فإن الكتاب أود أن أشكر لي بيونغ تشول ، تشوي هو صموئيل وفيليب لمساعدة قيمة. ونفذ هذا العمل في مسبك الجزيئية في مختبر لورانس بيركلي الوطني ، وهو مدعوم من قبل مكتب المفوضية ، والعلوم العلوم الأساسية للطاقة ، من وزارة الطاقة الأميركية بموجب العقد رقم DE - AC02 - 05CH11231 وزير الدفاع للحد من التهديد الوكالة تحت رقم العقد : IACRO - B0845281.

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ليس لدينا شيء في الكشف عنها.

وأهمية التطبيقات يصف هذا البروتوكول طريقة بسيطة وفعالة لتجميع وpeptoid مائي التجميع الذاتي للpeptoids في nanosheets. معظم المختبرات قادرة بسهولة من تجميع peptoids لأن المواد الرخيصة ، والخبرات والتقنيات الأساسية واضحة وتستخدم 4. وبالمثل ، فإن التجميع الذاتي للرقيقة جدا nanosheets العالية أمر يتطلب مجرد تكرار إمالة قارورة حل peptoid مائي مخفف 2. Peptoids هي مادة واعدة للبحوث الطبية البيولوجية وعلم النانو لأنها قوية ومرنة صناعيا بعد تسلسل محدد والانضباطي للغاية 5. وقد أثبتت Peptoids النشاط البيولوجي (6،7 المداواة ، والتشخيص 8 ، والتسليم بين الخلايا 9-10) وقابلة للطي في النانو الهرمية 3 ، 11-14. بسبب التركيب على وحدات ، واندماجي peptoid librويمكن تصنيعه بسهولة 15-19 الحمل وفرزهم لسلسلة واسعة من الأنشطة أو الممتلكات. على وجه الخصوص ، nanosheets بمثابة منصة المحتملة لعرض السقالات ثنائي الأبعاد ، mimetics الغشاء ، وأجهزة الاستشعار البيولوجية ، وتصنيع البروتين mimetics الجهاز. مع تسلسل مختلف ممكن عمليا لا ينضب ، عالم الأبحاث peptoid تتوسع بسرعة. المتغيرات في التوليف submonomer الصلبة مرحلة polypeptoids لأن القدرة على اختيار من لا يصدق أبجدية كبيرة ومتنوعة من مونومرات 20 ، الأسلوب submonomer يحتاج تعديلات في بعض الأحيان عن الحالات التي تكون فيها زيادة كفاءة اقتران كل خطوة من شأنها تحسين العائد الناتج الكلي. ادراج محمية سلاسل الجانب متجانسة يتطلب استخدام حمض الكلوروأسيتيك بدلا من حمض bromoacetic 21. تمديد أوقات التشرد والتعليم العاليوتستخدم عادة تركيزات الأمينات بعد حوالي 20 جوانات لمتواليات peptoid طويلة أو الأمينات أقل أليف النواة. تسخين وعاء التفاعل إلى 35 درجة مئوية ، وذلك باستخدام سفينة رد فعل المياه تغلف ، ويساعد على دفع التفاعل. لدرجة عالية من الأمينات المتقلبة مثل isopropylamine ، لا بد من توخي الحذر لتجنب التبخر. الأمينات في شكل ملح حمض الهيدروكلوريك ، مثل حمض الهيدروكلوريك بيتا ألانين تي بوتيل ، والحاجة ليكون حرا مقرها قبل أن يتم عرضه في رد فعل التشرد. ويمكن تحقيق هذا عن طريق إذابة أو تعليق أمين في DCM (~ 5G amine/25 مل DCM) ، وتحييد بمحلول متساوي المولية من هيدروكسيد الصوديوم المائي في قمع separatory. ويتم جمع طبقة DCM ويغسل طبقة مائية مع DCM إضافية. تجفف طبقات DCM مجتمعة أكثر من كبريتات الصوديوم وتصفيتها في أسفل القارورة الجولة ما قبل وزنه. إزالة المذيب عن طريق التبخر دوارة لتسفر عن النفط ، وتسجيل وزن المنتج. الزيتوز الخطوة الانقسام ، TFA كوكتيل الانقسام والانقسام الوقت يعتمد على عدد ومتنوعة لحماية الفئات المستخدمة. المبادئ التوجيهية للكوكتيلات تشطر تتشابه الانقسامات التقليدية deprotection الببتيد 1. عموما ، هناك حاجة الدقيقة 10 حضانات لمتواليات من دون حماية أو الجماعات أو متواليات مع عدد قليل من الجماعات عطوب حمض حماية عالية (مثل BOC ، trityl). وأوصى اثنين من حضانات ساعة لمتواليات أكثر صعوبة مع جماعات حماية (على سبيل المثال تي بوتيل استر ، MTR ، PBF) أو متواليات مع العديد من الجماعات لضمان حماية كاملة من كل deprotection السلسلة. والمنتجات الخام peptoid حل عموما في أسيتونتريل : 1:1 المياه (V / V) ، ولكن بنسب أعلى أسيتونتريل هي مشتركة مع سلاسل جنب مع hydrophobicity شامل عالية.

