Name: tape: a biodegradable hemostatic glue inspired by a ubiquitous compound in plants for surgical application
Uploaded: 2016-06-08T13:00:00.000+00:00
Duration: 8 min 40 s
Description: Watch this Scientific Journal Video about TAPE: A Biodegradable Hemostatic Glue Inspired by a Ubiquitous Compound in Plants for Surgical Application at JoVE.com

This study was supported by National Research Foundation of South Korea: Mid-career scientist grant (2014002855), and Ministry of Industry, Trade, and Natural Resources: World Premier Material Development Program. This work is also supported by in part by Center for Nature-inspired Technology (CNiT) in KAIST Institute for NanoCentury (KINC).

Tüm hayvan bakımı ve deneyler KAIST (Bilim ve Teknoloji Kore İleri Enstitüsü) tarafından sağlanan etik protokolü IRB (Institutional Review Board) göre yapılır. 1. ŞERİT Oluşumu <ol><li> Bir ta çözelti hazırlamak için, bir manyetik karıştırıcı üzerinde 4 mi boyutlu bir cam şişe yerleştirin ve bir karıştırma çubuğuna sahip olan damıtık su 1 ml. şişeye tannik asit 1 gr ve daha fazla 1 saat süre ile, 200 rpm&#39;de hafif karıştırma ile su içinde çözülür. TA, tamamen çözülür, karışım, kahverengi bir renk ile şeffaf hale gelir. </li><li> beyaz bir bulamaç yapmak için birkaç saniye girdap oluşturarak karıştırılarak damıtılmış su, 1 ml, ardından, PEG tozu, 1 g (4-kol, 10 kDa, ve doğrusal, 4.6 kDa) ilave edilerek, bir PEG solüsyonu hazırlayın. 10 dakika boyunca, 60 ° C&#39;de kuluçka, bu bulamacın tutun. Beyaz bir tamamen berrak hale gelinceye kadar. NOT: 10 kDa molekül ağırlığına sahip PEG erime noktası yaklaşık 55 olan- 60 ° C ve 4 kDa bir 53 olan - 58 ° C. 1 g kadar suda PEG yüksek bir konsantrasyon / ml berrak çözelti olarak elde edilebilir, böylece erimiş PEG suyla karışabilen olur. berrak bir PEG çözeltisi yüksek bir sıcaklıkta oluşturulduktan sonra, çözelti hala soğutulduktan sonra oda sıcaklığında stabildir. </li><li> PEG 329 ul ekle (4-kol, 10 kDa) çözeltisi 4.6 kDa olan bir doğrusal PEG durumunda adım 1.1 hazırlanan ta çözeltisi (1.2 671 ul bir PEG çözeltisi 311 ul ekleyin aşamasında hazırlanan bir mikro santrifüj tüpüne bir TA çözelti) 689 ul. Yavaşça homojen onları karıştırmak için dar bir spatula ile iki viskoz ve bal gibi çözümler karışır. DİKKAT: Her iki çözüm de oldukça viskoz, böylece bilim adamı yavaş ama yeterince yukarı çekin ve bir mikropipet ile çözüm aktarmanız gerekir. </li><li> sabit açılı rotor ile donatılmış bir santrifüjü içinde 3 dakika boyunca 12.300 x g&#39;de adım 1.3 elde edilen karışım dönerler. </li><li> dikkatle rembir mikropipet kullanarak mümkün olduğunca süpernatant kadar ove ve aşağı yerleşti ürünü toplamak: Bu tam olarak oluşmuş BANT olduğunu. . BANT oluştuktan sonra buzdolabında saklayın - birkaç hafta NOT (4 8 ° C): BANT gama radyasyonu ya da cerrahi uygulamalarda kullanım öncesi elektron ışın tedavisi ile sterilize edilebilir. </li></ol> TAPE yapışması Mukavemet 2. ölçümü <ol><li> Cilt dokusu üzerinde, tüm yağ çıkardıktan sonra biopsi darbe ucu ile kesilerek 6 mm bir çapa sahip olan domuz derisi doku iki adet hazırlayın. NOT: domuz deri dokusu sağlıklı domuz yan cilt elde edilmiştir ve Güney Kore Daejeon bulunan yerel et piyasadan satın alındı. </li><li> Her bir doku dış tarafına ticari siyanoakrilat yapıştırıcı uygulanır ve metal çubuk dokusu ekleyin. NOT: Metalik çubuk ek tanıtıcı yani dokular olarak kullanılırdoğrudan makine tarafından kavradı değil yeniden. Bu duruma göre, bu çekme makinesinin konfigürasyonunun aşağıdaki diğer malzemeler ile değiştirilebilir. </li><li> dokuda bir tarafında - TAPE bir damla uygulayın (6 mg TAPE bir damla yaklaşık 3). Daha sonra, Şekil 2A&#39;da görüldüğü gibi, bağlı oldukları şekilde eşit kendi iç tarafında iki doku arasındaki bir doku kullanılarak TAPE yayıldı. </li><li> Daha sonra el homojen karıştırın ve her doku ve TAPE arasındaki arayüzü en üst düzeye çıkarmak için birkaç kez takın ve dokuların iki tarafı ayırmak. </li><li> UTM ile çubuğun dikkatle kavrama her yan. yapışma gücü TAPE bağlı iki doku ayırmak için gereken kuvvet ile belirlenecektir. İlk olarak, 1 dakika için 20 N&#39;luk bir kuvvet uygulanır. Daha