer, 2005
\n Hipotonik ve hipertonik \u00e7\u00f6zeltiler Ozmotik \u015fok ile H\u00fccre par\u00e7alama H\u00fccre erimesi a\u00e7\u0131s\u0131ndan verimli ancak etkili h\u00fccresel kal\u0131nt\u0131lar\u0131 ortadan kald\u0131rmaz Vyavahare vd., 1997
\n Goissis vd., 2000
\n Dahl vd., 2003
\n Woods ve Gratzer, 2005
\n EDTA, EGTA \u0130ki de\u011ferli metal iyonlar\u0131 ba\u011flanan kenetleme maddeleri ile ECM h\u00fccre yap\u0131\u015fmas\u0131n\u0131 bozar Tipik enzimatik y\u00f6ntemlerle kullan\u0131lan hi\u00e7bir izoleye maruz kalmaz(\u00f6rne\u011fin, tripsin) Bader vd., 1998
\n Teebken vd., 2000
\n Gamba vd., 2002
\n Enzimatik
\n Tripsin C taraf\u0131ndaki Arg ve Lys peptit ba\u011flar\u0131n\u0131 b\u00f6ler Uzun s\u00fcreli maruz kalmada ECM yap\u0131s\u0131n\u0131 bozabilir, laminin, fibronektin, elastin ve GAGler kald\u0131r\u0131r Bader vd., 1998
\n Teebken vd., 2000
\n Gamba vd., 2002
\n Endon\u00fckleazlar Ribonukleotid ve deoksiribon\u00fckleotid zincirlerinin i\u00e7 ba\u011flar\u0131n hidrolizini katalize eder. Dokudan \u00e7\u0131kar\u0131lmas\u0131 zor ve ba\u011f\u0131\u015f\u0131kl\u0131k sistemini uyarabilir. Courtman vd., 1994
\n Dahl vd., 2003
\n Rieder vd., 2004
\n Woods ve Gratzer, 2005
\n Ekson\u00fckleazlar Ribonukleotid ve deoksiribon\u00fckleotid zincirlerinin terminal ba\u011flar\u0131n hidrolizini katalize eder. Gilbert vd., 2006
\n 3.1. Fiziksel Y\u00f6ntemler
\n Desel\u00fclerizasyonu kolayla\u015ft\u0131rmak i\u00e7in kullan\u0131labilen fiziksel y\u00f6ntemler donma, do\u011frudan bas\u0131n\u00e7, sonikasyon ve \u00e7alkalama i\u00e7erir. Bu y\u00f6ntemler aras\u0131nda en fazla tercih edilen y\u00f6ntem dondurma y\u00f6ntemidir. \u00d6zellikle bu y\u00f6ntemlerin tendon, ligment dokusu ile sinir dokusunda kullan\u0131ld\u0131\u011f\u0131 bilinmektedir (Jackson vd., 1990; 1991). Dondurma-\u00e7\u00f6zme (Freeze-Thaw) y\u00f6ntemi olarak adland\u0131r\u0131lan bu y\u00f6ntemde dokunun -86\u00b0C\u2019 ye dondurularak tekrar h\u0131zla 37\u00b0C\u2019ye getirilmesi i\u015flemidir. Y\u00f6ntem sonucunda ECM yap\u0131s\u0131nda minimal hasar s\u00f6z konusudur. H\u00fccresel protein ve molek\u00fclerin tam temizlenmesi i\u00e7in ek i\u015flemlere gereksinim duyulur. Ayr\u0131ca baz\u0131 dokularda tek dondurma- \u00e7\u00f6zme d\u00f6ng\u00fcs\u00fc genellikle yeterli olmamakta ve \u00e7oklu tekrar yap\u0131lmas\u0131 gerekmektedir (Gulati, 1988; Cortiella vd., 2010).
\n Direk bas\u0131n\u00e7 y\u00f6ntemi h\u00fccre lizizi i\u00e7in kullan\u0131lan di\u011fer bir fiziksel y\u00f6ntemdir. Ancak bu y\u00f6ntemin genelde ECM organizasyonu yo\u011fun olmayan dokularda (karaci\u011fer, akci\u011fer) etkili oldu\u011fu bildirilmi\u015ftir. Ayn\u0131 zamanda mekanik g\u00fc\u00e7, mesane ve ince ba\u011f\u0131rsakta kullan\u0131lan desel\u00fclerizasyon y\u00f6ntemlerindendir. Bu y\u00f6ntemler etkili olup ECM\u2019in \u00fc\u00e7 boyutlu do\u011fal yap\u0131s\u0131na minimal d\u00fczeyde hasar veren y\u00f6ntemlerdir (Yang vd., 2010).
