Name: dendrimer-based uneven nanopatterns to locally control surface adhesiveness: a method to direct chondrogenic differentiation
Uploaded: 2018-01-20T14:00:00
Description: Watch this Scientific Journal Video about Dendrimer-based Uneven Nanopatterns to Locally Control Surface Adhesiveness: A Method to Direct Chondrogenic Differentiation at JoVE.com

Yazarlar Oriol yazı tipi-Bach ve Albert G. Castaño ddk miktar onların yardım kabul edersiniz. Onlar da gelişmiş Digital mikroskobu birimi (IRB Barcelona) kendi tesislerinde video kaydetmek yazarlar izin Biyomedikal Araştırma Enstitüsü'nde kabul. Bu eser ağ Biyomedikal Araştırma Merkezi (CIBER tarafından), İspanya desteklenmiştir. CIBER bir girişim VI Ulusal ar-ge & ben planı 2008-2011, bulunan INGENIO 2010, Consolider programı, CIBER eylemleri ve Instituto de tarafından Salud Carlos III, Avrupa Bölgesel Kalkınma Fonu desteği ile finanse var. Bu eser üniversiteler ve araştırma departmanı yenilik, üniversiteler ve kurumsal Generalitat de Catalunya için komisyon tarafından desteklenen (2014 SGR 1442). Ayrıca OLIGOCODES (No projeler tarafından finanse edildi MAT2012-38573-C02) ve CTQ2013-41339-P, rekabet ve İspanyolca Ekonomi Bakanlığı tarafından Ayrıca INTERREG V-A İspanya-Portekiz 2014 2020 POCTEP (0245_IBEROS_1_E) için verilmiştir. C.R.P. İspanyolca Ekonomi Bakanlığı ve rekabet gücünü grant (No mali desteği kabul eder. IFI15/00151).

1. yüzey hazırlama
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1.4 x 1.1 cm Au(111) Mika yüzeyler üzerinde tavlama.
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<li>Cam seramik Ocak Au(111) substrat yerleştirin ve bir argon atmosfer altında soğutmak için belgili tanımlık substrate 3 dk. izin ver bir bütan alev ile TAV. Her Au(111) substrat için bu adımı yineleyin. Not: Au(111) yüzeylerde hemen tavlama sonra kullanılmalıdır.</li>
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Poli (L-laktik asit) hazırlanması (PLLA)-cam yüzeylerde kaplı.
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<li>Kesme ve cam s...

