Bu çalışma, mali İsviçre Ulusal Bilim Vakfı (- Akıllı Malzeme NRP 62) tarafından desteklenmiştir. Ayrıca B. Hanselmann, K. Kehl, U. Schütz ve B. LEUTHOLD desteği olduğunu kabul etti.

1.. Plazma başlatılan Polimerleşmesi <ol><li> Monomer çözeltisinin hazırlanması. <ol><li> 200 ml su içinde ve bir ayırma hunisinde heksan (100 mi) ile 3 kere yıkayın, HEMA (0,718 mol 100 mi) içinde çözülür. Sodyum klorid ile sulu faz Doymuş ve dietil eter (50 mi) ile HEMA ekstrakte edin. MgSO 4 üzerinde organik faz kurutulur ve çözücü madde vakum içinde (100 mbar, 40 ° C) çıkarın. Indirgenmiş basınç altında distile HEMA (15 mbar, 99 ° C). </li><li> Bölüm 1.1.1 &#39;de üretilen inhibitör içermeyen HEMA bir 0.62 M metanolik çözelti hazırlayın. Bir-boyunlu şişe içine, çözeltinin 30 ml dökün ve 1 saat için çözelti içinden Argon geçirmek suretiyle oksijen ortadan kaldırır. </li></ol></li><li> Yüzey indüklenen polimerizasyonu. <ol><li> Pozisyon yanyana, plazma odası, iki polikarbonat zarları (Şekil 2). Gaz fazı doğru işaret zarın parlak yan. </li><li> Ah, plazma odası geç5 dakika boyunca üksek vakum (20 mbar). Vakum vanasını kapatın ve bir argon ve oksijen gazı bağlı olan ve 15 sccm argon ve 2.5 sccm oksijen ile 2 saat boyunca bu karışımı ile tasfiye edilir odacığı, diğer valf, açın. </li><li> Plazmayı başlatmak ve (polikarbonat membran için: 12 W) istenen güce etkisini engeller ve plazma ile, 4 dakika için membranlar tedavi. Karşılık gelen valf, açılarak bölmenin ile monomer çözeltisi bağlayın. Plazma kapatın ve odasını tahliye. </li><li> Karşılık gelen valf, açılarak bölmenin ile monomer çözeltisi bağlayın ve bölmenin içine solüsyonu dökün. Zarlar monomer çözeltisi ile kaplı olduğundan emin olun. Argon bağlı vanasını açın ve 20 ° C (oda durumunu) 12 saat boyunca reaksiyon karışımının saklayın. </li><li> Monomer çözeltisi çıkarın. 5 dakika boyunca ultrasonik bir banyo içinde, metanol ile zarları yıkayın. Su ile yıkama işlemi tekrar edin. </li><li> In membran Kuruvakum altında 2 saat boyunca moleküler elekler üzerinde. </li></ol></li></ol> 2. Kaplamalı Membran Postmodification <ol><li> Tert-butylmetylether ve dimetil-aminopiridin (DMAP) (33 mg, 0.27 mmol), N, N-disikloheksilkarbodiimid (DCC) (0.27 mmol 55 mg); spirobenzopiran 1 ihtiva eden bir çözelti (Şekil 1) (0.27 mmol 100 mg) hazırlanması (TBME) (12 mi). </li><li> , Yuvarlak dipli bir şişeye bir karıştırma çubuğu ve koruma, bir koruyucu ızgara yerleştirin. Şişeyi kurutun ve argon ile şişe sel. </li><li> Kaplanmış zar, ardından şişeye, çözelti içine dökün. </li><li> 12 saat boyunca oda sıcaklığında yavaşça karıştırın. </li><li> Çözeltisi çıkarın ve 5 dakika boyunca ultrasonik bir banyoda tert-butylmetylether ile membran yıkayın. , Etanol ve su ile yıkama işlemini tekrarlayın. </li><li> 2 saat boyunca moleküler elekler üzerinde vakum içinde membran kurutun. </li></ol> 3. Yüzey Gerilimi Ölçümler <ol><li&gt; metal O-ring üzerinde standart bir bant ile membran Fix. O-halkası ile temas halinde değildir, zar yüzeyinde, bir kısmının üzerine nanopure, su (3.3 ul), bir damla yerleştirin. Membranın 5 farklı noktalarda temas açısı (CA) ölçün. </li><li> Numunelerin uzun süreli stabilitesini test etmek için, üç farklı 0 sonra membran noktalar, 1, 2, 3, 7, 14, ve 21 gün temas açılarını ölçmek. </li></ol> 4. Fotokromık Membranlarının geçirgenliği Testleri <ol><li> Su ile Franz difüzyon hücresi (12 mi) reseptör odası doldurun. </li><li> Bir Franz difüzyon hücresine membran düzeltildi. Zar reseptör haznesinin su ile temas halinde olduğundan emin olun. Sulu kafein çözeltisi (; 3.0 ml 20 mM) ile verici odası (membran üstünde odası) doldurun. Beyaz ışık verici bölmesi (Şekil 3) üst zarı ışın tedavisi. Reseptör hücre örnekleri (200 ul) toplamak; fo200 nm gözenek çapına sahip r parça kenarlı polikarbonat zarları, numuneleri her 10 dakikada bir toplar. </li><li> Adım 4.2 &#39;de anlatıldığı gibi deney tekrarlayın. ancak tüm geçirgenlik test esnasında UV ışığı (366 nm, 80 W / m 2) ile membran ışın. </li><li> Toplanan numunelerin kafein konsantrasyonunun belirlenmesi. <ol><li> UV / Vis spektrometre kullanılarak (ml 0.05 mg / ve 1.5 mM / L arasında) 15 farklı kafein konsantrasyonları ile bir kalibrasyon eğrisi çizilir. 293 nm &#39;de kalibre edin. </li><li> Kalibrasyon eğrisi kullanılarak toplanan numunelerin her konsantrasyonunu belirleyin. </li><li> Vs belirlenen konsantrasyonu çizilir. Toplanan numunelerin saati. Noktaları aracılığıyla doğrusal bir uyum yapmak ve yamaç Δc belirler. </li></ol></li></ol>