\n Hidrostatik bas\u0131n\u00e7 uygulama, sonikasyon ve ajitasyon (kar\u0131\u015ft\u0131rma, \u00e7alkalama) gibi teknikler genellikle tek ba\u015f\u0131na yeterli h\u00fccre par\u00e7alanmas\u0131 yapamazlar ancak enzim, hipertonik veya \u015felatlay\u0131c\u0131 ajanlarla kombine edildi\u011finde olduk\u00e7a ba\u015far\u0131l\u0131 sonu\u00e7lar al\u0131nabilmektedir (Schenke-Layland vd., 2003; Yang vd., 2010). Bu y\u00f6ntemin \u00f6nemli bir dezavantaj\u0131 etkili h\u00fccre lizizi s\u0131ras\u0131nda dokular\u0131n ultrastr\u00fckt\u00fcrel yap\u0131s\u0131nda ve bazal membran b\u00fct\u00fcnl\u00fc\u011f\u00fcnde hasar meydana gelmesidir (Hopkinson vd.,2008).
\n Is\u0131l olmayan geri d\u00f6n\u00fc\u015f\u00fcms\u00fcz elektroporasyon y\u00f6nteminde mikro saniyelik elektrik uyar\u0131lar\u0131 dokuya uygulanarak h\u00fccre membran\u0131nda olu\u015fan elektrik potansiyelinin bozulmas\u0131 sa\u011flanmakta ve membranda \u00e7ok k\u00fc\u00e7\u00fck delikler meydana getirilmektedir. Bu mikroporlar h\u00fccre hemostaz\u0131n\u0131 bozarak h\u00fccrede \u00f6l\u00fcme sebep olmaktad\u0131r. Ancak bu teknikte H\u00fccresizle\u015ftirme uygulanacak dokuya g\u00f6re kullan\u0131lan proplar\u0131n k\u00fc\u00e7\u00fck olmas\u0131, i\u015flemin \u00e7ok uzun zaman almas\u0131 ve in vivo ortamda yap\u0131lmas\u0131 en b\u00fcy\u00fck dezavantaj\u0131d\u0131r (Lee, 2005; Funamoto vd., 2010; \u00c7eviker vd., 2017).
\n 3.2. Kimyasal Y\u00f6ntemler
\n 3.2.1. Asit ve alkalin uygulamalar\u0131
\n Asit ve alkalin uygulamalar\u0131 desel\u00fclerizasyon protokollerinde h\u00fccrenin sitoplazmik komponentlerini \u00e7\u00f6zmek ve e\u015f zamanl\u0131 olarak RNA ve DNA gibi n\u00fckleik asitleri uzakla\u015ft\u0131rmak i\u00e7in kullan\u0131lmaktad\u0131r. Asetik asit, perasetik asit (PAA), hidroklorik asit, s\u00fclf\u00fcrik asit ve amonyum hidroksit h\u00fccre membran\u0131 ve intrasel\u00fcler organelleri etkili bir \u015fekilde par\u00e7alayan alkalin ve asit \u00f6rnekleridir (Yoo vd., 1998; Falke vd., 2003; Freytes vd., 2004).
\n Yap\u0131lan \u00e7al\u0131\u015fmalarda, domuz ince ba\u011f\u0131rsak submukozas\u0131 ve mesane tabakalar\u0131 yakla\u015f\u0131k %0,10-0,15 konsantrasyonda ki PAA ile desel\u00fclerize edilmi\u015ftir. Bu desel\u00fclerizasyon y\u00f6nteminin bu t\u00fcr ince ECM yap\u0131lar\u0131nda h\u00fccre materyallerinin ortadan kald\u0131r\u0131lmas\u0131nda y\u00fcksek bir etkinli\u011fe sahip oldu\u011fu, bunun yan\u0131 s\u0131ra mikroorganizmalar\u0131 etkileyerek ve mikrobiyal enzimleri okside ederek materyalin dezenfeksiyonunu da sa\u011flad\u0131\u011f\u0131 bildirilmi\u015ftir (Pruss vd., 1999; Hodde ve Hiles, 2002). \u0130nce ba\u011f\u0131rsak submukozas\u0131 ve mesane tabakalar\u0131nda PAA uygulamas\u0131 sonras\u0131nda immunohistolojik boyama y\u00f6ntemleri kullan\u0131larak kollajenin bir\u00e7ok tipi (I, III, IV, V,VI ve VII gibi) tan\u0131mlanm\u0131\u015ft\u0131r (Badylak vd., 1995; Brown vd., 2006). Ayn\u0131 zamanda PAA uygulamas\u0131 sonras\u0131nda ECM\u2019de do\u011fal yap\u0131daki glikozaminoglikanlar\u0131n korundu\u011fu bildirilmi\u015ftir (Hodde vd., 1996). Farkl\u0131 \u00e7al\u0131\u015fmalarda ise PAA uygulamas\u0131 sonras\u0131nda glikozaminoglikanlardan laminin ve fibronektinin ECM doku yap\u0131s\u0131nda kald\u0131\u011f\u0131 g\u00f6sterilmi\u015ftir (Hodde vd., 2002a; Brown vd., 2006). Transforme edici b\u00fcy\u00fcme fakt\u00f6r\u00fc beta, fibroblast b\u00fcy\u00fcme fakt\u00f6r\u00fc ve vask\u00fcler endoteliyal b\u00fcy\u00fcme fakt\u00f6r\u00fc gibi ECM\u2019de yer alan b\u00fcy\u00fcme fakt\u00f6rlerinin yap\u0131sal ve fonksiyonel yap\u0131lar\u0131n\u0131n bozulmadan kald\u0131\u011f\u0131 da di\u011fer \u00e7al\u0131\u015fma gruplar\u0131 taraf\u0131ndan bildirilmi\u015ftir (Voytik-Harbin vd., 1997; Hodde vd., 2001). PAA\u2019in biyolojik yap\u0131 malzemelerinin mekaniksel davran\u0131\u015flar\u0131na herhangi bir olumsuz etkisi olmad\u0131\u011f\u0131 da bildirimler aras\u0131ndad\u0131r (Freytes vd., 2004). \u0130nce ba\u011f\u0131rsak submukozas\u0131 ve mesane tabakalar\u0131 ile yap\u0131lan in vitro h\u00fccre k\u00fclt\u00fcr\u00fc \u00e7al\u0131\u015fmalar\u0131 da bu \u015fekilde elde edilen materyallerin kullan\u0131m a\u00e7\u0131s\u0131ndan m\u00fckemmele yak\u0131n bir doku iskelesi olduklar\u0131n\u0131 tekrarl\u0131 \u00e7al\u0131\u015fmalar ile g\u00f6stermi\u015ftir (Badylak vd., 1999; Hodde vd., 2002a; 2002b).
