نشكر BBSRC لتمويل على درجة الدكتوراه عن اللقاء السيد فريزر.

<ol>
	<li>Canton, I., McKean, R., Charnley, M., Blackwood, K., Fiorica, C., Ryan, A., MacNeil, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+an+Ibuprofen-releasing+biodegradable+PLA/PGA+electrospun+scaffold+for+tissue+regeneration.">Development of an Ibuprofen-releasing biodegradable PLA/PGA electrospun scaffold for tissue regeneration.</a> Biotechnology and bioengineering. 105, 396-408 (2010).</li><li>Blackwood, K., McKean, R., Canton, I., Freeman, C., Franklin, K., Cole, A., Brook, I., Farthing, P., Rimmer, S., Haycock, J., Ryan, A., MacNeil, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+biodegradable+electrospun+scaffolds+for+dermal+replacement.">Development of biodegradable electrospun scaffolds for dermal replacement.</a> Biomaterials. 29, 3091-3104 (2008).</li><li>Yang, F., Maurugan, R., Wang, S., Ramakrishna, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrospinning+of+nano/micro+scale+poly(L-lactic+acid)+aligned+fibers+and+their+potential+in+neural+tissue+engineering.">Electrospinning of nano/micro scale poly(L-lactic acid) aligned fibers and their potential in neural tissue engineering.</a> Biomaterials. 26, 2603-2610 (2005).</li><li>Sittichokechaiwut, A., Edwards, J. H., Scutt, A. M., Reilly, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Short+bouts+of+mechanical+loading+are+as+effective+as+dexamethasone+at+inducing+matrix+production+by+human+bone+marrow+mesenchymal+stem+cell.">Short bouts of mechanical loading are as effective as dexamethasone at inducing matrix production by human bone marrow mesenchymal stem cell.</a> Eur. Cell Mater. 20, 45-57 (2010).</li><li>Sill, T. J., von Recum, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrospinning:+applications+in+drug+delivery+and+tissue+engineering.">Electrospinning: applications in drug delivery and tissue engineering.</a> Biomaterials. 29 (13), 1989-2006 (2008).</li><li>Deitzel, J., Kleinmeyer, J., Harris, D., Beck Tan, N. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+processing+variables+on+the+morphology+of+electrospun+nanofibers+and+textiles.">The effect of processing variables on the morphology of electrospun nanofibers and textiles.</a> Polymer. 42, 261-272 (2001).</li><li>Fridrikh, S., Yu, J., Brenner, M., Rutledge, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlling+the+fiber+diameter+during+electrospinning.">Controlling the fiber diameter during electrospinning.</a> Physical review letters. 90, 1-4 (2003).</li><li>Fong, H., Chun, I., Reneker, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beaded+nanofibers+formed+during+electrospinning.">Beaded nanofibers formed during electrospinning.</a> Polymer. 40 (16), 4585-4592 (1999).</li><li>Selim, M., Bullock, A. J., Blackwood, K. A., Chapple, C. R., MacNeil, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developing+biodegradable+scaffolds+for+tissue+engineering+of+the+urethra.">Developing biodegradable scaffolds for tissue engineering of the urethra.</a> BJU Int. 107 (2), 296-302 (2010).</li><li>Tong, H. -. W., Wang, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+investigation+into+the+influence+of+electrospinning+parameters+on+the+diameter+and+alignment+of+poly(hydroxybutyrate-co-hydroxyvalerate)+fibers.">An investigation into the influence of electrospinning parameters on the diameter and alignment of poly(hydroxybutyrate-co-hydroxyvalerate) fibers.</a> Journal of Applied Polymer Science. 120 (3), 1694-1706 (2011).</li><li>Tong, H. -. W., Wang, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrospinning+of+poly(hydroxybutyrate-co-hydroxyvalerate)+fibrous+tissue+engineering+scaffolds+in+two+different+electric+fields.">Electrospinning of poly(hydroxybutyrate-co-hydroxyvalerate) fibrous tissue engineering scaffolds in two different electric fields.</a> Polymer Engineering &amp; Science. 51 (7), 1325-1338 (2011).</li><li>Retzepi, M., Donos, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guided+Bone+Regeneration:+biological+principle+and+therapeutic+applications.">Guided Bone Regeneration: biological principle and therapeutic applications.</a> Clinical oral implants research. 21, 567-576 (2010).</li><li>Moreau, J., Caccamese, J., Coletti, D., Sauk, J., Fisher, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+engineering+solutions+for+cleft+palates.">Tissue engineering solutions for cleft palates.</a> Journal of oral maxillofacial. 65, 2503-2511 (2007).</li><li>Yang, F., Both, S., Yang, X., Walboomers, X., Jansen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+an+electrospun+nano-apatite/PCL+composite+membrane+for+GTR/GBR+application.">Development of an electrospun nano-apatite/PCL composite membrane for GTR/GBR application.</a> Acta biomaterialia. 5, 3295-3304 (2009).</li><li>Yoshimoto, H., Shin, Y., Terai, H., Vacanti, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+biodegradable+nanofiber+scaffold+by+electrospinning+and+its+potential+for+bone+tissue+engineering.">A biodegradable nanofiber scaffold by electrospinning and its potential for bone tissue engineering.</a> Biomaterials. 24, 2077-2082 (2003).</li><li>Telemeco, T., Ayres, C., Bowlin, G., Wnek, G., Boland, E., Cohen, N., Baumgarten, C., Mathews, J., Simpson, D. <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+cellular+infiltration+into+tissue+engineering+scaffolds+composed+of+submicron+diameter+fibrils+produced+by+electrospinning.">Regulation of cellular infiltration into tissue engineering scaffolds composed of submicron diameter fibrils produced by electrospinning.</a> Acta biomaterialia. 1, 377-385 (2005).</li></ol>

