Biz uzman teknik yardım için Sonja Dufentester ve Elke Weber teşekkür ederiz. Biz tıp bilişimi destek el yazması ve Stefan Brückner düzeltiyorum için Faekah Gohar teşekkür ederim. Bu çalışma Else-Kröner-Fresenius-Stiftung (2012_A94) bir disiplinlerarası hibe ile desteklenmiştir. D. Bettenworth Tıp Fakültesi, Westfälische Wilhelms-Universität Münster bir araştırma bursu ile desteklenmiştir. M. Brückner Deutsche Forschungsgemeinschaft (DFG SFB1009B8) bir &quot;Gerok&quot; dönme pozisyonuna tarafından desteklenmiştir. Biz fare karikatür resmi için Heike Blum teşekkür ederim.

Tüm hayvan deneyleri Alman Hayvanları Koruma Kanunu&#39;na göre Landesamt für Natur, Umwelt und Verbraucherschutz (LANUV) tarafından onaylanmıştır. 1. Malzemeler ve Deney Düzeneği <ol><li> Hayvan bakım <ol><li> 20 ila 25 gr ağırlığında herhangi bir zorlanma kadın ya da erkek fareler kullanın ve yerel hayvan bakımı mevzuatına göre bunları barındıracak. </li><li> Kemirgenler için özel diyeti ile fareler Yem ve yonca-ücretsiz yemi uygulamak en az üç gün Endoluminal otomatik floresan en aza indirmek için sınavlara floresan önce. </li><li> Otoklavlanmış içme suyu ad libitum sağlayın. </li></ol></li><li> DDS ile indüklenen akut kolit indüksiyonu <ol><li> Su otoklavlanmış, 100 ml DSS, 3 g çözülerek çözelti: dekstran sülfat sodyum (DSS Da 36,000-50,000 moleküler ağırlık) (ağ / hac) bir% 3 hazırlayın. Fareye gerektiğinde özel içme suyu olarak, bu çözüm sunar ve fare / günde DSS çözeltisi 5 ml hesaplar. Yem devamDSS istenildiği 9 olmadan otoklavlanmalıdır su ile rol farenin. </li></ol></li><li> Kolorektal kanserin başlaması <ol><li> Mutajenik azoksimetan (AOM) çözündürün (kanser ve genetik hasara neden olabilir DİKKAT!) Steril izotonik tuz çözeltisi içinde 1 mg / ml bir nihai konsantrasyon elde edilmiştir. Vücut ağırlığı intraperitoneal olarak 1 ml şırınga (30 g), 10 kg ile 10 mg AOM tek bir doz uygulanır. </li><li> 21&#39;den% 3 ila 7 gün, 0 (ağ / hac), DSS, gün 14, tekrarlı devreler, gün 28-35 ve 42-49 gün ile (kontrol farelerinde hariç) Görev farenin enflamatuar tahrik kolorektal karsinogenezi tetiklemek için karıştırılabilir. Sadece bu zorluklara (ayrıntılı bir zaman çizelgesi için Şekil 4A) arasında Otoklavlanmış su ile fareler besleyin. Deney boyunca Otoklavlanmış su ile kontrol fareler besleyin. </li></ol></li><li> Floresan endoskopi hazırlanması (FE) <ol><li> Kullanım Flüoresanla Isothiocyanat (FITC) -, dekstran (molekül ağırlığı 70.000 Da moleküler FITC: Glukoz = 1: 250) için deterjanlarıdisplazi ilişkili vasküler desen görsel iyileştirme ile kolon adenomu avranış. </li><li> Intravenöz 5 dk floresan endoskopik muayene öncesi 100 ul PBS içinde seyreltilmiş 60 mg FITC-dekstran yönetmek. </li></ol></li><li> Anestezi <ol><li> (, Hacimce% 1.5-2 izofluran [2-kloro-2-(diflorometoksi) -1,1,1-trifloro-etan] 1,5 Lo 2 / dakika) ile anestezi sürekli izofluran kaynağını temin ederler. Sıkı anestezi kontrol etmek için bir yüz maskesi ile özel bir veteriner anestezi ekipman kullanın. </li></ol></li><li> Lavman hazırlanması <ol><li> Sıvı lavman 2 ml aşılamak (içindekiler: disodyumhidrojenfosfat (w /% 1.5 h) ve sodyum w / v) (% 11 dihidrojen-) kolona önemli dışkı yükleme şüphesi varsa görünümü gizleyebilir ki. </li></ol></li></ol> 2. Teknik Ekipmanlar <ol><li> Geliştirilen ve küçük hayvan endoskopi kullanımı onaylanmış bir veteriner endoskopik iş istasyonu kullanın. Wo bağlayınBir kamera birimi, ksenon ışık kaynağı, bir hava pompasına ve beyaz ışık endoskopi için bilinen bir PC monitörü rkstation. Sonra kamera ve minyatür sert teleskop (Şekil 5 1.9 mm dış çap, 10 cm uzunluğunda) bağlayın. </li><li> Biyopsi forseps veya enjeksiyon tüp uygulaması için çalışma kanalı (Şekil 5D) ile endoskop kılıfı kullanın. Tanısal kolonoskopi için çalışma kanalı olmadan kılıfı kullanın. </li><li> Kullanılmış izleyiciler uyarmak için floresan endoskopi için, ışık kaynağının yapılandırma ayarları (örneğin, FITC ile konjüge edilmiş dekstran 490 nm). Ayrıca, teleskop ve kamera (FITC-dekstran örneğin, 525 nm) arasında uygun bir Avanlanche entegre. </li><li> Biyopsisi almak için endoskop çalışma kanalına esnek biyopsi forseps (3 Charr., 28 cm) yerleştirin. </li><li> Topikal, intramukozal için çalışma kanalına esnek enjeksiyon tüp (0.96 mm) tanıtmak veya Endoluminaldiagnostik veya terapötik madde arasında dministration. </li><li> 42 ° C&#39;lik bir sıcaklık ile ısıtılabilen bir muayene masası kullanın. Bu farenin muayene sırasında hipotermik hale önler. </li></ol> Hayvanlar 3. Anestezi <ol><li> Küçük ama sızdırmaz kutu içinde fare ve yönetmek izofluran (% 100 (h / h),% 5 hacim / dak 3 L). Fare bilincini kaybedene kadar bekleyin. </li><li> Endoskopi için muayene