Biz onun tür destek için Prof JP Spatz (Biyofizik Kimya Bölümü, Heidelberg Üniversitesi ve Yeni Malzemeler Biyosistem Bölümü, Akıllı Sistemler Max Planck Enstitüsü, Stuttgart) teşekkür ederim. Max-Planck-Gesellschaft ve Deutsche Forschungsgemeinschaft (EAC-A DFG SFB/TR79.) Gelen mali destek de büyük ölçüde kabul edilmiştir.

1.. 11-Mercaptoundecanoyl-N-hidroksisüksinimid ester (MU-NHS) sentezi <ol><li> Oda sıcaklığında (RT) 40 ml diklorometan (pa), 10 ml aseton (PA) (dimetilamino) piridin 1 g 11-mercaptoundecanoic asit - bir 500 mg N-hidroksisüksinimid çözeltisi ve 30 mg 4 damla damla ilave edildi. </li><li> 0 ° C&#39;ye soğutun ve reaksiyon, damla damla 1.1 g N, (azot atmosferi altında) 10 ml diklorometan içinde N &#39;-disikloheksilkarbodiimid ekleyin. 1 saat boyunca düşük ısıda reaksiyonu Ol ve daha sonra gece boyunca oda sıcaklığında karıştırın. </li><li> Çökeltiyi filtre edin ve indirgenmiş basınç altında kurutun. 1:1 &#39;lik bir oranda petrol benzen ve etil asetat (pa) ile flaş kromatografisi ile ürünü saflaştırmak. </li></ol> 2. Homojen Altın Katmanlar hazırlanması <ol><li> Hassas mendil ile ve etil asetat (pa) ve metanol (pa) 1:1 &#39;lik bir karışımını içeren bir çözelti içinde 5 dakika boyunca ses dalgalarına maruz bunları temiz cam lameller. Rinse, metanol ile alt-tabakalar ve azot akışı altında kurutulur. </li><li> Püskürtmeli kaplama cihazı içinde temiz yüzeylerde yerleştirin. Odasını boşaltın. 60 saniye için püskürtme ile krom hedeften en üst krom oksit katmanı kaldırın. Metal kiriş ve bir 40 nm altın tabakası ile kaplanması, ardından bir 10 nm krom tabaka ile kaplama, onları altında numuneler. </li></ol> 3. BMP-2 yüzey immobilizasyonu <ol><li> Yaklaşık 1 mM&#39;lik nihai bir konsantrasyona kadar N, MU-NHS, N-dimethlyformamide (DMF) içinde eritin ve azot atmosferi altında 4 saat boyunca oda sıcaklığında MU-NHS çözeltisi içinde altın substratlar inkübe edin. </li><li> 2 dakika boyunca DMF içinde sonikasyon yüzeyler, DMF ve MeOH ile durulayın ve azot akışı altında kurutulur. </li><li> 100 ug / ml (-80 ° C&#39;de saklayın alikotları) &#39;lik bir konsantrasyonda steril 4 mM HCI rhBMP-2&#39;ye ait bir stok çözelti hazırlayın. Çalışma seyreltileri (3.5 ug / ml) için, PBS / NaCl (PBS, 1 M NaCl ihtiva eden) ve bir de stok solüsyonu seyreltikkullanımdan hemen önce pH 8.5 &#39;e djust. </li><li> Gece boyunca 4 ° C&#39;de rhBMP-2 çalışma çözeltisi içinde MU-NHS işlevselleştirilmiş yüzeyleri inkübe edin. Kuluçka Süpernatantı. Sonikasyon 2 dakika boyunca PBS / NaCI yüzeyler ve steril PBS / NaCI ile 3 kez yıkayın. </li></ol> 4. Yüzey Karakterizasyonu <ol><li> Antikorun spesifik olmayan bir şekilde emilmesi bloke etmek için,% 5 BSA ile inkübe yüzeyleri (w / v) bir anti-BMP-2 antikoru (1:100 içinde% 1 (ağırlık / hacim ile inkübasyon ve ardından oda sıcaklığında 1 saat için PBS solüsyonunda oda sıcaklığında 1 saat için) BSA / PBS çözeltisi). PBS ve 30 saniye süreyle sonikasyon ile iki kez yüzeyleri yıkayın. </li><li> Daha sonra PBS ile iki kere yıkayın, oda sıcaklığında 30 dakika için HRP ile konjüge edilmiş sekonder antikor (1:1000 içinde% 1 (w / v) BSA / PBS çözeltisi) ile alt-tabakaların inkübe edin. </li><li> Bir levha okuyucu ile 570 nm de HRP enzimatik aktivitesinin ölçülmesi için Ampliflu kırmızı deneyi kullanarak. </li></ol> 5. Biyolojik Aktivite Analizi <ol><li> 6-çukurlu pl içinde göz başına 1 x 10 tohum 5 fare C2C12 miyoblastlardır% 10 FBS,% 1 penisilin / streptomisin ve 37 ° C /% 5 CO2 seviyesinde 24 saat boyunca inkübe ile takviye piruvat içeren yüksek glukoz DMEM oluşan büyüme ortamı içinde Ateş,. </li><li> Hareketsiz BMP-2 dekore yüzeylere tohumlanmasından önce 3-5 saat boyunca serumsuz DMEM hücreleri açlıktan. </li><li> Kısa süreli sinyal indüksiyon araştırılması için, 200 ul taze serum içermeyen DMEM ile orta yerine ve hücreler üzerinde hareketsizleştirilmiş BMP-2 yüzeyleri yerleştirin. Yüzeyi çizmemek için ince uçlu cımbız ile ele alınmalıdır. </li><li> Yumuşak yüzeyleri çıkarın ve bir pipet ile ortam aspire, PBS ile iki kez hücreleri yıkayın. Hücre yanıtlarının analizi geçin. </li><li> Uzun süreli biyolojik aktivite, levha hücrelerin analizi için üzerine, altı gün boyunca düşük serum koşullarına (% 2 FBS) içinde 37 ° C /% 5 CO2 ile yüzeyler ve kültür onları. </li></ol>

