Name: the synthesis of rgd-functionalized hydrogels as a tool for therapeutic applications
Uploaded: 2016-10-07T14:00:00
Description: Watch this Scientific Journal Video about The Synthesis of RGD-functionalized Hydrogels as a Tool for Therapeutic Applications at JoVE.com

저자는 언어 편집을위한 유익한 토론을위한 교수 마우리 치오 MASI 미스 키아라 Allegretti에게 감사의 말씀을 전합니다. 저자의 연구는 반도 히오바니 Ricercatori 2010 (Ministero 델라 경례 GR-2010-2312573)에 의해 지원됩니다.

주 : 수신 한 화학 물질이 사용된다. 선형 RGD 구입되지만 FMOC 표준 고체상 펩타이드 합성 16, 19에 의해 제조 될 수있다. 용제는 분석 등급의이다. 투석 차단 동일 3,500 다 승 M과 멤브레인의 사용을 요구한다. 합성 된 화합물 1 400 메가 헤르츠 분광기하여 클로로포름에 기록 H NMR 스펙트럼 (CDCl3 중) 또는 용매와 같은 산화 중수소 (D 2 O)을 특징으로하고, 화학적 ?...

<ol>
	<li>Slaughter, B. V., Khurshid, S. S., Fisher, O. Z., Khademhosseini, A., Peppas, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hydrogels+in+Regenerative+Medicine.">Hydrogels in Regenerative Medicine.</a> Adv. Mater. 21 (32-33), 3307-3329 (2009).</li><li>Rossi, F., Perale, G., Papa, S., Forloni, G., Veglianese, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+options+for+drug+delivery+to+the+spinal+cord.">Current options for drug delivery to the spinal cord.</a> Expert Opin. Drug Deliv. 10 (3), 385-396 (2013).</li><li>Huebsch, 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=Harnessing+traction-mediated+manipulation+of+the+cell/matrix+interface+to+control+stem-cell+fate.">Harnessing traction-mediated manipulation of the cell/matrix interface to control stem-cell fate.</a> Nat. Mater. 9 (6), 518-526 (2010).</li><li>Mothe, A. J., Tam, R. Y., Zahir, T., Tator, C. H., Shoichet, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repair+of+the+injured+spinal+cord+by+transplantation+of+neural+stem+cells+in+a+hyaluronan-based+hydrogel.">Repair of the injured spinal cord by transplantation of neural stem cells in a hyaluronan-based hydrogel.</a> Biomaterials. 34 (15), 3775-3783 (2013).</li><li>Khetan, 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=Degradation-mediated+cellular+traction+directs+stem+cell+fate+in+covalently+crosslinked+three-dimensional+hydrogels.">Degradation-mediated cellular traction directs stem cell fate in covalently crosslinked three-dimensional hydrogels.</a> Nat. Mater. 12 (5), 458-465 (2013).</li><li>Ashley, G. W., Henise, J., Reid, R., Santi, D. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hydrogel+drug+delivery+system+with+predictable+and+tunable+drug+release+and+degradation+rates.">Hydrogel drug delivery system with predictable and tunable drug release and degradation rates.</a> Proc. Natl. Acad. Sci. U S A. 110 (6), 2318-2323 (2013).</li><li>Rossi, F., van Griensven, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polymer+Functionalization+as+a+Powerful+Tool+to+Improve+Scaffold+Performances.">Polymer Functionalization as a Powerful Tool to Improve Scaffold Performances.</a> Tissue Eng. Part A. 20 (15-16), 2043-2051 (2014).</li><li>Gould, S. T., Darling, N. J., Anseth, K. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small+peptide+functionalized+thiol-ene+hydrogels+as+culture+substrates+for+understanding+valvular+interstitial+cell+activation+and+de+novo+tissue+deposition.">Small peptide functionalized thiol-ene hydrogels as culture substrates for understanding valvular interstitial cell activation and de novo tissue deposition.</a> Acta Biomater. 8 (9), 3201-3209 (2012).</li><li>Azagarsamy, M. A., Anseth, K. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wavelength-Controlled+Photocleavage+for+the+Orthogonal+and+Sequential+Release+of+Multiple+Proteins.">Wavelength-Controlled Photocleavage for the Orthogonal and Sequential Release of Multiple Proteins.</a> Angew. Chem. Int. Edit. 52 (51), 13803-13807 (2013).</li><li>Larra&#241;eta, 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=Microwave-Assisted+Preparation+of+Hydrogel-Forming+Microneedle+Arrays+for+Transdermal+Drug+Delivery+Applications.">Microwave-Assisted Preparation of Hydrogel-Forming Microneedle Arrays for Transdermal Drug Delivery Applications.</a> Macromol. Mater. Eng. 300 (6), 586-595 (2015).</li><li>Cook, J. P., Goodall, G. W., Khutoryanskaya, O. V., Khutoryanskiy, V. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microwave-Assisted+Hydrogel+Synthesis:+A+New+Method+for+Crosslinking+Polymers+in+Aqueous+Solutions.">Microwave-Assisted Hydrogel Synthesis: A New Method for Crosslinking Polymers in Aqueous Solutions.</a> Macromol. Rapid Comm. 33 (4), 332-336 (2012).</li><li>Perale, G., 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=Multiple+drug+delivery+hydrogel+system+for+spinal+cord+injury+repair+strategies.">Multiple drug delivery hydrogel system for spinal cord injury repair strategies.</a> J. Control. Release. 159 (2), 271-280 (2012).</li><li>Rossi, F., Perale, G., Storti, G., Masi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Library+of+Tunable+Agarose+Carbomer-Based+Hydrogels+for+Tissue+Engineering+Applications:+The+Role+of+Cross-Linkers.">A Library of Tunable Agarose Carbomer-Based Hydrogels for Tissue Engineering Applications: The Role of Cross-Linkers.</a> J. Appl. Polym. Sci. 123 (4), 2211-2221 (2012).</li><li>Frith, J. 