<ol>
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Yazarlar ifşa hiçbir şey yok.

Plazma işlemi, iyonize argon neden olduğu mor bir gaz üretir. Bir turuncu renk bir sızıntı istenmeyen azot varlığını işaret eder. Plazma işlemi, sadece yüzeyde değil, aynı zamanda, kökler oluşturan membran 7,12 çözünen bir madde değildir. Çok fazla aşındırma, zarın geçirgenliğinin etkileyecek olan, önemli ölçüde gözenek çapına değiştirebilir. Sunulan kurulumunun kontrollü reaksiyon koşulları, plazma tarafından başlatılan aşılama işlemi tekrarlanabilirliğini artt?...

Malzemelerin plazma modifikasyonu birçok sanayi alanları önemli bir süreç haline gelmiştir. Malzemenin yığın özelliğini değiştirmeden yüzeyler ve yüzeylerin işlevsellik temizlemesi plazma işleme yüzey bilimi 1-8 önemli bir süreç haline gelmiştir. Polimerlerin plazma tedavi homolytic bağ bölünmesi ile sonuçlanır. Bunun anlamı, polimerik malzemeden bir kenar ve radikal zengin yüzeylerin oluşmasına yol açar. Oksijen molekülleri ihtiva eden plazma kullanılarak, yüzey oksijen açısından zengin hale gelir ve böylece daha hidrofilik 9-11. Bununla birlikte, yüzeylerin hidrofilliğini zaman 12 stabil değildir. Uzun süreli stabilitesini geliştirmek amacıyla, plazma ile muamele edilmiş yüzey, kimyasal olarak 13-15 sonra veya plazma işlemi sırasında modifiye edilebilir. Bu muamele normal olarak plazma işlemi sırasında gaz fazına reaktif bir monomer türü eklenmesi ile gerçekleştirilir, bu monomerler daha sonra polimerizepolimer yüzeyinin oluşturulan radikaller. Kimyasal muamele, uçucu olmayan bir monomer ile gerçekleştirilirse, polimer aşılama plazma modifikasyonundan sonra yer almak zorundadır. Kökleri yüzeyinde oluşan sonra kontrollü bir aşılama gerçekleştirmek için, plazma ayar kontrol edilen koşullar altında 12,16 çözelti içinde yüzeyden plazma tarafından başlatılan yüzey indüklenen polimerizasyonu sağlayan, tarif edilmektedir. Sunum parça kenarlı polimer membranların 12,17 değiştirilmesi üzerinde duruluyor. Bu membran yüzey gerilimini değiştirerek, geçirgenlik oranı, 12 değiştirilebilir. Bu temiz ve hızlı işlem polimer zar toplu özelliğini değiştirmeden tüm membran yüzeyini kaplayan çok ince bir tabakadan (&lt;5 nm) oluşturulmasını sağlar. Nedeniyle, plazma işlemi sırasında kenar için, parça kenarlı membranların gözenek çapı hafifçe 12 arttırır. Kenar oranı dependi olduğunung polimere ve doğrusal bir davranışa sahiptir. Reaktif işlevsel gruplarına sahip olan monomerler kullanırken, aşılı polimerler, daha da fonksiyonalize edilebilir. Bu, karboksilik asit ile fonksiyonelleştirilmiş spiropyran bir PHEMA-aşılı zarın postmodification ile gösterilmiştir. UV-ışını ile muamele zaman spiropyran bir merocyanine türlere dönüştürmek için bilinen, bu durum, bir fotokromık yüzey ile sonuçlanır. Spiropyran bir şekilde görünür ışık ile merocyanine form (Şekil 1) 18,19 irradyasyonu ile yeniden tesis edilebilir. Merocyanine formu spiropyran devlet daha fazla polar olduğu, kaplamanın yüzey gerilimi ışık 20 ile tetiklenebilir. Yüzey geriliminde değişim yönelik sulu çözeltiler, zarın geçirgenliği direnci etkilemektedir. Nasıl bu ışık duyarlı membranların geçirgenliği testleri gerçekleştirmek için set-up gösterilir ve geçirgenlik direnci önemli değişiklik (i azaltmak% 97 oranında n geçirgenlik direnci) gösterilmiştir. Bu tür bir zar, bir ilaç verme kurulumunda veya akıllı algılama sistemleri entegre edilebilir. <img alt="Şekil 1" fo:content-width="5in" fo:src="/files/ftp_upload/51680/51680fig1highres.jpg" src="/files/ftp_upload/51680/51680fig1.jpg" /> Spirobenzopiran bileşik 1 Şekil 1.. Photoisomerization.