masasının üzerine fare aktarın. Bir doz 100% v / v, 1.5 hacim,% / dk 1.5 L yüz maskesi ile izofluran teneffüs devam edin. Her zaman anestezi altında iken göz kuruluğunu önlemek için göz merhemi uygulanır. </li><li> Refleksleri kontrol ederek anestezi etkinliğini değerlendirmek. &#39;Refleks dönüş&#39; Giriş: yeterince anestezi ise, sırtında döşeme fare etrafında çevirmek gerekir. Kontrol &#39;ayak refleks&#39;: anestezi yumuşak bacak çekilmesi (evresine yol açmamalıdır hayvanın ayak arasındaki kısma, yeterli zamancerrahi tolerans). </li></ol> 4. Kolonoskopi <ol><li> Muayene masasına sırtını / anestezi fare eğilimli yatıyordu. </li><li> Önemli dışkı yükleme görünümü gizleyebilir olduğu şüphesi varsa kolona düğmeli kanül aracılığı ile lavman 2 ml uygulayın. Farenin lavman uygulanmasından sonra arınmak için bekleyin. Perforasyonunu önlemek için çok dikkatli endoskop yerleştirin. </li><li> Bunlardan biri hava pompasına bağlanır kılıfın her iki vanalarını açın. Havayı dağıtmak için parmağı ile diğer vana kapatılmalıdır. Özellikle biyopsi veya enjeksiyon durumunda, yavaş ve dikkatli bir şekilde, hava ile kolon şişirmek. </li><li> Perforasyon (makattan 4-5 cm) önlemek için sağ kolon eğrilme gibi sadece kadar endoskop ilerlemek. </li><li> Tanı kolonoskopi <ol><li> Endoskop geri çekerken, enflamatuar veya malign değişiklikler için mukozayı inceleyin. Bağırsak bütün çevresini değerlendirmek için geri yavaşça çekin. Intraluminal pa değerlendiringerektiği gibi uygun kurulmuş skorlama sistemleri kullanılarak patolojiler. </li><li> Yara alanların tekrarlayan görselleştirme sırasında görüntü alımı için aynı endoskopik pozisyonunu garanti etmek, kemirgen anüs ve mukozal lezyon arasındaki mesafeyi unutmayın. Ayrıca, endoskop ve görüntü alımı sırasında yara bölgesi arasında aynı mesafe elde etmek için bir aralayıcı olarak biyopsi forseps ucunu kullanın. Yara büyüklüğü 3 mm ihtiva endoskop kılıfın büyüklüğü ile ilgilidir. NOT: Önceki sınavlarda fotoğraf belgeleri ile optik karşılaştırıldığında aynı pozisyonda yerleştirin endoskop. Her biri aynı açı ve mesafe Tedbir lezyonlar endoskopik muayene kadar izleyin. </li></ol></li><li> Biyopsi işlemi <ol><li> Iki araştırıcının yardımı ile biyopsilerin alınması. Forseps ucu ikinci araştırmacıya monitörde görünür oluncaya kadar çalışma kanalına dikkatlice biyopsi forseps tanıtın. Bir dikkatle açık ve kapalı forsepsgeçersiz perforasyon. </li><li> Patolojinin sitesine forseps taşıyın. </li></ol></li><li> Enjeksiyon prosedürü <ol><li> Iki araştırıcının yardımı ile enjeksiyon işlemini gerçekleştirin. Tamamen bir madde ile ön-doldurma esnek enjeksiyon borusu (0.96 mm) tatbik edilecek. Kanül (30 G), ikinci araştırmacı için monitörde görünür oluncaya kadar çalışma kanalına tüp itin. Ince şırınga hazırlayın ve yavaşça teşhis ya da tedavi edici bir maddenin istenen miktarda ilaç. Enjeksiyon hacmi 50 ul yüksek olmalıdır. </li><li> 15-30 derecelik bir açıyla submocosa iğne takın. Mukoza yönünde eğim Yüz. Mukoza başarılı enjeksiyondan sonra bir karakteristik kaldırma işareti gösterir. </li></ol></li><li> Floresan endoskopi (FE) <ol><li> Intravenöz önce endoskopik muayene floresan 100 ul PBS içinde seyreltilmiş 60 mg FITC-dekstran yönetmek. </li><li> Enjeksiyonu arasındaki optimum zaman noktasını kontrol edinfloresan etiketli izleyici ve izleyici farmakoloji bağlıdır görüntüleme yöntemidir. Kullanılan tracer eksitasyon ve emisyon dalga boyunda uygun olarak bant geçiş filtresi sistemi ayarlarını yapılandırır. Mukoza yüzeyinin vasküler dağılımını değerlendirmek için hemen floresan boya intravenöz enjeksiyonundan sonra (örneğin, FITC) ile spesifik olmayan kan hacmi ajanları için floresan endoskopi gerçekleştirin. </li><li> Background-oranı daha iyi hedef sağlamak için hedeflenen izleyici ya da &#39;akıllı prob&#39; durumunda izleyici uygulamadan birkaç saat sonra görüntüleme düşünün. </li><li> Sonuçların fotoğraf ve video belgeleri gerçekleştirin. </li></ol></li></ol> 5. Post-kolonoskopi <ol><li> Boş bir kafes içinde fare ayırın ve çöp aspire gelen fareyi korumak için bir kağıt havlu üzerinde yatıyordu. Hipotermi önlemek için redlight lamba ile fare ısıtın. Fareyi gözlemleyin ve yeterli kavuşana kadar sahipsiz bırakmayınbilinç sternum yatma korumak için. Tamamen bilinçli kez, geri gelen kafesine fare yerleştirin. </li><li> Ve küçük ama sızdırmaz kutu bir deney, yerleştir sonunda CO 2 (% 100 (h / h), 100 hacimce% 3 L / dk) yönetmek. Fare tam bilincini kaybeder ve nefes durana kadar bekleyin. Boyun kırığı tarafından sevk fare. Karın laparotomi gerçekleştirin ve kolon eksplantasyona. Uzunlamasına kolon açın ve daha fazla histolojik veya moleküler değerlendirme için yıkayın. </li></ol>