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=An+injectable+hydrogel+incorporating+mesenchymal+precursor+cells+and+pentosan+polysulphate+for+intervertebral+disc+regeneration.">An injectable hydrogel incorporating mesenchymal precursor cells and pentosan polysulphate for intervertebral disc regeneration.</a> Biomaterials. 34 (37), 9430-9440 (2013).</li><li>Kolb, H. C., Finn, M. G., Sharpless, K. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Click+chemistry:+Diverse+chemical+function+from+a+few+good+reactions.">Click chemistry: Diverse chemical function from a few good reactions.</a> Angew. Chem. Int. Edit. 40 (11), (2001).</li><li>Sacchetti, A., Mauri, E., Sani, M., Masi, M., Rossi, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microwave-assisted+synthesis+and+click+chemistry+as+simple+and+efficient+strategy+for+RGD+functionalized+hydrogels.">Microwave-assisted synthesis and click chemistry as simple and efficient strategy for RGD functionalized hydrogels.</a> Tetrahedron Lett. 55 (50), 6817-6820 (2014).</li><li>Ossipov, D. A., Hilborn, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Poly(vinyl+alcohol)-based+hydrogels+formed+by+&amp;#34;click+chemistry&amp;#34.">Poly(vinyl alcohol)-based hydrogels formed by &amp;#34;click chemistry&amp;#34.</a> Macromolecules. 39 (5), 1709-1718 (2006).</li><li>Truong, V., Blakey, I., Whittaker, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hydrophilic+and+Amphiphilic+Polyethylene+Glycol-Based+Hydrogels+with+Tunable+Degradability+Prepared+by+&amp;#34;Click&amp;#34;+Chemistry.">Hydrophilic and Amphiphilic Polyethylene Glycol-Based Hydrogels with Tunable Degradability Prepared by &amp;#34;Click&amp;#34; Chemistry.</a> Biomacromolecules. 13 (12), 4012-4021 (2012).</li><li>Hou, R. Z., 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=New+synthetic+route+for+RGD+tripeptide.">New synthetic route for RGD tripeptide.</a> Prep. Biochem. Biotechnol. 36 (3), 243-252 (2006).</li><li>Rossi, F., Chatzistavrou, X., Perale, G., Boccaccini, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+and+Degradation+of+Agar-Carbomer+Based+Hydrogels+for+Tissue+Engineering+Applications.">Synthesis and Degradation of Agar-Carbomer Based Hydrogels for Tissue Engineering Applications.</a> J. Appl. Polym. Sci. 123 (1), 398-408 (2012).</li><li>Mauri, E., Rossi, F., Sacchetti, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tunable+drug+delivery+using+chemoselective+functionalization+of+hydrogels.">Tunable drug delivery using chemoselective functionalization of hydrogels.</a> Mater. Sci. Eng. C. 61, 851-857 (2016).</li><li>Joaquin, A., Peppas, N. A., Zoldan, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hydrogel+Polymer+Library+for+Developing+Induced+Pluripotent+Stem+Cell+Derived+Cardiac+Patches.">Hydrogel Polymer Library for Developing Induced Pluripotent Stem Cell Derived Cardiac Patches.</a> Tissue Eng. Part A. 20, S55-S55 (2014).</li><li>Rossi, F., 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=Tunable+hydrogel-Nanoparticles+release+system+for+sustained+combination+therapies+in+the+spinal+cord.">Tunable hydrogel-Nanoparticles release system for sustained combination therapies in the spinal cord.</a> Colloids Surf. B Biointerfaces. 108, 169-177 (2013).</li><li>Kolb, H. C., Sharpless, K. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+growing+impact+of+click+chemistry+on+drug+discovery.">The growing impact of click chemistry on drug discovery.</a> Drug Discov. Today. 8 (24), 1128-1137 (2003).</li><li>Ossipov, D. A., Yang, X., Varghese, O., Kootala, S., Hilborn, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modular+approach+to+functional+hyaluronic+acid+hydrogels+using+orthogonal+chemical+reactions.">Modular approach to functional hyaluronic acid hydrogels using orthogonal chemical reactions.</a> Chem. Commun. 46 (44), 8368-8370 (2010).</li><li>Anderson, S. B., Lin, C. C., Kuntzler, D. V., Anseth, K. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+performance+of+human+mesenchymal+stem+cells+encapsulated+in+cell-degradable+polymer-peptide+hydrogels.">The performance of human mesenchymal stem cells encapsulated in cell-degradable polymer-peptide hydrogels.</a> Biomaterials. 32 (14), 3564-3574 (2011).</li><li>Caron, I., 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=A+new+three+dimensional+biomimetic+hydrogel+to+deliver+factors+secreted+by+human+mesenchymal+stem+cells+in+spinal+cord+injury.">A new three dimensional biomimetic hydrogel to deliver factors secreted by human mesenchymal stem cells in spinal cord injury.</a> Biomaterials. 75, 135-147 (2016).</li><li>Lee, J. W., Kim, H., Lee, K. Y. <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+spacer+arm+length+between+adhesion+ligand+and+alginate+hydrogel+on+stem+cell+differentiation.">Effect of spacer arm length between adhesion ligand and alginate hydrogel on stem cell differentiation.</a> Carbohyd. Polym. 139, 82-89 (2016).</li><li>Liu, Y., Fan, Z., Wang, Y., Yu, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+Release+of+Low+Molecular+Protein+Insulin-like+Growth+Factor-1+through+Self-Assembling+Peptide+Hydrogel+with+Biotin+Sandwich+Approach.">Controlled Release of Low Molecular Protein Insulin-like Growth Factor-1 through Self-Assembling Peptide Hydrogel with Biotin Sandwich Approach.</a> J.Biomed. Eng. 32 (2), 387-392 (2015).</li></ol>