الإعلان عن أي تضارب في المصالح.

Electrospinning هو تقنية شعبية جدا لإنتاج السقالات لهندسة الأنسجة. 14، 15، 16 في حين أنه من السهل نسبيا لإنتاج السقالات electrospun الأساسية لاستخدام هذه التقنية التجريبية هي أيضا معقدة ومتعددة الأوجه مع العديد من المتغيرات. 6 هناك العديد من الدراسات التي تصف كيف يمكن لل...

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><td> اسم كاشف </td><td> شركة </td><td> فهرس العدد </td><td> تعليقات </td></tr><tr><td> بولي اللبنيك، شارك في حامض الجليكوليك </td><td> سيجما ألدريتش </td><td></td><td></td></tr><tr><td> حمض اللبن بولي </td><td> سيجما ألدريتش </td><td> 81273 </td><td> المتأصلة اللزوجة ~ 2.0dl / ز </td></tr><tr><td> بولي ε-caprolactone </td><td> سيجما ألدريتش </td><td></td><td></td></tr><tr><td> بولي البيتاهيدروكسي-CO-hydroxyvalerate 00:01 </td><td> Goodfellow </td><td> 578-446-59 </td><td> PHB88/PHV12 </td></tr><tr><td> ثنائي كلورو ميثان </td><td> سيغما الدريتش أو فيشر </td><td> 270997 أو D/1850/17 </td><td> &gt; 99.8٪ يحتوي على استقرار 50-150ppm أميلين </td></tr><tr><td> 50 البالونات الملونة متعددة </td><td> ويلكنسون في مخازن الأجهزة المحدودة </td><td> 0105790 </td><td></td></tr><tr><td> الماعز المضادة للأرنب مفتش (FC): FITC </td><td> AbDserotec </td><td> STAR121F </td><td></td></tr><tr><td> أرنب المضادة للبشرية ألفا الإيلاستين </td><td> AbDserotec </td><td> 4060-1060 </td><td></td></tr><tr><td> المسمار كاب GL45 PP 2 ميناء، PK / 2 </td><td> SLS </td><td> 1129750 </td><td></td></tr><tr><td> 4 &#39;،6-Diamidino-2-phenylindole dihydrochloride </td><td> سيجما ألدريتش </td><td> 32670 </td><td></td></tr><tr><td> CellTracker أخضر CMFDA </td><td> إينفيتروجن </td><td> C7025 </td><td></td></tr><tr><td> CellTracker أحمر CMTX </td><td> إينفيتروجن </td><td> C34552 </td><td></td></tr></tbody></table>