sonra, makine ile, 1 mm / dakika arasında bir oranda bir karşı yönde her çubuk çekin. kadar dokular tamamen ayrılır. NOT: Veriler bir kuvvet-mesafe olarak verilecektir (FD) eğrisi hareketi tarafından tespitHer bir çubuğun. </li><li> Yani, numune yüzey alanının adım 2.5 elde FD eğrisi gösterilen maksimum kuvvet (kN) bölünmesi ile TAPE yapışma gücünü hesaplamak, 3.14 x (0.003 m) 2. </li><li> suyun mevcudiyetinde yapışma gücünü kontrol için iki doku arasındaki ayrılmış alanda eden 20 ul ve hemen takın. makine ile, yine sıyrılma testi gerçekleştirin. </li></ol> 3. İn Vitro Parçalanma Testi <ol><li> Mikro santrifüj tüpünün bir kapak (D = 8 mm) kesilmiş ve W, C olarak tanımlamak için kapağı tartın. </li><li> TAPE 150 mg kapağını doldurun ve 0 W toplam başlangıç ​​ağırlığı olarak ayarlamak için tekrar hep birlikte tartın. DİKKAT: kap TAPE aşırı yüklemeyin. Bu adım 3.4 inkübasyon sırasında karıştırma işleminin elde edilen PBS tamponu akımı için fiziksel bir engel olması gibi TAPE yüksekliği, başlığın üst daha düşük olmalıdır. </li><li&gt; Bir hücre kültür şişesi içine bant içeren kap koyun (75 cm2), ve PBS tampon maddesi 50 ml (1 x, pH 7.4), hücre kültürü şişesine kapağa bandın tam olarak PBS tamponu batırılır, eklemek Şekil 3A&#39;da gösterilen (n = 5). </li><li> yumuşak karıştırma (50 rpm) ile fizyolojik koşullara benzer bir 37 ° C, orbital bir çalkalama inkübatöründe aşama 3.3&#39;te hazırlanan hücre kültürü şişesi, inkübe edin. DİKKAT: 50 rpm&#39;de karıştırma durumu tutun. Yüksek devir TAPE bir çöküş neden olabilir. </li><li> Her zaman noktasında, hücre kültürü şişesi TAPE kap almak ve daha sonra azot gazı üflenerek kurutun. Kalan TAPE içeren kap tartılır. W t her zaman noktasında ağırlığını ayarlayın. Tekrar taze PBS değiştirin ve her zaman noktasında W t ölçme sonra tekrar sallayın. </li><li> göreli kalan ağırlık (%) aşağıdaki denklem hesaplayın. Nispi kalan ağırlık (%) = (W t </sub&gt; - W, C) / (B 0 - W, C) x 100% </li></ol> TAPE 4. Hemostatik yeteneği NOT: Tüm hayvan deneyleri Sağlık ve Refah Kore Bakanlığı tarafından sağlanan yönergelere ve etik protokole uygun olarak yapılmalıdır. <ol><li> Ref 24 tarif edildiği gibi in vivo hemostatik yeteneğini değerlendirmek için, kanayan fare karaciğer modeli inceleyin. </li><li> (Grup başına n = 5) tiletamine-zolazepam (33,333 mg / kg) ve ksilazin (7,773 mg / kg) intraperitonal enjeksiyonu ile - beş fareleri (erkek 35 g, normal bir ICR fare, 6 hafta, 30) anestezisi. Uygun anesthetization doğrulamak için, hayvanın pençe hafifçe tutam ve hiçbir hareket hayvan cerrahisi yapmak için yeterli anestezi olduğunu gösterir vb, pençe çekilmesi gibi hareketleri gözlemleyin. </li><li> , Hayvanın gözlerinin kuruluğunu önlemek bir altında yeterince süre gözlerine veteriner merhem uygulamak içinestezia. orta hat insizyon yoluyla karaciğer Açığa ve kanama ikna etmek için bir 18 G iğne ile karaciğer delin. </li><li> steril gazlı bez ile akan kanın çıkarın ve kanama sitesinde hemen TAPE veya fibrin yapıştırıcı (pozitif kontrol) 100 ul koydu. NOT: Daha başka sütür nedeniyle yara dokuları üzerinde çok kan dayanıklı yapıştırıcı özellikleri TAPE uygulandıktan sonra ihtiyaç vardır. Negatif kontrol için, bir tedavi kanama bölgesinde meydana gelir. </li><li> Her durumda, hasar sitesinden kan toplamak için karaciğer altına bilinen kütleye sahip bir filtre kağıdı koyun. Taze bir 4 kez her 30 saniye (yani., 2 dk) ile kağıdı değiştirin. </li><li> her 30 saniyede toplanan her filtre kağıdı üzerinde emilen kan kitle ölçün. Hayvan deneyden sonra, CO 2 boğulma ötenazi yoluyla fareler kurban. </li></ol>

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The authors have nothing to disclose.

Bir bitkiden türetilmiş polifenolik bileşik, TA suya dayanıklı moleküler etkileşim esinlenerek hemostatik yapıştırıcı adlı TAPE tamamen yeni bir sınıf gelişti. TA belirgin nedeniyle, anti-oksidan, anti-bakteriyel, anti-mutajenik ve anti-kanserojen özellikleri dikkat çekti temsili hidrolize olabilen tanen olduğunu. herhangi bir başka kimyasal sentetik prosedürler olmadan santrifüj ardından iki sulu çözeltilerin sadece bir adım karıştırma gibi TAPE yapma süreci, s...