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Yazarlar ifşa hiçbir şey yok.

Epitelyal yara iyileşmesi devam eden bir süreçtir. Mide-bağırsak mukozasına olan yüzey hücrelerinin sürekli fizyolojik pullanarak 16 epitel hücrelerinin yenilenmesini gerektiren sık ortaya çıkar. Sonuç olarak, yara iyileşmesinde bozulma, gastrointestinal ülser ve 17 anastomoz kaçağı 18 dahil olmak üzere çeşitli hastalıklar üzerinde çok büyük bir etkisi vardır. Epitel iyileşmesini uyarmak için moleküler arka plan değerlendirilmesi yanı sıra potansiyel ila...

Hayvan modelleri büyük ölçüde sayısız bağırsak patolojileri anlayışımızı zenginleştirilmiş. Laboratuvar faresi (Mus musculus) nedeniyle bol genetik ve genomik bilgiye biyomedikal araştırma bir başbakan hayvan modeli olarak ortaya çıktı ve transgenik ve nakavt suşları kolayca kullanılabilir etti. Anlayış hastalık patogenezini artırmaya ek olarak, hayvan modelleri de önemlisi ilaç adayları gibi klinik öncesi teşhis ya da tedavi müdahale test edilmesi için kullanılır. Ancak, insan hastalığı taklit fare modellerinin çeşitliliği rağmen, rutin hasta bakımında kullanılan birçok tanısal ve girişimsel seçenekleri fareler için mevcut değildir. Buna göre, gözetim stratejiler genellikle dolaylı gözlemlere sınırlıdır kemirgen hastalığı veya terapötik müdahalelerin etkisinin seyrini izlemek ya da ölüm sonrası analizleri yayınlamak için. Non-invaziv girişimler izleme fareler için hastalık aktivitesi endeksleri gibi canlılık mevcut iken, qukilo kaybı veya kazancı, kan, idrar ve dışkı antification bu sadece dolaylı göstergeleri ve bireyler arası değişkenliğin tarafından önyargılı, analiz eder. Ayrıca, otopsi tekrarlı zaman noktalarında boyuna gözlemler önlemek analiz eder. Sofistike görüntüleme teknikleri sadece son 1,2 getirilmiştir farelerde hastalık aktivitesini izlemek için. Bu görüntüleme teknikleri tekrarlanan analizler için izin olsa da, onlar doğrudan mukozal görselleştirme izin veya biyopsi satın alma veya ilaç adayları topikal ve intramukozal uygulama olarak tanısal veya tedavi edici girişimler izin vermez, bağırsak üzerinde bir açıklayıcı ve genellikle özensiz bir görünüm sağlar. Son zamanlarda, canlı farelerde kullanım için yüksek çözünürlüklü endoskopi sistemleri 3,4 geliştirilmiştir. İlk defa bu endoskopik teknikler, yara iyileşmesi veya bağırsak iltihabı sağlayan obj olarak Endoluminal kolon hastalık patolojilerinin doğrudan görselleştirme izintekrarlanan zaman noktalarında aynı hayvanın uzunlamasına çalışmalar sağlayan ci, gerçek zamanlı durum. Yanı sıra tekil bir fare tekrarlanmıştır biyopsiler izin vermek veya endoskopik tedavi edici sistemler aynı zamanda ilgi alanı, bir maddenin doğrudan uygulanmasını sağlayan bir ayrı tümör veya lokalize iltihabı etkilemek için kullanılabilir. Tedavi ve kontrol maddeleri ilgi alanına doğrudan teslim edilebilir, bunun için bireyler arası değişkenliğin hariç, aynı fare gerçekleştirilebilir. Bu sistemler artık bu tür kolit şiddeti (MEICS) 5-7 kemirgen endoskopik indeksi gibi kolon iltihabı, yara iyileşmesi, laparoskopik karaciğer biyopsisi ve çeşitli skorlama sistemleri kullanılarak karaciğer tümörleri 8 ve tümör gelişiminin ortotopik indüksiyon değerlendirilmesi için istihdam edilmiştir. Kolon duvarının kalınlaşması, vasküler desen değişiklikleri, fibrin varlığı, MUCOS bir ayrıntı: MEICS iltihabı değerlendirmek için beş parametreden oluşmaktadıral yüzey, ve dışkı kıvamı. Bu protokolde, bağırsak yara iyileşmesi, enflamasyon ve kolon kanseri olan kemirgen modellerinde katı endoskopi kullanılmasını tarif eder. Öncelikle, biz yara iyileşmesi ve inflamatuvar barsak endoskopik değerlendirme yanı sıra kolit aktivite boyuna değerlendirilmesi ve kemirgen kolon karsinogenezi çalışma göstermektedir. Kemirgen endoskopi tanımlayıcı kullanımının ötesinde, biz biyopsisi almak için endoskopik enstrümantasyon kullanımı hakkında ayrıntılı yönergeler sağlar ve farklı ilgi bileşenleri (örneğin, ilaç adayları veya tümör hücrelerinin) topikal ve intramukozal uygulama. Son olarak, kolorektal tümörlerin ortamda gelişmiş moleküler görüntüleme teknikleri kullanılır sıçangil floresan endoskopi kullanımını göstermektedir.