postproduction processing of electrospun fibres for tissue engineering

Electrospinning هو وسيلة شائعة الاستخدام وتنوعا لإنتاج السقالات (القابلة للتحلل في كثير من الأحيان) لهندسة الأنسجة 3D. 1، 2، 3 العديد من الأنسجة في الجسم الحي الخضوع لانتفاخ ذو محورين بدرجات متفاوتة مثل الجلد، الكلمة، والمثانة والحوض حتى الحنك الصلب والأطفال تنمو. في إنتاج السقالات لهذه الأغراض هناك حاجة لتطوير السقالات من خصائص النشاط الحيوي المناسب (سواء كانت تتحقق من دون أو مع الخلايا) والتي تكون معقمة للاستعمال السريري. التركيز في هذه الورقة ليست هي الطريقة لوضع معايير electrospinning الأساسية (كما أن هناك دراسات واسعة النطاق في electrospinning)، ولكن حول كيفية تعديل نسج مرحلة ما بعد الإنتاج السقالات لجعلها صالحة للأغراض هندسة الأنسجة - هنا سماكة، الخواص الميكانيكية والتعقيم (مطلوب ل الاستخدام السريري) وتعتبر ونحن وصف أيضا كيف يمكن زرعها الخلايا على السقالات وتعرض للسلالة ذو محورين إلى حالة منهم لتطبيقات محددة. Electrospinning يميل الى انتاج صفائح رقيقة، وجامع electrospinning يصبح المغلفة مع عزل الألياف يصبح موصل الفقراء من هذا القبيل أن الألياف لم تعد الودائع على ذلك. وبالتالي نحن تصف النهج لإنتاج الهياكل سمكا بواسطة الحرارة أو البخار الصلب زيادة قوة السقالات ولكن ليس بالضرورة مرونة. يوصف أيضا الغزل متتابعة من السقالات من البوليمرات المختلفة لتحقيق السقالات المعقدة. يمكن أن يؤثر سلبا على منهجيات تعقيم قوة ومرونة من السقالات. قارنا ثلاث طرق لآثارها على خصائص النشاط الحيوي على السقالات electrospun من حامض اللبنيك، شارك في الجليكوليك بولي (PLGA). يوصف التصوير من الخلايا في السقالات وتقييم إنتاج البروتينات خارج الخلية (ECM) مصفوفة من الخلايا في السقالات. ويمكن زراعة الخلايا في المختبر على السقالات سقالة تحسين قوة ومرونة ولكن النسيج literatu الهندسةإعادة يدل على أن الخلايا غالبا ما تفشل لانتاج ECM المناسبة عند تربيتها في ظروف ثابتة. هناك بعض النظم التجارية المتاحة التي تسمح واحد لخلايا الثقافة على السقالات في ظل أنظمة تكييف ديناميكي - مثال واحد هو Electroforce بوس 3100 والتي يمكن استخدامها لممارسة برنامج تكييف على الخلايا في السقالات التي عقدت باستخدام قبضة الميكانيكية داخل وسائل الإعلام غرفة مليئة 4 يوصف هذا النهج إلى ثقافة مفاعل حيوي ميزانية خلية لتشويه للرقابة في 2 الأبعاد. نظهر أن يمكن تحريض الخلايا لإنتاج الإيلاستين في ظل هذه الظروف. أخيرا وصفت تقييم خصائص النشاط الحيوي من السقالات المجهزة تربيتها مع أو من دون الخلايا.

Watch this Scientific Journal Video about Postproduction Processing of Electrospun Fibres for Tissue Engineering at JoVE.com

Postproduction Processing of Electrospun Fibres for Tissue Engineering

يمكن معالجة السقالات Electrospun مرحلة ما بعد الانتاج لتطبيقات هندسة الأنسجة. نحن هنا وصف أساليب الغزل السقالات المعقدة (من خلال الدوران على التوالي)، لجعل سمكا السقالات (بواسطة متعددة الطبقات باستخدام الحرارة أو الصلب بخار)، لتحقيق عقم (الإنتاج العقيم أو التعقيم مرحلة ما بعد الإنتاج)، وخصائص النشاط الحيوي لتحقيق مناسب.