<ol>
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Yazarlar ifşa hiçbir şey yok.

Bu protokolde, biyoaktif rhBMP-2 ile fonksiyonelleştirilmiş yüzeylerin hazırlanmasını tarif etmektedir. , RhBMP-2 protein 2) kovalent immobilizasyon altın yüzeyi üzerinde bir çift fonksiyonlu bağlayıcı, kendi kendine bir araya tek tabaka (SAM) 1) başlangıç ​​sistemi: Bu yaklaşım, iki adım içermektedir. Önceki çalışmada, iki fonksiyonlu linker ile büyüme faktörü bağlayıcı etkili doğrulanmış ve yüzey hareketsizleştirilmiş rhBMP-2 biyolojik aktivitesini muhafaza 19 olduğ...

Kemik morfogenetik protein 2 (BMP-2), transforme edici büyüme faktörü (TGF-β) ailesi ve de novo kemik oluşumunun indükleyici hem de embriyonik gelişim ve yetişkin sırasında çeşitli dokuların düzenleyicisi olarak işlev görür 1-3 homeostasis bir üyesidir. Biyolojik olarak etkin olan homodimerik BMP-2 proteininin her bir monomer son derece tüm BMP 4 korunan bir &quot;sistein düğüm&quot; motifi içerir. Yedinci sistein iki monomer 5,6 arasında moleküller arası bir bağ oluşturan, dimerizasyon katılır ise yedi sistein kalıntılarının altı, her bir monomerin kararlı moleküller arası disülfid bağı oluşturur. Bu yüksek ölçüde korunmuş sistein düğüm BMP-2 proteininin üç boyutlu yapısını tanımlar ve bu ısı, denatüranlar ve asidik pH 7-9 karşı direnç gibi eşsiz özelliklerini belirler. BMP-2, böylece sinyal iletiminin uyarılması, serin / treonin kinaz zar reseptörlerine bağlanan <sup&gt; 10-12. Reseptör oligomerizasyon moduna bağlı olarak, farklı bir sinyal yollar aktive edilir: a Smad-bağımlı yolu Smad karmaşık nükleer translokasyonu ve transkripsiyon aktivasyonu reseptör fosforilasyon ile aktif hale sonuçları ise bir Smad-bağımsız bir sinyal kaskadı, p38 aracılığı ile sinyalizasyon alkalin fosfataz, indüklenmesine yol açacaktır gibi farklılaşma inhibitörü (Id) 12-14 gibi belirli hedef genler,. Kemik, BMP-2, böylece iyileşmesi ve kemik de novo oluşumunu teşvik edici, osteoblastlar içine mezenkimal kök hücrelerin farklılaşmasını indükler. Şu anda, rekombinant BMP-2 sitelerinin kırık iyileşmesini artırmak için klinik olarak uygulanır ifade edilmiştir. Kemik doku mühendisliğinde yaygın bir strateji, lokal verme sistemleri ile karşılaştırıldığında daha az invaziv bir enjekte edilebilir büyüme faktörleri, kullanılmasıdır. Bununla birlikte, in vivo çalışmalarda ve klinik uygulamalar göstermiştir ki, sahip olduğu kısa biyolojik yarı-ömür, spesifik olmayan locAdalet ve Hafıza ve BMP-2 hızlı lokal temizleme çeşitli lokal ve sistemik sorunlara ektopik 15 yol açabilir. Bu nedenle, etkili bir sunum, malzeme içinde veya üzerinde BMP-2&#39;nin sürüklenmesini veya immobilizasyon elde etmek için, hedef bölgede, yerel ve sürekli uygulama için de gereklidir. Sürekli teslim gibi fiziksel olarak yakalanmalarında, adsorpsiyon ya da iyon kompleks 16 gibi kovalent olmayan bir tutma yaklaşımlar ile elde edilebilir. Bununla birlikte, bilinen bu moleküllerin 17 denatürasyonda yüzeylerine proteinlerin spesifik olmayan adsorpsiyon olabilir neden olur. Büyüme faktörlerinin bağlanması kovalent için, desteklerin farklı son on yıl içinde geliştirilmiştir. Örneğin, proteinin amino ya da karboksil grupları hedefleyen iki işlevli bağlama moleküllerinin kullanılması, zorunlu olarak hareketsiz hale elde etmek için protein modifikasyonu gerektirmeyen yaklaşımın bir türüdür. Aslında, ise protein modifikasyonu, protein yönünü kontrol etme avantajını sunmaktadıryapay etki, peptit etiketleri ve bölgeye özgü zincirlerinin giriş büyüme biyolojik aktivitesi 17 faktörleri değiştirebilir. Bu durumda, bağlı destek malzemesi ile etkileşim için denatürasyon aşmak için, yüzeyler istenilen faktörü 18 bağlanması ve ardından bir bağlayıcı molekülün bir kendi kendine monte tek tabaka (SAM) ile, örneğin, önceden işlevselleştirilebilir. Bu serbest amin kalıntıları hedefleyerek kovalent bir yüzey üzerine BMP-2 kılınması için bir SAM tabanlı bir yaklaşım kullandık ve hareketsizleştirilmiş protein kısa ve uzun süreli biyolojik aktiviteye 19 hem de muhafaza göstermiştir. Bu protokol, hücre membranında meydana gelir ve osteojenik sinyalizasyon sorumlu hücre içi sinyal düzenleyen mekanizmaları üzerinde in vitro çalışmalar için hücrelere BMP-2 sunmak için basit ve verimli bir yol sağlamaktadır.