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><td> اسم الكاشف </td><td> شركة </td><td> فهرس العدد </td><td> تعليقات </td></tr><tr><td> ثنائي ميثيل الفورماميد </td><td> EMD </td><td> EM - DX1726P - 1 </td><td> 99 + ٪ </td></tr><tr><td> N - methylpyrrolidinone </td><td> BDH </td><td> BDH1141 - 4LP </td><td> 99 ٪ </td></tr><tr><td> Bromoacetic حمض </td><td> العضوية Acros </td><td> 200000-106 </td><td> 99 ٪ </td></tr><tr><td> 4 - Methylpiperidine </td><td> سيغما الدريتش </td><td> M73206 </td><td> 96 ٪ </td></tr><tr><td> N ، N&#39; - diisopropylcarbodiimide </td><td> الكيميائي امبكس </td><td> 001100 </td><td> 99.5 ٪ </td></tr><tr><td> ثنائي كلورو ميثان </td><td> EMD </td><td> EMD - DX0835 </td><td> ACS الصف </td></tr><tr><td> أسيتونتريل </td><td> EMD </td><td> EM - AX0145P - 1 </td><td> 99.8 ٪ </td></tr><tr><td> Trifluoroacetic حمض </td><td> سيغما الدريتش </td><td> T6508 </td><td> 99 ٪ </td></tr><tr><td> Triisopropylsilane </td><td> سيغما الدريتش </td><td> 233781 --10G </td><td> لTFA تشطر </td></tr><tr><td> 1،2 - ثنائي كلورو إيثان </td><td> JT بيكر </td><td> JTH076 - 33 </td><td> لsiliconization من أوعية التفاعل الزجاج </td></tr><tr><td> الفينيثيلامين </td><td> سيغما الدريتش </td><td> 407267 - 100ML </td><td> &gt; 99.5 ٪ مسعور الجانب سلسلة الأمينات </td></tr><tr><td> BOC - الإيثيلنديامين </td><td> CNH تكنولوجيز </td><td> C - 1112 </td><td> الموجبة الجانب سلسلة الأمينات </td></tr><tr><td> تي بوتيل بيتا ألانين حمض الهيدروكلوريك </td><td> الكيميائي امبكس الدولية </td><td> 04407 </td><td> أنيوني الجانب سلسلة الأمينات </td></tr><tr><td> α - Cyano - 4 - hydroxycinnamic حمض </td><td> سيغما الدريتش </td><td> C8982 - 10X10MG </td><td> لمصفوفة MALDI </td></tr><tr><td> النيل الأحمر </td><td> سيغما الدريتش </td><td> 19123 - 10MG </td><td> لمضان التصوير </td></tr><tr><td> Dichlorodimethylsilane </td><td> سيغما الدريتش </td><td> 80430 - F - 500G </td><td> لsiliconization من أوعية التفاعل الزجاج </td></tr><tr><td> PP المحتوى المتاح frittedالتلال </td><td> تطبيق