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			<td height="21" style="height:21px;width:469px;">Name</td>
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			<td style="width:255px;">Comment</td>
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			<td height="21" style="height:21px;width:469px;">Reagents</td>
			<td style="width:423px;"></td>
			<td style="width:320px;"></td>
			<td></td>
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			<td height="20" style="height:20px;">Alfalfa-free diet</td>
			<td>Harlan Laboritories, Madison, USA</td>
			<td style="width:320px;">2014</td>
			<td></td>
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			<td height="20" style="height:20px;">Azoxymethane (AOM)</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
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			<td></td>
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			<td height="21" style="height:21px;">Dextran sulphate sodium (DSS)</td>
			<td>TdB Consulatancy, Uppsala, Sweden</td>
			<td>DB001</td>
			<td></td>
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			<td height="20" style="height:20px;width:469px;">Eosin</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">E 4382</td>
			<td></td>
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			<td height="20" style="height:20px;width:469px;">Ethylenediaminetetraacetic acid (EDTA)&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">E 9884</td>
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			<td height="20" style="height:20px;">Falcon Tube 50ml</td>
			<td>BD Biosciences, Erembodegem, Belgium</td>
			<td style="width:320px;">352070</td>
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			<td height="20" style="height:20px;">Florene 100V/V</td>
			<td>Abbott, Wiesbaden, Germany</td>
			<td style="width:320px;">B506</td>
			<td></td>
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			<td height="20" style="height:20px;">Fluorescein-Isothiocyanat (FITC)&#8211;dextrane&#160;</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">FD4-250MG</td>
			<td></td>
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			<td height="21" style="height:21px;width:469px;">Haematoxylin&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">HHS32-1L</td>
			<td></td>
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			<td height="21" style="height:21px;width:469px;">Isopentane (2- Methylbutane)</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">M32631-1L</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">Methylene blue</td>
			<td style="width:423px;">Merck, Darmstadt, Germany</td>
			<td style="width:320px;">1159430025</td>
			<td></td>
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			<td height="21" style="height:21px;width:469px;">O.C.T. Tissue Tek compound&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:423px;">Sakura, Zoeterwonde, Netherlands</td>
			<td style="width:320px;">4583</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Omnican F - canula</td>
			<td style="width:423px;">Braun, Melsungen, Germany</td>
			<td style="width:320px;">9161502</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Phosphate buffered saline, PBS</td>
			<td style="width:423px;">Lonza, Verviers, Belgium</td>
			<td style="width:320px;">4629</td>
			<td></td>
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		<tr height="20">
			<td height="20" style="height:20px;">Sodium Chloride 0,9%</td>
			<td>Braun, Melsungen, Germany</td>
			<td style="width:320px;">5/12211095/0411</td>
			<td></td>
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		<tr height="20">
			<td height="20" style="height:20px;">Standard diet</td>
			<td>Altromin, Lage, Germany</td>
			<td style="width:320px;">1320</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tissue-Tek Cryomold</td>
			<td>Sakura, Leiden, Netherlands</td>
			<td style="width:320px;">4566</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Vitro &#8211; Clud&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:423px;">R. Langenbrinck, Teningen, Germany</td>
			<td style="width:320px;">04-0002&#160;</td>
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			<td height="21" style="height:21px;">Equipment</td>
			<td></td>
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			<td height="20" style="height:20px;">AIDA Control</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20 096020</td>
			<td></td>
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			<td height="20" style="height:20px;">Bandpass filter</td>
			<td>Semrock, Rochester, USA</td>
			<td style="width:320px;">HC 716/40</td>
			<td></td>
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		<tr height="20">
			<td height="20" style="height:20px;">Bandpass filter</td>
			<td>Semrock, Rochester, USA</td>
			<td style="width:320px;">HC 809/81</td>
			<td></td>
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		<tr height="20">
			<td height="20" style="height:20px;">Biopsy Forceps, 3 Fr., 28cm</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">61071ZJ</td>
			<td></td>
		</tr>
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			<td height="20" style="height:20px;">Dell Monitor</td>
			<td>Dell, Frankfurt am Main, Germany</td>
			<td style="width:320px;">U2412Mb</td>
			<td></td>
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			<td height="20" style="height:20px;">Examination Sheath, 9 Fr.</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">61029D</td>
			<td></td>
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			<td height="20" style="height:20px;">Examination Sheath, 9 Fr.</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">61029C</td>
			<td></td>
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			<td height="20" style="height:20px;">Fiber Optic Light Cable, 3.5mm</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">69495NL</td>
			<td></td>
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			<td height="20" style="height:20px;">Fluorescein Blue Filter System</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20100032</td>
			<td></td>
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			<td height="20" style="height:20px;">Fluorescein Barrier Filter</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20100033</td>
			<td></td>
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			<td height="20" style="height:20px;">Foot switch</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20010430</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">HOPKINS Telescope, 1.9mm, Length 10cm</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">1830231</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">SCB D-light P&#160;</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20 133720</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">SCB tricam SL II</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20 2230 20</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tubing set instruments VETPUMP II</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">69811</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tricam PDD PAL</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">20221037</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">UniVet Porta</td>
			<td>Groppler Medizintechnik, Deggendorf, Germany</td>
			<td style="width:320px;">BKGM 0451</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Vetpump 2</td>
			<td>Karl Storz - Endoskope, Tuttlingen, Germany</td>
			<td style="width:320px;">69321620</td>
			<td></td>
		</tr>
	</tbody>
</table>