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	<li>Jiang, F., H&#246;rber, H., Howard, J., M&#252;ller, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assembly+of+collagen+into+microribbons:+Effects+of+pH+and+electrolytes.">Assembly of collagen into microribbons: Effects of pH and electrolytes.</a> J. Struct. Biol. 148 (3), 268-278 (2004).</li><li>Smith, M. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Force-induced+unfolding+of+fibronectin+in+the+extracellular+matrix+of+living+cells.">Force-induced unfolding of fibronectin in the extracellular matrix of living cells.</a> PLoS Biol. 5 (10), 2243-2254 (2007).</li><li>Little, W. C., Smith, M. L., Ebneter, U., Vogel, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assay+to+mechanically+tune+and+optically+probe+fibrillar+fibronectin+conformations+from+fully+relaxed+to+breakage.">Assay to mechanically tune and optically probe fibrillar fibronectin conformations from fully relaxed to breakage.</a> Matrix Biol. 27 (5), 451-461 (2008).</li><li>Christman, K. L., Enriquez-Rios, V. D., Maynard, H. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanopatterning+proteins+and+peptides.">Nanopatterning proteins and peptides.</a> Soft Matter. 2, 928-939 (2006).</li><li>Falconnet, D., Csucs, G., Grandin, H. M., Textor, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surface+engineering+approaches+to+micropattern+surfaces+for+cell-based+assays.">Surface engineering approaches to micropattern surfaces for cell-based assays.</a> Biomaterials. 27 (16), 3044-3063 (2006).</li><li>Arnold, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activation+of+integrin+function+by+nanopatterned+adhesive+interfaces.">Activation of integrin function by nanopatterned adhesive interfaces.</a> ChemPhysChem. 5 (3), 383-388 (2004).</li><li>Cavalcanti-Adam, E. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lateral+spacing+of+integrin+ligands+influences+cell+spreading+and+focal+adhesion+assembly.">Lateral spacing of integrin ligands influences cell spreading and focal adhesion assembly.</a> Eur. J. Cell. Biol. 85 (3-4), 219-224 (2006).</li><li>Cavalcanti-Adam, E. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+spreading+and+focal+adhesion+dynamics+are+regulated+by+spacing+of+integrin+ligands.">Cell spreading and focal adhesion dynamics are regulated by spacing of integrin ligands.</a> Biophys. J. 92 (8), 2964-2974 (2007).</li><li>Arnold, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+interactions+with+hierarchically+structured+nano-patterned+adhesive+surfaces.">Cell interactions with hierarchically structured nano-patterned adhesive surfaces.</a> Soft Matter. 5 (1), 72-77 (2009).</li><li>Malmstr&#246;m, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Large+area+protein+patterning+reveals+nanoscale+control+of+focal+adhesion+development.">Large area protein patterning reveals nanoscale control of focal adhesion development.</a> Nano Lett. 10 (2), 686-694 (2010).</li><li>Deeg, J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+local+versus+global+ligand+density+on+cellular+adhesion.">Impact of local versus global ligand density on cellular adhesion.</a> Nano Lett. 11 (4), 1469-1476 (2011).</li><li>Medda, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigation+of+early+cell&#8211;surface+interactions+of+human+mesenchymal+stem+cells+on+nanopatterned+&#946;-type+titanium-niobium+alloy+surfaces.">Investigation of early cell&#8211;surface interactions of human mesenchymal stem cells on nanopatterned &#946;-type titanium-niobium alloy surfaces.</a> Interface Focus. 4, 20130046 (2014).</li><li>Wang, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+RGD+nanospacing+on+differentiation+of+stem+cells.">Effect of RGD nanospacing on differentiation of stem cells.</a> Biomaterials. 34 (12), 2865-2874 (2013).</li><li>Wang, X., Ye, K., Li, Z. H., Yan, C., Ding, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adhesion,+proliferation,+and+differentiation+of+mesenchymal+stem+cells+on+RGD+nanopatterns+of+varied+nanospacings.">Adhesion, proliferation, and differentiation of mesenchymal stem cells on RGD nanopatterns of varied nanospacings.