Geçmiş on yılda, çabaları nedeniyle cerrahi tedaviler sırasında kullanımı ve düşük doku invazivliğinde onların kolaylık biyolojik olarak parçalanabilen / biyo-emilebilir yapıştırıcılar ile yaraları kapatmak için mevcut cerrahi sütür ve zımba yerine yapılmıştır. Ticari olarak temin edilebilen doku yapıştırıcılar, dört tip olarak sınıflandırılırlar: (1) siyanoakrilat türevleri 1, (2) 2,3 trombin polimerlerin fibrinojenin fibrine gelen enzimatik dönüşümü ile oluşan fibrin tutkalları, (3) bu tür kimyasal olarak ya da fiziksel olarak protein-esaslı malzemeler çapraz bağlanmış albümini ve / ya da jelatin 4,5, ve (4) bir sentetik polimer bazlı olanlar 6. birçok klinik uygulamalarda kullanılmaktadır olmasına rağmen, tüm yapıştırıcılar, kendi içsel dezavantajları ve bunların yaygın kullanımına engel olabilir dezavantajları var. Siyanoakrilat bazlı yapıştırıcılar dokulara yüksek yapışma gücünü gösterir, ama onların zehirli yan ürünler gibi bozunma sırasında oluşan siyanoasetat ve formaldehit gibi, sık sık işaret nedeninflamatuar tepkilerin 7 ificant derece. Jelatin bazlı yapıştırıcıların 8 sığır, tavuk, domuz ve balık gibi fibrin yapıştırıcılar ve hayvanlar için insan kanı plazmasından: Fibrin yapıştırıcılar ve albümin veya jelatin bazlı malzemeler gibi hayvansal kaynaklardan virüsler gibi enfeksiyöz bileşenlerin iletimi ile ilgili güvenlik sorunları var. Bir kaç sentetik polimer bazlı yapıştırıcılar Federal İlaç İdaresi (FDA) tarafından onaylanmış olmasına rağmen, sentetik polimerlerin yapılmış en yapıştırıcılar üretim süreci adımlarını en aza indirmek ve biyouyumluluk 9 ulaşmada zorluklar yaşamaya devam. En önemlisi, tüm yapıştırıcılar ıslak mendil 10 zayıf mekanik ve yapışma mukavemeti muzdarip. Son zamanlarda, deniz midye 11-13, kertenkeleleri 14, midye 15 kertenkele ve Endoparazitik solucanlar 16 esinlenerek biomimetic doku yapıştırıcıları nedeniyle ayarlanabilir mekanik ve mevcut tıbbi yapıştırıcı olarak umut verici alternatifler ortaya çıkmaktadırlarbiyouyumluluk ile yapışkan özellikleri. Onlar ticari ürünler 17 haline gelmeden önce Ancak, bu güne kadar hala ele alınması gereken konular vardır. Burada, bir bitki-türevi yapıştırıcı molekülü, tanik asit (TA), ve bir biyo-inert bir polimer poli (etilen glikol) (PEG) arasındaki moleküller arası hidrojen bağları yoluyla hazırlanır BANT da adlandırılır tutkal tamamen yeni türden bir rapor Adından da anlaşılacağı gibi. TA yayg bitkilerin ikincil metabolizma sırasında bulunan temsili hidrolize olabilen tanen olduğunu. Bu, anti-mutajenik ve anti-kanserojen özellikleri nedeniyle, anti-oksidan çok ilgi çekmektedir ve poli (N -isopropylacrylamide) (PNIPAM) ve poli (N birçok polimer ile supramoleküler etkileşimlerine dahil olduğu gösterilmiştir - vinilpirolidon) (PVPON), katman-katman (Lbl) filmleri 18-20 ve ilaç salan mikrokapsüller 21-23 oluşturulur. Bu çalışmada, biz ta verimli bir şekilde hareket edebilir keşfetmeksuya dayanıklı yapıştırıcı fonksiyonel kısım, TAPE tıbbi yapıştırıcı oluşturmak için. Ta basit karıştırma ile, non-kirlenme polimeri, PEG ile supramoleküler yapışkan hale 2.5 kat ticari fibrin yapıştırıcı ile karşılaştırıldığında artan yapışma gücü ve bu yapışma da suyun varlığında, bağlanma ve ayrılma kadar 20 döngü boyunca devam edildi . Onun hemostatik yeteneği in vivo bir karaciğer kanaması modeli üzerinde test edilmiş ve birkaç saniye içinde kanamayı durdurmak için iyi bir hemostatik yeteneğini gösterdi. TAPE biyo-esinlenmiş yaklaşımlarla mevcut sorunlarının sakıncaları çözme içine yeni anlayış ortaya çıkarabilir ilk bitkisel kaynaklı yapıştırıcı olarak ilgili alandaki önemli bir anlamı vardır. Biz de nedeniyle basit hazırlama yöntemi, ölçeklenebilirlik, ayarlanabilir biyodegradasyon oranı gibi muko-yapıştırıcılar, ilaç salan yamalar, yara bakımı pansuman, ve diğerleri gibi tıp ve eczacılık uygulamalarında çeşitli TAPE yaygın kullanımını bekliyoruz yanı sıra yüksek ıslak dayanıklı Kuvvetli Yapıştırıcıiyon özellikleri.