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	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">Name of Reagent/ Equipment</td>
			<td style="width:284px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">2-Hydroxyethyl methacrylate, 97%</td>
			<td style="width:284px;">Sigma-Aldrich</td>
			<td style="width:119px;">128635</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">Hexane 99%</td>
			<td style="width:284px;">Biosolve</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">Magnesium sulfate (MgSO4, anhydrous)</td>
			<td style="width:284px;">Sigma-Aldrich</td>
			<td>M7506</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">Methanol, 99%&#160;</td>
			<td style="width:284px;">Sigma-Aldrich</td>
			<td style="width:119px;">14262</td>
			<td>dried over molecular sieves</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">N,N-Dicylcohexylcarbodiimide, 99%</td>
			<td style="width:284px;">Sigma-Aldrich</td>
			<td style="width:119px;">D8002</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">Dimethyl aminopyridine, 99%</td>
			<td style="width:284px;">Sigma-Aldrich</td>
			<td style="width:119px;">107700</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">Tert-butylmethylether, 98%</td>
			<td style="width:284px;">Fluka</td>
			<td style="width:119px;">306975</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">Polycarbonate membrane</td>
			<td style="width:284px;">Whatman</td>
			<td style="width:119px;"></td>
			<td>Nanopore Track Etched (TE) (1.0 &#956;m, 0.2 &#956;m, 0.1 &#956;m, 50 nm, 30 nm and 15 nm pore diameter; 47 mm or 25 mm membrane diameter)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">Caffeine (reagent plus)</td>
			<td style="width:284px;">Sigma-Aldrich</td>
			<td style="width:119px;">C0750&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">Franz diffusion cell (12 ml)</td>
			<td style="width:284px;">SES-Analysesysteme</td>
			<td style="width:119px;">6C010015</td>
			<td>15mm unjacheted Franz Cell, 12 ml Receptor volume, Flat ground, clear glass, stirbat and clamp</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">UV-Lamp</td>
			<td style="width:284px;"></td>
			<td style="width:119px;"></td>
			<td>UV irradiation (366 nm, 15 W/m2)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">White light lamp</td>
			<td style="width:284px;"></td>
			<td style="width:119px;"></td>
			<td>White light irradiation (500 W bulb)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">UV/Vis spectrophotometer</td>
			<td style="width:284px;">Varian 50Bio/50MPR</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:373px;">Polyester membranes</td>
			<td style="width:284px;">Sterlitech</td>
			<td style="width:119px;">PET0225100</td>
			<td>Polyester Membrane Filters, 0.2 &#956;m pore diameter, 25 mmm diameter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">Polyvinylidene fluoride membranes</td>
			<td style="width:284px;">Millipore</td>
			<td style="width:119px;"></td>
			<td>PVDF Membranes Durapore (0.22 &#956;m pore diameter; 47 mm membrane diameter)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">Argon (99.9995%)</td>
			<td style="width:284px;">Alphagaz</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">Dressler Cesar RF Power Generator</td>
			<td style="width:284px;"></td>
			<td style="width:119px;"></td>
			<td>Plasma chamber setup</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">MKS Multi Gas Controller 647C</td>
			<td style="width:284px;"></td>
			<td style="width:119px;"></td>
			<td>Plasma chamber setup</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">MKS Mass-Flow controllers</td>
			<td style="width:284px;"></td>
			<td style="width:119px;"></td>
			<td>Plasma chamber setup</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">Vacuubrand RE 2.5 rotary vane vacuum pump</td>
			<td style="width:284px;"></td>
			<td style="width:119px;"></td>
			<td>Plasma chamber setup</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">Contact angle measurement device Kr&#252;ss G10</td>
			<td style="width:284px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:373px;">Balances Mettler Toledo AB204-S and Mettler ME30</td>
			<td style="width:284px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