The PAA post-polymerization modification with alkyne moieties and the RGD functionalization with the azide group guarantee the formation of a stable bond between the polymer and the peptide. Indeed, triazole serves as a rigid linking unit among the carbon atoms, attached to the 1,4 positions of the 1,2,3-triazole ring and it cannot be cleaved hydrolytically or otherwise. In addition, triazole is extremely difficult to oxidize and reduce, unlike other cyclic structures such as benzenoids and related aromatic heterocycles<...

하이드로 겔은 천연 또는 합성, 및 독특한 입체 구조 특징 친수성 가교 중합체에 의해 형성되는 삼차원 네트워크이다. 이 장치는 약물 전달, 조직 공학, 유전자 사업자와 스마트 센서 1, 2의 생물 의학 분야에서 점점 더 매력적이다. 실제로, 높은 수분 함량뿐만 아니라 유동 학적 및 기계적 특성들에게 연조직 미세 환경을 모방 수용성 사이토킨 또는 성장 인자의 전달을 위해 그들에게 효과적인 도구를 만들기에 적합한 후보를 만든다. 가장 유망한 사용 중 하나는 세포 및 생물 활성 화합물을 담지 주사 생체 같다. 생체 외 및 생체 내 실험 3,4에서 관찰 하이드로 겔은 잡고 정확하게 생리 관련 방식으로 줄기 세포 규제 신호를 제공함으로써 세포 생존 및 제어 줄기 세포 운명을 향상시킬 수 있습니다. 이것의 주요 장점은 가능성신체를 통해 모든 마이그레이션 및 대상 목표 (5)를 잃고, 순환 토런트로 지역과 extravasates 잎 세포의 양을 최소화 (현장에서) 접종의 영역 내 주입 된 세포를 유지합니다. 삼차원 네트워크 하이드로 겔의 안정성은 고분자 사슬 6 간의 공유 결합 또는 응집력에 의해 형성의 가교 부위에 기인한다. 이 프레임 워크에서는 직교 선택적 화학 고분자 사슬에 적용이 하이드로 겔 공연 (7)을 개선 할 수있는 다목적 도구입니다. 실제로, 적절한 화학기를 갖는 중합체의 변형은 세포 생존 및 조직 형성에서의 사용을 향상시키기 위해 적절한 화학적, 물리적 및 기계적 성질을 제공하도록 도울 수있다. 동일한 방식으로, 기술 중 RGD 펩티드의 사용은 세포 부착 및 생존율의 개선을 허용 겔 매트릭스 내에 세포 성장 인자를로드한다. RGD는 구성된 트리 펩티드이다지금까지 아르기닌, 글리신, 아스파르트 산, 가장 효과적인 인해 종종 하나 이상의 세포 부착 수용체를 처리 할 수있는 능력 및 세포 정박 동작 생존 8,9에 생물학적 충격 트리 펩티드를 사용. 본 연구에서는 RGD-작용 하이드로 겔의 합성은 친절 세포 미세 환경에 대한 충분한 생화학 적 특성을 특징으로 네트워크를 설계하는 목적으로 연구되고있다. 하이드로 젤 합성 마이크로파 방사선의 사용은 부반응을 최소화하고, 종래의 열 공정 (10)에 비해 짧은 시간에 높은 반응 속도 및 수율을 얻을 수있는 간단한 방법을 제공한다. 이 방법은 중합체의 상호 작용 및 반응 시스템 (11) 내의 유기 용매의 부재에 의한 정제 단계 및 수율 멸균 하이드로 겔을 필요로하지 않는다. 따라서, 어떤 모드 때문에 고분자 네트워크에 연결 RGD의 높은 비율을 보장합니다ifications 겔 형성에 관련된 고분자 화학 그룹으로 요구된다. 카르복실기, PAA와 카보 머, 하이드 록실 그룹, PEG에서 아가 로스에서 중축 합 반응을 통해 하이드로 겔 입체 구조를 야기. 