</a> Organogenesis. 9 (4), 280-286 (2013).</li><li>Wang, X., Li, S. Y., Yan, C., Liu, P., Ding, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fabrication+of+RGD+micro/nanopattern+and+corresponding+study+of+stem+cell+differentiation.">Fabrication of RGD micro/nanopattern and corresponding study of stem cell differentiation.</a> Nano Lett. 15 (3), 1457-1467 (2015).</li><li>Li, Z. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+RGD+nanospacing+on+chondrogenic+differentiation+of+mesenchymal+stem+cells.">Effects of RGD nanospacing on chondrogenic differentiation of mesenchymal stem cells.</a> J. Mater. Chem. B. 3 (12), 5197-5209 (2015).</li><li>Stephanopoulos, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioactive+DNA-peptide+nanotubes+enhance+the+differentiation+of+neural+stem+cells+into+neurons.">Bioactive DNA-peptide nanotubes enhance the differentiation of neural stem cells into neurons.</a> Nano Lett. 15 (1), 603-609 (2015).</li><li>Rolland, O., Turrin, C. O., Caminade, A. M., Majoral, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dendrimers+and+nanomedicine:+Multivalency+in+action.">Dendrimers and nanomedicine: Multivalency in action.</a> New J. Chem. 33, 1809-1824 (2009).</li><li>Saovapakhiran, A., D&#8217;Emanuele, A., Attwood, D., Penny, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surface+modification+of+PAMAM+dendrimers+modulates+the+mechanism+of+cellular+internalization.">Surface modification of PAMAM dendrimers modulates the mechanism of cellular internalization.</a> Bioconjug. Chem. 20 (4), 693-701 (2009).</li><li>Albertazzi, L., Fernandez-Villamarin, M., Riguera, R., Fernandez-Megia, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peripheral+functionalization+of+dendrimers+regulates+internalization+and+intracellular+trafficking+in+living+cells.">Peripheral functionalization of dendrimers regulates internalization and intracellular trafficking in living cells.</a> Bioconjug. Chem. 23 (5), 1059-1068 (2012).</li><li>Mikhail, A. S., Jones, K. S., Sheardown, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dendrimer+grafted+cell+adhesion+peptide-modified+PDMS.">Dendrimer grafted cell adhesion peptide-modified PDMS.</a> Biotechnol. Prog. 24 (4), 938-944 (2008).</li><li>Kino-oka, M., Kim, J., Kurisaka, K., Kim, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preferential+growth+of+skeletal+myoblasts+and+fibroblasts+in+co-culture+on+a+dendrimer-immobilized+surface.">Preferential growth of skeletal myoblasts and fibroblasts in co-culture on a dendrimer-immobilized surface.</a> J. Biosci. Bioeng. 115 (1), 96 (2013).</li><li>Kim, M. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphological+regulation+and+aggregate+formation+of+rabbit+chondrocytes+on+dendrimer+immobilized+surfaces+with+D-glucose+display.">Morphological regulation and aggregate formation of rabbit chondrocytes on dendrimer immobilized surfaces with D-glucose display.</a> J. Biosci. Bioeng. 107 (2), 196-205 (2009).</li><li>Lomba, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+adhesion+on+surface+patterns+generated+by+the+photocrosslinking+of+hyperbranched+polyesters+with+a+trisdiazonium+salt.">Cell adhesion on surface patterns generated by the photocrosslinking of hyperbranched polyesters with a trisdiazonium salt.</a> React. Funct. Polym. 73 (3), 499-507 (2013).</li><li>Maheshwari, G., Brown, G., Lauffenburger, D. A., Wells, A., Griffith, L. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+adhesion+and+motility+depend+on+nanoscale+RGD+clustering.">Cell adhesion and motility depend on nanoscale RGD clustering.</a> J. Cell Sci. 113 (Pt 10), 1677-1686 (2000).</li><li>Kim, M. H., Kino-oka, M., Kawase, M., Yagi, K., Taya, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synergistic+effect+of+D-glucose+and+epidermal+growth+factor+display+on+dynamic+behaviors+of+human+epithelial+cells.">Synergistic effect of D-glucose and epidermal growth factor display on dynamic behaviors of human epithelial cells.