<table><tbody><tr><td>Tannic acid</td><td>Sigma-aldrich</td><td>403040</td><td></td></tr><tr><td>Poly(ethylene oxide), 4-arm, hydroxy terminated</td><td>Aldrich</td><td>565709</td><td>Averge Mn ~ 10,000</td></tr><tr><td>Poly(ethylene glycol)</td><td>Aldrich</td><td>373001</td><td>Average Mn 4,600</td></tr><tr><td>Biopsy punch</td><td>Miltex</td><td>33-36</td><td>Diameter = 6 mm</td></tr><tr><td>Aron Alpha</td><td>Toagosei Co., Ltd.</td><td></td><td>Instant glue</td></tr><tr><td>Universal testing machine (UTM)</td><td>Instron</td><td>5583</td><td></td></tr><tr><td>Microcentrifuge tubes</td><td>SPL life science</td><td>60015</td><td>1.5 ml</td></tr><tr><td>Petri dish</td><td>SPL life science</td><td>10090</td><td>90 x 15 mm</td></tr><tr><td>Sodium phosphate monobasic</td><td>Sigma</td><td>S5011</td><td>1x PBS ingredient</td></tr><tr><td>Sodium phosphate dibasic</td><td>Sigma</td><td>S5136</td><td>1x PBS ingredient</td></tr><tr><td>Sodium chloride</td><td>Duchefa biochemie</td><td>S0520.5000</td><td>1x PBS ingredient</td></tr><tr><td>Incubating shaker</td><td>Lab companion</td><td>SIF6000R</td><td></td></tr><tr><td>ICR mice</td><td>Orient bio</td><td>Normal ICR mouse</td><td>6 weeks, 30 - 35 g, male</td></tr><tr><td>Tiletamine-zolazepam (Zoletil 50)</td><td>Virbac</td><td></td><td></td></tr><tr><td>Zylazine (Rompun)</td><td>Bayer</td><td></td><td></td></tr><tr><td>PrecisionGlide&lt;sup&gt;TM&lt;/sup&gt; needle (18 G)</td><td>BD</td><td>302032</td><td>18 G</td></tr><tr><td>Filter paper</td><td>Whatman</td><td>1001 125</td><td>Diameter = 125 mm</td></tr><tr><td>Parafilm</td><td>Bemis Flexible Pakaging</td><td>PM996</td><td></td></tr></tbody></table>

tape: a biodegradable hemostatic glue inspired by a ubiquitous compound in plants for surgical application

Bu video, ticari doku yapıştırıcıları daha etkili bir hemostatik yeteneği ve daha suya dayanıklı yapışma gücüne sahiptir biyobozunur cerrahi yapıştırıcı hazırlanması için basit protokol tanımlamaktadır. Tıbbi yapıştırıcılar nedeniyle minimal invaziv özelliğe sahip kullanımı onların kolaylık sütür ve zımba potansiyel alternatif araçları gibi büyük ilgi çekmiştir. biyo-odaklı malzemeler durumunda örneğin, fibrin tutkalları ve siyanoakrilat bazlı malzemeler ticari olarak temin edilebilen, daha çok organik moleküller ya da karmaşık bir protein saflaştırma yöntemleri kimyasal sentezler, bir dizi ihtiyaç dahil olmak üzere doku yapıştırıcı geliştirmek için çeşitli protokoller, olmasına rağmen (yani, fibrin yapıştırıcı). Ayrıca, cerrahi yapıştırıcı geliştirme yıkma muhafaza nedeniyle vücudun ıslak bir ortamda iyi bir performans sağlamada zorluklar hala bir meydan okuma ise yüksek yapışma özellikleri sergileyen. Bir hazırlamak için yeni bir yöntem gösterilmektedirbitkisel kökenli bir ıslak-dirençli bir yapıştırıcı molekülün fiziksel karıştırma sonrasında oluşturulan bir su ile karışmayan supramoleküler agrega ağırlığı bazlı ayırma ile, bant olarak bilinen tıbbi yapıştırıcı, T, bir CID (TA), ve iyi bilinen Annic biyopolimer, poli (etilen) glikol (PEG) içerebilir. Bizim yaklaşımla, BANT su varlığında ticari fibrin yapıştırıcı daha 2.5 kat yüksek yapışma gücü gösterir. Ayrıca, BANT fizyolojik koşullarda biyolojik olarak parçalanabilir ve doku kanamaya karşı güçlü bir hemostatik yapıştırıcı olarak kullanılabilir. Böyle muko-yapışması için polimerler, ilaç deposu, ve diğerleri gibi tıbbi ortamlarda ve ilaç verme uygulamalarında, çeşitli TAPE yaygın kullanımı bekliyoruz.

1 lik hacim oranında (Şekil 1A): BANT TA (1 ug / ml damıtılmış su içinde) ve 2 PEG (1 ug / ml damıtılmış su içinde) içeren iki sulu solüsyonun karışımını santrifüjden sonra yerleşir bir supramoleküler toplamıdır. karışım oranı yüksek yapışma mukavemeti ulaşmada önemli bir faktördür; BANT 2 ile oluşturulur: 1 olduğunda karıştırma oranında, ta 25 adet hidroksil grubu (-OH) ile 20 tane fazla en yüksek moleküller arası hidrojen ba?...