preparation of light-responsive membranes by a combined surface grafting and postmodification process

Ticari olarak temin edilebilen parça kenarlı bir polimer membran yüzey gerilimini değiştirmek amacıyla, bir yüzey tarafından başlatılan polimerizasyonun bir prosedür sunulmuştur. Membran yüzeyinden polimerizasyon 2-hidroksietil metakrilat (HEMA) bir metanolik çözeltisi ile zar yüzeyi reaksiyona sokulması ve ardından, zarın plazma işlemi ile indüklenir. Özel dikkat önce yüzeyde polimerizasyona plazma muamelesi için işlem parametreleri verilir. Örneğin, membran farklı (örneğin, polyester, polikarbonat, poliviniliden florid) üzerinde plazma muamele etkisi incelenmiştir. Bundan başka, yüzey-aşılı membran zamana bağımlı stabilite, temas açısı ölçümleri ile gösterilmiştir. Bu şekilde poli (2-hidroksietil metakrilat) (PHEMA) aşılama zaman, yüzey daha fazla arzu edilen maddenin bir karboksilik asit fonksiyonu ile, polimerin alkol grubu esterlenmesi ile değiştirilebilir.Bu reaksiyonlar, bu nedenle, membran yüzeyinde fonksiyonelleştirilmesi için kullanılabilir. Örneğin, membran yüzey gerilimi değiştirilebilir ya da sunulan ışık yanıt gibi arzu edilen bir işlevsellik sokulabilir. Bu, ışık duyarlı zara açan bir karboksilik asit fonksiyonalize spirobenzopiran birimi ile PHEMA reaksiyona sokulması ile gösterilir. Çözücünün seçimi postmodification aşamada önemli bir rol oynar ve bu yazıda daha ayrıntılı olarak tartışılmıştır. Bu tür işlevselleştirilmiş membran geçirgenliği ölçümleri, harici bir ışık kaynağı olan bir Franz hücre kullanılarak gerçekleştirilir. UV yelpazesi için görünür gelen ışığın dalga boyunu değiştirerek, sulu çözeltiler kafein geçirgenlik bir değişiklik gözlenmiştir.

Etch oranı, farklı zaman sonra membran tartı ile takip edilebilir. Şekil 4&#39;ten görülebileceği gibi, etch oranı poliester, poliviniliden florür takip eder ve polikarbonat kütle kaybı karşı etch zaman doğrusal bağıntı eğiminden belirlenebilir doğrusal etch oranı, membranlar. Şekil 4&#39;te gösterildiği gibi, polikarbonat membran her üç polimer zarların düşük etch hızını gösterir. Aşındırmanın bir sonucu gözenek çapı değişimidir. Yakla...

Watch this Scientific Journal Video about Preparation of Light-responsive Membranes by a Combined Surface Grafting and Postmodification Process at JoVE.com

Preparation of Light-responsive Membranes by a Combined Surface Grafting and Postmodification Process

Bir plazma kaynaklı polimerizasyon prosedürü polimer zarlar üzerinde yüzey başlatılan polimerizasyonu için açıklanmıştır. Fotokromık maddeler ile aşılanmış polimerin daha ileri postmodification ışığa duyarlı membran geçirgenliği ölçümleri iletken bir protokol ile sunulmuştur.