المعالجة بعد الإنتاج من الألياف Electrospun للهندسة الأنسجة

Electrospinning is a commonly used and versatile method to produce  scaffolds (often biodegradable) for 3D tissue engineering.1, 2, 3 Many tissues in vivo ...

postproduction-processing-of-electrospun-fibres-for-tissue-engineering

Research

JoVE Journal

Bioengineering

18.0K Views.  University of Sheffield. يمكن معالجة السقالات Electrospun مرحلة ما بعد الانتاج لتطبيقات هندسة الأنسجة. نحن هنا وصف أساليب الغزل السقالات المعقدة (من خلال الدوران على التوالي)، لجعل سمكا السقالات (بواسطة متعددة الطبقات باستخدام الحرارة أو الصلب بخار)، لتحقيق عقم (الإنتاج العقيم أو التعقيم مر?...

Video: Postproduction Processing of Electrospun Fibres for Tissue Engineering

Fabrication of Biologically Derived Injectable Materials for Myocardial Tissue Engineering

This protocol provides methods for the preparation of an injectable extracellular matrix (ECM) gel for myocardial tissue engineering applications. Briefly, decellularized tissue is lyophilized, milled, enzymatically digested, and then brought to physiological pH. The lyophilization removes all water content from the tissue, resulting in dry ECM that can be ground into a fine powder with a small mill. After milling, the ECM powder is digested with pepsin to form an injectable matrix. After adjustment to pH 7.4, the liquid matrix material can be injected into the myocardium. Results of previous characterization assays have shown that matrix gels produced from decellularized pericardial and myocardial tissue retain native ECM components, including diverse proteins, peptides and glycosaminoglycans. Given the use of this material for tissue engineering, in vivo characterization is especially useful; here, a method for performing an intramural injection into the left ventricular (LV) free wall is presented as a means of analyzing the host response to the matrix gel in a small animal model. Access to the chest cavity is gained through the diaphragm and the injection is made slightly above the apex in the LV free wall. The biologically derived scaffold can be visualized by biotin-labeling before injection and then staining tissue sections with a horse radish peroxidase-conjugated neutravidin and visualizing via diaminobenzidine (DAB) staining. Analysis of the injection region can also be done with histological and immunohistochemical staining. In this way, the previously examined pericardial and myocardial matrix gels were shown to form fibrous, porous networks and promote vessel formation within the injection region.

Methods for preparing an injectable matrix gel from decellularized tissue and injecting it into rat myocardium in vivo are described.

تصنيع مواد حقن المشتقة بيولوجيا للهندسة الأنسجة عضلة القلب

This protocol provides methods for the preparation of an injectable extracellular matrix (ECM) gel for myocardial tissue engineering applications.  ...

Elastomeric PGS Scaffolds in Arterial Tissue Engineering

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and increasing obesity1. Currently, bypass surgery using autologous vessels, allografts, and synthetic grafts are known as a commonly used for arterial substitutes2. However, these grafts have limited applications when an inner diameter of arteries is less than 6 mm due to low availability, thrombotic complications, compliance mismatch, and late intimal hyperplasia3,4. To overcome these limitations, tissue engineering has been successfully applied as a promising alternative to develop small-diameter arterial constructs that are nonthrombogenic, robust, and compliant. Several previous studies have developed small-diameter arterial constructs with tri-lamellar structure, excellent mechanical properties and burst pressure comparable to native arteries5,6. While high tensile strength and burst pressure by increasing collagen production from a rigid material or cell sheet scaffold, these constructs still had low elastin production and compliance, which is a major problem to cause graft failure after implantation. Considering these issues, we hypothesized that an elastometric biomaterial combined with mechanical conditioning would provide elasticity and conduct mechanical signals more efficiently to vascular cells, which increase extracellular matrix production and support cellular orientation.
The objective of this report is to introduce a fabrication technique of porous tubular scaffolds and a dynamic mechanical conditioning for applying them to arterial tissue engineering. We used a biodegradable elastomer, poly (glycerol sebacate) (PGS)7 for fabricating porous tubular scaffolds from the salt fusion method. Adult primary baboon smooth muscle cells (SMCs) were seeded on the lumen of scaffolds, which cultured in our designed pulsatile flow bioreactor for 3 weeks. PGS scaffolds had consistent thickness and randomly distributed macro- and micro-pores. Mechanical conditioning from pulsatile flow bioreactor supported SMC orientation and enhanced ECM production in scaffolds. These results suggest that elastomeric scaffolds and mechanical conditioning of bioreactor culture may be a promising method for arterial tissue engineering.