<table border="1">
	<tbody>
		<tr>
			<td>Name of Material/ Equipment</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments/Description</td>
		</tr>
		<tr>
			<td>N-hydroxysuccinimide</td>
			<td>Sigma-Aldrich</td>
			<td>130672</td>
			<td></td>
		</tr>
		<tr>
			<td>4-(dimethylamino)pyridin</td>
			<td>Sigma-Aldrich</td>
			<td>522805</td>
			<td></td>
		</tr>
		<tr>
			<td>Acetone</td>
			<td>AppliChem</td>
			<td>A2282</td>
			<td></td>
		</tr>
		<tr>
			<td>11-mercaptoundecanoic acid</td>
			<td>Sigma-Aldrich</td>
			<td>674427</td>
			<td></td>
		</tr>
		<tr>
			<td>Dichlormethane</td>
			<td>Merck</td>
			<td>106050</td>
			<td></td>
		</tr>
		<tr>
			<td>N,N&#39;-dicyclohexylcarbodiimide</td>
			<td>Sigma-Aldrich</td>
			<td>D80002</td>
			<td></td>
		</tr>
		<tr>
			<td>Petroleum benzene</td>
			<td>Merck</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Glass coverslips</td>
			<td>Carl Roth</td>
			<td>M 875</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethylacetate</td>
			<td>AppliChem</td>
			<td>A3550</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Carl Roth</td>
			<td>4627</td>
			<td></td>
		</tr>
		<tr>
			<td>N,N-dimethylformamide</td>
			<td>Carl Roth</td>
			<td>T921</td>
			<td></td>
		</tr>
		<tr>
			<td>rhBMP-2</td>
			<td>R&#38;D Systems</td>
			<td>355-BM</td>
			<td>Carrier-free; expressed in E.coli</td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>PAA</td>
			<td>H15-002</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Carl Roth</td>
			<td>HN00.2</td>
			<td></td>
		</tr>
		<tr>
			<td>Poly(dimethyl siloxane) (PDMS)</td>
			<td>Dow Corning</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sylgard 184 silicone elastomer kit</td>
			<td>Dow Corning</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-rhBMP-2</td>
			<td>Sigma</td>
			<td>B9553</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat anti-mouse IgG-HRP</td>
			<td>Santa Cruz</td>
			<td>sc-2005</td>
			<td>Secondary antibody</td>
		</tr>
		<tr>
			<td>Ampliflu Red assay</td>
			<td>Sigma</td>
			<td>90101</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s Modified Eagle Medium (DMEM) (1x), liquid</td>
			<td>Gibco</td>
			<td>41966</td>
			<td>High glucose</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum (FBS)</td>
			<td>Sigma</td>
			<td>F7524</td>
			<td>Sterile filtered, cell culture tested</td>
		</tr>
		<tr>
			<td>Pen/Strep</td>
			<td>Gibco</td>
			<td>15140</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin 0.05% (1x) with EDTA 4Na</td>
			<td>Gibco</td>
			<td>25300</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycine (0.1 M)</td>
			<td>Riedel-de Ha&#235;n</td>
			<td>33226</td>
			<td></td>
		</tr>
		<tr>
			<td>IGEPAL CA-630 (1%)</td>
			<td>Sigma</td>
			<td>I8896</td>
			<td>Lysis buffer (ALP assay)19</td>
		</tr>
		<tr>
			<td>Magnesium chloride (MgCl2)(1 mM)</td>
			<td>Carl Roth</td>
			<td>HNO3.2</td>
			<td></td>
		</tr>
		<tr>
			<td>Zinc chloride (ZnCl2) (1 mM)</td>
			<td>Carl Roth</td>
			<td>3533.1</td>
			<td></td>
		</tr>
		<tr>
			<td>p-nitrophenylphosphate (pNPP)</td>
			<td>Sigma</td>
			<td>S0942</td>
			<td>Phosphatase substrate</td>
		</tr>
		<tr>
			<td>Anti-mysin heavy chain (MHC)</td>
			<td>Developmental Studies Hybridoma Bank, University of Iowa</td>
			<td>MF20</td>
			<td>Monoclonal antibody</td>
		</tr>
		<tr>
			<td>Alexa Fluor 488 Goat anti-mouse IgG</td>
			<td>Invitrogen</td>
			<td>A11001</td>
			<td></td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>Sigma</td>
			<td>D9542</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Equipment</td>
		</tr>
		<tr>
			<td>Ultrsonic bath (Sonorex Super RK 102H), Frequency 35 kHz</td>
			<td>BANDELIN electronic GmbH &#38; Co. KG</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>MED 020 Sputtercoating system</td>
			<td>BAL-TEC AG</td>
			<td></td>
			<td>Coating conditions 
			Cr: 120 mA, 1.3 x 10-2 mbar, 30 sec 
			Au: 60 mA, 5.0 x 10-2 mbar, 45 sec</td>
		</tr>
		<tr>
			<td>Tecan Infinite M200 Plate reader</td>
			<td>Tecan</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

