Yazarlar, Ulusal Sağlık ve Avustralya Tıbbi Araştırma Konseyi (NHMRC) (: 632.512 Proje Hibe ID) fon desteği kabul etmek istiyorum. Louise Dunn Ulusal Kalp Vakfı Kariyer Geliştirme Bursu tarafından NHMRC Erken Kariyer Bursu ve Christina Bursill tarafından desteklenmiştir.

1. Ateller ve Tıkayıcı Pansuman hazırlanması <ol><li> 0.5 mm kalınlığında silikon kaplama ve kullanım makas veya silikon diskler oluşturmak için bir biyopsi yumruk 10 mm daireler Anahat. </li><li> Merkezi 10 mm dairenin ortasında bir 5 mm biyopsi yumruk ve bir atel olarak kullanılacak disk gibi-bir &quot;halka&quot; oluşturmak için bir delik oluşturmak için sıkıca bastırın. </li><li> Bu Opsite ve dairesel pansuman oluşturmak için kullanımı makas gibi şeffaf bir kapalı pansuman 10 mm daireler Anahat. </li></ol> 2. Deney Hayvanları <ol><li> Yapılacak tüm deneyler için Hayvan Etik Kurulu onayı alın. </li><li> Ticari bir damızlık (örneğin Jackson Laboratories) 8 haftalık (22-26 gram) erkek C57BL/6J fareler kullanın. </li><li> 21 standart koşullar altında farenin Ol ° C ve yiyecek ve suya serbest erişim ile, 12 saat ışık-karanlık çevrimi. </li><li> (İsteğe bağlı): Diyabet bir bolus intraperitoneal inje tarafından 6 ila 7 haftalık farelerde neden olabilirDüzenli kan şekeri izleme (AccuChek glucometer) ile teyit hiperglisemi ile 165 mg / kg streptozotosin (sitrat tamponu, pH 4,5) bir ction,. Diyabetik farelerde poliüri olabilir ve böylece yatak ıslaklık ortadan kaldırmak için ve ağırlıkları yakından izlenmelidir daha sık değiştirilmesi gerekebilir. </li></ol> 3. Anestezi ve Operatif Hazırlık <ol><li> % 100 oksijen izofluran% 5 (akış hızı 1 L / dk) ile genel anestezi neden ve izofluran% 1-3 ile anestezi korumak. </li><li> Bastırılır fare derin pedal refleksleri olun ve yüzüstü pozisyonda fare yerleştirin. </li><li> Daha da geri ve iki kürek kemikleri arasında 3 cm boyun tabanından kesme makineleri ile kürk kaldırarak ameliyat bölgesi hazırlayın. </li><li> Tüy dökücü krem ​​hafif bir uygulama 2 dakika daha uzun süre uygulanabilir. Islak gazlı bezlerden tüm krem ​​sağlamak için kullanılabilir ve kalan kürk kaldırılır. </li><li> Bir ile cilt silinalkol sürüntü ve% 10 iki uygulama povidon-iyot (Betadine) ve fare asmak. </li></ol> 4. Eksizyon ve Yara splintleme <ol><li> Fare yara iyileşme modeli şematik gösterimi Şekil 1&#39;de sunulmuştur. </li><li> (Şekil 2A-2B) omuz hizasında fare orta hattın her iki tarafında yara için iki adet dairesel model anahat steril bir 4 mm biyopsi yumruk kullanın. </li><li> Anahat ve istemsiz, carnosus (Şekil 2e) de dahil olmak üzere deri altı doku (Şekil 2c-2d), boyunca devam eden tam kat yara oluşturmak için iris makası ortasında cilt kaldırmak için dişli forseps kullanın ve tüketim dairesel parça doku. </li><li> Orta hat (Şekil 2f) diğer tarafında yara için işlemi tekrarlayın. </li><li> Silikon atel her taraftan plastik