언급 된 중합체는 척수 손상 수리먼트 (12) 하이드로 겔의 합성에 사용된다. 이러한 장치는 이전에보고 된 생체 적합성뿐만 아니라 많은 생체 조직들 및 요변 특성에 유사한 기계적 물리 화학적 특성을 보여, 13, 14 일. 또한, 그들은 주입의 영역에서, 현장에서 현지화 된 상태로 유지됩니다. 이 연구에서, PAA의 카르복실기 알킨 잔기 (도 1)를 수정하고 RGD 지드 화합물 구조 (CH 2) N으로 준비된 화합물과 트리 펩티드 단기 -NH (2)의 반응을 이용한 합성 - N 3 (&lt;강한&gt; 그림 2). 그 후, 수정 된 PAA는 CuAAC 클릭 반응 15-17 (그림 3)를 통해 RGD-아 지드 유도체와 반응한다. 구리 (I) 촉매를 사용하여 반응 속도 및 선택성 둘 다의 주요 개선 리드. CuAAC 반응은 유기 합성에서 폭넓게 중합체 과학 분야에서 사용된다. 이는 작용기 고효율 및 높은 내성을 결합하고, 유기 용매의 사용에 의해 영향을받지 않는다. 높은 선택성, 빠른 반응 시간 및 간단한 정제 과정은 원하는 부분 (18)을 이식 성 중합체, 블록 공중 합체 또는 사슬의 획득을 허용한다. 이 클릭 전략은 가능한 최종 생화학 응용 프로그램에 따라 물리 화학적 특성을 사용자 정의 할 수 중합 후 폴리머를 수정할 수 있습니다. CuAAC 실험 조건은 쉽게 (구리의 산화가 최소한으로 발생할 수있는 반면, 반응이 물에 민감하다) 재현, 그리고 자연이다졸 형성된 제품의 안정성을 보장한다. 구리 금속의 사용으로 인해 세포에 대한 잠재적 인 독성 효과 및 생물학적 미세 환경에서 중요한 점으로 간주 될 수 있지만, 투석 촉매 잔기의 완전한 제거를 허용하도록 정제 방법으로 사용된다. 마지막으로, PAA는 RGD 세포 또는 약물 담체로서 이러한 시스템의 잠재적 기능을 확인하기 위해 조사되는 하이드로 겔의 합성 (도 4) 및 생성 된 네트워크의 물리 화학적 특성에 사용되는 변성. <img alt="그림 1" src="/files/ftp_upload/54445/54445fig1.jpg" /> 그림 1 : PAA는 알킨의 합성을 수정 알킨 그룹과 PAA의 작용의 방식을,. &quot;N&quot;은 카르복실기가 propargylamine와 반응에 단량체를 나타냅니다. <a href="http://ecsource.jove.com/files/ftp_upload/54445/54445fig1large.jpg" target="_blank">보려면 여기를 클릭하십시오</a>이 그림의 더 큰 버전. <img alt="그림 2" src="/files/ftp_upload/54445/54445fig2.jpg" /> 그림 2 :.. RGD-아 지드 합성 RGD-아 지드 유도체의 합성 <a href="http://ecsource.jove.com/files/ftp_upload/54445/54445fig2large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 3" src="/files/ftp_upload/54445/54445fig3.jpg" /> 그림 3 : 반응을 클릭 제도 RGD-아 지드 유도체 및 알킨-PAA 사이 클릭 반응의.. <a href="http://ecsource.jove.com/files/ftp_upload/54445/54445fig3large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 4" src="/files/ftp_upload/54445/54445fig4.jpg" /> 그림 4 : 히드로 겔 SYNThesis. RGD 작용 하이드로 겔 합성 절차. <a href="http://ecsource.jove.com/files/ftp_upload/54445/54445fig4large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a>