covalent binding of bmp-2 on surfaces using a self-assembled monolayer approach

Kemik morfogenetik protein 2 (BMP-2), kemik dokusunun hücre dışı matris içinde gömülü bir büyüme faktörüdür. Bu şekilde iyileşme ve de novo kemik oluşumunu stimüle osteoblastlar içine mezenkimal hücre farklılaşmasının tetikleyici olarak BMP-2 davranır. Iskeleleri ile birlikte rekombinant insan BMP-2 (rhBMP-2) klinik kullanımı sunum moduna göre, son tartışmalar yükseltti ve miktarı teslim edilecek. Burada sunulan protokol hücreler üzerinde in vitro çalışmalar için BMP-2 sunmak için basit ve verimli bir yol sağlamaktadır. Bir heterobifonksiyonel bağlayıcının bir öz monte tek tabaka oluşturmak üzere nasıl tarif ve rhBMP-2 kovalent immobilizasyon elde etmek için daha sonraki bağlanma aşamasını göstermektedir. Bu yaklaşımla, bu proteinin biyolojik aktivitesini muhafaza ederken, BMP-2 sürekli bir sunum elde etmek mümkündür. Aslında, BMP-2 yüzey hareketsiz spesifik reklamlar önleyerek hedef inceleme sağlar:orption, büyüme faktörü miktarının azaltılması ve süre, en önemlisi de, yüzeyden kontrolsüz salınmasını engellemektedir. Hücreler, kovalent olarak hareketsizleştirilmiş rhBMP-2 sunulması yüzeylere maruz kaldıkları zaman, BMP-2 tetiklediği hem kısa hem de uzun süreli sinyal olaylarının BMP-2 ile uyarılması, hücre tepkileri ile ilgili in vitro çalışmalar için bu yaklaşım uygun hale yer alıyor.

Bir biyolojik belirsiz fakat kimyasal akort sistemi sağlar beri bizim kurulum, altın yardımcı madde olarak seçildi. Dahası, kendi kendine bir araya tek katmanlarının uygulanması pek çok avantaj içerir: bunların fonksiyonel uç-gruplar ayrıca modifiye edilebilir ise SAMs kendiliğinden, metaller ve az kusurları ile bir şekilde tek tabakaları üzerinde kendi &quot;baş grupları&quot; ile adsorbe. Böylece kontrollü ama son derece uyarlanabilir şekilde 20 arabirimi özelliklerini uyarlamak iç...

Watch this Scientific Journal Video about Covalent Binding of BMP-2 on Surfaces Using a Self-assembled Monolayer Approach at JoVE.com

Covalent Binding of BMP-2 on Surfaces Using a Self-assembled Monolayer Approach

Bu yüzeylerde BMP-2 etkin hareketsiz hale getirmeye yönelik bir metodu tarif etmektedir. Yaklaşımımız serbest amin kalıntısı ile BMP-2 bağlanması kovalent elde etmek için bir kendi kendine monte tek tabaka oluşumuna dayanır. Bu yöntem, hücre membranında sinyalini incelemek için yararlı bir araçtır.