koruyucu kaplama çıkarın. </li><li> Cyanoacryla uygulayınte yapıştırıcı (örneğin Super Glue veya Krazy Tutkal) bir silikon atel bir tarafa. </li><li> Merkezi yara üzerinde atel (Şekil 2g) ve çapa konumlandırma (Şekil 2 saat) sağlamak için kesintiye 6-0 naylon sütür ile atel. </li><li> Diğer yara üzerindeki splintleme işlemi tekrarlayın. </li><li> Bu zaman noktasında gerekli ise, bir yara (Şekil 2i) ve diğer araç kontrolü için, test edilecek olan tedavi edici bileşik (100 ul) uygulanır. </li><li> Şeffaf bir oklüzif sargı (örneğin, OpSite) (Şekil 2J-2k) ile yara kapsamaktadır. </li><li> Bir cetvel atel altına yerleştirilir ve bir fotomikrograf (2l Şekil) alınır. </li></ol> 6. Ameliyat sonrası Yönetimi <ol><li> Carprofen (5 mg / kg) ameliyat sonrası ağrı kesici için alt deri enjeksiyon yoluyla günde bir kez uygulanır. </li><li> Ameliyat hayvan ayrı ayrı kafeslerde tutuldu ve ısı m muhafaza edildikten sonraats tamamen iyileşti kadar. </li><li> Ağrı ve kilo kaybı bulguları için günde iki kez hayvan izleyin. Biz ağrı veya kilo kaybı hiçbir brüt davranış görüntüler görülmektedir. </li></ol> 7. Yara Ölçme ve Tedavi <ol><li> Yara, günlük olarak ölçülebilir. </li><li> % 5 izofluran gaz (akış hızı 1 L / dk) ile genel anestezi neden, ve sonra fare derin duyu refleksleri sağlamak% 1-3 izofluran kullanılarak bastırılır. </li><li> Forseps ile yavaşça geri kabuğu tıkayıcı pansuman. </li><li> Yara çapının ölçülmesi için cerrahi kumpas kullanarak. Biz X, Y ve Z ekseni (Şekil 2m-2o) yanı sıra ileride (Şekil 2 p) için bir fotomikrograf boyunca üç ölçümün ortalama almak. </li><li> İsteğe bağlı: Şu anda hayvan bir lazer Doppler görüntüleme (Şekil 2q) kullanarak kan perfüzyon için değerlendirilebilir. </li><li> Tedavi edici bileşik ve araç kontrolü arasında yeniden uygulama performans olabilir,Bu noktada med. </li><li> Temiz bir şeffaf kapalı pansuman sonra yeniden uygulanır ve tamamen iyileşti kadar hayvanların sıcak tutulur. </li><li> Not: atel düzgün güvenli değilse yara kolayca sözleşme olacaktır (Şekil 2r). </li></ol> 8. Histolojik Analizi <ol><li> Anestezi aşırı dozda 10 gün sonra fareler Euthanize. </li><li> Forseps kullanarak ve bir neşter bıçak sıyırın dikkatle atel dikişler çıkarın ve. </li><li> Çevresinde ve yara alanı altında geniş bir tam eksizyon oluşturmak ve fosfat içinde% 4 paraformaldehit içinde doku inkübe iris makası kullanarak 4 ° C&#39;da gece boyunca (PBS), tamponlu tuzlu su. </li><li> ° C sonra -80 Ekim bileşik ve dondurularak doku ve yerine dikkatle dabbing aşırı çözeltinin ayrılması 4 ° C de 24 saat daha süreyle PBS çözeltisi içinde% 17 sukroz doku transferi </li><li> Hematoksilin ve eozin boyama, yara yapısı ve epitel boşluğu görselleştirmek için kullanılabilir. Neovascularisation kılcal sayısı (Von Willebrand faktörü kullanılarak) belirlemek için immünohistokimyasal analizleri ile tespit edilebilir. </li></ol>