فصل </td><td> 2416 </td><td> 6 مل البولي بروبلين مع خرطوشة عيار 20 ملم PE فريت </td></tr><tr><td> 3 الطريق المتاح luer محول </td><td> كول Parmer </td><td> 31200-80 </td><td> محبس لتجميع المتاح دليل عاء التفاعل </td></tr><tr><td> Luer قفل حلقة </td><td> كول Parmer </td><td> 45503-19 </td><td> ¼ &quot;يليق المتاح دليل التوليف عاء التفاعل </td></tr><tr><td> Luer التجهيزات </td><td> كول Parmer </td><td> 45500-20 </td><td> ¼ &quot;يليق المتاح دليل التوليف عاء التفاعل </td></tr><tr><td> PP المتاح pipets </td><td> VWR </td><td> 16001-194 </td><td> لنقل TFA </td></tr><tr><td> Luer المحاقن البلاستيكية قفل </td><td> العلم الوطني </td><td> S7515 - 5 </td><td> 6 مل المحاقن </td></tr><tr><td> 1 قنينة زجاجية الدرام </td><td> VWR </td><td> 66011-041 </td><td> مع الفينول مصبوب سقف المسمار مع البولي فينيل التي تواجهها الخطوط الملاحية المنتظمة اللب </td></tr><tr><td> 20 مل scintillation فيال </td><td> VWR </td><td> 66022-060 </td><td> مع غطاء PP المرفقة واللب احباط بطانة </td></tr><tr><td> آمن لاصقة هل ختم </td><td> Invitrogen </td><td> S - 24736 </td><td> لمضان التصوير </td></tr><tr><td> الشرائح الزجاجية </td><td> الإلكترون المجهري العلوم </td><td> 63411 </td><td> لمضان التصوير </td></tr><tr><td> تغطية زلة </td><td> VWR </td><td> 48366-067 </td><td> لمضان التصوير </td></tr><tr><td> 4 رقاقة سيليكون &quot; </td><td> تيد بيلا </td><td> 16007 </td><td> قبل الزهر في رقائق ملم 5x7 </td></tr><tr><td> 0.45 فلتر </td><td> VWR ، Acrodisc </td><td> 28143-924 </td><td> لهبلك. السليكوون الغشاء </td></tr><tr><td> Agarose </td><td> دينار بحريني </td><td> 212272 </td><td> لمضان التصوير </td></tr><tr><td> SPE فراغ المنوع </td><td> سيغما الدريتش </td><td> 57044 </td><td> مثال متعددة فراغ SPE </td></tr><tr><td> Fritted الزجاج السفينة </td><td> الآس الزجاج </td><td> 6402-12 </td><td> المسامية Cفريت </td></tr><tr><td> البلازما الأنظف / معقم </td><td> Harrick البلازما </td><td> PDC - 32G </td><td> مثال على نظافة البلازما لإعداد رقائق السيليكون لوزارة شؤون المرأة </td></tr></tbody></table>