Watch this Scientific Journal Video about TAPE: A Biodegradable Hemostatic Glue Inspired by a Ubiquitous Compound in Plants for Surgical Application at JoVE.com

TAPE: A Biodegradable Hemostatic Glue Inspired by a Ubiquitous Compound in Plants for Surgical Application

Biz etkin bir hemostatik yeteneğine sahip biyobozunur tıbbi tutkal hazırlamak için basit protokol açıklar. ŞERİT tannik asit karıştırılması ile hazırlanan bir suyla karışmayan supramoleküler agrega olan ticari fibrin yapıştırıcı ile karşılaştırıldığında 2.5 kat daha fazla suya dayanıklı yapışma, sonuçta, her yerde bulunan bir bileşik, bitkilerde bulunan ve poli (etilen) glikol.

BANT: Cerrahi Uygulama için Bitkilerde bir Ubiquitous Bileşik esinlenerek bir Biyobozunur Hemostatik Tutkal

This video describes the simplest protocol for preparing biodegradable surgical glue that has an effective hemostatic ability and greater water-resistant ...

tape-biodegradable-hemostatic-glue-inspired-ubiquitous-compound

Research

JoVE Journal

Bioengineering

14.1K Görüntüleme.  Korea Advanced Institute of Science and Technology (KAIST).  Biz etkin bir hemostatik yeteneğine sahip biyobozunur tıbbi tutkal hazırlamak için basit protokol açıklar. ŞERİT tannik asit karıştırılması ile hazırlanan bir suyla karışmayan supramoleküler agrega olan ticari fibrin yapıştırıcı ile karşılaştırıldığında 2.5 kat daha fazla suya dayanıklı yapışma, sonuçta, her yerde bulunan bir bileşik, bitkilerde bulunan ve poli (etilen) glikol. 

Video: TAPE: A Biodegradable Hemostatic Glue Inspired by a Ubiquitous Compound in Plants for Surgical Application

Engineering Biological-Based Vascular Grafts Using a Pulsatile Bioreactor

Much effort has been devoted to develop and advance the methodology to regenerate functional small-diameter arterial bypasses. In the physiological environment, both mechanical and chemical stimulation are required to maintain the proper development and functionality of arterial vessels1,2.
Bioreactor culture systems developed by our group are designed to support vessel regeneration within a precisely controlled chemo-mechanical environment mimicking that of native vessels. Our bioreactor assembly and maintenance procedures are fairly simple and highly repeatable3,4. Smooth muscle cells (SMCs) are seeded onto a tubular polyglycolic acid (PGA) mesh that is threaded over compliant silicone tubing and cultured in the bioreactor with or without pulsatile stimulation for up to 12 weeks. There are four main attributes that distinguish our bioreactor from some predecessors. 1) Unlike other culture systems that simulate only the biochemical surrounding of native blood vessels, our bioreactor also creates a physiological pulsatile environment by applying cyclic radial strain to the vessels in culture. 2) Multiple engineered vessels can be cultured simultaneously under different mechanical conditions within a controlled chemical environment. 3) The bioreactor allows a mono layer of endothelial cells (EC) to be easily coated onto the luminal side of engineered vessels for animal implantation models. 4) Our bioreactor can also culture engineered vessels with different diameter size ranged from 1 mm to 3 mm, saving the effort to tailor each individual bioreactor to fit a specific diameter size. 
The engineered vessels cultured in our bioreactor resemble native blood vessels histologically to some degree. Cells in the vessel walls express mature SMC contractile markers such as smooth muscle myosin heavy chain (SMMHC)3. A substantial amount of collagen is deposited within the extracellular matrix, which is responsible for ultimate mechanical strength of the engineered vessels5. Biochemical analysis also indicates that collagen content of engineered vessels is comparable to that of native arteries6. Importantly, the pulsatile bioreactor has consistently regenerated vessels that exhibit mechanical properties that permit successful implantation experiments in animal models3,7. Additionally, this bioreactor can be further modified to allow real-time assessment and tracking of collagen remodeling over time, non-invasively, using a non-linear optical microscopy (NLOM)8. To conclude, this bioreactor should serve as an excellent platform to study the fundamental mechanisms that regulate the regeneration of functional small-diameter vascular grafts.

Our group has developed a bioreactor culture system that mimics the physiological pulsatile stresses of the cardiovascular system to regenerate implantable small-diameter vascular grafts.

Pulsatif Biyoreaktör kullanarak Mühendislik Biyolojik Tabanlı Vasküler Greftler

Much effort has been devoted to develop and advance the methodology to regenerate functional small-diameter arterial bypasses. In the physiological ...

Biyomühendislik

A Chitosan Based, Laser Activated Thin Film Surgical Adhesive, 'SurgiLux': Preparation and Demonstration