<ol>
	<li>Sen, C. K., 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=Human+skin+wounds:+a+major+and+snowballing+threat+to+public+health+and+the+economy.">Human skin wounds: a major and snowballing threat to public health and the economy.</a> Wound Repair. 17, 763-771 (2009).</li><li>Sen, C. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wound+healing+essentials:+let+there+be+oxygen.">Wound healing essentials: let there be oxygen.</a> Wound Repair Regen. 17, 1-18 (2009).</li><li>Gurtner, G. C., Werner, S., Barrandon, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wound+repair+and+regeneration.">Wound repair and regeneration.</a> Nature. 453, 314-321 (2008).</li><li>Lindblad, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Considerations+for+selecting+the+correct+animal+model+for+dermal+wound-healing+studies.">Considerations for selecting the correct animal model for dermal wound-healing studies.</a> J. Biomater. Sci. Polym. Ed. 19, 1087-1096 (2008).</li><li>Grose, R., Werner, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wound-healing+studies+in+transgenic+and+knockout+mice.">Wound-healing studies in transgenic and knockout mice.</a> Mol. Biotechnol. 28, 147-166 (2004).</li><li>Reid, R. R., Said, H. K., Mogford, J. E., Mustoe, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+future+of+wound+healing:+pursuing+surgical+models+in+transgenic+and+knockout+mice.">The future of wound healing: pursuing surgical models in transgenic and knockout mice.</a> J. Am. Coll. Surg. 199, 578-585 (2004).</li><li>Fang, R. C., Mustoe, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+wound+healing:+utility+in+transgenic+mice.">Animal models of wound healing: utility in transgenic mice.</a> J. Biomater. Sci. Polym. Ed. 19, 989-1005 (2008).</li><li>Wong, V. W., Sorkin, M., Glotzbach, J. P., Longaker, M. T., Gurtner, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surgical+approaches+to+create+murine+models+of+human+wound+healing.">Surgical approaches to create murine models of human wound healing.</a> J. Biomed. Biotechnol. 2011, 969618 (2011).</li><li>Galiano, R. D., Michaels, J. t., Dobryansky, M., Levine, J. P., Gurtner, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+and+reproducible+murine+model+of+excisional+wound+healing.">Quantitative and reproducible murine model of excisional wound healing.</a> Wound Repair. 12, 485-492 (2004).</li><li>Galiano, R. D., 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=Topical+vascular+endothelial+growth+factor+accelerates+diabetic+wound+healing+through+increased+angiogenesis+and+by+mobilizing+and+recruiting+bone+marrow-derived+cells.">Topical vascular endothelial growth factor accelerates diabetic wound healing through increased angiogenesis and by mobilizing and recruiting bone marrow-derived cells.</a> The American Journal of Pathology. 164, 1935-1947 (2004).</li><li>Thangarajah, H., 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=The+molecular+basis+for+impaired+hypoxia-induced+VEGF+expression+in+diabetic+tissues.">The molecular basis for impaired hypoxia-induced VEGF expression in diabetic tissues.</a> Proceedings of the National Academy of Sciences of the United States of America. 106, 13505-13510 (2009).</li><li>Raza, A., Bayles, C., Biebel, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigation+of+Bacterial+Growth+and+Moisture+Handling+Properties+of+Transparent+Dressings:+3M+Tegaderm+Transparent+Dressing,+3M+Tegaderm+HP+Transparent+Dressing,+and+Opsite+IV3000+Transparent+Dressing.">Investigation of Bacterial Growth and Moisture Handling Properties of Transparent Dressings: 3M Tegaderm Transparent Dressing, 3M Tegaderm HP Transparent Dressing, and Opsite IV3000 Transparent Dressing.</a> Smith and Nephew Report. , (1998).</li><li>Chung, T. Y., Peplow, P. V., Baxter, G. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Testing+photobiomodulatory+effects+of+laser+irradiation+on+wound+healing:+development+of+an+improved+model+for+dressing+wounds+in+mice.">Testing photobiomodulatory effects of laser irradiation on wound healing: development of an improved model for dressing wounds in mice.</a> Photomed. Laser Surg. 28, 589-596 (2010).</li></ol>