murine endoscopy for in vivo multimodal imaging of carcinogenesis and assessment of intestinal wound healing and inflammation

Fare modeller yaygın insan hastalıkların oluş çalışma ve klinik öncesi tanı usullerinin yanı sıra terapötik girişimlerin değerlendirmek için kullanılır. Bununla birlikte, patolojik değişimlerin geçerli değerlendirme genellikle histolojik analizini gerektirir ve ex vivo olarak gerçekleştirilen zaman, hayvanın ölümü gerektirir. Bu nedenle, geleneksel deneysel ortamlarda, birey-içi izlem muayeneleri nadiren mümkündür. Dolayısıyla, canlı farelerde kemirgen endoskopi ortaya çıkışı ile mide-bağırsak mukozası ve aynı zamanda görsel değişiklikler izlemek için prosedürü tekrarlayın hem ilk kez araştırmacılar sağlar. In vivo kemirgen endoskopi birçok uygulamaları, bağırsak iltihabı veya yara iyileşmesini okuyan defalarca mukozal biyopsilerin dahil olmak üzere mevcut ve yerel minyatür enjeksiyon kataterlerinden tanısal veya terapötik ajanlar yönetmek. En son, moleküler görüntüleme genişletmiştir tanısal görüntüleme moBelirli photoprobes kullanarak farklı hedef moleküllerin saptanmasına olanak tanıyan spesifik, şekilleri. Sonuç olarak, kemirgen endoskopi in vivo görüntüleme tanı deneysel için yeni bir son teknoloji olarak ortaya çıkmıştır ve önemli ölçüde çeşitli alanlarda preklinik araştırma etkileyebilir.

Bağırsak yara iyileşmesi in vivo izleme (Şekil 1A 1 mm&#39;ye eşit) rutin endoskopi sırasında, mukozal yaralar 3 Fransızca çapında minyatür biyopsi forseps mekanik oluşturuldu. Daha sonra, yara iyileşmesi, günlük muayene, endoskopik olarak izlenip, örneğin resim düzenleme yazılımı kullanarak kalıntı yara bölgesinde ölçülmesiyle nicelleştirilmiştir, İmageJ (Şekil 1B). Zaman içinde, bireysel yara kapatma k?...

Watch this Scientific Journal Video about Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation at JoVE.com

Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation

Murin için Endoskop<em&gt; In Vivo</em&gt; Karsinojenez ve bağırsak Değerlendirilmesi Multimodaller Görüntüleme Yara İyileşmesi ve İltihap

Küçük hayvan görüntüleme teknikleri seri tetkikler ve in vivo terapötik müdahaleyi sağlar. Son zamanlarda, uygulamaların kapsamı önemli genişletmiştir ve şu anda kolon tümör gelişiminin değerlendirilmesi, yara iyileşmesi ve enflamasyon izleme içerir. Bu protokol kemirgen endoskopi bu çeşitli potansiyel uygulamaları göstermektedir.

Mouse models are widely used to study pathogenesis of human diseases and to evaluate diagnostic procedures as well as therapeutic interventions ...

murine-endoscopy-for-vivo-multimodal-imaging-carcinogenesis

Research

JoVE Journal

Medicine

Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation

18.5K Görüntüleme.  University Hospital Münster.  Küçük hayvan görüntüleme teknikleri seri tetkikler ve in vivo terapötik müdahaleyi sağlar. Son zamanlarda, uygulamaların kapsamı önemli genişletmiştir ve şu anda kolon tümör gelişiminin değerlendirilmesi, yara iyileşmesi ve enflamasyon izleme içerir. Bu protokol kemirgen endoskopi bu çeşitli potansiyel uygulamaları göstermektedir. 