Bir Kombine Yüzey Tırnak ve Postmodification Süreci Işık duyarlı Membranların Hazırlanması

In order to modify the surface tension of commercial available track-edged polymer membranes, a procedure of surface-initiated polymerization is ...

preparation-light-responsive-membranes-combined-surface-grafting

Research

JoVE Journal

Bioengineering

11.7K Görüntüleme.  Empa, Swiss Federal Laboratories for Materials Science and Technology.  Bir plazma kaynaklı polimerizasyon prosedürü polimer zarlar üzerinde yüzey başlatılan polimerizasyonu için açıklanmıştır. Fotokromık maddeler ile aşılanmış polimerin daha ileri postmodification ışığa duyarlı membran geçirgenliği ölçümleri iletken bir protokol ile sunulmuştur. 

Video: Preparation of Light-responsive Membranes by a Combined Surface Grafting and Postmodification Process

Measurement of Aggregate Cohesion by Tissue Surface Tensiometry

Rigorous measurement of intercellular binding energy can only be made using methods grounded in thermodynamic principles in systems at equilibrium. We have developed tissue surface tensiometry (TST) specifically to measure the surface free energy of interaction between cells. The biophysical concepts underlying TST have been previously described in detail1,2. The method is based on the observation that mutually cohesive cells, if maintained in shaking culture, will spontaneously assemble into clusters. Over time, these clusters will round up to form spheres. This rounding-up behavior mimics the behavior characteristic of liquid systems. Intercellular binding energy is measured by compressing spherical aggregates between parallel plates in a custom-designed tissue surface tensiometer. The same mathematical equation used to measure the surface tension of a liquid droplet is used to measure surface tension of 3D tissue-like spherical aggregates. The cellular equivalent of liquid surface tension is intercellular binding energy, or more generally, tissue cohesivity. Previous studies from our laboratory have shown that tissue surface tension (1) predicts how two groups of embryonic cells will interact with one another1-5, (2) can strongly influence the ability of tissues to interact with biomaterials6, (3) can be altered not only through direct manipulation of cadherin-based intercellular cohesion7, but also by manipulation of key ECM molecules such as FN8-11 and 4) correlates with invasive potential of lung cancer12, fibrosarcoma13, brain tumor14 and prostate tumor cell lines15. In this article we will describe the apparatus, detail the steps required to generate spheroids, to load the spheroids into the tensiometer chamber, to initiate aggregate compression, and to analyze and validate the tissue surface tension measurements generated.

We describe a method of measuring binding energy, expressible as tissue surface tension, between cells within 3D tissue-like aggregates. Differences in tissue surface tension have been demonstrated to correlate with invasiveness of lung, muscle, and brain tumors, and are fundamental determinants of establishing spatial relationships between different cell types.

Doku Yüzey tensiometresi Agrega Uyum ölçümü

Rigorous measurement of intercellular binding energy can only be made using methods grounded in thermodynamic principles in systems at equilibrium.  We ...

Biyomühendislik

Self-reporting Scaffolds for 3-Dimensional Cell Culture

Culturing cells in 3D on appropriate scaffolds is thought to better mimic the in vivo microenvironment and increase cell-cell interactions. The resulting 3D cellular construct can often be more relevant to studying the molecular events and cell-cell interactions than similar experiments studied in 2D. To create effective 3D cultures with high cell viability throughout the scaffold the culture conditions such as oxygen and pH need to be carefully controlled as gradients in analyte concentration can exist throughout the 3D construct. Here we describe the methods of preparing biocompatible pH responsive sol-gel nanosensors and their incorporation into poly(lactic-co-glycolic acid) (PLGA) electrospun scaffolds along with their subsequent preparation for the culture of mammalian cells. The pH responsive scaffolds can be used as tools to determine microenvironmental pH within a 3D cellular construct. Furthermore, we detail the delivery of pH responsive nanosensors to the intracellular environment of mammalian cells whose growth was supported by electrospun PLGA scaffolds. The cytoplasmic location of the pH responsive nanosensors can be utilized to monitor intracellular pH (pHi) during ongoing experimentation.

Biocompatible pH responsive sol-gel nanosensors can be incorporated into poly(lactic-co-glycolic acid) (PLGA) electrospun scaffolds. The produced self-reporting scaffolds can be used for in&#160;situ monitoring of microenvironmental conditions whilst culturing cells upon the scaffold. This is beneficial as the 3D cellular construct can be monitored in real-time without disturbing the experiment.