<table border="0" cellpadding="0" cellspacing="0" width="521">
	<tbody>
		<tr height="67">
			<td height="67" style="height:67px;width:183px;">Poly(acrylic acid) solution average Mw ~ 100,000, 35 wt % in H2O</td>
			<td style="width:108px;">Sigma Aldrich</td>
			<td style="width:87px;">523925</td>
			<td style="width:145px;">CAS 9003-01-4</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Poly(ethylene glycol) 2,000</td>
			<td>Sigma Aldrich</td>
			<td>84797</td>
			<td>CAS 25322-68-3</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Carbomeer 974P</td>
			<td>Fagron</td>
			<td>1387083</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Agarose&#160;</td>
			<td>Invitrogen Corp.</td>
			<td style="width:87px;">16500-500</td>
			<td>UltraPure Agarose</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">RGD peptide</td>
			<td style="width:108px;">abcam</td>
			<td>ab142698</td>
			<td style="width:145px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">4-azidobutanoic acid</td>
			<td style="width:108px;">Aurum Pharmatech</td>
			<td style="width:87px;">Z-2421</td>
			<td>&#160;CAS 54447-68-6</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Oxalyl chloride</td>
			<td style="width:108px;">Sigma Aldrich</td>
			<td>O8801</td>
			<td>CAS 79-37-8</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:183px;">Propargylamine hydrochloride 95%</td>
			<td>Sigma Aldrich</td>
			<td>P50919</td>
			<td>CAS 15430-52-1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:183px;">Copper(I) iodide</td>
			<td>Sigma Aldrich</td>
			<td>3140</td>
			<td>CAS 7681-65-4</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium ascorbate</td>
			<td>Sigma Aldrich</td>
			<td>Y0000039</td>
			<td>CAS 134-03-2</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Phosphate buffered saline</td>
			<td>Sigma Aldrich</td>
			<td>P4417</td>
			<td></td>
		</tr>
		<tr height="85">
			<td height="85" style="height:85px;">Dialysis Membrane</td>
			<td style="width:108px;">Spectrum Laboratories, Inc.</td>
			<td>132725</td>
			<td style="width:145px;">Spectra/Por 3 Dialysis Membrane&#160; Standard RC Tubing&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; MWCO: 3,5 kD</td>
		</tr>
	</tbody>
</table>