solid-phase submonomer synthesis of peptoid polymers and their self-assembly into highly-ordered nanosheets

Peptoids هي فئة جديدة من بيوميمتيك ، غير الطبيعية تسلسل الخاصة التي تقاوم التحلل البروتيني heteropolymers ، المعرض نشاط بيولوجي محتمل ، وأضعاف النانو في أعلى الترتيب. يشبه في بنيته الببتيدات ، وبولي peptoids N - استبداله glycines ، حيث تعلق على الجانب سلاسل النيتروجين بدلا من الكربون ألفا. تخفيف بهم من التوليف والتنوع الهيكلي يسمح اختبار مبادئ التصميم الأساسية لمحرك دي نوفو تصميم وهندسة المواد الجديدة النشطة بيولوجيا وذات البنية النانومترية. هنا ، هو تقديم دليل بسيط peptoid بروتوكول التوليف التي تسمح للتوليف polypeptoids سلسلة طويلة (تصل إلى 50mers) في غلة ممتازة. مطلوبة المعدات الأساسية فقط ، وتقنيات بسيطة (مثل نقل السائل ، والترشيح) ، والكواشف المتاحة تجاريا ، مما يجعل peptoids إضافة إلى مجموعة أدوات للوصول العديد من الباحثين. ويزرع في العمود الفقري peptoid one مونومر في وقت تIA submonomer الأسلوب الذي يتكون من مرحلتين مونومر دورة بالإضافة إلى ذلك : أسيلة والتشريد. أولا ، وحامض bromoacetic تفعيلها في الموقع مع N ، N&#39; - diisopropylcarbodiimide acylates راتنج متجهة الأمينات الثانوية. الثانية ، والتشريد محب النواة من بروميد من الأمينات الأولية التالي لتقديم سلسلة من الجانبين. وكرر دورة على مرحلتين حتى يتم الوصول إلى طول السلسلة المطلوب. كفاءة اقتران هذه الدورة على مرحلتين يتجاوز عادة 98 ٪ ويمكن تجميع peptoids طالما 50 البقايا. سلاسل متنوعة الانضباطي للغاية ودقيقة وقابلة للتحقيق كيميائيا مع أسلوب submonomer كما يمكن أن المئات من الأمينات الأولية متوفرة بسهولة إدراجها مباشرة. Peptoids آخذة في الظهور كمادة بيوميمتيك تنوعا للبحوث nanobioscience لما لها من قوة الاصطناعية ، والمرونة ، ويأمر على المستوى الذري. للطي من سلسلة واحدة ، انفورما ، amphiphilicوقد تجلى مؤخرا نشوئها الغنية polypeptoid الى nanosheet العالية المطلوبة. هذا هو peptoid 36 مير التي تتكون من المونومرات ثلاثة فقط متاحة تجاريا : مسعور ، وأنيوني الموجبة. ودفن مسعور سلاسل الجانب الفنيل في حين أن جوهر nanosheet أمين الأيونية وسلاسل محاذاة الجانب الكربوكسيل على وجوه ماء. وnanosheets peptoid بمثابة منصة المحتملة لmimetics الغشاء ، mimetics البروتين ، وتصنيع الجهاز ، وأجهزة الاستشعار. ووصف الطرق لتخليق peptoid ، وتشكيل ورقة ، والتصوير المجهري وتوفر طريقة بسيطة لتمكين التصاميم nanosheet peptoid المستقبل.

Watch this Scientific Journal Video about Solid-phase Submonomer Synthesis of Peptoid Polymers and their Self-Assembly into Highly-Ordered Nanosheets at JoVE.com

Solid-phase Submonomer Synthesis of Peptoid Polymers and their Self-Assembly into Highly-Ordered Nanosheets

ويرد بسيطة وعامة التوليف peptoid الطريقة اليدوية التي تنطوي على المعدات والكواشف الأساسية المتاحة تجاريا ، مما يمكن توليفها peptoids بسهولة في معظم المختبرات. يوصف التوليف ، وتنقية وتوصيف ل36mer peptoid amphiphilic ، وكذلك التجميع الذاتي حيز العالية أمر nanosheets.

الصلبة مرحلة تجميع Submonomer من البوليمرات Peptoid واحترامهن الجمعية ، في الارتحال التي تأمر Nanosheets

Peptoids are a novel class of biomimetic, non-natural, sequence-specific heteropolymers that resist proteolysis, exhibit potent biological activity, and ...

solid-phase-submonomer-synthesis-peptoid-polymers-their-self-assembly

Research

JoVE Journal

Bioengineering

29.2K Views.  Lawrence Berkeley National Laboratory. ويرد بسيطة وعامة التوليف peptoid الطريقة اليدوية التي تنطوي على المعدات والكواشف الأساسية المتاحة تجاريا ، مما يمكن توليفها peptoids بسهولة في معظم المختبرات. يوصف التوليف ، وتنقية وتوصيف ل36mer peptoid amphiphilic ، وكذلك التجميع الذاتي حيز العالية أمر nanosheets.

Video: Solid-phase Submonomer Synthesis of Peptoid Polymers and their Self-Assembly into Highly-Ordered Nanosheets