</a> J. Biosci. Bioeng. 104 (5), 428-431 (2007).</li><li>Pericet-Camara, R., Cahill, B. P., Papastavrou, G., Borkovec, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nano-patterning+of+solid+substrates+by+adsorbed+dendrimers.">Nano-patterning of solid substrates by adsorbed dendrimers.</a> Chem. Commun. 3, 266-268 (2007).</li><li>Lagunas, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Large-scale+dendrimer-based+uneven+nanopatterns+for+the+study+of+local+arginine&#8211;glycine&#8211;aspartic+acid+(RGD)+density+effects+on+cell+adhesion.">Large-scale dendrimer-based uneven nanopatterns for the study of local arginine&#8211;glycine&#8211;aspartic acid (RGD) density effects on cell adhesion.</a> Nano Res. 7 (3), 399-409 (2014).</li><li>Lagunas, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tailoring+RGD+local+surface+density+at+the+nanoscale+toward+adult+stem+cell+chondrogenic+commitment.">Tailoring RGD local surface density at the nanoscale toward adult stem cell chondrogenic commitment.</a> Nano Res. , (2017).</li><li>Prats-Alfonso, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effective+and+Versatile+Strategy+for+the+Total+Solid-Phase+Synthesis+of+Alkanethiols+for+Biological+Applications.">Effective and Versatile Strategy for the Total Solid-Phase Synthesis of Alkanethiols for Biological Applications.</a> Eur. J. Org. Chem. 2013 (7), 1233-1239 (2013).</li><li>Zhu, Y. B., Gao, C. Y., Liu, X. Y., He, T., Shen, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immobilization+of+biomacromolecules+onto+aminolyzed+poly(L-lactic+acid)+toward+acceleration+of+endothelium+regeneration.">Immobilization of biomacromolecules onto aminolyzed poly(L-lactic acid) toward acceleration of endothelium regeneration.</a> Tissue Eng. 10 (1-2), 53-61 (2004).</li><li>G&#252;ell, A., D&#237;ez-P&#233;rez, I., Gorostiza, P., Sanz, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+reliable+probes+for+electrochemical+tunneling+spectroscopy.">Preparation of reliable probes for electrochemical tunneling spectroscopy.</a> Anal. Chem. 76 (17), 5218-5222 (2004).</li><li>Bobick, B. E., Chen, F. H., Le, A. M., Tuan, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+the+chondrogenic+phenotype+in+culture.">Regulation of the chondrogenic phenotype in culture.</a> Birth Defects Res. C Embryo Today. 87 (4), 351-371 (2009).</li><li>De Lise, A. M., Fisher, L., Tuan, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+interactions+and+signaling+in+cartilage+development.">Cellular interactions and signaling in cartilage development.</a> Osteoarthritis Cartilage. 8 (5), 309-334 (2000).</li><li>Kosher, R. A., Kulyk, W. M., Gay, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Collagen+gene+expression+during+limb+cartilage+differentiation.">Collagen gene expression during limb cartilage differentiation.</a> J. Cell Biol. 102 (4), 1151-1156 (1986).</li><li>Biebricher, A., Paul, A., Tinnefeld, P., Golzhauser, A., Sauer, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+three-dimensional+immobilization+of+biomolecules+on+chemically+patterned+surfaces.">Controlled three-dimensional immobilization of biomolecules on chemically patterned surfaces.</a> J. Biotechnol. 112 (1-2), 97-107 (2004).</li><li>Tinazli, A., Piehler, J., Beuttler, M., Guckenberger, R., Tamp&#233;, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Native+protein+nanolithography+that+can+write,+read+and+erase.">Native protein nanolithography that can write, read and erase.</a> Nat. Nanotechnol. 2, 220-225 (2007).</li><li>Oberhansl, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Facile+Modification+of+Silica+Substrates+Provides+a+Platform+for+Direct-Writing+Surface+Click+Chemistry.">Facile Modification of Silica Substrates Provides a Platform for Direct-Writing Surface Click Chemistry.</a> Small. 8 (4), 541-545 (2012).</li></ol>