the synthesis of rgd-functionalized hydrogels as a tool for therapeutic applications

The use of polymers as biomaterials has provided significant advantages in therapeutic applications. In particular, the possibility to modify and functionalize polymer chains with compounds that are able to improve biocompatibility, mechanical properties, or cell viability allows the design of novel materials to meet new challenges in the biomedical field. With the polymer functionalization strategies, click chemistry is a powerful tool to improve cell-compatibility and drug delivery properties of polymeric devices. Similarly, the fundamental need of biomedicine to use sterile tools to avoid potential adverse-side effects, such as toxicity or contamination of the biological environment, gives rise to increasing interest in the microwave-assisted strategy.
The combination of click chemistry and the microwave-assisted method is suitable to produce biocompatible hydrogels with desired functionalities and improved performances in biomedical applications. This work aims to synthesize RGD-functionalized hydrogels. RGD (arginylglycylaspartic acid) is a tripeptide that can mimic cell adhesion proteins and bind to cell-surface receptors, creating a hospitable microenvironment for cells within the 3D polymeric network of the hydrogels. RGD functionalization occurs through Huisgen 1,3-dipolar cycloaddition. Some PAA carboxyl groups are modified with an alkyne moiety, whereas RGD is functionalized with azido acid as the terminal residue of the peptide sequence. Finally, both products are used in a copper catalyzed click reaction to permanently link the peptide to PAA. This modified polymer is used with carbomer, agarose and polyethylene glycol (PEG) to synthesize a hydrogel matrix. The 3D structure is formed due to an esterification reaction involving carboxyl groups from PAA and carbomer and hydroxyl groups from agarose and PEG through microwave-assisted polycondensation. The efficiency of the gelation mechanism ensures a high degree of RGD functionalization. In addition, the procedure to load therapeutic compounds or biological tools within this functionalized network is very simple and reproducible.

n은 카르복실기, 아민과 반응 단량체 라벨도 1에 나타내었다 같이 PAA의 알킨 유도체를 효율적으로, 폴리 아크릴산 및 propargylamine에서 합성된다. 제품의 아이덴티티는 1 H-NMR 분광법으로 확인 하였다. (5) 삼중 결합 변성 PAA의 1H-NMR 스펙트럼을 나타낸다. <img alt="그림 5" src="/files/ftp_upload/54445/544...

Watch this Scientific Journal Video about The Synthesis of RGD-functionalized Hydrogels as a Tool for Therapeutic Applications at JoVE.com

The Synthesis of RGD-functionalized Hydrogels as a Tool for Therapeutic Applications

We present a protocol for the synthesis of RGD-functionalized hydrogels as devices for cell and drug delivery. The procedure involves copper catalyzed alkyne-azide cycloaddition (CuAAC) between alkyne-modified polyacrylic acid (PAA) and a RGD-azide derivative. The hydrogels are formed using microwave-assisted polycondensation and their physicochemical properties are investigated.

치료 응용 프로그램을위한 도구로 RGD-작용 히드로 겔의 합성

The use of polymers as biomaterials has provided significant advantages in therapeutic applications. In particular, the possibility to modify and ...