Shape Memory Polymers for Active Cell Culture

Shape memory polymers (SMPs) are a class of "smart" materials that have the ability to change from a fixed, temporary shape to a pre-determined permanent shape upon the application of a stimulus such as heat1-5. In a typical shape memory cycle, the SMP is first deformed at an elevated temperature that is higher than its transition temperature, Ttrans [either the melting temperature (Tm) or the glass transition temperature (Tg)]. The deformation is elastic in nature and mainly leads to a reduction in conformational entropy of the constituent network chains (following the rubber elasticity theory). The deformed SMP is then cooled to a temperature below its Ttrans while maintaining the external strain or stress constant. During cooling, the material transitions to a more rigid state (semi-crystalline or glassy), which kinetically traps or "freezes" the material in this low-entropy state leading to macroscopic shape fixing. Shape recovery is triggered by continuously heating the material through Ttrans under a stress-free (unconstrained) condition. By allowing the network chains (with regained mobility) to relax to their thermodynamically favored, maximal-entropy state, the material changes from the temporary shape to the permanent shape.
Cells are capable of surveying the mechanical properties of their surrounding environment6. The mechanisms through which mechanical interactions between cells and their physical environment control cell behavior are areas of active research. Substrates of defined topography have emerged as powerful tools in the investigation of these mechanisms. Mesoscale, microscale, and nanoscale patterns of substrate topography have been shown to direct cell alignment, cell adhesion, and cell traction forces7-14. These findings have underscored the potential for substrate topography to control and assay the mechanical interactions between cells and their physical environment during cell culture, but the substrates used to date have generally been passive and could not be programmed to change significantly during culture. This physical stasis has limited the potential of topographic substrates to control cells in culture.
Here, active cell culture (ACC) SMP substrates are introduced that employ surface shape memory to provide programmed control of substrate topography and deformation. These substrates demonstrate the ability to transition from a temporary grooved topography to a second, nearly flat memorized topography. This change in topography can be used to control cell behavior under standard cell culture conditions.

A method for developing cell culture substrates with the ability to change topography during culture is described. The method makes use of smart materials known as shape memory polymers that have the ability to memorize a permanent shape. This concept is adaptable to a wide range of materials and applications.

البوليمرات الذاكرة شكل خلية للثقافة أحدث

Shape memory polymers (SMPs) are a class of "smart" materials that have the ability to change from a fixed, temporary shape to a pre-determined permanent ...

Directed Cellular Self-Assembly to Fabricate Cell-Derived Tissue Rings for Biomechanical Analysis and Tissue Engineering

Each year, hundreds of thousands of patients undergo coronary artery bypass surgery in the United States.1 Approximately one third of these patients do not have suitable autologous donor vessels due to disease progression or previous harvest. The aim of vascular tissue engineering is to develop a suitable alternative source for these bypass grafts. In addition, engineered vascular tissue may prove valuable as living vascular models to study cardiovascular diseases. Several promising approaches to engineering blood vessels have been explored, with many recent studies focusing on development and analysis of cell-based methods.2-5 Herein, we present a method to rapidly self-assemble cells into 3D tissue rings that can be used in vitro to model vascular tissues.
To do this, suspensions of smooth muscle cells are seeded into round-bottomed annular agarose wells. The non-adhesive properties of the agarose allow the cells to settle, aggregate and contract around a post at the center of the well to form a cohesive tissue ring.6,7 These rings can be cultured for several days prior to harvesting for mechanical, physiological, biochemical, or histological analysis. We have shown that these cell-derived tissue rings yield at 100-500 kPa ultimate tensile strength8 which exceeds the value reported for other tissue engineered vascular constructs cultured for similar durations (&lt;30 kPa).9,10 Our results demonstrate that robust cell-derived vascular tissue ring generation can be achieved within a short time period, and offers the opportunity for direct and quantitative assessment of the contributions of cells and cell-derived matrix (CDM) to vascular tissue structure and function.

This article outlines a versatile method to create cell-derived tissue rings by cellular self-assembly. Smooth muscle cells seeded into ring-shaped agarose wells aggregate and contract to form robust three-dimensional (3D) tissues within 7 days. Millimeter-scale tissue rings are conducive to mechanical testing and serve as building blocks for tissue assembly.

توجه الخلوي الذاتي الجمعية الى افتعال خواتم نسيج الخلايا المستمدة من النشاط الحيوي لتحليل وهندسة الأنسجة

Each year, hundreds of thousands of patients undergo coronary artery bypass surgery in the United States.1 Approximately one third of these patients do ...