Açıklanan protokol geliştirilmesi sırasında birkaç kritik adım düşünülmelidir. İlk tarama prob mikroskobu teknikleri ile nanopattern karakterizasyonu anlamına gelir. Nanopatterns görselleştirmek için nerede desenlendirme üretilen Yüzey pürüzlülük değerinin altında civarındadır dendrimers ortalama çapı olması gerekir (şekil 1B) STM tarafından ölçülen 4 – 5 nm. Ayrıca, yüksek çözünürlüklü STM görüntüleme için bu durumda Au(111) iletken yüzeylerde sınırlıd...

Burada biz Nano yerel adhesiveness kontrolünü sağlayan hücre kültür yüzeyler elde etmek için bir basit ve çok yönlü dendrimer tabanlı nanopatterning açıklayınız. 1,2,3 ve hücre adezyon yüzeyleri nanopatterning yapışma4' eilgili hücresel gereksinimleri içine derin anlayışlar sağlamıştır ECM organizasyon ayrıntılarını Nano bildirilmiştir, 5. Micellar nanopatterns litografi tabanlı kullanarak deneyler ortaya bir eşik değeri yaklaşık 70 nm RGD peptid nanospacing, hücre adezyon önemli ölçüde bu değer6,7,8 gecikmiş için ,9. Bu çalışmalar aynı zamanda büyük etkisi yerel küresel ligand yoğunluğu daha hücre adezyon9,10,11vurgulanır.
Morfogenez sırasında çevreye olan hücre etkileşimleri son karmaşık doku yapılar oluşturulmuştur kadar hangi devam ilk farklılaşma olayları tetikler. Bu çerçevede, nanopatterned yüzeyler ilk hücre yüzeyine etkileşimler morfogenez üzerinde etkisi mücadele için kullanılmaktadır. Litografi tabanlı RGD nanopatterns 68 yanal bir boşluk ile β-türü Ti-40Nb nm alaşımlar RGD nanospacings 95 ve 150 nm arasında bir farklılaşma geliştirmek iken sigara için kabul edilen kök hücreler12, farklılaşmamış fenotip korumak için yardım Adipojenik/Osteojenik13,14,15 chondrogenic kader ve16doğru Mezenkimal Kök hücre (MSCs). Ayrıca, sinyal bileşenlerle değiştiren Self montaj oluştururlar hücre adezyon ve farklılaşma Nano mimari düzenleme, sinyal gönderme ipuçları17sağlayarak doğrudan gösterilmiştir. Bu bağlamda, dendrimers yüzeyler üzerine onların dış küre18,19,20 moieties hücre etkileşim ile birikimi hücre adezyon21,22eğitim için kullanılan, Morfoloji23,24ve göç olayları25,26. Yine de, bu çalışmalarda yüzey karakterizasyonu eksikliği zor dendrimer yüzey yapılandırma ve hücre yanıt arasında herhangi bir ilişki kurmak için yapar.
Ne zaman dendrimers düşük ücret yüzeylere çözümleri ile düşük iyonik güçlü absorbe Dendrimer nanopatterns sıvı gibi sipariş ve tanımlanmış aralığı ile elde edilebilir. 27 bu özellik temelinde, burada büyük ölçekli düzensiz nanopatterns RGD functionalized dendrimers Nano yönetilsin yerel RGD yüzey yoğunluğu düşük ücret yüzeylerde, elde etmek için bir yöntem mevcut. Su temas açısı (CA), x-ışını photoelectron spektroskopisi (XPS) ve tarama prob mikroskobu teknikleri (STM ve AFM nanopatterns) gösteri çözümünde ilk dendrimer konsantrasyon değiştirme yerel ligand yoğunluğu ayarlanabilir. Yerel RGD yüzey yoğunluğu AFM görüntülerden en az interparticle mesafeler tarafından olasılık kontur haritaları sayısal ve hücre deneyleri ile ilişkili. Diğer nanopatterning teknikleri4ile karşılaştırıldığında, nanopatterning dendrimer tabanlı basit ve kolayca böylece hücre kültür uygulamaları ile tam olarak uyumlu olmak büyük yüzey alanlarını ölçeklendirilebilir. Nanopatterns biyoaktif yüzeylerde hücre adezyon28 ve yetişkin insan MSCs29chondrogenic indüksiyon yerel RGD yüzey yoğunluğu etkisini değerlendirmek için kullanılır. Bizim sonuçlar RGD dendrimer tabanlı nanopatterns hücre büyümesini sürdürmek ve hücre adezyon tarafından yüksek yerel RGD yüzey yoğunluğu takviye edilmiştir gösterir. Farklılaşma deneylerde MSC yoğunlaşma ve erken chondrogenic farklılaşma hücre yüzeyler için ara adhesiveness tercih. Hangi dendrimer ile çevre grupları değiştirilebilir kolaylığı nedeniyle, burada açıklanan yöntemi daha fazla konsantrasyon bağımlı etkileri hücrelerde sarfetmek diğer ECM ligandlar için genişletilebilir.