\n 3.2.2. \u0130yonik olmayan deterjanlar
\n \u0130yonik olmayan deterjanlar desel\u00fclerizasyon protokollerinde yayg\u0131n olarak kullan\u0131lan bir y\u00f6ntemdir. Bunun en \u00f6nemli nedeni g\u00f6receli olarak di\u011fer y\u00f6ntemlere g\u00f6re doku yap\u0131s\u0131 \u00fczerine daha hafif bir etkisi olmas\u0131d\u0131r. \u0130yonik olmayan deterjanlar\u0131n lipid-lipid ve lipid-protein etkile\u015fimini bozdu\u011fu ancak protein-protein ba\u011flant\u0131s\u0131n\u0131 etkilemeyerek doku ya da organdaki proteinin fonksiyonel konformasyonunu bozmad\u0131\u011f\u0131 bildirilmi\u015ftir (Seddon vd., 2004).
\n Triton X-100 desel\u00fclerizasyon protokol\u00fcnde en yayg\u0131n olarak kullan\u0131lan iyonik deterjanlar aras\u0131ndad\u0131r. Dokunun Triton X-100\u2019e maruz b\u0131rak\u0131lma s\u00fcresi birka\u00e7 saatten 14 g\u00fcne kadar de\u011fi\u015fmektedir (Cartmell ve Dunn, 2000; Dahl vd., 2003; Lin vd., 2004; Woods ve Gratzer, 2005). Dokular\u0131n Triton X-100 ile muamele edilerek desel\u00fclerize edilmesi ile \u00e7ok farkl\u0131 sonu\u00e7lar elde edilmektedir. Kalp kapak\u00e7\u0131\u011f\u0131n\u0131n 24 saat Triton X-100 ile desel\u00fclerize edilmesi ile kapak\u00e7\u0131k yap\u0131s\u0131n\u0131n korunabildi\u011fi ve t\u00fcm h\u00fccre membranlar\u0131n\u0131n uzakla\u015ft\u0131r\u0131labildi\u011fi, kom\u015fu miyokardiyum ve aortik duvarda ise h\u00fccresel materyallere rastlanabildi\u011fi bir di\u011fer \u00e7al\u0131\u015fma ile bildirilmi\u015ftir (Grauss vd., 2005). Farkl\u0131 gruplar\u0131n \u00e7al\u0131\u015fmalar\u0131nda ise d\u00f6rt saatin \u00fczerindeki Triton X-100 muamelesi ile kan damarlar\u0131, tendon ve ligamentlerde h\u00fccresel kal\u0131nt\u0131lar\u0131n tamamen dokudan uzakla\u015ft\u0131r\u0131lamad\u0131\u011f\u0131 g\u00f6sterilmi\u015ftir. T\u00fcm dokularda histolojik boyama sonucunda h\u00fccre membranlar\u0131na rastlanm\u0131\u015f ve \u00f6n \u00e7apraz ba\u011flar\u0131nda yap\u0131lan immunohistolojik boyama sonucunda bir h\u00fccre iskeleti proteini olan vimentine rastland\u0131\u011f\u0131 a\u00e7\u0131klanm\u0131\u015ft\u0131r (Cartmell ve Dunn, 2000; Dahl vd., 2003; Woods ve Gratzer 2005).
\n 3.2.3. \u0130yonik deterjanlar
\n \u0130yonik deterjanlar, sitoplazmik ve h\u00fccre membran\u0131n\u0131n \u00e7\u00f6z\u00fcnmesinde etkili bir y\u00f6ntemdir, ancak protein-protein etkile\u015fimini bozarak protein denat\u00fcrasyonuna neden olmaktad\u0131r (Seddon vd. 2004). Bilinen en yayg\u0131n iyonik deterjanlar; sodyum dodesil s\u00fclfat (SDS) ve sodyum deoksikolat ile Triton X-200\u2019d\u00fcr (Rieder vd., 2004; Chen vd., 2004; Hudson vd., 2004; Lin vd., 2004; Woods ve Gratzer, 2005).