Optical Control of Living Cells Electrical Activity by Conjugated Polymers

Hybrid interfaces between organic semiconductors and living tissues represent a new tool for in-vitro and in-vivo applications. In particular, conjugated polymers display several optimal properties as substrates for biological systems, such as good biocompatibility, excellent mechanical properties, cheap and easy processing technology, and possibility of deposition on light, thin and flexible substrates. These materials have been employed for cellular interfaces like neural probes, transistors for excitation and recording of neural activity, biosensors and actuators for drug release. Recent experiments have also demonstrated the possibility to use conjugated polymers for all-optical modulation of the electrical activity of cells. Several in-vitro study cases have been reported, including primary neuronal networks, astrocytes and secondary line cells. Moreover, signal photo-transduction mediated by organic polymers has been shown to restore light sensitivity in degenerated retinas, suggesting that these devices may be used for artificial retinal prosthesis in the future. All in all, light sensitive conjugated polymers represent a new approach for optical modulation of cellular activity.
In this work, all the steps required to fabricate a bio-polymer interface for optical excitation of living cells are described. The function of the active interface is to transduce the light stimulus into a modulation of the cell membrane potential. As a study case, useful for in-vitro studies, a polythiophene thin film is used as the functional, light absorbing layer, and Human Embryonic Kidney (HEK-293) cells are employed as the biological component of the interface. Practical examples of successful control of the cell membrane potential upon stimulation with light pulses of different duration are provided. In particular, it is shown that both depolarizing and hyperpolarizing effects on the cell membrane can be achieved depending on the duration of the light stimulus. The reported protocol is of general validity and can be straightforwardly extended to other biological preparations.

A method for stimulation of in-vitro cell cultures electrical activity with visible light, based on the use of organic semiconducting polymers is described.

مراقبة البصرية من الخلايا الحية النشاط الكهربائي من قبل مترافق البوليمرات

Hybrid interfaces between organic semiconductors and living tissues represent a new tool for in-vitro and in-vivo applications. In particular, conjugated ...

Fabrication of a Dipole-assisted Solid Phase Extraction Microchip for Trace Metal Analysis in Water Samples

This paper describes a fabrication protocol for a dipole-assisted solid phase extraction (SPE) microchip available for trace metal analysis in water samples. A brief overview of the evolution of chip-based SPE techniques is provided. This is followed by an introduction to specific polymeric materials and their role in SPE. To develop an innovative dipole-assisted SPE technique, a chlorine (Cl)-containing SPE functionality was implanted into a poly(methyl methacrylate) (PMMA) microchip. Herein, diverse analytical techniques including contact angle analysis, Raman spectroscopic analysis, and laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) analysis were employed to validate the utility of the implantation protocol of the C-Cl moieties on the PMMA. The analytical results of the X-ray absorption near-edge structure (XANES) analysis also demonstrated the feasibility of the Cl-containing PMMA used as an extraction medium by virtue of the dipole-ion interactions between the highly electronegative C-Cl moieties and the positively charged metal ions.

The fabrication protocol of a dipole-assisted solid phase extraction microchip for the trace metal analysis is presented.

تلفيق من ساعد ثنائي القطب-الصلبة المرحلة استخراج رقاقة لتتبع تحليل المعادن في عينات المياه

This paper describes a fabrication protocol for a dipole-assisted solid phase extraction (SPE) microchip available for trace metal analysis in water ...

Synthesis of Gold Nanoparticle Integrated Photo-responsive Liposomes and Measurement of Their Microbubble Cavitation upon Pulse Laser Excitation

Photo-responsive nanoparticles (NPs) have received considerable attention because of their potential in providing spatial, temporal, and dosage control over the drug release. However, most of the relevant technologies are still in the development process and are unprocurable by clinics. Here, we describe a facile fabrication of these photo-responsive NPs with commercially available gold NPs and thermo-responsive liposomes. Calcein is used as a model drug to evaluate the encapsulation efficiency and the release kinetic profile upon heat/light stimulation. Finally, we show that this photo-triggered release is due to the membrane disruption caused by microbubble cavitation, which can be measured with hydrophone.

This protocol describes a simple preparation method for gold nanoparticle integrated photo-responsive liposomes with the commercially available materials. It also shows how to measure the microbubble cavitation process of the synthesized liposomes upon the treatment of pulsed laser.

توليف الذهب الجسيمات النانوية الليبوزومات المتكاملة صور متجاوبة وقياس تلك Microbubble التجويف على نبض الليزر الإثارة

Photo-responsive nanoparticles (NPs) have received considerable attention because of their potential in providing spatial, temporal, and dosage control ...