Video: Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation

In vivo Near Infrared Fluorescence (NIRF) Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading to an accumulation of lipid-laden intra-luminal plaque, smooth muscle cell proliferation and progressive narrowing of the vessel lumen. The formation of such vulnerable plaques prone to rupture underlies the majority of cases of acute myocardial infarction. The complex molecular and cellular inflammatory cascade is orchestrated by the recruitment of T lymphocytes and macrophages and their paracrine effects on endothelial and smooth muscle cells.1
Molecular imaging in atherosclerosis has evolved into an important clinical and research tool that allows in vivo visualization of inflammation and other biological processes. Several recent examples demonstrate the ability to detect high-risk plaques in patients, and assess the effects of pharmacotherapeutics in atherosclerosis.4 While a number of molecular imaging approaches (in particular MRI and PET) can image biological aspects of large vessels such as the carotid arteries, scant options exist for imaging of coronary arteries.2 The advent of high-resolution optical imaging strategies, in particular near-infrared fluorescence (NIRF), coupled with activatable fluorescent probes, have enhanced sensitivity and led to the development of new intravascular strategies to improve biological imaging of human coronary atherosclerosis.
Near infrared fluorescence (NIRF) molecular imaging utilizes excitation light with a defined band width (650-900 nm) as a source of photons that, when delivered to an optical contrast agent or fluorescent probe, emits fluorescence in the NIR window that can be detected using an appropriate emission filter and a high sensitivity charge-coupled camera. As opposed to visible light, NIR light penetrates deeply into tissue, is markedly less attenuated by endogenous photon absorbers such as hemoglobin, lipid and water, and enables high target-to-background ratios due to reduced autofluorescence in the NIR window. Imaging within the NIR 'window' can substantially improve the potential for in vivo imaging.2,5
Inflammatory cysteine proteases have been well studied using activatable NIRF probes10, and play important roles in atherogenesis. Via degradation of the extracellular matrix, cysteine proteases contribute importantly to the progression and complications of atherosclerosis8. In particular, the cysteine protease, cathepsin B, is highly expressed and colocalizes with macrophages in experimental murine, rabbit, and human atheromata.3,6,7 In addition, cathepsin B activity in plaques can be sensed in vivo utilizing a previously described 1-D intravascular near-infrared fluorescence technology6, in conjunction with an injectable nanosensor agent that consists of a poly-lysine polymer backbone derivatized with multiple NIR fluorochromes (VM110/Prosense750, ex/em 750/780nm, VisEn Medical, Woburn, MA) that results in strong intramolecular quenching at baseline.10 Following targeted enzymatic cleavage by cysteine proteases such as cathepsin B (known to colocalize with plaque macrophages), the fluorochromes separate, resulting in substantial amplification of the NIRF signal. Intravascular detection of NIR fluorescence signal by the utilized novel 2D intravascular NIRF catheter now enables high-resolution, geometrically accurate in vivo detection of cathepsin B activity in inflamed plaque.
In vivo molecular imaging of atherosclerosis using catheter-based 2D NIRF imaging, as opposed to a prior 1-D spectroscopic approach,6 is a novel and promising tool that utilizes augmented protease activity in macrophage-rich plaque to detect vascular inflammation.11,12 The following research protocol describes the use of an intravascular 2-dimensional NIRF catheter to image and characterize plaque structure utilizing key aspects of plaque biology. It is a translatable platform that when integrated with existing clinical imaging technologies including angiography and intravascular ultrasound (IVUS), offers a unique and novel integrated multimodal molecular imaging technique that distinguishes inflammatory atheromata, and allows detection of intravascular NIRF signals in human-sized coronary arteries.

In vivo Near Infrared Fluorescence NIRF Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

We detail a new near-infrared fluorescence (NIRF) catheter for 2-dimensional intravascular molecular imaging of plaque biology in vivo. The NIRF catheter can visualize key biological processes such as inflammation by reporting on the presence of plaque-avid activatable and targeted NIR fluorochromes. The catheter utilizes clinical engineering and power requirements and is targeted for application in human coronary arteries. The following research study describes a multimodal imaging strategy that utilizes a novel in vivo intravascular NIRF catheter to image and quantify inflammatory plaque in proteolytically active inflamed rabbit atheromata.

İnflamatuar Plak Kızılötesi Floresans (NIRF) İntravasküler Moleküler Görüntüleme, Multimodal Yaklaşım civarında in vivo Ateroskleroz Görüntüleme

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading ...

Tıp

Functional Assessment of Intestinal Motility and Gut Wall Inflammation in Rodents: Analyses in a Standardized Model of Intestinal Manipulation

Inflammation of the gastrointestinal tract is a common reason for a variety of human diseases. Animal research models are critical in investigating the complex cellular and molecular of intestinal pathology. Although the tunica mucosa is often the organ of interest in many inflammatory diseases, recent works demonstrated that the muscularis externa (ME) is also a highly immunocompetent organ that harbours a dense network of resident immunocytes.1,2 These works were performed within the standardized model of intestinal manipulation (IM) that leads to inflammation of the bowel wall, mainly limited to the ME. Clinically this inflammation leads to prolonged intestinal dysmotility, known as postoperative ileus (POI) which is a frequent and unavoidable complication after abdominal surgery.3 The inflammation is characterized by liberation of proinflammatory mediators such as IL-64 or IL-1&beta; or inhibitory neurotransmitters like nitric oxide (NO).5 Subsequently, tremendous numbers of immunocytes extravasate into the ME, dominated by polymorphonuclear neutrophils (PMN) and monocytes and finally maintain POI.2 Lasting for days, this intestinal paralysis leads to an increased risk of aspiration, bacterial translocation and infectious complications up to sepsis and multi organ failure and causes a high economic burden.6
In this manuscript we demonstrate the standardized model of IM and in vivo assessment of gastrointestinal transit (GIT) and colonic transit. Furthermore we demonstrate a method for separation of the ME from the tunica mucosa followed by immunological analysis, which is crucial to distinguish between the inflammatory responses in these both highly immunoactive bowel wall compartments. All analyses are easily transferable to any other research models, affecting gastrointestinal function.