\n SDS, dokudan h\u00fccresel elemanlar\u0131n uzakla\u015ft\u0131r\u0131lmas\u0131nda \u00e7ok etkili bir y\u00f6ntemdir. SDS\u2019\u0131 di\u011fer deterjanlardan ay\u0131ran \u00f6zellik, sitoplazmik proteinlerden vimentin dahil t\u00fcm h\u00fccresel kal\u0131nt\u0131lar\u0131n tamamen ortadan kald\u0131r\u0131lmas\u0131n\u0131 sa\u011flamas\u0131d\u0131r (Woods ve Gratzer, 2005). SDS, ECM\u2019in do\u011fal doku yap\u0131s\u0131n\u0131 bozarak glikozaminoglikan konsantrasyonunda azalmaya ve kollajen birlikteli\u011finin bozulmas\u0131na neden olmaktad\u0131r. Ancak SDS ile dokudan kollajen uzakla\u015ft\u0131r\u0131lmas\u0131 ger\u00e7ekle\u015fmemektedir (Faulk vd., 2014).
\n Sodyum deoksikolat da ayn\u0131 \u015fekilde h\u00fccresel kal\u0131nt\u0131lar\u0131 uzakla\u015ft\u0131rmada \u00e7ok etkili bir y\u00f6ntem olmas\u0131yla birlikte doku yap\u0131s\u0131na SDS\u2019e g\u00f6re \u00e7ok daha fazla zarar vermektedir. Bu y\u00f6ntemin tek ba\u015f\u0131na kullan\u0131ld\u0131\u011f\u0131 herhangi bir rapor bulunmad\u0131\u011f\u0131ndan, dokudan elde edilen ECM\u2019de sodyum deoksikolat\u0131n etkisinin ne kadar oldu\u011fu bilinmemektedir. Sodyum deoksikolat sinir dokusu desel\u00fclerizasyonunda \u00e7ok say\u0131da zwitteriyonik deterjan ile kombin bir \u015fekilde kullan\u0131ld\u0131\u011f\u0131, kompleks desel\u00fclerizasyonun bu kombinasyona Triton X-100\u2019\u00fcn de eklenmesi ile ba\u015far\u0131ld\u0131\u011f\u0131 a\u00e7\u0131klanm\u0131\u015ft\u0131r (Hudson vd., 2004).
\n 3.2.4. Zwitteriyonik deterjanlar
\n Zwitteriyonik deterjanlar hem iyonik hem de iyonik olmayan deterjan \u00f6zelli\u011fi g\u00f6sterirler. Bu deterjanlar\u0131n protein denat\u00fcre etme g\u00fcc\u00fc iyonik olmayan deterjanlara g\u00f6re \u00e7ok daha fazlad\u0131r (Seddon vd., 2004). Zwitteriyonik deterjanlardan 3-[(3-kolamidopropil)-dimetilamonyo]-1-propansulfonat (CHAPS) kan damarlar\u0131n\u0131n desel\u00fclerizasyonunda (Dahl vd., 2003), s\u00fclfobetain-10 (SB-10) ve s\u00fclfobetain-16 (SB16) ise sinir desel\u00fclerizasyonunda kullan\u0131lmaktad\u0131r. CHAPS ile muamele edilen arter dokusunun histolojik incelemesinde kollajen ve elastin morfolojisinin normal oldu\u011fu ve do\u011fal damar yap\u0131s\u0131ndaki kollajen yap\u0131s\u0131n\u0131n korundu\u011fu g\u00f6sterilmi\u015ftir. Ayn\u0131 \u00e7al\u0131\u015fma ile periferal sinir desel\u00fclerizasyonunda SB-10 ve SB-16\u2019n\u0131n iyonik bir deterjan olan Triton X-200 ile kombin edilerek kullan\u0131ld\u0131\u011f\u0131 bildirilmi\u015ftir. Bu kombinasyon uygulamas\u0131n\u0131n sinir ECM yap\u0131s\u0131na Triton X-100 ve sodyum deoksikolat\u0131n birlikte uygulanmas\u0131ndan daha az zarar verdi\u011fi de \u00e7al\u0131\u015fmalar sonu\u00e7lar\u0131n\u0131n aras\u0131ndad\u0131r (Hudson vd., 2004).