The overall goal of this methodology is to synthesize hydrogels for tissue engineering applications very easily. This method can help answer key questions in the tissue engineering field, such as the possibility to maintain high cell viability in three dimensional cell culture systems. The main advantage of this technique is that is using quick reaction, we can easily modify hydrogel systems to improve their ability to maintain cell viable.The implications of this technique extend toward the therapy of spinal cord injury because the cells loaded within the scaffold can rebuild the damaged tissue. For this method can provide significant insight into cell therapies as regenerative medicine and their implication in many methodologies. It can also be applied in several other systems like bone, heart, and cartilage.To synthesize the RGD-azide derivative, dissolve 50 milligrams of RGD in one milliliter of one molar sodium hydroxide. Then dissolve 24 milligrams of 4-azidobutanoyl chloride in two milliliters of tetrahydrofuran. Add all of the RGD solution into the 4-azidobutanoyl chloride solution at zero degrees Celsius dropwise.Return to room temperature and stir overnight. Add one milliliter of one molar HCL. Remove the solvent under reduced pressure using a rotary evaporator.Characterize the obtained product by proton nuclear magnetic resonance spectroscopy, dissolving the sample in deuterium oxide or D2O. To perform the PAA alkine modification, dissolve 200 milligrams of 35%in weight PAA solution in 15 milliliters of distilled water. Add 15.4 milligrams of propargylamine hydrochloride.Then dissolve 42.8 milligrams of HOBt in 14 milliliters of a one to one ratio of acetonytryle distilled water solution by heating to 50 degrees Celsius. Add all of the HOBt solution to the PAA solution at room temperature. Next, add 53.6 milligrams of EDC to the reaction mixture.Use one molar HCL to adjust the PH to 5.5 and stir the reaction system overnight at room temperature. Next, dissolve 11.2 grams of sodium chloride in two liters of distilled water. And then add 200 microliters of 37%weight by weight HCL.Dialyze the solution using a membrane with a molecular weight cutoff of 3.5 kilodaltons. Perform dialysis for three days. Change the dialysis solution daily with two liters of freshly prepared distilled water containing 200 microliters of 37%weight by weight HCL.Store the final solution at minus 80 degrees Celsius. Characterize the functionalized polymer by proton nuclear magnetic resonance spectroscopy, dissolving the sample in D2O. To synthesize the PAA-RGD polymer, dissolve 78 milligrams of PAA modified alkine in 10 milliliters of distilled water.Then dissolve 25 milligrams of the RGD-azide derivative in five milliliters of tetrahydrafuran. Add all of the RGD solution to the polymeric solution. Next, add 2.2 milligrams of copper iodide.And 2.2 milligrams of sodium ascorbate. Reflux the resulting mixture overnight at 60 degrees Celsius with stirring. Cool the mixture to 25 degrees Celsius.To dialyze the solution, dissolve 11.2 grams of sodium chloride in two liters of distilled water and add 200 microliters of 37 percent weight by weight HCL. Perform dialysis for three days. Change the dialysis solution daily with two liters of freshly prepared distilled water containing 200 microliters of 37%weight by weight HCL.Then place the final solution at minus 80 degrees Celsius. Characterize and store the resulting PAA-RGD polymer as done for its constituents. Blend 40 milligrams of carbomer and 10 milligrams of the functionalized PAA in nine milliters of phosphate-buffered saline at room temperature until complete dissolution.Add 400 milligrams of PEG to the solution and keep stirring for 45 minutes. Stop the stirring and allow the system to settle for 30 minutes. Then use one molar sodium hydroxide to adjust the PH to 7.4.To five milliliters of the obtained mixture, add 25 milligrams of agarose powder. Irradiate the system with microwave radiation at 500 watts for 30 seconds to one minute until boiling. And electromagnetically heat up the solution to 80 degrees Celsius.Leave the mixture exposed to room temperature until its temperature decreases to 50 degrees Celsius. Then add five milliliters of PBS to obtain a solution at a one to one volumetric ratio. Prepare a 12-multiwell plate containing steel cylinders with a diameter of 1.1 centimeters.Take 500 microliters aliquots from the solution and place them into each steel cylinder. Leave the plate at rest for 45 minutes until complete gelification of the system. Remove the cylinders using stainless steel forceps to obtain the hydrogels.See the text protocol for characterizing the physicochemical properties of the RGD-functionalized hydrogels. Carbomer 974P, PAA-RGD, and PEG, together with agarose, take part in a statistical random polycondensation after microwave irradiation. Hydroxyl groups contained in PEG and agarose react with carboxyl groups present in Carbomer 947P and PAA-RGD to form ACPEG-RGD hydrogels.Considering rheological properties, the storage modulus was found to be approximately one order of magnitude higher than the loss modulus, indicating an elastic rather than viscous material. Moreover, it is clearly visible that hydrogel behavior is dominated at low strain values by the elastic modulus. Scanning electron microscopy shows the morphology of an RGD functionalized hydrogel sample and a hydrogel without functionalization.Both hydrogels revealed that they possess a highly entangled structure of interconnected pores, with some bigger pores containing small pores and some fibular networks on the pore walls. The similar properties indicate that the presence of RGD does not alter the polymer network. After this development, this technique paved the way for research in the field of tissue engineering to explore the stability of these cell culture systems in several other diseases.After watching this video, you should have a good understanding of how to functionalize hydrogels. Improving their ability to maintain viable stem cells.