3-Boyutlu Hücre Kültürü için Self-raporlama iskeleleri

Culturing cells in 3D on appropriate scaffolds is thought to better mimic the in vivo microenvironment and increase cell-cell interactions. The resulting ...

Generation and Grafting of Tissue-engineered Vessels in a Mouse Model

The construction of vascular conduits is a fundamental strategy for surgical repair of damaged and injured vessels resulting from cardiovascular diseases. The current protocol presents an efficient and reproducible strategy in which functional tissue engineered vessel grafts can be generated using partially induced pluripotent stem cell (PiPSC) from human fibroblasts. We designed a decellularized vessel scaffold bioreactor, which closely mimics the matrix protein structure and blood flow that exists within a native vessel, for seeding of PiPSC-endothelial cells or smooth muscle cells prior to grafting into mice. This approach was demonstrated to be advantageous because immune-deficient mice engrafted with the PiPSC-derived grafts presented with markedly increased survival rate 3 weeks after surgery. This protocol represents a valuable tool for regenerative medicine, tissue engineering and potentially patient-specific cell-therapy in the near future.

Here, we present a protocol to generate tissue engineered vessel grafts that are functional for grafting into mice by double seeding partially induced pluripotent stem cell (PiPSC) - derived smooth muscle cells and PiPSC - derived endothelial cells on a decellularized vessel scaffold bioreactor.

Bir Fare Modeli Doku mühendisliği Kapların Üretimi ve Tırnak

The construction of vascular conduits is a fundamental strategy for surgical repair of damaged and injured vessels resulting from cardiovascular diseases. ...

Adapting the Electrospinning Process to Provide Three Unique Environments for a Tri-layered In Vitro Model of the Airway Wall

Electrospinning is a highly adaptable method producing porous 3D fibrous scaffolds that can be exploited in in vitro cell culture. Alterations to intrinsic parameters within the process allow a high degree of control over scaffold characteristics including fiber diameter, alignment and porosity. By developing scaffolds with similar dimensions and topographies to organ- or tissue-specific extracellular matrices (ECM), micro-environments representative to those that cells are exposed to in situ can be created. 
The airway bronchiole wall, comprised of three main micro-environments, was selected as a model tissue. Using decellularized airway ECM as a guide, we electrospun the non-degradable polymer, polyethylene terephthalate (PET), by three different protocols to produce three individual electrospun scaffolds optimized for epithelial, fibroblast or smooth muscle cell-culture. Using a commercially available bioreactor system, we stably co-cultured the three cell-types to provide an in vitro model of the airway wall over an extended time period.
This model highlights the potential for such methods being employed in in vitro diagnostic studies investigating important inter-cellular cross-talk mechanisms or assessing novel pharmaceutical targets, by providing a relevant platform to allow the culture of fully differentiated adult cells within 3D, tissue-specific environments.

Adapting the Electrospinning Process to Provide Three Unique Environments for a Tri-layered In Vitro Model of the Airway Wall

Advancements in biomaterial technologies enable the development of three-dimensional multi-cell-type constructs. We have developed electrospinning protocols to produce three individual scaffolds to culture the main structural cells of the airway to provide a 3D in vitro model of the airway bronchiole wall.

Elektrospinning Süreci Uyarlama bir Tri-katmanlı için üç Benzersiz ortamlar sağlamak için<em&gt; In Vitro</em&gt; Havayolu Wall Modeli

Electrospinning is a highly adaptable method producing porous 3D fibrous scaffolds that can be exploited in in vitro cell culture. Alterations to ...

Qualitative Characterization of the Aqueous Fraction from Hydrothermal Liquefaction of Algae Using 2D Gas Chromatography with Time-of-flight Mass Spectrometry

Two-dimensional gas chromatography coupled with time-of-flight mass spectrometry is a powerful tool for identifying and quantifying chemical components in complex mixtures. It is often used to analyze gasoline, jet fuel, diesel, bio-diesel and the organic fraction of bio-crude/bio-oil. In most of those analyses, the first dimension of separation is non-polar, followed by a polar separation. The aqueous fractions of bio-crude and other aqueous samples from biofuels production have been examined with similar column combinations. However, sample preparation techniques such as derivatization, solvent extraction, and solid-phase extraction were necessary prior to analysis. In this study, aqueous fractions obtained from the hydrothermal liquefaction of algae were characterized by two-dimensional gas chromatography coupled with time-of-flight mass spectrometry without prior sample preparation techniques using a polar separation in the first dimension followed by a non-polar separation in the second. Two-dimensional plots from this analysis were compared with those obtained from the more traditional column configuration. Results from qualitative characterization of the aqueous fractions of algal bio-crude are discussed in detail. The advantages of using a polar separation followed by a non-polar separation for characterization of organics in aqueous samples by two-dimensional gas chromatography coupled with time-of-flight mass spectrometry are highlighted.