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		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Glass micro slides, plain</td>
			<td style="width:169px;">Corning</td>
			<td style="width:307px;">2947-75x25</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Deionized water</td>
			<td style="width:169px;">Millipore</td>
			<td style="width:307px;"></td>
			<td>18M&#937; cm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Ethanol 96%</td>
			<td style="width:169px;">PanReac</td>
			<td style="width:307px;">131085.1212</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:504px;">L-Lactide/DL-Lactide copolymer</td>
			<td style="width:169px;">Corbion</td>
			<td style="width:307px;"></td>
			<td>95/05 molar ratio</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:504px;">1,4 - dioxane</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:307px;">296309-1L</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:504px;">Silicone oil, high temperature</td>
			<td style="width:169px;">Acros Organics</td>
			<td style="width:307px;">174665000</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:504px;">Spinner</td>
			<td style="width:169px;">Laurell</td>
			<td style="width:307px;">WS-650MZ-23NPP/Lite</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:504px;">Tissue culture laminar flow hood</td>
			<td style="width:169px;">Telstar</td>
			<td style="width:307px;">Bio II Advance</td>
			<td>Class II biological safety cabinet</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Filter unit</td>
			<td style="width:169px;">Millex-GP</td>
			<td style="width:307px;">SLGP033RB</td>
			<td>0.22 &#181;m</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Syringe 10 mL</td>
			<td style="width:169px;">Discardit</td>
			<td style="width:307px;">309110</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Atomic Force microscope</td>
			<td style="width:169px;">Veeco Instruments</td>
			<td style="width:307px;">Dimension 3000 AFM instrument</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:504px;">Silicon AFM probes</td>
			<td style="width:169px;">Budget Sensors</td>
			<td style="width:307px;">Tap300AI-G</td>
			<td>Resonant Freq. 300 kHz, k = 40 N/m</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:504px;">Scanning tunneling microscope</td>
			<td style="width:169px;">Molecular Imaging</td>
			<td style="width:307px;">PicoSPM microscope</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:504px;">Pt0.8:Ir0.2 wire</td>
			<td style="width:169px;">Advent</td>
			<td style="width:307px;">PT671012</td>
			<td>Diameter 0.25 mm</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:504px;">WSxM 4.0 software</td>
			<td style="width:169px;">Nanotec electronica</td>
			<td style="width:307px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Optical contact angle (CA) system</td>
			<td style="width:169px;">Dataphysics</td>
			<td style="width:307px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">SCA20 software</td>
			<td style="width:169px;">Dataphysics</td>
			<td style="width:307px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:504px;">X-ray photoelectron spectrometer</td>
			<td style="width:169px;">Physical Electronics</td>
			<td style="width:307px;">Perkin-Elmer PHI 5500 Multitechnique System</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Fibronectin from bovine plasma</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:307px;">F1141-1MG</td>
			<td>1.0 mL solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Dulbecco&#39;s Phosphate Buffer Saline (DPBS)</td>
			<td style="width:169px;">Gibco</td>
			<td style="width:307px;">21600-10</td>
			<td>Powder</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Mouse embryo fibroblasts</td>
			<td style="width:169px;">ATCC</td>
			<td style="width:307px;">ATCC CRL-1658</td>
			<td>NIH/3T3</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:504px;">Dulbecco&#39;s modified eagle medium (DMEM) liquid high glucose</td>
			<td style="width:169px;">Gibco</td>
			<td style="width:307px;">11960044</td>
			<td>liquid high glucose, no glutamine, 500 mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Fetal Bovine Serum (FBS)</td>
			<td style="width:169px;">Gibco</td>
			<td style="width:307px;">16000044</td>
			<td>500 mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">L-Glutamine</td>
			<td style="width:169px;">Invitrogen</td>
			<td style="width:307px;">25030</td>
			<td>200 mM (100X)</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:504px;">Penicillin-streptomycin</td>
			<td style="width:169px;">Invitrogen</td>
			<td style="width:307px;">15140</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:504px;">Sodium pyruvate</td>
			<td style="width:169px;">Invitrogen</td>
			<td style="width:307px;">11360039</td>
			<td>100 mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">T75 culture flasks</td>
			<td style="width:169px;">Nunclon</td>
			<td style="width:307px;">156499</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:504px;">Trypsin</td>
			<td style="width:169px;">Life Technologies</td>
			<td style="width:307px;">25200072</td>
			<td>0,25% EDTA</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Centrifuge</td>
			<td style="width:169px;">Hermle Labortechnik</td>
			<td style="width:307px;">Z 206 A</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:504px;">Non-tissue culture treated plate, 12 well</td>
			<td style="width:169px;">Falcon</td>
			<td style="width:307px;">351143</td>
			<td>Non-adherent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Adipose-derived hMSCs</td>
			<td style="width:169px;">ATCC</td>
			<td style="width:307px;">ATCC PCS-500-011</td>
			<td>Cell vial 1 mL</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">MSC basal medium</td>
			<td style="width:169px;">ATCC</td>
			<td style="width:307px;">ATCC PCS-500-030</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">MSC growth kit</td>
			<td style="width:169px;">ATCC</td>
			<td style="width:307px;">ATCC PCS-500-040</td>
			<td>Low serum</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Chondrocyte differentiation tool</td>
			<td style="width:169px;">ATCC</td>
			<td style="width:307px;">ATCC PCS-500-051</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Formalin solution</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:307px;">HT5011-15ML</td>
			<td>neutral buffered, 10%</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:504px;">Ammonium chloride</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:307px;">A9434-500G</td>
			<td>for molecular biology, suitable for cell culture, &#8805;99.5%</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Saponin</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:307px;">47036-50G-F</td>
			<td>for molecular biology, used as non-ionic surfactant, adjuvant</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:504px;">Bovine Serum Albumin (BSA)</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:307px;">A3059-50G</td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:504px;">Rabbit monoclonal [Y113] anti-paxillin antibody</td>
			<td style="width:169px;">Abcam</td>
			<td style="width:307px;">ab32084</td>
			<td>Diluted 1:200</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Mouse monoclonal [1F5] anti-collagen alpha-1 XX chain&#160;</td>
			<td style="width:169px;">Acris Antibodies</td>
			<td style="width:307px;">AM00212PU-N</td>
			<td>Diluted: 1:400</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:504px;">Alexa Fluor 488-conjugated goat anti-mouse IgG (H+L) secondary antibody</td>
			<td style="width:169px;">Invitrogen</td>
			<td style="width:307px;">A10667</td>
			<td>2 mg/mL. Diluted 1:1000</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Alexa Fluor 568-conjugated goat anti-rabbit IgG (H+L) secondary antibody</td>
			<td style="width:169px;">Invitrogen</td>
			<td style="width:307px;">A11036</td>
			<td>2 mg/mL. Diluted 1:1000</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:504px;">Hoechst 33342</td>
			<td style="width:169px;">Thermo Fisher</td>
			<td style="width:307px;">H3570 10ML</td>
			<td>10 mg/mL. Diluted 1:1000</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">Cover glass 24x24 mm</td>
			<td style="width:169px;">Deltalab</td>
			<td style="width:307px;">D102424</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:504px;">Fluoromount</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:307px;">F4680-25ML</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:504px;">Epifluorescence Microscope</td>
			<td style="width:169px;">Nikon</td>
			<td style="width:307px;">Eclipse E1000 upright microscope</td>
			<td>with a CCD camera</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:504px;">Confocal Microscope</td>
			<td style="width:169px;">Leica Microsystems</td>
			<td style="width:307px;"></td>
			<td>Leica SPE Upright Confocal Microscope</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">ImageJ 1.50g freeware</td>
			<td style="width:169px;">http://imgej.nih.gov/ij</td>
			<td style="width:307px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">MATLAB software</td>
			<td style="width:169px;">The MATHWORKS, Inc.</td>
			<td style="width:307px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:504px;">OriginPro 8.5 software&#160;</td>
			<td style="width:169px;">OriginLab Coorporation</td>
			<td style="width:307px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