High-throughput Identification of Bacteria Repellent Polymers for Medical Devices

Medical devices are often associated with hospital-acquired infections, which place enormous strain on patients and the healthcare system as well as contributing to antimicrobial resistance. One possible avenue for the reduction of device-associated infections is the identification of bacteria-repellent polymer coatings for these devices, which would prevent bacterial binding at the initial attachment step. A method for the identification of such repellent polymers, based on the parallel screening of hundreds of polymers using a microarray, is described here. This high-throughput method resulted in the identification of a range of promising polymers that resisted binding of various clinically relevant bacterial species individually and also as multi-species communities. One polymer, PA13 (poly(methylmethacrylate-co-dimethylacrylamide)), demonstrated significant reduction in attachment of a number of hospital isolates when coated onto two commercially available central venous catheters. The method described could be applied to identify polymers for a wide range of applications in which modification of bacterial attachment is important.

A high-throughput microarray method for the identification of polymers which reduce bacterial surface binding on medical devices is described.

عالية الإنتاجية تحديد البكتيريا البوليمرات طارد للمستلزمات الطبية

Medical devices are often associated with hospital-acquired infections, which place enormous strain on patients and the healthcare system as well as ...

Fabricating Degradable Thermoresponsive Hydrogels on Multiple Length Scales via Reactive Extrusion, Microfluidics, Self-assembly, and Electrospinning

While various smart materials have been explored for a variety of biomedical applications (e.g., drug delivery, tissue engineering, bioimaging, etc.), their ultimate clinical use has been hampered by the lack of biologically-relevant degradation observed for most smart materials. This is particularly true for temperature-responsive hydrogels, which are almost uniformly based on polymers that are functionally non-degradable (e.g., poly(N-isopropylacrylamide) (PNIPAM) or poly(oligoethylene glycol methacrylate) (POEGMA)). As such, to effectively translate the potential of thermoresponsive hydrogels to the challenges of remote-controlled or metabolism-regulated drug delivery, cell scaffolds with tunable cell-material interactions, theranostic materials with the potential for both imaging and drug delivery, and other such applications, a method is required to render the hydrogels (if not fully degradable) at least capable of renal clearance following the required lifetime of the material. To that end, this protocol describes the preparation of hydrolytically-degradable hydrazone-crosslinked hydrogels on multiple length scales based on the reaction between hydrazide and aldehyde-functionalized PNIPAM or POEGMA oligomers with molecular weights below the renal filtration limit. Specifically, methods to fabricate degradable thermoresponsive bulk hydrogels (using a double barrel syringe technique), hydrogel particles (on both the microscale through the use of a microfluidics platform facilitating simultaneous mixing and emulsification of the precursor polymers and the nanoscale through the use of a thermally-driven self-assembly and cross-linking method), and hydrogel nanofibers (using a reactive electrospinning strategy) are described. In each case, hydrogels with temperature-responsive properties similar to those achieved via conventional free radical cross-linking processes can be achieved, but the hydrazone cross-linked network can be degraded over time to re-form the oligomeric precursor polymers and enable clearance. As such, we anticipate these methods (which may be generically applied to any synthetic water-soluble polymer, not just smart materials) will enable easier translation of synthetic smart materials to clinical applications.

Protocols are described for the fabrication of degradable thermoresponsive hydrogels based on hydrazone cross-linking of polymeric oligomers on the bulk scale, microscale, and nanoscale, the latter for preparation of both gel nanoparticles and nanofibers.

اختﻻق الهلاميات المائية ثيرموريسبونسيفي القابلة للتحلل في جداول طول متعددة عن طريق رد الفعل البثق، ميكروفلويديكس، التجميع الذاتي، واليكتروسبينينج

While various smart materials have been explored for a variety of biomedical applications (e.g., drug delivery, tissue engineering, bioimaging, etc.), ...