Research

JoVE Journal

Bioengineering

Fluorescence-quenching of a Liposomal-encapsulated Near-infrared Fluorophore as a Tool for In Vivo Optical Imaging

Fluorescence-quenching of a Liposomal-encapsulated Near-infrared Fluorophore as a Tool for In Vivo Optical Imaging

위한 도구로 리포좀 캡슐화 된 근적외선 형광의 형광 담금질<em&gt; 생체</em&gt; 광학 이미징

Optical imaging offers a wide range of diagnostic modalities and has attracted a lot of interest as a tool for biomedical imaging. Despite the enormous ...

연구

생체공학

Solid Lipid Nanoparticles (SLNs) for Intracellular Targeting Applications

Solid Lipid Nanoparticles SLNs for Intracellular Targeting Applications

고체 지질 나노 입자 (SLNs) 세포 내 타겟팅을위한 응용 프로그램

Nanoparticle-based delivery vehicles have shown great promise for intracellular targeting applications, providing a mechanism to specifically alter ...

Light-mediated Formation and Patterning of Hydrogels for Cell Culture Applications

세포 배양 응용 프로그램에 대한 히드로 겔의 빛 매개 형성 및 패터닝

Click chemistries have been investigated for use in numerous biomaterials applications, including drug delivery, tissue engineering, and cell culture. In ...

Tunable Hydrogels from Pulmonary Extracellular Matrix for 3D Cell Culture

3 차원 세포 배양을위한 폐 세포 외 매트릭스에서 조정 히드로 겔

Here we present a method for establishing multiple component cell culture hydrogels for in vitro lung cell culture. Beginning with healthy en bloc lung ...

Synthesis of Hydrogels with Antifouling Properties As Membranes for Water Purification

물 정화를위한 막으로 방오 특성을 가진 히드로 겔의 합성

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

DNA Nanotubes as a Versatile Tool to Study Semiflexible Polymers

Semiflexible 고분자 공부를 다양 한 도구로 DNA 나노튜브

Mechanical properties of complex, polymer-based soft matter, such as cells or biopolymer networks, can be understood in neither the classical frame of ...

Ultrathin Porated Elastic Hydrogels As a Biomimetic Basement Membrane for Dual Cell Culture

듀얼에 대 한 생체 모방 지하실 멤브레인으로 ultrathin Porated 탄성 Hydrogels 세포 배양

The basement membrane is a critical component of cellular bilayers that can vary in stiffness, composition, architecture, and porosity. In vitro studies ...

Production of Elastin-like Protein Hydrogels for Encapsulation and Immunostaining of Cells in 3D

캡슐화 및 3d에서 셀의 Immunostaining에 대 한 엘라 스 틴 같은 단백질 Hydrogels의 생산

Two-dimensional (2D) tissue culture techniques have been essential for our understanding of fundamental cell biology. However, traditional 2D tissue ...

Synthesis of Near-Infrared Emitting Gold Nanoclusters for Biological Applications

생물학적 응용 을 위한 근적외선 방출 금 나노클러스터의 합성

Over the past decade, fluorescent gold nanoclusters (AuNCs) have witnessed growing popularity in biological applications and enormous efforts have been ...

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

암 세포 배양에서 수집된 매체에서 액체 크로마토그래피 질량 분광법에 의해 분석된 선택된 Kynurenines의 동시 정량화

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