\n 3.2.5. Trib\u00fctil fosfat (TBF)
\n Son zamanlarda yap\u0131lan \u00e7al\u0131\u015fmalarda TBF\u2019\u0131n kozmotropik ajan olarak tendon ve ligament greftlerinin desel\u00fclerizasyonunda da kullan\u0131ld\u0131\u011f\u0131 bildirilmi\u015ftir. TBF uygulamas\u0131 ile s\u0131\u00e7an kuyruk tendonundan t\u00fcm h\u00fccresel kal\u0131nt\u0131lar\u0131n uzakla\u015ft\u0131r\u0131ld\u0131\u011f\u0131, ancak kemik ligament ba\u011flanma noktalar\u0131nda bu uzakla\u015ft\u0131rma i\u015fleminde sorunlar\u0131n g\u00f6zlendi\u011fi a\u00e7\u0131klanm\u0131\u015ft\u0131r. Ayn\u0131 \u00e7al\u0131\u015fmalarda, TBF uygulamas\u0131 ile tendondan izole edilen kollajen fibrillerinin gerilme g\u00fcc\u00fcnde herhangi bir kay\u0131p g\u00f6zlenmedi\u011fi de bildirilmi\u015ftir (Cartmell ve Dunn, 2000; Woods ve Gratzer, 2005). TBF\u2019\u0131n kozmotropik ajan olarak desel\u00fclerizasyon i\u015fleminden elde edilen ECM\u2019in mekaniksel davran\u0131\u015f\u0131na minimal bir etki yapt\u0131\u011f\u0131 i\u00e7in \u00fcmit verici bir y\u00f6ntem oldu\u011fu belirtilmi\u015ftir (Woods ve Gratzer, 2005).
\n 3.2.6. Hipotonik ve hipertonik sol\u00fcsyonlar
\n Deiyonize su ya da d\u00fc\u015f\u00fck iyonik i\u00e7erikli sol\u00fcsyonlar gibi hipotonik ve hipertonik sol\u00fcsyonlarla yap\u0131lan osmotik \u015fok uygulamas\u0131 organ ve dokularda h\u00fccre lizisine neden olmaktad\u0131r (Goissis vd., 2000; Dahl vd., 2003; Woods ve Gratzer, 2005). S\u0131ras\u0131yla 11 saatlik hipotonik sol\u00fcsyon ve 11 saatlik hipertonik sol\u00fcsyon muamelesi sonucunda h\u00fccre lizisinin ger\u00e7ekle\u015fti\u011fi ancak dokulardan ortaya \u00e7\u0131kan h\u00fccre kal\u0131nt\u0131lar\u0131n\u0131n tamamen uzakla\u015ft\u0131r\u0131lamad\u0131\u011f\u0131 bildirilmi\u015ftir (Dahl vd., 2003). Ayn\u0131 \u00e7al\u0131\u015fmada bu y\u00f6nteme ek olarak, h\u00fccresel kal\u0131nt\u0131lar\u0131n tamamen uzakla\u015ft\u0131r\u0131lmas\u0131 i\u00e7in biyolojik ve kimyasal uygulamalar\u0131n gereklili\u011fine de\u011finilmi\u015ftir.
\n 3.2.7. \u015eelatlay\u0131c\u0131 deterjanlar
\n Etilendiamin tetraasetik asit (EDTA) ve etilen glikol bistetraasetik asit (EGTA) gibi \u015felatlay\u0131c\u0131 ajanlar halka \u015feklinde kompleks bir molek\u00fcler yap\u0131ya sahip olup merkezi metal iyonlara kararl\u0131 bir \u015fekilde ba\u011flanarak bu iyonlar\u0131n izole edilebilmesini sa\u011flamaktad\u0131rlar. H\u00fccre-ECM adezyonunda g\u00f6revli \u00e7ift de\u011ferlikli katyonlar\u0131n (\u00d6rne\u011fin; Ca+2 ve Mg+2) ba\u011flanmas\u0131yla, bu ajanlar dokudan h\u00fccresel materyallerin uzakla\u015ft\u0131r\u0131lmas\u0131n\u0131 sa\u011flamaktad\u0131r. EDTA genellikle tripsin kombinasyonu ile kullan\u0131lan bir y\u00f6ntem olarak bildirilmi\u015ftir (Khorramirouz vd., 2014).
\n 3.2.8. Alkoller ve di\u011fer ajanlar
\n \u0130zopropanol, aseton, etanol, metanol ve gliserol gibi ajanlar h\u00fccrelerde dehidrasyon ve liziz yaparak etkilidir. \u00d6zellikle dokunun kalsifikasyonuna sebep olan fosfolipidlerin uzakla\u015ft\u0131r\u0131lmas\u0131nda alkoller lipaz gibi enzimatik ajanlardan daha \u00e7ok etkilidir. Dikkat edilmesi gereken husus alkollerin proteinlerde \u00e7\u00f6kmeye sebep olmas\u0131ndan dolay\u0131 ECM yap\u0131s\u0131nda hasara yol a\u00e7mas\u0131d\u0131r (Cole, 1984; Gorschewsky vd., 2005).
\n Aseton \u00f6zellikle lipidlerin ECM yap\u0131s\u0131ndan uzakla\u015ft\u0131r\u0131lmas\u0131nda kullan\u0131labilir ancak alkollere benzer \u015fekilde dokunun protein yap\u0131s\u0131nda bozulmaya sebep olabilir (Gilbert vd., 2006).