Kovalent Öz-monte tabakalı Yaklaşım Kullanarak Yüzeyler üzerinde BMP-2 bağlanması

Bone morphogenetic protein 2 (BMP-2) is a growth factor embedded in the extracellular matrix of bone tissue. BMP-2 acts as trigger of mesenchymal cell ...

covalent-binding-bmp-2-on-surfaces-using-self-assembled-monolayer

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Unit 1

Covalent Bonding and Structure

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13.8K Görüntüleme.  University of Heidelberg.  Bu yüzeylerde BMP-2 etkin hareketsiz hale getirmeye yönelik bir metodu tarif etmektedir. Yaklaşımımız serbest amin kalıntısı ile BMP-2 bağlanması kovalent elde etmek için bir kendi kendine monte tek tabaka oluşumuna dayanır. Bu yöntem, hücre membranında sinyalini incelemek için yararlı bir araçtır. 

Video: Covalent Binding of BMP-2 on Surfaces Using a Self-assembled Monolayer Approach

Stabilizing Hepatocellular Phenotype Using Optimized Synthetic Surfaces

Currently, one of the major limitations in cell biology is maintaining differentiated cell phenotype. Biological matrices are commonly used for culturing and maintaining primary and pluripotent stem cell derived hepatocytes. While biological matrices are useful, they permit short term culture of hepatocytes, limiting their widespread application. We have attempted to overcome the limitations using a synthetic polymer coating. Polymers represent one of the broadest classes of biomaterials and possess a wide range of mechanical, physical and chemical properties, which can be fine-tuned for purpose. Importantly, such materials can be scaled to quality assured standards and display batch-to-batch consistency. This is essential if cells are to be expanded for high through-put screening in the pharmaceutical testing industry or for cellular based therapy. Polyurethanes (PUs) are one group of materials that have shown promise in cell culture. Our recent progress in optimizing a polyurethane coated surface, for long-term culture of human hepatocytes displaying stable phenotype, is presented and discussed.

This article will focus on developing polymer coated surfaces for long-term, stable culture of stem cell derived human hepatocytes.

Optimize Sentetik Yüzeyler Hepatosellüler Fenotip kullanma Stabilize

Currently, one of the major limitations in cell biology is maintaining differentiated cell phenotype. Biological matrices are commonly used for culturing ...

Kimya

An Inverse Analysis Approach to the Characterization of Chemical Transport in Paints

The ability to directly characterize chemical transport and interactions that occur within a material (i.e., subsurface dynamics) is a vital component in understanding contaminant mass transport and the ability to decontaminate materials. If a material is contaminated, over time, the transport of highly toxic chemicals (such as chemical warfare agent species) out of the material can result in vapor exposure or transfer to the skin, which can result in percutaneous exposure to personnel who interact with the material. Due to the high toxicity of chemical warfare agents, the release of trace chemical quantities is of significant concern. Mapping subsurface concentration distribution and transport characteristics of absorbed agents enables exposure hazards to be assessed in untested conditions. Furthermore, these tools can be used to characterize subsurface reaction dynamics to ultimately design improved decontaminants or decontamination procedures. To achieve this goal, an inverse analysis mass transport modeling approach was developed that utilizes time-resolved mass spectroscopy measurements of vapor emission from contaminated paint coatings as the input parameter for calculation of subsurface concentration profiles. Details are provided on sample preparation, including contaminant and material handling, the application of mass spectrometry for the measurement of emitted contaminant vapor, and the implementation of inverse analysis using a physics-based diffusion model to determine transport properties of live chemical warfare agents including distilled mustard (HD) and the nerve agent VX.

In this paper, a procedure for quantifying the mass transport parameters of chemicals in various materials is presented. This process involves employing an inverse-analysis based diffusion model to vapor emission profiles recorded by real-time, mass spectrometry in high vacuum.

Boyalar Kimyasal Ulaştırma Karakterizasyonu bir Ters Analizi Yaklaşımı

The ability to directly characterize chemical transport and interactions that occur within a material (i.e., subsurface dynamics) is a vital component in ...