A two-dimensional gas chromatography-time-of-flight mass spectrometry method is described for characterization of the aqueous fraction of bio-crude produced from hydrothermal liquefaction of algae. This protocol can also be employed to analyze the aqueous fraction of liquid products from fast pyrolysis, catalytic fast pyrolysis, catalytic deoxygenation and hydro-treating.

Zaman-of-flight Kütle Spektrometre ile 2D Gaz Kromatografisi Kullanarak Alg Hidrotermal Sıvılaşma gelen Sulu Kesir Nitel Karakterizasyonu

Two-dimensional gas chromatography coupled with time-of-flight mass spectrometry is a powerful tool for identifying and quantifying chemical components in ...

Automated Robotic Dispensing Technique for Surface Guidance and Bioprinting of Cells

This manuscript describes the introduction of cell guidance features followed by the direct delivery of cells to these features in a hydrogel bioink using an automated robotic dispensing system. The particular bioink was selected as it allows cells to sediment towards and sense the features. The dispensing system bioprints viable cells in hydrogel bioinks using a backpressure assisted print head. However, by replacing the print head with a sharpened stylus or scalpel, the dispensing system can also be employed to create topographical cues through surface etching. The stylus movement can be programmed in steps of 10 &#181;m in the X, Y and Z directions. The patterned grooves were able to orientate mesenchymal stem cells, influencing them to adopt an elongated morphology in alignment with the grooves&#39; direction. The patterning could be designed using plotting software in straight lines, concentric circles, and sinusoidal waves. In a subsequent procedure, fibroblasts and mesenchymal stem cells were suspended in a 2% gelatin bioink, for bioprinting in a backpressure driven extrusion printhead. The cell bearing bioink was then printed using the same programmed coordinates used for the etching. The bioprinted cells were able to sense and react to the etched features as demonstrated by their elongated orientation along the direction of the etched grooves.

This protocol describes a bioprinting methodology using an automated robotic depositing system that incorporates etched topographical guidance cues with the precision deposition of a cell bearing hydrogel bioink. The printed cells are directly delivered to the etched features and are able to sense and orientate with them.

Hücre Yüzey Rehberlik ve Bioprinting için Otomatik Robotik Dağıtım Tekniği

This manuscript describes the introduction of cell guidance features followed by the direct delivery of cells to these features in a hydrogel bioink using ...

Synthesis of Hydrogels with Antifouling Properties As Membranes for Water Purification

Hydrogels have been widely utilized to enhance the surface hydrophilicity of membranes for water purification, increasing the antifouling properties and thus achieving stable water permeability through membranes over time. Here, we report a facile method to prepare hydrogels based on zwitterions for membrane applications. Freestanding films can be prepared from sulfobetaine methacrylate (SBMA) with a crosslinker of poly(ethylene glycol) diacrylate (PEGDA) via photopolymerization. The hydrogels can also be prepared by impregnation into hydrophobic porous supports to enhance the mechanical strength. These films can be characterized by attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) to determine the degree of conversion of the (meth)acrylate groups, using goniometers for hydrophilicity and differential scanning calorimetry (DSC) for polymer chain dynamics. We also report protocols to determine the water permeability in dead-end filtration systems and the effect of foulants (bovine serum albumin, BSA) on membrane performance.

This paper reports practical methods to prepare hydrogels in freestanding films and impregnated membranes and to characterize their physical properties, including water transport properties.

Su Arıtma Membranlar olarak Zehirli Özelliklere Sahip Hidrojeller Sentezi

Hydrogels have been widely utilized to enhance the surface hydrophilicity of membranes for water purification, increasing the antifouling properties and ...