Yazarlar ifşa hiçbir şey yok.

Bu deri yara iyileşmesinin deneysel fare modelidir. Bu modelin önemli bir özelliği, insanlarda oluşan işlemi için benzer hale reepitelizasyon ve yeni doku oluşumu meydana gelebilir, böylece yara büzülmesini önlemek için silikon splintlerin kullanılmasıdır. Bu model, çok yönlü ve fizyolojik ve patofizyolojik (örneğin, diyabet) ortamlarda yara iyileşmesi değerlendirmek için de kullanılabilir. Bu model aynı zamanda ekonomik bir ortamda potansiyel yara iyileşmesi veya anjiyojenez tedavi d...

Bozulmuş yara iyileşmesi önemli morbidite ve tüm dünyada ölüm sorumludur, bu diyabet 1,2 olanlar için geçerlidir. İnsanlarda, yara iyileşmesi 3 örtüşen önemli var olduğu, süreçlerin bir süreklilik olduğunu. Hemen sonra yaralama, inflamatuvar süreçleri başlatılır. Inflamatuar hücreler hücre çoğalması, göç ve anjiyogenez süreçleri teşvik faktörler bırakın. Reepitelizasyon ve yeni doku oluşumu sonra bu kollajen gibi matriks proteinlerinin apoptoz ve yeniden organizasyon hem de gerektirir yeniden bir aşaması vardır. Yara iyileşmesinin karmaşıklığı şu in vitro çoğaltılması ve bu hayvan modellerinin kullanılması gerekmektedir edilemez. Bugüne kadar, yara iyileşmesi çalışmaları iyileşme süreçleri eşdeğer ve insanlar için benzer olmasını sağlamak için, bu tür domuz gibi büyük hayvan modelleri ile sınırlı kalmıştır. Ancak, büyük anima kullanarakbu tür çalışmalar için ls evine zor ve her zaman pratik 4 değildir olabilir. Laboratuvar fare kolayca genetik 5-7 mekanik soruşturma için manipüle edilebilir ekonomik bir hayvan modeli temsil eder. Ancak, fare yaralar daralma 8 sürecinde öncelikle nedeniyle, insan için farklı iyileşir. Bu da, kısmen, insanlarda büyük ölçüde eksik olan istemsiz carnosus adlandırılan geniş bir deri altı çizgili kas tabakası kaynaklanmaktadır. Farelerde, bu kas tabakası deriye bağımsız olarak derin kas hareket etmesine olanak sağlar ve cilt aşağıdaki yaralama hızlı daralma sorumludur. Bu sınırlamayı aşmak için, sıçangil yara iyileşmesi bir splintin kullanımı (Şekil 1) 8,9 insan yara iyileşmesi çoğaltmak için adapte edilebilir. Bu videoda yeniden epithelializa insan süreçleri yara daralma ortadan kaldırır ve daha yakından yaklaşık olarak atel fare yara modeli göstermektion ve yeni doku oluşumu. Bu modelde panniculus carnosus içeren iki tam kat eksizyon fare orta hat her iki tarafında sırtı, bir oluşturulur. Bir silikon atel yapıştırıcı yardımı ile yara etrafına yerleştirilen ve atel sonra kesilen dikişlerle sabitlenir. Her fare böylece hayvan sayısını azaltarak, bir yara tedavi ve diğer araç kontrol alma ile, kendi kontrol gibi davranır. Topikal uygulama sonrası, şeffaf bir kapalı pansuman uygulanır. Daha da topikal uygulamalar ve / veya yara bölgesinde 10,11 ölçümü için gerekli zaman sargı kaldırılabilir. De deneylerin tamamlanması, yara kapama, morfolojik mimari ve neovaskülarizasyon derecesi immünohistokimyasal olarak değerlendirilebilir. Bu ekonomik ve model yapmak kolay da diyabet veya diğer patofizyolojilere bağlamında yara iyileşmesi değerlendirmek için kullanılabilir.