Postoperative ileus (POI) is a complication of abdominal surgery leading to increased morbidity and a prolonged hospital stay. Because prophylactic or therapeutic strategies are lacking intensified research is necessary. Therefore we established a standardized and feasible mouse model to investigate the pathophysiology of POI and to study potential therapeutic options.

Bağırsak Manipülasyon bir Standart Model Analizleri: Kemirgenler Barsak Motilite ve Gut Duvar İnflamasyon İşlevsel Değerlendirilmesi

Inflammation of the gastrointestinal tract is a common reason for a variety of human diseases. Animal research models are critical in investigating the ...

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

Yara Şifa murin Modeli

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive nephropathy induced by unilateral ureteric obstruction (UUO), which can either be irreversible (UUO) or reversible (R-UUO). In the irreversible UUO model, the ureter may be obstructed for variable periods of time in order to induce increasingly severe renal inflammation and interstitial fibrotic scarring. In the reversible R-UUO model the ureter is obstructed to induce hydronephrosis, tubular dilation and inflammation. After a suitable period of time the ureteric obstruction is then surgically reversed by anastomosis of the severed previously obstructed ureter to the bladder in order to allow complete decompression of the kidney and restoration of urinary flow to the bladder. The irreversible UUO model has been used to investigate various aspects of renal inflammation and scarring including the pathogenesis of disease and the testing of potential anti-inflammatory or anti-fibrotic therapies. The more challenging model of R-UUO has been used by some investigators and does offer significant research potential as it allows the study of inflammatory and immune processes and tissue remodeling in an injured and scarred kidney following the removal of the injurious stimulus. As a result, the R-UUO model offers investigators the opportunity to explore the resolution of kidney inflammation together with key aspects of tissue repair. These experimental models are of relevance to human disease as patients often present with obstruction of the renal tract that requires decompression and are commonly left with significant residual kidney impairment that has no current treatment options and may lead to eventual end stage kidney failure.

The murine model of irreversible unilateral ureteric obstruction (UUO) is presented together with the model of reversible UUO in which the ureteric obstruction is relieved by anastomosis of the severed ureter into the bladder. These models enable the study of renal inflammation and scarring as well as tissue remodeling.

Geri dönüşümsüz ve Tersinir Tek taraflı Üreterik Obstrüksiyon bir fare modeli

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive ...

Calcification of Vascular Smooth Muscle Cells and Imaging of Aortic Calcification and Inflammation

Cardiovascular disease is the leading cause of morbidity and mortality in the world. Atherosclerotic plaques, consisting of lipid-laden macrophages and calcification, develop in the coronary arteries, aortic valve, aorta, and peripheral conduit arteries and are the hallmark of cardiovascular disease. In humans, imaging with computed tomography allows for the quantification of vascular calcification; the presence of vascular calcification is a strong predictor of future cardiovascular events. Development of novel therapies in cardiovascular disease relies critically on improving our understanding of the underlying molecular mechanisms of atherosclerosis. Advancing our knowledge of atherosclerotic mechanisms relies on murine and cell-based models. Here, a method for imaging aortic calcification and macrophage infiltration using two spectrally distinct near-infrared fluorescent imaging probes is detailed. Near-infrared fluorescent imaging allows for the ex vivo quantification of calcification and macrophage accumulation in the entire aorta and can be used to further our understanding of the mechanistic relationship between inflammation and calcification in atherosclerosis. Additionally, a method for isolating and culturing animal aortic vascular smooth muscle cells and a protocol for inducing calcification in cultured smooth muscle cells from either murine aortas or from human coronary arteries is described. This in vitro method of modeling vascular calcification can be used to identify and characterize the signaling pathways likely important for the development of vascular disease, in the hopes of discovering novel targets for therapy.

Vascular calcification is an important predictor of and contributor to human cardiovascular disease. This protocol describes methods for inducing calcification of cultured primary vascular smooth muscle cells and for quantifying calcification and macrophage burden in animal aortas using near-infrared fluorescence imaging.

Aort Kalsifikasyon ve Enflamasyon Vasküler düz kas hücrelerinin ve Görüntüleme Kalsifikasyon

Cardiovascular disease is the leading cause of morbidity and mortality in the world.  Atherosclerotic plaques, consisting of lipid-laden macrophages and ...

Multimodal Quantitative Phase Imaging with Digital Holographic Microscopy Accurately Assesses Intestinal Inflammation and Epithelial Wound Healing

The incidence of inflammatory bowel disease, i.e., Crohn&#39;s disease and Ulcerative colitis, has significantly increased over the last decade. The etiology of IBD remains unknown and current therapeutic strategies are based on the unspecific suppression of the immune system. The development of treatments that specifically target intestinal inflammation and epithelial wound healing could significantly improve management of IBD, however this requires accurate detection of inflammatory changes. Currently, potential drug candidates are usually evaluated using animal models in vivo or with cell culture based techniques in vitro. Histological examination usually requires the cells or tissues of interest to be stained, which may alter the sample characteristics and furthermore, the interpretation of findings can vary by investigator expertise. Digital holographic microscopy (DHM), based on the detection of optical path length delay, allows stain-free quantitative phase contrast imaging. This allows the results to be directly correlated with absolute biophysical parameters. We demonstrate how measurement of changes in tissue density with DHM, based on refractive index measurement, can quantify inflammatory alterations, without staining, in different layers of colonic tissue specimens from mice and humans with colitis. Additionally, we demonstrate continuous multimodal label-free monitoring of epithelial wound healing in vitro, possible using DHM through the simple automated determination of the wounded area and simultaneous determination of morphological parameters such as dry mass and layer thickness of migrating cells. In conclusion, DHM represents a valuable, novel and quantitative tool for the assessment of intestinal inflammation with absolute values for parameters possible, simplified quantification of epithelial wound healing in vitro and therefore has high potential for translational diagnostic use.