Sutures are a 4,000 year old technology that remain the 'gold-standard' for wound closure by virtue of their repair strength (~100 KPa). However, sutures can act as a nidus for infection and in many procedures are unable to effect wound repair or interfere with functional tissue regeneration.1 Surgical glues and adhesives, such as those based on fibrin and cyanoacrylates, have been developed as alternatives to sutures for the repair of such wounds. However, current commercial adhesives also have significant disadvantages, ranging from viral and prion transfer and a lack of repair strength as with the fibrin glues, to tissue toxicity and a lack of biocompatibility for the cyanoacrylate based adhesives. Furthermore, currently available surgical adhesives tend to be gel-based and can have extended curing times which limit their application.2 Similarly, the use of UV lasers to facilitate cross-linking mechanisms in protein-based or albumin 'solders' can lead to DNA damage while laser tissue welding (LTW) predisposes thermal damage to tissues.3 Despite their disadvantages, adhesives and LTW have captured approximately 30% of the wound closure market reported to be in excess of US $5 billion per annum, a significant testament to the need for sutureless technology.4
In the pursuit of sutureless technology we have utilized chitosan as a biomaterial for the development of a flexible, thin film, laser-activated surgical adhesive termed 'SurgiLux'. This novel bioadhesive uses a unique combination of biomaterials and photonics that are FDA approved and successfully used in a variety of biomedical applications and products. SurgiLux overcomes all the disadvantages associated with sutures and current surgical adhesives (see Table 1).
In this presentation we report the relatively simple protocol for the fabrication of SurgiLux and demonstrate its laser activation and tissue weld strength. SurgiLux films adhere to collagenous tissue without chemical modification such as cross-linking and through irradiation using a comparatively low-powered (120 mW) infrared laser instead of UV light. Chitosan films have a natural but weak adhesive attraction to collagen (~3 KPa), laser activation of the chitosan based SurgiLux films emphasizes the strength of this adhesion through polymer chain interactions as a consequence of transient thermal expansion.5 Without this 'activation' process, SurgiLux films are readily removed.6-9 SurgiLux has been tested both in vitro and in vivo on a variety of tissues including nerve, intestine, dura mater and cornea. In all cases it demonstrated good biocompatibility and negligible thermal damage as a consequence of irradiation.6-10

The fabrication of a novel, flexible thin film surgical adhesive from FDA approved ingredients, chitosan and indocyanine green is described. Bonding of this adhesive to collagenous tissue through a simple activation process with a low-powered infra-red laser is demonstrated.

Bir Kitosan Bazlı, Lazer Aktif İnce Film Cerrahi Yapıştırıcı, &#39;SurgiLux&#39;: Hazırlık ve Gösteri

Sutures are a 4,000 year old technology that remain the 'gold-standard' for wound closure by virtue of their repair strength (~100 KPa). However, sutures ...

Engineering a Bilayered Hydrogel to Control ASC Differentiation

Natural polymers over the years have gained more importance because of their host biocompatibility and ability to interact with cells in vitro and in vivo. An area of research that holds promise in regenerative medicine is the combinatorial use of novel biomaterials and stem cells. A fundamental strategy in the field of tissue engineering is the use of three-dimensional scaffold (e.g., decellularized extracellular matrix, hydrogels, micro/nano particles) for directing cell function. This technology has evolved from the discovery that cells need a substrate upon which they can adhere, proliferate, and express their differentiated cellular phenotype and function 2-3. More recently, it has also been determined that cells not only use these substrates for adherence, but also interact and take cues from the matrix substrate (e.g., extracellular matrix, ECM)4. Therefore, the cells and scaffolds have a reciprocal connection that serves to control tissue development, organization, and ultimate function. Adipose-derived stem cells (ASCs) are mesenchymal, non-hematopoetic stem cells present in adipose tissue that can exhibit multi-lineage differentiation and serve as a readily available source of cells (i.e. pre-vascular endothelia and pericytes). Our hypothesis is that adipose-derived stem cells can be directed toward differing phenotypes simultaneously by simply co-culturing them in bilayered matrices1. Our laboratory is focused on dermal wound healing. To this end, we created a single composite matrix from the natural biomaterials, fibrin, collagen, and chitosan that can mimic the characteristics and functions of a dermal-specific wound healing ECM environment.

This protocol focuses on utilizing the inherent ability of stem cells to take cue from their surrounding extracellular matrix and be induced to differentiate into multiple phenotypes. This methods manuscript extends our description and characterization of a model utilizing a bilayered hydrogel, composed of PEG-fibrin and collagen, to simultaneously co-differentiate adipose-derived stem cells1.

ASC farklılaşma kontrol için bir iki tabakalı Hidrojel mühendislik

Natural polymers over the years have gained more importance because of their host biocompatibility and ability to interact with cells in vitro and in ...

Fabrication and Application of Rose Bengal-chitosan Films in Laser Tissue Repair

Photochemical tissue bonding (PTB) is a sutureless technique for tissue repair, which is achieved by applying a solution of rose bengal (RB) between two tissue edges1,2. These are then irradiated by a laser that is selectively absorbed by the RB. The resulting photochemical reactions supposedly crosslink the collagen fibers in the tissue with minimal heat production3. In this report, RB has been incorporated in thin chitosan films to fabricate a novel tissue adhesive that is laser-activated. Adhesive films, based on chitosan and containing ~0.1 wt% RB, are fabricated and bonded to calf intestine and rat tibial nerves by a solid state laser (&lambda;=532 nm, Fluence~110 J/cm2, spot size~0.5 cm). A single-column tensiometer, interfaced with a personal computer, is used to test the bonding strength. The RB-chitosan adhesive bonds firmly to the intestine with a strength of 15 &plusmn; 6 kPa, (n=30). The adhesion strength drops to 2 &plusmn; 2 kPa (n=30) when the laser is not applied to the adhesive. The anastomosis of tibial nerves can be also completed without the use of sutures. A novel chitosan adhesive has been fabricated that bonds photochemically to tissue and does not require sutures.

Sutures are usually needed to repair tissue during surgical procedures. However, their application can be problematic as they are invasive and may damage tissue. The fabrication and application methods of a novel tissue adhesive are here reported. This adhesive film is laser-activated and does not require the use of sutures.