Elastomeric PGS scaffolds with vascular smooth muscle cells cultured in a pulsatile flow bioreactor may lead to promising small-diameter arterial constructs with native ECM production in a relatively short culture period.

PGS السقالات المرنة في هندسة الأنسجة الشرياني

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and ...

Encapsulation of Cardiomyocytes in a Fibrin Hydrogel for Cardiac Tissue Engineering

Culturing cells in a three dimensional hydrogel environment is an important technique for developing constructs for tissue engineering as well as studying cellular responses under various culture conditions in vitro. The three dimensional environment more closely mimics what the cells observe in vivo due to the application of mechanical and chemical stimuli in all dimensions 1. Three-dimensional hydrogels can either be made from synthetic polymers such as PEG-DA 2 and PLGA 3 or a number of naturally occurring proteins such as collagen 4, hyaluronic acid 5 or fibrin 6,7. Hydrogels created from fibrin, a naturally occurring blood clotting protein, can polymerize to form a mesh that is part of the body's natural wound healing processes 8. Fibrin is cell-degradable and potentially autologous 9, making it an ideal temporary scaffold for tissue engineering.
Here we describe in detail the isolation of neonatal cardiomyocytes from three day old rat pups and the preparation of the cells for encapsulation in fibrin hydrogel constructs for tissue engineering. Neonatal myocytes are a common cell source used for in vitro studies in cardiac tissue formation and engineering 4. Fibrin gel is created by mixing fibrinogen with the enzyme thrombin. Thrombin cleaves fibrinopeptides FpA and FpB from fibrinogen, revealing binding sites that interact with other monomers 10. These interactions cause the monomers to self-assemble into fibers that form the hydrogel mesh. Because the timing of this enzymatic reaction can be adjusted by altering the ratio of thrombin to fibrinogen, or the ratio of calcium to thrombin, one can injection mold constructs with a number of different geometries 11,12. Further we can generate alignment of the resulting tissue by how we constrain the gel during culture 13.
After culturing the engineered cardiac tissue constructs for two weeks under static conditions, the cardiac cells have begun to remodel the construct and can generate a contraction force under electrical pacing conditions 6. As part of this protocol, we also describe methods for analyzing the tissue engineered myocardium after the culture period including functional analysis of the active force generated by the cardiac muscle construct upon electrical stimulation, as well as methods for determining final cell viability (Live-Dead assay) and immunohistological staining to examine the expression and morphology of typical proteins important for contraction (Myosin Heavy Chain or MHC) and cellular coupling (Connexin 43 or Cx43) between myocytes.

We describe the isolation of neonatal cardiomyocytes and the preparation of the cells for encapsulation in fibrin hydrogel constructs for tissue engineering. We describe methods for analyzing the tissue engineered myocardium after the culture period including active force generated upon electrical stimulation and cell viability and immunohistological staining.

تغليف الخلايا العضلية في هيدروجيل الفايبرين للهندسة الأنسجة القلبية

Culturing cells in a three dimensional hydrogel environment is an important technique for developing constructs for tissue engineering as well as studying ...

Capillary Force Lithography for Cardiac Tissue Engineering

Cardiovascular disease remains the leading cause of death worldwide1. Cardiac tissue engineering holds much promise to deliver groundbreaking medical discoveries with the aims of developing functional tissues for cardiac regeneration as well as in vitro screening assays. However, the ability to create high-fidelity models of heart tissue has proven difficult. The heart&#8217;s extracellular matrix (ECM) is a complex structure consisting of both biochemical and biomechanical signals ranging from the micro- to the nanometer scale2. Local mechanical loading conditions and cell-ECM interactions have recently been recognized as vital components in cardiac tissue engineering3-5.
A large portion of the cardiac ECM is composed of aligned collagen fibers with nano-scale diameters that significantly influences tissue architecture and electromechanical coupling2. Unfortunately, few methods have been able to mimic the organization of ECM fibers down to the nanometer scale. Recent advancements in nanofabrication techniques, however, have enabled the design and fabrication of scalable scaffolds that mimic the in vivo structural and substrate stiffness cues of the ECM in the heart6-9.
Here we present the development of two reproducible, cost-effective, and scalable nanopatterning processes for the functional alignment of cardiac cells using the biocompatible polymer poly(lactide-co-glycolide) (PLGA)8 and a polyurethane (PU) based polymer. These anisotropically nanofabricated substrata (ANFS) mimic the underlying ECM of well-organized, aligned tissues and can be used to investigate the role of nanotopography on cell morphology and function10-14.
Using a nanopatterned (NP) silicon master as a template, a polyurethane acrylate (PUA) mold is fabricated. This PUA mold is then used to pattern the PU or PLGA hydrogel via UV-assisted or solvent-mediated capillary force lithography (CFL), respectively15,16. Briefly, PU or PLGA pre-polymer is drop dispensed onto a glass coverslip and the PUA mold is placed on top. For UV-assisted CFL, the PU is then exposed to UV radiation (&#955; = 250-400 nm) for curing. For solvent-mediated CFL, the PLGA is embossed using heat (120 &#176;C) and pressure (100 kPa). After curing, the PUA mold is peeled off, leaving behind an ANFS for cell culture. Primary cells, such as neonatal rat ventricular myocytes, as well as human pluripotent stem cell-derived cardiomyocytes, can be maintained on the ANFS2.