\n 3.3. Biyolojik Y\u00f6ntemler
\n 3.3.1. Enzimler
\n N\u00fckleaz, tripsin, kollajenaz, lipaz, dispaz, termolizin ve \u03b1-galaktozidaz H\u00fccresizle\u015ftirme i\u015fleminde kullan\u0131lan enzimlerdir. \u00d6zellikle h\u00fccre art\u0131klar\u0131n\u0131n ve istenmeyen ECM yap\u0131 elemanlar\u0131n\u0131n se\u00e7ici olarak temizlenmesinde faydal\u0131d\u0131r. Ancak sadece enzimatik y\u00f6ntem h\u00fccrenin tamamen temizlenmesinde tam etkili olmad\u0131\u011f\u0131 gibi kullan\u0131lan enzim art\u0131klar\u0131 da tekrar h\u00fccrelendirme i\u015fleminde sorun yaratabilir veya kendileri imm\u00fcn sistemi aktive edebilir. DNaz ve RNaz gibi n\u00fckleazlar h\u00fccre lizizi sonras\u0131nda ortaya \u00e7\u0131kan DNA ve RNA n\u00fckleik asitlerinin ve n\u00fckleotidlerinin par\u00e7alanmas\u0131n\u0131 sa\u011flar (Elder vd., 2010; Funamoto vd., 2010; Peterson vd., 2010; Yang vd., 2010). Benzonaz gibi endon\u00fckleazlar ve k\u0131s\u0131tlamas\u0131 olmayan (non-restriction) endon\u00fckleazlar ekzon\u00fckleazlara g\u00f6re DNA par\u00e7alanmas\u0131nda \u00e7ok daha etkilidir (Cole, 1984; Peterson vd., 2010).
\n Serin proteaz olan Tripsin, enzimatik h\u00fccresizle\u015ftirmede s\u0131k\u00e7a kullan\u0131lan enzimatik ajand\u0131r. Tripsin ile yap\u0131lan h\u00fccresizle\u015ftirmede ECM yap\u0131s\u0131nda bulunan GAG\u2019lar\u0131n korunmas\u0131na ra\u011fmen kollajen ve elastinin tripsine diren\u00e7lerinin az olmas\u0131, tripsinin dikkatli kullan\u0131lmas\u0131n\u0131 gerektirir (Yang vd.,2010; Crapo vd., 2011). Di\u011fer bir dezavantaj\u0131 ise etki s\u00fcresinin uzun olmas\u0131d\u0131r. \u00d6zellikle kal\u0131n dokularda bu s\u00fcre \u00e7ok daha fazla artar. Ancak bu \u015fekilde kal\u0131n dokular\u0131n h\u00fccresizle\u015ftirilmesinde, di\u011fer ajanlar\u0131n daha derine penetre olabilmeleri i\u00e7in tripsin kullan\u0131m\u0131 ka\u00e7\u0131n\u0131lmaz olabilir (Xu vd., 2014).
\n Kollajenazlar, \u00f6zellikle kollajenin gerekli olmad\u0131\u011f\u0131 ECM olan dokular\u0131n h\u00fccresizle\u015ftirilmesinde kullan\u0131labilir. Lipazlar ise lipidlerin uzakla\u015ft\u0131r\u0131lmas\u0131nda etkilidir ancak t\u00fcm lipidlerin uzakla\u015ft\u0131r\u0131lmas\u0131nda yaln\u0131z ba\u015f\u0131na yetersizdir (Flynn, 2010; Brown vd., 2011). Ksenojenik dokular\u0131n h\u00fccre y\u00fczeylerinde bulunan immunojenik h\u00fccre y\u00fczey antijeni olan galaktoz-\u03b1-(1,3)-galaktoz (Gal epitop) uzakla\u015ft\u0131r\u0131lmas\u0131 i\u00e7in \u03b1-galaktozidaz kullan\u0131labilir (Xu vd., 2008).
\n 3.3.2. Non-enzimatik ajanlar
\n \u015eelasyon yap\u0131c\u0131 ajanlar olan etilen diamin tetra asetik asit (EDTA) ve etilen glikol tetra asetik asit (EGTA) metal iyonlar\u0131n\u0131 uzakla\u015ft\u0131rarak h\u00fccrelerin ECM proteinlerinden uzakla\u015ft\u0131r\u0131lmas\u0131nda kullan\u0131l\u0131r (Klebe, 1974; Gailit, 1988). Ayr\u0131ca yap\u0131sal olarak protein- protein ili\u015fkisinin de bozulmas\u0131na sebep olur. Bu ajanlar yaln\u0131z ba\u015f\u0131na, \u00e7alkalama yap\u0131lsa dahi y\u00fczeysel h\u00fccrelerin yok edilmesinde bile etkili olamaz. Dolay\u0131s\u0131 ile genellikle y\u00f6ntemlerde tripsin ya da deterjanlar ile kombine halde kullan\u0131lmal\u0131d\u0131r (Sasaki vd., 2009; Funamoto vd., 2010; Gui vd., 2010; Reing vd., 2010; Crapo vd., 2011; Xu vd., 2014).