dendrimer-based uneven nanopatterns to locally control surface adhesiveness: a method to direct chondrogenic differentiation

Hücresel yapışma ve farklılaşma hücre dışı Matriks (ECM) bileşenleri, nano eğilim tarafından büyük bir etkiye sahip yerel konsantrasyonları ile klimalı. Burada büyük ölçekli düzensiz nanopatterns arginin-glisin-aspartik asit (RGD) elde etmek için bir yöntem mevcut-yerel RGD Nano yönetilsin functionalized dendrimers yüzey yoğunluğu. Nanopatterns dendrimers ilk farklı konsantrasyonlarda çözümlerinden yüzey adsorpsiyon tarafından kurulur ve su temas açısı tarafından (CA), x-ışını photoelectron spektroskopisi (XPS), ile karakterizedir ve tarama prob mikroskobu teknikleri gibi tünelleme mikroskobu (STM) ve Atomik Kuvvet Mikroskobu (AFM) tarama. RGD yerel yüzey yoğunluğu en az interparticle mesafeler aracılığıyla olasılık kontur haritaları AFM görüntüleri kullanarak ve sonra hücre adezyon yanıt ve farklılaşma ile ilişkili ölçülür. Nanopatterning yöntemi burada sunulan geniş yüzey alanları için basit bir şekilde ölçeklendirilebilir basit bir işlemdir. Bu nedenle hücre kültür protokolleri ile tamamen uyumlu ve konsantrasyon bağımlı etkileri hücrelerde sarfetmek diğer ligandlar uygulanabilir.