In this protocol, we demonstrate the fabrication of biomimetic cardiac cell culture substrata made from two distinct polymeric materials using capillary force lithography. The described methods provide a scalable, cost-effective technique to engineer the structure and function of macroscopic cardiac tissues for in vitro and in vivo applications.

الشعرية قوة الطباعة الحجرية لهندسة الأنسجة القلبية

Cardiovascular disease remains the leading cause of death worldwide1. Cardiac tissue engineering holds much promise to deliver groundbreaking medical ...

Tissue Engineering of a Human 3D in vitro Tumor Test System

Cancer is one of the leading causes of death worldwide. Current therapeutic strategies are predominantly developed in 2D culture systems, which inadequately reflect physiological conditions in vivo. Biological 3D matrices provide cells an environment in which cells can self-organize, allowing the study of tissue organization and cell differentiation. Such scaffolds can be seeded with a mixture of different cell types to study direct 3D cell-cell-interactions. To mimic the 3D complexity of cancer tumors, our group has developed a 3D in vitro tumor test system.	
	Our 3D tissue test system models the in vivo situation of malignant peripheral nerve sheath tumors (MPNSTs), which we established with our decellularized porcine jejunal segment derived biological vascularized scaffold (BioVaSc). In our model, we reseeded a modified BioVaSc matrix with primary fibroblasts, microvascular endothelial cells (mvECs) and the S462 tumor cell line. For static culture, the vascular structure of the BioVaSc is removed and the remaining scaffold is cut open on one side (Small Intestinal Submucosa SIS-Muc). The resulting matrix is then fixed between two metal rings (cell crowns).	
	Another option is to culture the cell-seeded SIS-Muc in a flow bioreactor system that exposes the cells to shear stress. Here, the bioreactor is connected to a peristaltic pump in a self-constructed incubator. A computer regulates the arterial oxygen and nutrient supply via parameters such as blood pressure, temperature, and flow rate. This setup allows for a dynamic culture with either pressure-regulated pulsatile or constant flow.	
	In this study, we could successfully establish both a static and dynamic 3D culture system for MPNSTs. The ability to model cancer tumors in a more natural 3D environment will enable the discovery, testing, and validation of future pharmaceuticals in a human-like model.

Tissue Engineering of a Human 3D in vitro Tumor Test System

Methods to create human 3D tumor tissues as test systems are described. These technologies are based on a decellularized Biological Vascularized Scaffold (BioVaSc), primary human cells and a tumor cell line, which can be cultured under static as well as under dynamic conditions in a flow bioreactor.

هندسة الأنسجة ل3D الإنسان<em&gt; في المختبر</em&gt; نظام ورم اختبار

Cancer is one of the leading causes of death worldwide.  Current therapeutic strategies are predominantly developed in 2D culture  systems, which ...