Surface Engineering of Pancreatic Islets with a Heparinized StarPEG Nanocoating

Cell surface engineering can protect implanted cells from host immune attack. It can also reshape cellular landscape to improve graft function and survival post-transplantation. This protocol aims to achieve surface engineering of pancreatic islets using an ultrathin heparin-incorporated starPEG (Hep-PEG) nanocoating. To generate the Hep-PEG nanocoating for pancreatic islet surface engineering, heparin succinimidyl succinate (Heparin-NHS) was first synthesized by modification of its carboxylate groups using N-(3-dimethylamino propyl)-N'-ethyl carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). The Hep-PEG mixture was then formed by crosslinking of the amino end-functionalized eight-armed starPEG (starPEG-(NH2)8) and Heparin-NHS. For islet surface coating, mouse islets were isolated via collagenase digestion and gradient purification using Histopaque. Isolated islets were then treated with ice cold Hep-PEG solution for 10 min to allow covalent binding between NHS and the amine groups of islet cell membrane. Nanocoating with the Hep-PEG incurs minimal alteration to islet size and volume and heparinization of the islets with Hep-PEG may also reduce instant blood-mediated inflammatory reaction during islet transplantation. This "easy-to-adopt" approach is mild enough for surface engineering of living cells without compromising cell viability. Considering that heparin has shown binding affinity to multiple cytokines, the Hep-PEG nanocoating also provides an open platform that enables incorporation of unlimited functional biological mediators and multi-layered surfaces for living cell surface bioengineering.

This protocol aims to achieve surface engineering of pancreatic islets using a heparin-incorporated starPEG nanocoating via pseudo-bioorthogonal chemistry between the N-hydroxysuccinimide groups of the nanocoating and the amine groups of islet cell membrane.

Heparinized StarPEG Nanocoating ile pankreas adacıkları yüzey Mühendisliği

Cell surface engineering can protect implanted cells from host immune attack. It can also reshape cellular landscape to improve graft function and ...

Detection of Endotoxin in Nano-formulations Using Limulus Amoebocyte Lysate (LAL) Assays

When present in pharmaceutical products, a Gram-negative bacterial cell wall component endotoxin (often also called lipopolysaccharide) can cause inflammation, fever, hypo- or hypertension, and, in extreme cases, can lead to tissue and organ damage that may become fatal. The amounts of endotoxin in pharmaceutical products, therefore, are strictly regulated. Among the methods available for endotoxin detection and quantification, the Limulus Amoebocyte Lysate (LAL) assay is commonly used worldwide. While any pharmaceutical product can interfere with the LAL assay, nano-formulations represent a particular challenge due to their complexity. The purpose of this paper is to provide a practical guide to researchers inexperienced in estimating endotoxins in engineered nanomaterials and nanoparticle-formulated drugs. Herein, practical recommendations for performing three LAL formats including turbidity, chromogenic and gel-clot assays are discussed. These assays can be used to determine endotoxin contamination in nanotechnology-based drug products, vaccines, and adjuvants.

Detection of Endotoxin in Nano-formulations Using Limulus Amoebocyte Lysate LAL Assays

Detection of endotoxins in engineered nanomaterials represents one of the grand challenges in the field of nanomedicine. Here, we present a case study that describes the framework composed of three different LAL formats to estimate potential endotoxin contamination in nanoparticles.

Nano-formülasyonları endotoksin tespiti Limulus Amoebocyte Lysate (LAL) deneyleri kullanarak

When present in pharmaceutical products, a Gram-negative bacterial cell wall component endotoxin (often also called lipopolysaccharide) can cause ...

Wet-spinning-based Molding Process of Gelatin for Tissue Regeneration

This article presents an inexpensive method to fabricate gelatin, as a natural polymer, into monofilament fibers or other appropriate forms. Through the wet spinning method, gelatin fibers are produced by smooth extrusion in a suitable coagulation medium. To increase the functional surface of these gelatin fibers and their ability to mimic the features of tissues, gelatin can be molded into a tube form by referring to this concept. Examined by in vitro and in vivo tests, the gelatin tubes demonstrate a great potential for application in tissue engineering. Acting as a suitable filling gap material, gelatin tubes can be used to substitute the tissue in the damaged area (e.g., in the nervous or cardiovascular system), as well as to promote regeneration by providing a direct replacement of stem cells and neural circuitry. This protocol provides a detailed procedure for creating a biomaterial based on a natural polymer, and its implementation is expected to greatly benefit the development of correlative natural polymers, which help to realize tissue regeneration strategies.

We developed and describe a protocol based on the wet spinning concept, for the construction of gelatin-based biomaterials used for the application of tissue engineering.

Islak eğirme esaslı kalıplama işlemi doku rejenerasyonu için jelatin

This article presents an inexpensive method to fabricate gelatin, as a natural polymer, into monofilament fibers or other appropriate forms. Through the ...