\n Fenilmetils\u00fclfonil florid (PMSF), aprotonin ve l\u00f6peptin gibi serin proteaz inhibit\u00f6rleri ECM\u2019de olu\u015fmas\u0131 muhtemel istenmeyen zararlar\u0131 engeller (Gorschewsky vd., 2005; Gilbert vd., 2008; Alhamdani vd., 2010; Gui vd., 2010; Reing vd., 2010). Aksi takdirde h\u00fccre \u00f6l\u00fcm\u00fc s\u0131ras\u0131nda h\u00fccre i\u00e7erisinde ki proteazlar sal\u0131nabilir.
\n Penisilin, streptomisin, amfoterisin B ve sodyum asid gibi antibiyotikler ve antimikotikler H\u00fccresizle\u015ftirme s\u00fcresince mikrobiyal kontaminasyonu engellemek i\u00e7in kullan\u0131l\u0131r fakat biyolojik iskelelerde tekrar h\u00fccrelendirme s\u0131ras\u0131nda potansiyel bir engel olu\u015fturabilir (Crapo vd., 2011).
\n 3.4. Proteaz inhibit\u00f6rleri
\n Desel\u00fclerizasyon protokolleri s\u0131ras\u0131nda hasar g\u00f6ren h\u00fccrelerden \u00e7ok say\u0131da proteaz sal\u0131nabilmektedir. Uzun s\u00fcreli kimyasal uygulamalar\u0131 sonras\u0131nda ortamda beliren proteazlar ECM\u2019in do\u011fal yap\u0131s\u0131na zarar verebilir. Bu nedenle dokular\u0131n desel\u00fclerizasyonu i\u00e7in kullan\u0131lan sol\u00fcsyonlar\u0131n i\u00e7ine proteaz inhibit\u00f6rlerinden fenil metil s\u00fclfonil florid, aprotinin veya l\u00f6peptin eklenebilmektedir (Fermor vd., 2015). Tampon sol\u00fcsyonunun pH\u2019\u0131 7-8 aras\u0131nda tutulursa yine proteaz aktivitesi durur. Bunlara ek olarak lizis sol\u00fcsyonunun s\u0131cakl\u0131\u011f\u0131n\u0131n ve s\u00fcresinin kontrol\u00fc de proteaz aktivitesini s\u0131n\u0131rlamaktad\u0131r (James, 1978).
\n 3.5. Antibiyotikler
\n Uzun s\u00fcreli kimyasal desel\u00fclerizasyon uygulamalar\u0131 sonras\u0131nda kar\u015f\u0131la\u015f\u0131lan bir di\u011fer sorun, elde edilen ECM materyalinin kontaminasyonuna yol a\u00e7acak bakteri varl\u0131\u011f\u0131n\u0131n ortaya \u00e7\u0131kma ihtimalidir. Bu nedenle uygulanan bir\u00e7ok protokolde kullan\u0131lan desel\u00fclerizasyon sol\u00fcsyonlar\u0131 i\u00e7ine penisilin, streptomisin veya amfoterisin-B eklenmektedir (Affonso da Costa vd., 2004; Hilbert vd., 2004; Woods ve Gratzer, 2005).
\n 3.6. Desel\u00fclerizasyon Sonucu Art\u0131k Kimyasallar\u0131n Uzakla\u015ft\u0131r\u0131lmas\u0131
\n Yukar\u0131da a\u00e7\u0131klanan desel\u00fclerizasyon y\u00f6ntemleri, h\u00fccrelere zarar verme yetenekleri nedeniyle kullan\u0131lan \u00e7ok \u00e7e\u015fitli kimyasallar\u0131 i\u00e7erir. E\u011fer kimyasallar, muamele sonras\u0131nda y\u00fcksek konsantrasyonlarda dokuda kal\u0131rsa, iskele in vivo yerle\u015ftirildi\u011finde konak h\u00fccreleri i\u00e7in toksik olaca\u011f\u0131 muhtemeldir. Desel\u00fclerize edilmi\u015f iskelet materyalinde kal\u0131nt\u0131 kimyasallar\u0131n varl\u0131\u011f\u0131n\u0131 \u00f6l\u00e7mek i\u00e7in analizlerin geli\u015ftirilmesine ihtiya\u00e7 vard\u0131r. Benzer \u015fekilde, yukar\u0131da tarif edilen baz\u0131 i\u015flemler s\u0131\u011f\u0131r kaynaklar\u0131ndan (yani, DNaz, RNaz, tripsin) t\u00fcretilmi\u015f enzimleri de i\u00e7erir (Gilbert vd., 2006).
\n 4. DESEL\u00dcLER\u0130ZASYON Y\u00d6NTEMLER\u0130N\u0130N DOKULARDA KULLANIMI
\n Her doku t\u00fcr\u00fc spesifik bir yap\u0131ya ve farkl\u0131 bile\u015fime sahiptir, bu nedenle her doku t\u00fcr\u00fc i\u00e7in yaln\u0131zca bir standart protokol\u00fcn kullan\u0131lmas\u0131 m\u00fcmk\u00fcn de\u011fildir. Uygulanacak desel\u00fclerizasyon y\u00f6nteminin se\u00e7imi, i\u015flem g\u00f6recek doku t\u00fcr\u00fcne ve doku kompozisyonuna ba\u011fl\u0131d\u0131r (Hrebikova vd., 2015). Tablo 2\u2019de baz\u0131 doku ve organlara uygun desel\u00fclerizasyon y\u00f6ntemleri verilmi\u015ftir.