Electrospun Fibrous Scaffolds of Poly(glycerol-dodecanedioate) for Engineering Neural Tissues From Mouse Embryonic Stem Cells

For tissue engineering applications, the preparation of biodegradable and biocompatible scaffolds is the most desirable but challenging task. &#160;Among the various fabrication methods, electrospinning is the most attractive one due to its simplicity and versatility. Additionally, electrospun nanofibers mimic the size of natural extracellular matrix ensuring additional support for cell survival and growth. This study showed the viability of the fabrication of long fibers spanning a larger deposit area for a novel biodegradable and biocompatible polymer named&#160;poly(glycerol-dodecanoate) (PGD)1 by using a newly designed collector for electrospinning. PGD exhibits unique elastic properties with similar mechanical properties to nerve tissues, thus it is suitable for neural tissue engineering applications. The synthesis and fabrication set-up for making fibrous scaffolding materials was simple, highly reproducible, and inexpensive. In biocompatibility testing, cells derived from mouse embryonic stem cells could adhere to and grow on the electrospun PGD fibers. In summary, this protocol provided a versatile fabrication method for making PGD electrospun fibers to support the growth of mouse embryonic stem cell derived neural lineage cells.

Electrospun Fibrous Scaffolds of Polyglycerol-dodecanedioate for Engineering Neural Tissues From Mouse Embryonic Stem Cells

Synthesis and fabrication of electrospun long fibers spanning a larger deposit area via a newly designed collector from a novel biodegradable polymer named poly(glycerol-dodecanoate) (PGD) was reported. The fibers were able to support the growth of cells derived from mouse pluripotent stem cells.

Electrospun الليفية سقالات بولي (الجلسرين-dodecanedioate) للهندسة الأنسجة العصبية من الخلايا الجذعية الجنينية الماوس

For tissue engineering applications, the preparation of biodegradable and biocompatible scaffolds is the most desirable but challenging task. &#160;Among ...

Engineering 3D Cellularized Collagen Gels for Vascular Tissue Regeneration

Synthetic materials are known to initiate clinical complications such as inflammation, stenosis, and infections when implanted as vascular substitutes. Collagen has been extensively used for a wide range of biomedical applications and is considered a valid alternative to synthetic materials due to its inherent biocompatibility (i.e., low antigenicity, inflammation, and cytotoxic responses). However, the limited mechanical properties and the related low hand-ability of collagen gels have hampered their use as scaffold materials for vascular tissue engineering. Therefore, the rationale behind this work was first to engineer cellularized collagen gels into a tubular-shaped geometry and second to enhance smooth muscle cells driven reorganization of collagen matrix to obtain tissues stiff enough to be handled.
The strategy described here is based on the direct assembling of collagen and smooth muscle cells (construct) in a 3D cylindrical geometry with the use of a molding technique. This process requires a maturation period, during which the constructs are cultured in a bioreactor under static conditions (without applied external dynamic mechanical constraints) for 1 or 2 weeks. The &#8220;static bioreactor&#8221; provides a monitored and controlled sterile environment (pH, temperature, gas exchange, nutrient supply and waste removal) to the constructs. During culture period, thickness measurements were performed to evaluate the cells-driven remodeling of the collagen matrix, and glucose consumption and lactate production rates were measured to monitor the cells metabolic activity. Finally, mechanical and viscoelastic properties were assessed for the resulting tubular constructs. To this end, specific protocols and a focused know-how (manipulation, gripping, working in hydrated environment, and so on) were developed to characterize the engineered tissues.

In this work, we present a technique for the rapid fabrication of living vascular tissues by direct culturing of collagen, smooth muscle cells and endothelial cells. In addition, a new protocol for the mechanical characterization of engineered vascular tissues is described.

الهندسة 3D Cellularized الهلام الكولاجين لتجديد الأنسجة الوعائية

Synthetic materials are known to initiate clinical complications such as inflammation, stenosis, and infections when implanted as vascular substitutes. ...

Preparation of Thermoresponsive Nanostructured Surfaces for Tissue Engineering

Thermoresponsive poly(N-isopropylacrylamide) (PIPAAm)-immobilized surfaces for controlling cell adhesion and detachment were fabricated by the Langmuir-Schaefer method. Amphiphilic block copolymers composed of polystyrene and PIPAAm (St-IPAAms) were synthesized by reversible addition-fragmentation chain transfer (RAFT) radical polymerization. A chloroform solution of St-IPAAm molecules was gently dropped into a Langmuir-trough apparatus, and both barriers of the apparatus were moved horizontally to compress the film to regulate its density. Then, the St-IPAAm Langmuir film was horizontally transferred onto a hydrophobically modified glass substrate by a surface-fixed device. Atomic force microscopy images clearly revealed nanoscale sea-island structures on the surface. The strength, rate, and quality of cell adhesion and detachment on the prepared surface were modulated by changes in temperature across the lower critical solution temperature range of PIPAAm molecules. In addition, a two-dimensional cell structure (cell sheet) was successfully recovered on the optimized surfaces. These unique PIPAAm surfaces may be useful for controlling the strength of cell adhesion and detachment.