Biz Nano (şekil 1) ele alınması gereken yüzey adhesiveness sağlayan bir nanopatterning yöntemi mevcut. RGD-Cys-D1 kimyasal yapısını şekil 1Aile gösterilir. Dendrimers elektrik iletken Au(111) yüzeylerde yüksek çözünürlük STM karakterizasyonu için desenli. Çözüm (en çok 10-5% w/w) konsantrasyonlarda düşük dendrimer 4-5 nm çapında (şekil 1B), daha yükse...

Watch this Scientific Journal Video about Dendrimer-based Uneven Nanopatterns to Locally Control Surface Adhesiveness: A Method to Direct Chondrogenic Differentiation at JoVE.com

Dendrimer-based Uneven Nanopatterns to Locally Control Surface Adhesiveness: A Method to Direct Chondrogenic Differentiation

Dendrimer tabanlı yerel arginin-glisin-aspartik asit (RGD) yüzey yoğunluğu Nano yönetilsin düzensiz nanopatterns elde etmek için bir yöntem açıklanan ve hücre adezyon ve chondrogenic farklılaşması çalışma için uygulanmıştır.

Düzensiz Nanopatterns Dendrimer tabanlı yerel olarak kumanda yüzeyi Adhesiveness için: Chondrogenic farklılaşma yönlendirmek için bir yöntem

Cellular adhesion and differentiation is conditioned by the nanoscale disposition of the extracellular matrix (ECM) components, with local concentrations ...

Research

JoVE Journal

Bioengineering

Measurement of Aggregate Cohesion by Tissue Surface Tensiometry

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 ...

Design of a Cyclic Pressure Bioreactor for the Ex Vivo Study of Aortic Heart Valves

Design of a Cyclic Pressure Bioreactor for the Ex Vivo Study of Aortic Heart Valves

Devirli Basınç Biyoreaktör Tasarım Ex vivo</emAort Kalp Vanalar> Eğitim

The aortic valve, located between the left ventricle and the aorta, allows for unidirectional blood flow, preventing backflow into the ventricle. Aortic ...

Engineering a Bilayered Hydrogel to Control ASC Differentiation

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 ...

A Toolkit to Enable Hydrocarbon Conversion in Aqueous Environments

Sulu Ortamlarda Hidrokarbon Dönüşüm Enable A Toolkit

This work puts forward a toolkit that enables the conversion of alkanes by Escherichia coli and presents a proof of principle of its applicability. The ...

A Novel Method for Localizing Reporter Fluorescent Beads Near the Cell Culture Surface for Traction Force Microscopy

Çekme Kuvveti Mikroskopi Hücre Kültürü Yüzey civarında yerelleştirme raportör flöresan Beads için Yeni Bir Yöntem

PA gels have long been used as a platform to study cell traction forces due to ease of fabrication and the ability to tune their elastic properties. When ...

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

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

An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry

Multispectral görüntüleme akışı sitometrisi kullanarak In vitro micronucleus assay gerçekleştirmek için otomatik bir yöntem

The in vitro micronucleus (MN) assay is often used to evaluate cytotoxicity and genotoxicity but scoring the assay via manual microscopy is laborious and ...

Experimental Methods to Study Human Postural Control

İnsan Postural Kontrolünü İncelemek İçin Deneysel Yöntemler

Many components of the nervous and musculoskeletal systems act in concert to achieve the stable, upright human posture. Controlled experiments accompanied ...

Simultaneous Quantification of Selected Kynurenines Analyzed by Liquid Chromatography-Mass Spectrometry in Medium Collected from Cancer Cell Cultures

Kanser Hücre Kültürlerinden Toplanan Ortamda Sıvı Kromatografi-Kütle Spektrometresi ile Analiz Edilen Seçilmiş Kynureninlerin Eşzamanlı Nicelemesi

The kynurenine pathway and the tryptophan catabolites called kynurenines have received increased attention for their involvement in immune regulation and ...

Single-Cell Optical Action Potential Measurement in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

İnsan kaynaklı Pluripotent Kök Hücre Türevli Kardiyomiyositlerde Tek Hücreli Optik Eylem Potansiyeli Ölçümü

Conventional intracellular microelectrode techniques to quantify cardiomyocyte electrophysiology are extremely complex, labor intensive, and typically ...