Facile Preparation of Internally Self-assembled Lipid Particles Stabilized by Carbon Nanotubes

We present a facile method to prepare nanostructured lipid particles stabilized by carbon nanotubes (CNTs). Single-walled (pristine) and multi-walled (functionalized) CNTs are used as stabilizers to produce Pickering type oil-in-water (O/W) emulsions. Lipids namely, Dimodan U and Phytantriol are used as emulsifiers, which in excess water self-assemble into the bicontinuous cubic Pn3m phase. This highly viscous phase is fragmented into smaller particles using a probe ultrasonicator in presence of conventional surfactant stabilizers or CNTs as done here. Initially, the CNTs (powder form) are dispersed in water followed by further ultrasonication with the molten lipid to form the final emulsion. During this process the CNTs get coated with lipid molecules, which in turn are presumed to surround the lipid droplets to form a particulate emulsion that is stable for months. The average size of CNT-stabilized nanostructured lipid particles is in the submicron range, which compares well with the particles stabilized using conventional surfactants. Small angle X-ray scattering data confirms the retention of the original Pn3m cubic phase in the CNT-stabilized lipid dispersions as compared to the pure lipid phase (bulk state). Blue shift and lowering of the intensities in characteristic G and G&#39; bands of CNTs observed in Raman spectroscopy characterize the interaction between CNT surface and lipid molecules. These results suggest that the interactions between the CNTs and lipids are responsible for their mutual stabilization in aqueous solutions. As the concentrations of CNTs employed for stabilization are very low and lipid molecules are able to functionalize the CNTs, the toxicity of CNTs is expected to be insignificant while their biocompatibility is greatly enhanced. Hence the present approach finds a great potential in various biomedical applications, for instance, for developing hybrid nanocarrier systems for the delivery of multiple functional molecules as in combination therapy or polytherapy.

We report on a smart application of carbon nanotubes for kinetic stabilization of lipid particles that contain self-assembled nanostructures in their cores. The preparation of lipid particles requires rather low concentrations of carbon nanotubes permitting their use in biomedical applications such as drug delivery.

Karbon Nanotüpler tarafından Stabilize İçeriden Kendinden monte Lipid Parçacıklar uyduruk hazırlanması

We present a facile method to prepare nanostructured lipid particles stabilized by carbon nanotubes (CNTs). Single-walled (pristine) and multi-walled ...

Highly Stable, Functional Hairy Nanoparticles and Biopolymers from Wood Fibers: Towards Sustainable Nanotechnology

Nanoparticles, as one of the key materials in nanotechnology and nanomedicine, have gained significant importance during the past decade. While metal-based nanoparticles are associated with synthetic and environmental hassles, cellulose introduces a green, sustainable alternative for nanoparticle synthesis. Here, we present the chemical synthesis and separation procedures to produce new classes of hairy nanoparticles (bearing both amorphous and crystalline regions) and biopolymers based on wood fibers. Through periodate oxidation of soft wood pulp, the glucose ring of cellulose is opened at the C2-C3 bond to form 2,3-dialdehyde groups. Further heating of the partially oxidized fibers (e.g., T = 80 &#176;C) results in three products, namely fibrous oxidized cellulose, sterically stabilized nanocrystalline cellulose (SNCC), and dissolved dialdehyde modified cellulose (DAMC), which are well separated by intermittent centrifugation and co-solvent addition. The partially oxidized fibers (without heating) were used as a highly reactive intermediate to react with chlorite for converting almost all aldehyde to carboxyl groups. Co-solvent precipitation and centrifugation resulted in electrosterically stabilized nanocrystalline cellulose (ENCC) and dicarboxylated cellulose (DCC). The aldehyde content of SNCC and consequently surface charge of ENCC (carboxyl content) were precisely controlled by controlling the periodate oxidation reaction time, resulting in highly stable nanoparticles bearing more than 7 mmol functional groups per gram of nanoparticles (e.g., as compared to conventional NCC bearing &#60;&#60; 1 mmol functional group/g). Atomic force microscopy (AFM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) attested to the rod-like morphology. Conductometric titration, Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR), dynamic light scattering (DLS), electrokinetic-sonic-amplitude (ESA) and acoustic attenuation spectroscopy shed light on the superior properties of these nanomaterials.

Synthesis schemes to prepare highly stable wood fiber-based hairy nanoparticles and functional cellulose-based biopolymers have been detailed.

Son derece kararlı, Fonksiyonel Tüylü Nanoparçacıklar ve Ahşap Elyaf Biyopolimerler: Sürdürülebilir Nanoteknoloji Doğru

Nanoparticles, as one of the key materials in nanotechnology and nanomedicine, have gained significant importance during the past decade. While ...

Microfluidic-based Synthesis of Covalent Organic Frameworks (COFs): A Tool for Continuous Production of COF Fibers and Direct Printing on a Surface

Covalent Organic Frameworks (COFs) are a class of porous covalent materials which are frequently synthesized as unprocessable crystalline powders. The first COF was reported in 2005 with much effort centered on the establishment of new synthetic routes for its preparation. To date, most available synthetic methods for COF synthesis are based on bulk mixing under solvothermal conditions. Therefore, there is increasing interest in developing systematic protocols for COF synthesis that provide for fine control over reaction conditions and improve COF processability on surfaces, which is essential for their use in practical applications. Herein, we present a novel microfluidic-based method for COF synthesis where the reaction between two constituent building blocks, 1,3,5-benzenetricarbaldehyde (BTCA) and 1,3,5-tris(4-aminophenyl)benzene (TAPB), takes place under controlled diffusion conditions and at room temperature. Using such an approach yields sponge-like, crystalline fibers of a COF material, hereafter called MF-COF. The mechanical properties of MF-COF and the dynamic nature of the approach allow the continuous production of MF-COF fibers and their direct printing onto surfaces. The general method opens new potential applications requiring advanced printing of 2D or 3D COF structures on flexible or rigid surfaces.