Nanoscaled sea-island surfaces composed of thermoresponsive block copolymers were fabricated by the Langmuir-Schaefer method for controlling spontaneous cell adhesion and detachment. Both the preparation of the surface and the adhesion and detachment of cells on the surface were visualized.

إعداد Thermoresponsive ذات البنية النانومترية السطوح للهندسة الأنسجة

Thermoresponsive poly(N-isopropylacrylamide) (PIPAAm)-immobilized surfaces for controlling cell adhesion and detachment were fabricated by the ...

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and trade one defect for another. Tissue engineering holds the promise to address this increasing demand. However, most tissue engineering strategies fail to generate stable and functional tissue substitutes because of poor vascularization. This paper focuses on an in vivo tissue engineering chamber model of intrinsic vascularization where a perfused artery and a vein either as an arteriovenous loop or a flow-through pedicle configuration is directed inside a protected hollow chamber. In this chamber-based system angiogenic sprouting occurs from the arteriovenous vessels and this system attracts ischemic and inflammatory driven endogenous cell migration which gradually fills the chamber space with fibro-vascular tissue. Exogenous cell/matrix implantation at the time of chamber construction enhances cell survival and determines specificity of the engineered tissues which develop. Our studies have shown that this chamber model can successfully generate different tissues such as fat, cardiac muscle, liver and others. However, modifications and refinements are required to ensure target tissue formation is consistent and reproducible. This article describes a standardized protocol for the fabrication of two different vascularized tissue engineering chamber models in vivo.

Tissue Engineering by Intrinsic Vascularization in an In Vivo Tissue Engineering Chamber

This is a guideline for constructing in vivo vascularized tissue using a microsurgical arteriovenous loop or a flow-through pedicle configuration inside a tissue engineering chamber. The vascularized tissues generated can be employed for organ regeneration and replacement of tissue defects, as well as for drug testing and disease modeling.

هندسة الأنسجة عن طريق الجوهرية الأوعية الدموية في<em&gt; في فيفو</em&gt; غرفة هندسة الأنسجة

In reconstructive surgery, there is a clinical need for an alternative to the current methods of autologous reconstruction which are complex, costly and ...

A Protocol for Decellularizing Mouse Cochleae for Inner Ear Tissue Engineering

In mammals, mechanosensory hair cells that facilitate hearing lack the ability to regenerate, which has limited treatments for hearing loss. Current regenerative medicine strategies have focused on transplanting stem cells or genetic manipulation of surrounding support cells in the inner ear to encourage replacement of damaged stem cells to correct hearing loss. Yet, the extracellular matrix (ECM) may play a vital role in inducing and maintaining function of hair cells, and has not been well investigated. Using the cochlear ECM as a scaffold to grow adult stem cells may provide unique insights into how the composition and architecture of the extracellular environment aids cells in sustaining hearing function. Here we present a method for isolating and decellularizing cochleae from mice to use as scaffolds accepting perfused adult stem cells. In the current protocol, cochleae are isolated from euthanized mice, decellularized, and decalcified. Afterward, human Wharton&#39;s jelly cells (hWJCs) that were isolated from the umbilical cord were carefully perfused into each cochlea. The cochleae were used as bioreactors, and cells were cultured for 30 days before undergoing processing for analysis. Decellularized cochleae retained identifiable extracellular structures, but did not reveal the presence of cells or noticeable fragments of DNA. Cells perfused into the cochlea invaded most of the interior and exterior of the cochlea and grew without incident over a duration of 30 days. Thus, the current method can be used to study how cochlear ECM affects cell development and behavior.

The goal of this protocol is to demonstrate an effective method to decellularize and decalcify mouse cochleae for utilization as scaffolds for tissue engineering applications.

بروتوكول ديسيلولاريزينج كوكليا الماوس لهندسة الأنسجة الإذن الداخلية

In mammals, mechanosensory hair cells that facilitate hearing lack the ability to regenerate, which has limited treatments for hearing loss. Current ...