<table border="0">
<tbody>
 <tr>
 <td>Name of Reagent/Material</td>
 <td>Company</td>
 <td>Catalog Number</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Press-to-seal silicone sheeting 0.5 mm thick</td>
 <td>Invitrogen</td>
 <td>P18178</td>
 <td>Cut into "donuts" with external diameter of 1cm external, 0.5 cm internal diameter</td>
 </tr>
 <tr>
 <td>Biopsy punch 5 mm</td>
 <td>Steifel</td>
 <td>BC-B1-0500</td>
 <td>To outline wound area to be excised</td>
 </tr>
 <tr>
 <td>Vannas scissors 8.5 cm curved</td>
 <td>World Precision Instruments </td>
 <td>501232</td>
 <td>For wound incision and excision</td>
 </tr>
 <tr>
 <td>Dumonte #7b forceps, 11 cm</td>
 <td>World Precision Instruments</td>
 <td>501302</td>
 <td>To grip skin when creating incision and excising skin </td>
 </tr>
 <tr>
 <td>Graefe forceps, serrated 10cm</td>
 <td>World Precision Instruments</td>
 <td>14142</td>
 <td>To help attach silicone splint to skin</td>
 </tr>
 <tr>
 <td>Needle holder, smooth jaws, curved, 12.5 cm</td>
 <td>World Precision Instruments</td>
 <td>14132</td>
 <td></td>
 </tr>
 <tr>
 <td>Malis forceps, smooth, straight, 12 cm</td>
 <td>Codman and Shurtleff, Inc (J&amp;J)</td>
 <td>80-1500</td>
 <td>To suture the silicon rings to the skin</td>
 </tr>
 <tr>
 <td>Ruler, 0.5 mm gradation</td>
 <td>n/a</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Calipers 0.25 mm gradation</td>
 <td>Duckworth and Kent</td>
 <td>9-653</td>
 <td>To measure wound area</td>
 </tr>
 <tr>
 <td>Opsite FlexiFix transparent adhesive film. 10 cm x 1 m</td>
 <td>Smith &amp; Nephew</td>
 <td>66030570</td>
 <td></td>
 </tr>
 <tr>
 <td>Rimadyl (Carprofen)</td>
 <td>Pfizer</td>
 <td>462986</td>
 <td></td>
 </tr>
 </tbody>
</table>

murine model of wound healing

Yara iyileşmesi ve onarım insan hayatta meydana gelen en karmaşık biyolojik süreçleri bulunmaktadır. Yaralanma sonrası, birden fazla biyolojik yollar aktif hale gelir. Örneğin diyabetli hastalarda görülür Engelli yara iyileşmesi, bu amputasyon gibi ciddi olumsuz sonuçlara yol açabilir. Yara onarımı teşvik yeni ilaçlar geliştirmek için giderek artan bir ivme nedenle, var. Bunların test gibi genellikle pratik değildir domuz gibi büyük hayvan modellerinde sınırlı kalmıştır. Bu ekonomik ve mekanik soruşturma için izin verir genetik manipülasyon, mükellef olarak fareler, ideal klinik öncesi modeli temsil eder. Ancak, öncelikle daralma yoluyla gerçekleşir olarak bir fare iyileşme insanlarınkine temelden farklıdır sarılır. Bizim fare modeli yara etrafında bir atel dahil ederek bu üstesinden gelir. Yara splintleme olarak, onarım işlemi yakından ayna epitelizasyon, hücre çoğalması ve anjiyogenez, ardından bağlıdırinsan yara iyileşmesinin biyolojik prosesler. Tutarlılık ve bakım gerektiren iken, bu fare modeli karmaşık cerrahi teknikleri içerir ve örneğin, damar gelişimini inhibe edebilir ya da iltihap umut vaat eden maddelerin güçlü bir test edilmesine izin vermektedir. Iki yara hazırlanır Dahası, her bir fare, test bileşiği ve aynı hayvan üzerinde araç kontrolü, hem de uygulama sağlayan, kendi kontrolü olarak görev yapar. Sonuç olarak, biz insanların bu karşılaştırılabilir yara iyileşmesi, pratik bir,-öğrenmek için kolay ve güçlü modeli göstermektedir.

(Gün 0 çap / gün x çap) - bir yara kapatma kısmının eğri yaranın ortalama çapının hesaplanması ve bir yüzde, yani, 100 gibi sonuçlar ifade ile belirlenir. Bu deneyde, bir tedavi bileşiği (veya taşıyıcı ortam kontrol) yaraya günlük olarak uygulandı. Terapötik bileşim büyük ölçüde hızlandırılmış yara kapatma kısmının (Şekil 3). Bu atel çıkarıldıktan hızlı yara daralma (Şekil 2r) yol ve insanlarda görülen yara iyileşmesinin desen u...

Watch this Scientific Journal Video about Murine Model of Wound Healing at JoVE.com

Murine Model of Wound Healing

Fizyolojik ve patofizyolojik ortamlarda terapötik bileşiklerini değerlendirmek için de kullanılabilir deri yara iyileşmesinin Bir murin modeli.

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. ...