Microfluidic-based Synthesis of Covalent Organic Frameworks COFs: A Tool for Continuous Production of COF Fibers and Direct Printing on a Surface

We present a novel microfluidic-based method for synthesis of covalent organic frameworks (COFs). We demonstrate how this approach can be used to produce continuous COF fibers, and also 2D or 3D COF structures on surfaces.

Kovalent Organik Çerçevelerin Mikroakışkan Temelli Sentezi (COF&#39;ler): Bir Yüzey Üzerinde COF Liflerinin Sürekli Üretimi ve Doğrudan Baskı için Bir Aracı

Covalent Organic Frameworks (COFs) are a class of porous covalent materials which are frequently synthesized as unprocessable crystalline powders. The ...

Light-driven Molecular Motors on Surfaces for Single Molecular Imaging

The design and synthesis of a synthetic system that aims for the direct visualization of a synthetic rotary motor at the single molecule level on surfaces are demonstrated. This work requires careful design, considerable synthetic effort, and proper analysis. The rotary motion of the molecular motor in solution is shown by 1H NMR and UV-vis absorption spectroscopy techniques. In addition, the method to graft the motor onto an amine-coated quartz is described. This method helps to gain more insight into molecular machines.

The manuscript describes how to synthesize and graft a molecular motor on surfaces for single molecular imaging.

Işık odaklı moleküler motorlar yüzeylerde tek moleküler görüntüleme için

The design and synthesis of a synthetic system that aims for the direct visualization of a synthetic rotary motor at the single molecule level on surfaces ...

Analysis of Complex Molecules and Their Reactions on Surfaces by Means of Cluster-Induced Desorption/Ionization Mass Spectrometry

Desorption/Ionization Induced by Neutral SO2 Clusters (DINeC) is employed as a very soft and efficient desorption/ionization technique for mass spectrometry (MS) of complex molecules and their reactions on surfaces. DINeC is based on a beam of SO2 clusters impacting on the sample surface at low cluster energy. During cluster-surface impact, some of the surface molecules are desorbed and ionized via dissolvation in the impacting cluster; as a result of this dissolvation-mediated desorption mechanism, low cluster energy is sufficient and the desorption process is extremely soft. Both surface adsorbates and molecules of which the surface is composed of can be analyzed. Clear and fragmentation-free spectra from complex molecules such as peptides and proteins are obtained. DINeC does not require any special sample preparation, in particular no matrix has to be applied. The method yields quantitative information on the composition of the samples; molecules at a surface coverage as low as 0.1 % of a monolayer can be detected. Surface reactions such as H/D exchange or thermal decomposition can be observed in real-time and the kinetics of the reactions can be deduced. Using a pulsed nozzle for cluster beam generation, DINeC can be efficiently combined with ion trap mass spectrometry. The matrix-free and soft nature of the DINeC process in combination with the MSn capabilities of the ion trap allows for very detailed and unambiguous analysis of the chemical composition of complex organic samples and organic adsorbates on surfaces.

Neutral SO2 clusters of low kinetic energy (&#60; 0.8 eV/constituent) are used to desorb complex surface molecules such as peptides or lipids for further analysis by means of mass spectrometry using an ion trap mass spectrometer. No special sample preparation is required, and real-time observation of reactions is possible.

Kümelenme Sonucu Desorpsiyon/İyonlaşma Kütle Spektrometresi Yoluyla Karmaşık Moleküllerin ve Yüzeylerdeki Reaksiyonlarının Analizi

Desorption/Ionization Induced by Neutral SO2 Clusters (DINeC) is employed as a very soft and efficient desorption/ionization technique for mass ...

Covalent Attachment of Single Molecules for AFM-based Force Spectroscopy

Atomic force microscopy (AFM)-based single molecule force spectroscopy is an ideal tool for investigating the interactions between a single polymer and surfaces. For a true single molecule experiment, covalent attachment of the probe molecule is essential because only then can hundreds of force-extension traces with one and the same single molecule be obtained. Many traces are in turn necessary to prove that a single molecule alone is probed. Additionally, passivation is crucial for preventing unwanted interactions between the single probe molecule and the AFM cantilever tip as well as between the AFM cantilever tip and the underlying surface. The functionalization protocol presented here is reliable and can easily be applied to a variety of polymers. Characteristic single molecule events (i.e., stretches and plateaus) are detected in the force-extension traces. From these events, physical parameters such as stretching force, desorption force and desorption length can be obtained. This is particularly important for the precise investigation of stimuli-responsive systems at the single molecule level. As exemplary systems poly(ethylene glycol) (PEG), poly(N-isopropylacrylamide) (PNiPAM) and polystyrene (PS) are stretched and desorbed from SiOx (for PEG and PNiPAM) and from hydrophobic self-assembled monolayer surfaces (for PS) in aqueous environment.