Fabrikasyon ve Lazer Doku Onarımı Rose Bengal-kitosan Film Uygulamaları

Photochemical tissue bonding (PTB) is a sutureless technique for tissue repair, which is achieved by applying a solution of rose bengal (RB) between two ...

Pluripotent Stem Cell Derived Cardiac Cells for Myocardial Repair

Human induced pluripotent stem cells (hiPSCs) must be fully differentiated into specific cell types before administration, but conventional protocols for differentiating hiPSCs into cardiomyocytes (hiPSC-CMs), endothelial cells (hiPSC-ECs), and smooth muscle cells (SMCs) are often limited by low yield, purity, and/or poor phenotypic stability. Here, we present novel protocols for generating hiPSC-CMs, -ECs, and -SMCs that are substantially more efficient than conventional methods, as well as a method for combining cell injection with a cytokine-containing patch created over the site of administration. The patch improves both the retention of the injected cells, by sealing the needle track to prevent the cells from being squeezed out of the myocardium, and cell survival, by releasing insulin-like growth factor (IGF) over an extended period. In a swine model of myocardial ischemia-reperfusion injury, the rate of engraftment was more than two-fold greater when the cells were administered with the cytokine-containing patch comparing to the cells without patch, and treatment with both the cells and the patch, but not with the cells alone, was associated with significant improvements in cardiac function and infarct size.

We present three novel and more efficient protocols for differentiating human induced pluripotent stem cells into cardiomyocytes, endothelial cells, and smooth muscle cells and a delivery method that improves the engraftment of transplanted cells by combining cell injection with patch-mediated cytokine delivery.

Pluripotent Kök Hücre Miyokard Onarım Kardiyak Hücreleri Türetilmiş

Human induced pluripotent stem cells (hiPSCs) must be fully differentiated into specific cell types before administration, but conventional protocols for ...

Gelişim Biyolojisi

Characterization of Leukocyte-platelet Rich Fibrin, A Novel Biomaterial

Autologous platelet concentrates represent promising innovative tools in the field of regenerative medicine and have been extensively used in oral surgery. Unlike platelet rich plasma (PRP) that is a gel or a suspension, Leukocyte-Platelet Rich Fibrin (L-PRF) is a solid 3D fibrin membrane generated chair-side from whole blood containing no anti-coagulant. The membrane has a dense three dimensional fibrin matrix with enriched platelets and abundant growth factors. L-PRF is a popular adjunct in surgeries because of its superior handling characteristics as well as its suturability to the wound bed. The goal of the study is to demonstrate generation as well as provide detailed characterization of relevant properties of L-PRF that underlie its clinical success.

Leucocyte-Platelet Rich Fibrin (L-PRF) represents an FDA cleared preparation of autologous platelet concentrates that possesses unique fibrin architecture, enriched platelets and abundant growth factors. Here, we present a protocol for chair-side generation of L-PRF as well as evaluate its mechanical properties including uniaxial testing and suture retention strength testing.

Lökosit-trombosit Zengin Fibrin, Yeni Bir Biyomateryalinin Karakterizasyonu

Autologous platelet concentrates represent promising innovative tools in the field of regenerative medicine and have been extensively used in oral ...

Polyelectrolyte Complex for Heparin Binding Domain Osteogenic Growth Factor Delivery

During reconstructive bone surgeries, supraphysiological amounts of growth factors are empirically loaded onto scaffolds to promote successful bone fusion. Large doses of highly potent biological agents are required due to growth factor instability as a result of rapid enzymatic degradation as well as carrier inefficiencies in localizing sufficient amounts of growth factor at implant sites. Hence, strategies that prolong the stability of growth factors such as BMP-2/NELL-1, and control their release could actually lower their efficacious dose and thus reduce the need for larger doses during future bone regeneration surgeries. This in turn will reduce side effects and growth factor costs. Self-assembled PECs have been fabricated to provide better control of BMP-2/NELL-1 delivery via heparin binding and further potentiate growth factor bioactivity by enhancing in vivo stability. Here we illustrate the simplicity of PEC fabrication which aids in the delivery of a variety of growth factors during reconstructive bone surgeries.

Self-assembled polyelectrolyte complexes (PEC) fabricated from heparin and protamine were deposited on alginate beads to entrap and regulate the release of osteogenic growth factors. This delivery strategy enables a 20-fold reduction of BMP-2 dose in spinal fusion applications. This article illustrates the benefits and fabrication of PECs.

Heparin Binding Domain osteojenik Büyüme Faktörü Teslim polielektrolit Kompleksi

During reconstructive bone surgeries, supraphysiological amounts of growth factors are empirically loaded onto scaffolds to promote successful bone ...

A Hydrogel Construct and Fibrin-based Glue Approach to Deliver Therapeutics in a Murine Myocardial Infarction Model.