\n Tablo 2. Baz\u0131 Doku ve Organlara G\u00f6re Desel\u00fclerizasyon Y\u00f6ntemleri
\n Doku ve Organ T\u00fcr\u00fc Desel\u00fclerizasyon Y\u00f6ntemi
\n \u0130nce tabakal\u0131 dokular (perikard) 1) Dondurmak
\n 2) Ozmotik Sol\u00fcsyonlar
\n 3) Enzim
\n 4) Kimyasal
\n Daha kal\u0131n tabakal\u0131 dokular (dermis) 1) Dondurmak
\n 2) Enzim
\n 3) Alkol
\n 4) Kimyasal
\n Amorf dokular 1) Dondurmak
\n 2) Mekanik Bozulma
\n 3) Alkol
\n 4) Enzimatik
\n 5) Kimyasal
\n Organ desel\u00fclerizasyonu
\n (Trake vb.) 1) Dondurmak
\n 2) Ozmotik Sol\u00fcsyonlar
\n 3) Deterjan
\n 4) Enzimatik
\n 5) Ozmotik \u00c7\u00f6zeltiler
\n Organ desel\u00fclerizasyonu
\n (karaci\u011fer vb.) 1) Dondurmak
\n 2) Ozmotik Sol\u00fcsyonlar
\n 3) Deterjan
\n 5. SONU\u00c7
\n Doku m\u00fchendisli\u011fi ve rejeneratif t\u0131pta h\u00fccre d\u0131\u015f\u0131 matris (ECM), desel\u00fclerizasyon i\u015fleminden t\u00fcretilmi\u015f bir biyolojik iskele olarak doku ve organlarda kullan\u0131labilir. Desel\u00fclerizasyonun as\u0131l amac\u0131, ECM\u2019nin yap\u0131sal ve fonksiyonel proteinler, glikozaminoglikanlar, b\u00fcy\u00fcme fakt\u00f6rleri vb. yer alan \u00fc\u00e7 boyutlu yap\u0131s\u0131n\u0131n korunmas\u0131d\u0131r. Desel\u00fclerizasyon s\u00fcrecinin etkinli\u011fi, doku yo\u011funlu\u011fu ve organizasyonu, h\u00fccre giderme y\u00f6ntemi, biyolojik bile\u015fenler ve hedef klinik uygulamalar gibi \u00e7e\u015fitli fakt\u00f6rlere ba\u011fl\u0131d\u0131r. Desel\u00fclerizasyon y\u00f6ntemlerinde sadece tek tip y\u00f6ntem kullanmak uygun olmay\u0131p; fiziksel, kimyasal ve enzimatik y\u00f6ntemlerin kombinasyonu h\u00fccre \u00e7\u0131kar\u0131m\u0131 ile ilgili daha iyi bir sonu\u00e7 sa\u011flamaktad\u0131r. Optimal desel\u00fclerizasyon y\u00f6nteminin geli\u015ftirilmesi b\u00fcy\u00fck \u00e7abalar gerektirir ve hala doku m\u00fchendisli\u011fi ve rejeneratif t\u0131pta \u00e7al\u0131\u015fmalar devam etmektedir.
\n Te\u015fekk\u00fcrler; 4894-YL1-17 No`lu Proje ile \u00e7al\u0131\u015fmam\u0131z\u0131 maddi olarak destekleyen S\u00fcleyman Demirel \u00dcniversitesi Bilimsel Ara\u015ft\u0131rma Projeleri Y\u00f6netim Birimi Ba\u015fkanl\u0131\u011f\u0131\u2019na te\u015fekk\u00fcr ederiz.
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\u00d6ZET Desel\u00fclerize edilmi\u015f doku ve organlar \u00e7e\u015fitli doku m\u00fchendisli\u011fi ve rejeneratif t\u0131p uygulamalar\u0131nda ba\u015far\u0131yla kullan\u0131lmaktad\u0131r. Ekstrasel\u00fcler matristen elde edilen biyolojik bir iskele, i\u015flem g\u00f6recek dokudan h\u00fccreleri etkin bir \u015fekilde ortadan kald\u0131ran \u00e7e\u015fitli desel\u00fclerizasyon y\u00f6ntemleri ile \u00fcretilebilir. Kullan\u0131lan desel\u00fclerizasyon y\u00f6ntemleri dokular\u0131n Read More …<\/a><\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[189,1],"tags":[],"class_list":["post-429814","post","type-post","status-publish","format-standard","hentry","category-turkish-essays","category-uncategorized"],"_links":{"self":[{"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/posts\/429814","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/comments?post=429814"}],"version-history":[{"count":0,"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/posts\/429814\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/media?parent=429814"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/categories?post=429814"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/tags?post=429814"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}