Covalent attachment of probe molecules to atomic force microscopy (AFM) cantilever tips is an essential technique for the investigation of their physical properties. This allows us to determine the stretching force, desorption force and length of polymers via AFM-based single molecule force spectroscopy with high reproducibility.

AFM tabanlı Kuvvet Spektroskopisi için Tek Moleküllerin Kovalent Eki

Atomic force microscopy (AFM)-based single molecule force spectroscopy is an ideal tool for investigating the interactions between a single polymer and ...

Synthesis of pH Dependent Pyrazole, Imidazole, and Isoindolone Dipyrrinone Fluorophores using a Claisen-Schmidt Condensation Approach

Methine-bridged conjugated bicyclic aromatic compounds are common constituents of a range of biologically relevant molecules such as porphyrins, dipyrrinones, and pharmaceuticals. Additionally, restricted rotation of these systems often results in highly to moderately fluorescent systems as observed in 3H,5H-dipyrrolo[1,2-c:2',1'-f]pyrimidin-3-ones, xanthoglows, pyrroloindolizinedione analogs, BODIPY analogs, and the phenolic and imidazolinone ring systems of Green Fluorescent Protein (GFP). This manuscript describes an inexpensive and operationally simple method of performing a Claisen-Schmidt condensation to generate a series of fluorescent pH dependent pyrazole/imidazole/isoindolone dipyrrinone analogs. While the methodology illustrates the synthesis of dipyrrinone analogs, it can be translated to produce a wide range of conjugated bicyclic aromatic compounds. The Claisen-Schmidt condensation reaction utilized in this method is limited in scope to nucleophiles and electrophiles that are enolizable under basic conditions (nucleophile component) and non-enolizable aldehydes (electrophile component). Additionally, both the nucleophilic and electrophilic reactants must contain functional groups that will not inadvertently react with hydroxide. Despite these limitations, this methodology offers access to completely novel systems that can be employed as biological or molecular probes.

The Claisen-Schmidt condensation reaction is an important methodology for the generation of methine-bridged conjugated bicyclic aromatic compounds. Through utilizing a base-mediated variant of the aldol reaction, a range of fluorescent and/or biologically relevant molecules can be accessed through a generally inexpensive and operationally simple synthetic approach.

Claisen-Schmidt Yoğuşma Yaklaşımı Kullanılarak pH Bağımlı Pyrazole, Imidazol ve İzinolon Dipyrrinone Flor Floroforlarının Sentezi

Methine-bridged conjugated bicyclic aromatic compounds are common constituents of a range of biologically relevant molecules such as porphyrins, ...

Analysis of Organochlorine Pesticides in a Soil Sample by a Modified QuEChERS Approach Using Ammonium Formate

Currently, the QuEChERS method represents the most widely used sample preparation protocol worldwide for analyzing pesticide residues in a broad variety of matrices both in official and non-official laboratories. The QuEChERS method using ammonium formate has previously proven to be advantageous compared to the original and the two official versions. On the one hand, the simple addition of 0.5 g of ammonium formate per gram of sample is sufficient to induce phase separation and achieve good analytical performance. On the other hand, ammonium formate reduces the need for maintenance in routine analyses. Here, a modified QuEChERS method using ammonium formate was applied for the simultaneous analysis of organochlorine pesticide (OCP) residues in agricultural soil. Specifically, 10 g of the sample was hydrated with 10 mL of water and then extracted with 10 mL of acetonitrile. Next, phase separation was carried out using 5 g of ammonium formate. After centrifugation, the supernatant was subjected to a dispersive solid-phase extraction clean-up step with anhydrous magnesium sulfate, primary-secondary amine, and octadecylsilane. Gas chromatography-mass spectrometry was used as the analytical technique. The QuEChERS method using ammonium formate is demonstrated as a successful alternative for extracting OCP residues from a soil sample.

The present protocol describes the utilization of ammonium formate for phase partitioning in QuEChERS, together with gas chromatography-mass spectrometry, to successfully determine organochlorine pesticide residues in a soil sample.

Bir Toprak Numunesindeki Organoklorin Pestisitlerinin Amonyum Format Kullanarak Modifiye QuEChERS Yaklaşımı ile Analizi

Currently, the QuEChERS method represents the most widely used sample preparation protocol worldwide for analyzing pesticide residues in a broad variety ...