Accurate assessment of anti-inflammatory effects is of utmost importance for the evaluation of potential new drugs for the treatment of inflammatory bowel disease. Digital holographic microscopy provides assessment of inflammation in murine and human colonic tissue samples as well as automated multimodal evaluation of epithelial wound healing in vitro.

Dijital Holografik Mikroskop ile multimodal Kantitatif Faz Görüntüleme Doğru Bağırsak İltihabı ve Epitel Yara Şifa Değerlendirdi

The incidence of inflammatory bowel disease, i.e., Crohn&#39;s disease and Ulcerative colitis, has significantly increased over the last decade. The ...

Fluorescence-mediated Tomography for the Detection and Quantification of Macrophage-related Murine Intestinal Inflammation

Murine models of disease are indispensable to scientific research. However, many diagnostic tools such as endoscopy or tomographic imaging are not routinely employed in animal models. Conventional experimental readouts often rely on post mortem and ex vivo analyses, which prevent intra-individual follow-up examinations and increase the number of study animals needed. Fluorescence-mediated tomography enables the non-invasive, repetitive, quantitative, three-dimensional assessment of fluorescent probes. It is highly sensitive and permits the use of molecular makers, which allows for the specific detection and characterization of distinct molecular targets. In particular, targeted probes represent an innovative tool for analyzing gene activation and protein expression in inflammation, autoimmune disease, infection, vascular disease, cell migration, tumorigenesis, etc. In this article, we provide step-by-step instructions on this sophisticated imaging technology for the in vivo detection and characterization of inflammation (i.e., F4/80-positive macrophage infiltration) in a widely used murine model of intestinal inflammation. This technique might also be used in other research areas, such as immune cell or stem cell tracking.

Target-specific probes represent an innovative tool for analyzing molecular mechanisms, such as protein expression in various types of disease (e.g., inflammation, infection, and tumorigenesis). In this study, we describe a quantitative three-dimensional tomographic assessment of intestinal macrophage infiltration in a murine model of colitis using F4/80-specific fluorescence-mediated tomography.

Floresan-aracılı tomografi algılama ve fare bağırsak iltihabı makrofaj ile ilgili miktar

Murine models of disease are indispensable to scientific research. However, many diagnostic tools such as endoscopy or tomographic imaging are not ...

Microscopy Based Methods for the Assessment of Epithelial Cell Migration During In Vitro Wound Healing

Cell migration is a mandatory aspect for wound healing. Creating artificial wounds on research animal models often results in costly and complicated experimental procedures, while potentially lacking in precision. In vitro culture of epithelial cell lines provides a suitable platform for researching the cell migratory behavior in wound healing and the impact of treatments on these cells. The physiology of epithelial cells is often studied in non-confluent conditions; however, this approach may not resemble natural wound healing conditions. Disrupting the epithelium integrity by mechanical means generates a realistic model, but may impede the application of molecular techniques. Consequently, microscopy based techniques are optimal for studying epithelial cell migration in vitro. Here we detail two specific methods, the artificial wound scratch assay and the artificial migration front assay, that can obtain quantitative and qualitative data, respectively, on the migratory performance of epithelial cells.

Microscopy Based Methods for the Assessment of Epithelial Cell Migration During  In Vitro Wound Healing

This manuscript describes how incision-like lesions made on cultured epithelial cell monolayers conveniently model wound healing in vitro, allowing for imaging by confocal or laser scanning microscopy, and which can provide high-quality quantitative and qualitative data for studying both cell behavior and the mechanisms involved in migration.

Mikroskopi Vitro yara iyileşmesi sırasında epitel hücre göç değerlendirilmesi için yöntemleri temel

Cell migration is a mandatory aspect for wound healing. Creating artificial wounds on research animal models often results in costly and complicated ...

Corneal Epithelial Abrasion with Ocular Burr As a Model for Cornea Wound Healing

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to injury. In fact, the most common type of eye injury found in clinic is a corneal abrasion. Here, we utilize an ocular burr to induce an abrasion resulting in removal of the corneal epithelium in vivo on anesthetized mice. This method allows for targeted and reproducible epithelial disruption, leaving other areas intact. In addition, we describe the visualization of the abraded epithelium with fluorescein staining and provide concrete advice on how to visualize the abraded cornea. Then, we follow the timeline of wound healing 0, 18, and 72 h after abrasion, until the wound is re-epithelialized. The epithelial abrasion model of corneal injury is ideal for studies on epithelial cell proliferation, migration and re-epithelialization of the corneal layers. However, this method is not optimal to study stromal activation during wound healing, because the ocular burr does not penetrate to the stromal cell layers. This method is also suitable for clinical applications, for example, pre-clinical test of drug effectiveness.

This protocol describes a method to inflict an abrasion to the ocular surface of the mouse, and to follow the wound healing process thereafter. The protocol takes advantage of an ocular burr to partially remove the surface epithelium of the eye in anaesthetized mice.

Kornea epitel aşınma oküler Burr ile bir Model olarak kornea için yara iyileşme

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to ...

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.