Aqueous Droplets Used as Enzymatic Microreactors and Their Electromagnetic Actuation

For the successful implementation of microfluidic reaction systems, such as PCR and electrophoresis, the movement of small liquid volumes is essential. In conventional lab-on-a-chip-platforms, solvents and samples are passed through defined microfluidic channels with complex flow control installations. The droplet actuation platform presented here is a promising alternative. With it, it is possible to move a liquid drop (microreactor) on a planar surface of a reaction platform (lab-in-a-drop). The actuation of microreactors on the hydrophobic surface of the platform is based on the use of magnetic forces acting on the outer shell of the liquid drops which is made of a thin layer of superhydrophobic magnetite particles. The hydrophobic surface of the platform is needed to avoid any contact between the liquid core and the surface to allow a smooth movement of the microreactor. On the platform, one or more microreactors with volumes of 10 &#181;L can be positioned and moved simultaneously. The platform itself consists of a 3 x 3 matrix of electrical double coils which accommodate either neodymium or iron cores. The magnetic field gradients are automatically controlled. By variation of the magnetic field gradients, the microreactors' magnetic hydrophobic shell can be manipulated automatically to move the microreactor or open the shell reversibly. Reactions of substrates and corresponding enzymes can be initiated by merging the microreactors or bringing them into contact with surface immobilized catalysts.

Lab-in-a-drop reaction systems allow the versatile implementation of complex reactions in a microfluidic scale. An automated actuation platform consisting of a 3 x 3 matrix of electromagnetic coils was developed and successfully used to merge two 10 &#181;L microreactors and thereby initiate an enzymatic reaction in the resulting liquid marbles.

قطرات مائي يستخدم الانزيمية ميكروريكتورس وما يشتغل الكهرومغناطيسي

For the successful implementation of microfluidic reaction systems, such as PCR and electrophoresis, the movement of small liquid volumes is essential. In ...

Synthesis of Cationized Magnetoferritin for Ultra-fast Magnetization of Cells

Many important biomedical applications, such as cell imaging and remote manipulation, can be achieved by labeling cells with superparamagnetic iron oxide nanoparticles (SPIONs). Achieving sufficient cellular uptake of SPIONs is a challenge that has traditionally been met by exposing cells to elevated concentrations of SPIONs or by prolonging exposure times (up to 72 hr). However, these strategies are likely to mediate toxicity. Here, we present the synthesis of the protein-based SPION magnetoferritin as well as a facile surface functionalization protocol that enables rapid cell magnetization using low exposure concentrations. The SPION core of magnetoferritin consists of cobalt-doped iron oxide with an average particle diameter of 8.2 nm mineralized inside the cavity of horse spleen apo-ferritin. Chemical cationization of magnetoferritin produced a novel, highly membrane-active SPION that magnetized human mesenchymal stem cells (hMSCs) using incubation times as short as one minute and iron concentrations as lows as 0.2 mM.

A protocol for the synthesis and cationization of cobalt-doped magnetoferritin is presented, as well as a method to rapidly magnetize stem cells with cationized magnetoferritin.

توليف Cationized Magnetoferritin للمغنطة فائقة السرعة من الخلايا

Many important biomedical applications, such as cell imaging and remote manipulation, can be achieved by labeling cells with superparamagnetic iron oxide ...

Engineering Molecular Recognition with Bio-mimetic Polymers on Single Walled Carbon Nanotubes

Semiconducting single-wall carbon nanotubes (SWNTs) are a class of optically active nanomaterial that fluoresce in the near infrared, coinciding with the optical window where biological samples are most transparent. Here, we outline techniques to adsorb amphiphilic polymers and polynucleic acids onto the surface of SWNTs to engineer their corona phases and create novel molecular sensors for small molecules and proteins. These functionalized SWNT sensors are both biocompatible and stable. Polymers are adsorbed onto the nanotube surface either by direct sonication of SWNTs and polymer or by suspending SWNTs using a surfactant followed by dialysis with polymer. The fluorescence emission, stability, and response of these sensors to target analytes are confirmed using absorbance and near-infrared fluorescence spectroscopy. Furthermore, we demonstrate surface immobilization of the sensors onto glass slides to enable single-molecule fluorescence microscopy to characterize polymer adsorption and analyte binding kinetics.

We present a protocol for engineering the corona phase of near infrared fluorescent single walled carbon nanotubes (SWNTs) using amphiphilic polymers and DNA to develop sensors for molecular targets without known recognition elements.

الاعتراف الجزيئية والهندسة مع البوليمرات الحيوية المحاكاة على واحدة المسورة الكربون الأنابيب النانومترية

Semiconducting single-wall carbon nanotubes (SWNTs) are a class of optically active nanomaterial that fluoresce in the near infrared, coinciding with the ...