The murine MI model is widely recognized in the field of cardiovascular disease, and has consistently been used as a first step to test the efficacy of treatments in vivo1. The traditional, established protocol has been further fine-tuned to minimize the damage to the animal. Notably, the pectoral muscle layers are teased away rather than simply cut, and the thoracotomy is approached intercostally as opposed to breaking the ribs in a sternotomy, preserving the integrity of the ribcage. With these changes, the overall stress on the animal is decreased. 
Stem cell therapies aimed to alleviate the damage caused by MIs have shown promise over the years for their pro-angiogenic and anti-apoptotic benefits. Current approaches of delivering cells to the heart surface typically involve the injection of the cells either near the damaged site, within a coronary artery, or into the peripheral blood stream2-4. While the cells have proven to home to the damaged myocardium, functionality is limited by their poor engraftment at the site of injury, resulting in diffusion into the blood stream5. This manuscript highlights a procedure that overcomes this obstacle with the use of a cell-encapsulated hydrogel patch. The patch is fabricated prior to the surgical procedure and is placed on the injured myocardium immediately following the occlusion of the left coronary artery. To adhere the patch in place, biocompatible external fibrin glue is placed directly on top of the patch, allowing for it to dry to both the patch and the heart surface. This approach provides a novel adhesion method for the application of a delicate cell-encapsulating therapeutic construct.

This protocol aims to alleviate the limitation of poor cell engraftment for stem cell treatment of myocardial infarctions through the use of a hydrogel system and a fibrin-based glue. With this approach, cell-to-tissue contact post-infarction can be maintained, increasing the therapeutic potential of beneficial agents at the site of injury.

Bir Hidrojel Construct ve Fare Miyokard Enfarktüsü Modeli Therapeutics sunun için Tutkal Yaklaşım Fibrin tabanlı.

The murine MI model is widely recognized in the field of cardiovascular disease, and has consistently been used as a first step to test the efficacy of ...

Tıp

Synthesis of Strong Adhesive Hydrogel, Gelatin O-Nitrosobenzaldehyde

Adhesive materials have become popular biomaterials in the field of biomedical and tissue engineering. In our previous work, we presented a new material - gelatin o-nitrosobenzaldehyde (gelatin-NB) - which is mainly used for tissue regeneration and has been validated in animal models of corneal injury and inflammatory bowel disease. This is a novel hydrogel formed by modifying biological gelatin with o-nitrosobenzaldehyde (NB). Gelatin-NB was synthesized by activating the carboxyl group of NB-COOH and reacting with gelatin through 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). The obtained compound was purified to generate the final product, which can be stably stored for at least 18 months. NB has a strong adhesion to -NH2 on the tissue, which can form many C = N bonds, thus increasing the adhesion of gelatin-NB to the tissue interface. The preparation process comprises steps for the synthesis of the NB-COOH group, modification of the group, synthesis of gelatin-NB, and purification of the compound. The goal is to describe the specific synthesis process of gelatin-NB in detail and to demonstrate the application of gelatin-NB to damage repair. Moreover, the protocol is presented to further strengthen and expand the nature of the material produced by the scientific community for more applicable scenarios.

The protocol presented here shows the synthesis of a strong adhesive hydrogel gelatin o-nitrosobenzaldehyde (gelatin-NB). Gelatin-NB has rapid and efficient tissue adhesion ability, which can form a strong physical barrier to protect wound surfaces, so it is expected to be applied to the field of injury repair biotechnology.

Güçlü yapışkan hidrojel, jelatin o-nitrosobenzaldehit sentezi

Adhesive materials have become popular biomaterials in the field of biomedical and tissue engineering. In our previous work, we presented a new material - ...

Occlusion of the Great and Small Saphenous Vein Using Copolymeric Glue Based on N-Butyl Cyanoacrylate and Methacryloxy Sulfolane

We present the preliminary results of a longitudinal observational study aimed at evaluating the effectiveness and safety at different short- and long-term follow-ups of vascular occlusion of the great saphenous vein (GSV) and small saphenous vein (SSV) affected by severe pathological reflux, using an innovative modified cyanoacrylate surgical glue composed of N-butyl cyanoacrylate and methacryloxy sulfolane (NBCA+MS). Ninety patients, prospectively recruited for 1 year, underwent the study with EcoColor-Doppler (ECD) to evaluate the maximum diameters of the GSV and SSV in the orthostatic position and the reflux time (RT). An RT greater than 0.5 s was considered pathologic. Clinical, etiology, anatomy, and pathophysiology (CEAP) assessment was used for the complete evaluation of each patient in the study. All the patients were treated by NBCA+MS glue to obtain vein occlusion and observed before treatment (baseline; T0), within 6 h after treatment (T1), 1 month after treatment (T2), 3 months after treatment (T3), 6 months after treatment (T4), and 1 year after treatment (T5). Chi-square (&#967;) analysis was performed to evaluate the effectiveness and safety of the treatment. All the patients participated in the entire duration of the study. Complete occlusion was maintained in 100% of patients at T1, 98.9% at T2 and T3, and 97.8% at T4 and T5 (p &lt; 0.001). None of the patients suffered from post-surgical thrombosis. No blue hyperpigmentation, or paresthesia was observed during the entire observation period. Immediately after treatment, 7.7% of patients needed painkillers; 1 week after treatment, 100% of patients returned to normal life. Vascular occlusion of the great or small saphenous vein using NBCA+MS glue is a safe procedure with persistent benefits after a 1 year follow-up. This procedure can be performed with local anesthesia, allowing a quick return to normal life. Thanks to its low invasiveness, the treatment is not painful.

Here, we present a protocol to treat the great saphenous vein (GSV) and the small saphenous vein (SSV) affected by severe pathological reflux, using an original sclerosing and embolizing cyanoacrylate-based glue composed of N-butyl cyanoacrylate and methacryloxy sulfolane (NBCA+MS).

BANT: Cerrahi Uygulama için Bitkilerde bir Ubiquitous Bileşik esinlenerek bir Biyobozunur Hemostatik Tutkal

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