Research

JoVE Journal

Medicine

63.6K Görüntüleme.  The Heart Research Institute. Fizyolojik ve patofizyolojik ortamlarda terapötik bileşiklerini değerlendirmek için de kullanılabilir deri yara iyileşmesinin Bir murin modeli.

Video: Murine Model of Wound Healing

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

Mouse models are widely used to study pathogenesis of human diseases and to evaluate diagnostic procedures as well as therapeutic interventions preclinically. However, valid assessment of pathological alterations often requires histological analysis, and when performed ex vivo, necessitates death of the animal. Therefore in conventional experimental settings, intra-individual follow-up examinations are rarely possible. Thus, development of murine endoscopy in live mice enables investigators for the first time to both directly visualize the gastrointestinal mucosa and also repeat the procedure to monitor for alterations. Numerous applications for in vivo murine endoscopy exist, including studying intestinal inflammation or wound healing, obtaining mucosal biopsies repeatedly, and to locally administer diagnostic or therapeutic agents using miniature injection catheters. Most recently, molecular imaging has extended diagnostic imaging modalities allowing specific detection of distinct target molecules using specific photoprobes. In conclusion, murine endoscopy has emerged as a novel cutting-edge technology for diagnostic experimental in vivo imaging and may significantly impact on preclinical research in various fields.

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

Small animal imaging techniques allow serial diagnostic examinations and therapeutic interventions in vivo. Recently, the scope of applications has significantly widened and currently includes assessment of colonic tumor development, wound healing and monitoring of inflammation. This protocol illustrates these diverse potential applications of murine endoscopy.

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

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

Tıp

A Mouse Fetal Skin Model of Scarless Wound Repair

Early in utero, but not in postnatal life, cutaneous wounds undergo regeneration and heal without formation of a scar. Scarless fetal wound healing occurs across species but is age dependent. The transition from a scarless to scarring phenotype occurs in the third trimester of pregnancy in humans and around embryonic day 18 (E18) in mice. However, this varies with the size of the wound with larger defects generating a scar at an earlier gestational age. The emergence of lineage tracing and other genetic tools in the mouse has opened promising new avenues for investigation of fetal scarless wound healing. However, given the inherently high rates of morbidity and premature uterine contraction associated with fetal surgery, investigations of fetal scarless wound healing in vivo require a precise and reproducible surgical model. Here we detail a reliable model of fetal scarless wound healing in the dorsum of E16.5 (scarless) and E18.5 (scarring) mouse embryos.

During mammalian development, early gestational skin wounds heal without a scar. Here we detail a reliable and reproducible model of fetal scarless wound healing in the cutaneous dorsum of E16.5 (scarless) and E18.5 (scarring) mouse embryos.

Scarless Yara Repair Fare Fetal Cilt Modeli

Early in utero, but not in postnatal life, cutaneous wounds undergo regeneration and heal without formation of a scar. Scarless fetal wound healing occurs ...

Demonstration of the Rat Ischemic Skin Wound Model

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, and unrelieved pressure due to immobility. Animal models have been developed that attempt to mimic these conditions for the purpose of furthering our understanding of the complexity of chronic wounds. The model described herein is a rat ischemic skin flap model that permits a prolonged reduction of blood flow resulting in wounds that become ischemic and resemble a chronic wound phenotype (reduced vascularization, increased inflammation and delayed wound closure). It consists of a bipedicled dorsal flap with 2 ischemic wounds placed centrally and 2 non-ischemic wounds lateral to the flap as controls. A novel addition to this ischemic skin flap model is the placement of a silicone sheet beneath the flap that functions as a barrier and a splint to prevent revascularization and reduce contraction as the wounds heal. Despite the debate of using rats for wound healing studies due to their quite distinct anatomic and physiologic differences compared to humans (i.e., the presence of a panniculus carnosus muscle, short life-span, increased number of hair follicles, and their ability to heal infected wounds) the modifications employed in this model make it a valuable alternative to previously developed ischemic skin flap models.

The rat, due to its size, availability, and rather docile behavior, has been utilized as a research model for many years. The goal of this protocol is to utilize the rat as an ischemic skin wound healing model to provide valuable insight into the pathophysiology of chronic wounds.

Sıçan İskemik Cilt Yara Modeli Gösteri

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, 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 ...

Creation and Transplantation of an Adipose-derived Stem Cell (ASC) Sheet in a Diabetic Wound-healing Model

Artificial skin has achieved considerable therapeutic results in clinical practice. However, artificial skin treatments for wounds in diabetic patients with impeded blood flow or with large wounds might be prolonged. Cell-based therapies have appeared as a new technique for the treatment of diabetic ulcers, and cell-sheet engineering has improved the efficacy of cell transplantation. A number of reports have suggested that adipose-derived stem cells (ASCs), a type of mesenchymal stromal cell (MSC), exhibit therapeutic potential due to their relative abundance in adipose tissue and their accessibility for collection when compared to MSCs from other tissues. Therefore, ASCs appear to be a good source of stem cells for therapeutic use. In this study, ASC sheets from the epididymal adipose fat of normal Lewis rats were successfully created using temperature-responsive culture dishes and normal culture medium containing ascorbic acid. The ASC sheets were transplanted into Zucker diabetic fatty (ZDF) rats, a rat model of type 2 diabetes and obesity, that exhibit diminished wound healing. A wound was created on the posterior cranial surface, ASC sheets were transplanted into the wound, and a bilayer artificial skin was used to cover the sheets. ZDF rats that received ASC sheets had better wound healing than ZDF rats without the transplantation of ASC sheets. This approach was limited because ASC sheets are sensitive to dry conditions, requiring the maintenance of a moist wound environment. Therefore, artificial skin was used to cover the ASC sheet to prevent drying. The allogenic transplantation of ASC sheets in combination with artificial skin might also be applicable to other intractable ulcers or burns, such as those observed with peripheral arterial disease and collagen disease, and might be administered to patients who are undernourished or are using steroids. Thus, this treatment might be the first step towards improving the therapeutic options for diabetic wound healing.

Creation and Transplantation of an Adipose-derived Stem Cell ASC Sheet in a Diabetic Wound-healing Model

Adipose-derived stem cells (ASCs) are easily isolated and harvested from the fat of normal rats. ASC sheets can be created using cell-sheet engineering and can be transplanted into Zucker diabetic fatty rats exhibiting full-thickness skin defects with exposed bone and then covered with a bilayer of artificial skin.

Oluşturma ve diyabetik yara iyileşmesi manken bir yağ elde edilen kök hücre (ASC) Sheet nakli

Artificial skin has achieved considerable therapeutic results in clinical practice. However, artificial skin treatments for wounds in diabetic patients ...

Development of a Direct Pulp-capping Model for the Evaluation of Pulpal Wound Healing and Reparative Dentin Formation in Mice

Dental pulp is a vital organ of a tooth fully protected by enamel and dentin. When the pulp is exposed due to cariogenic or iatrogenic injuries, it is often capped with biocompatible materials in order to expedite pulpal wound healing. The ultimate goal is to regenerate reparative dentin, a physical barrier that functions as a "biological seal" and protects the underlying pulp tissue. Although this direct pulp-capping procedure has long been used in dentistry, the underlying molecular mechanism of pulpal wound healing and reparative dentin formation is still poorly understood. To induce reparative dentin, pulp capping has been performed experimentally in large animals, but less so in mice, presumably due to their small sizes and the ensuing technical difficulties. Here, we present a detailed, step-by-step method of performing a pulp-capping procedure in mice, including the preparation of a Class-I-like cavity, the placement of pulp-capping materials, and the restoration procedure using dental composite. Our pulp-capping mouse model will be instrumental in investigating the fundamental molecular mechanisms of pulpal wound healing in the context of reparative dentin in vivo by enabling the use of transgenic or knockout mice that are widely available in the research community.

We describe a step-by-step method of performing direct pulp capping on mice teeth for the evaluation of pulpal wound healing and reparative dentin formation in vivo.

Pulpa Değerlendirilmesi için Doğrudan Selüloz-kapatma Modelinin Geliştirilmesi Yara İyileşmesi ve Farelerde Onarım dentin oluşumu

Dental pulp is a vital organ of a tooth fully protected by enamel and dentin. When the pulp is exposed due to cariogenic or iatrogenic injuries, it is ...

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 ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

İzometrik Contractility ölçüm tel Myography kullanarak fare mezenterik arter

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

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.