Bioinspired Synthesis of In-Situ Gelling Biomaterials

原位凝胶生物材料的仿生合成

• 批准号: 6610841
• 负责人: Phillip B Messersmith
• 金额: $ 29.95万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6610841
• 关键词: SDS polyacrylamide gel electrophoresis; bioengineering /biomedical engineering; biomaterial compatibility; biomaterial development /preparation; biomaterials; calcium ion; charge coupled device camera; chondrocytes; combinatorial chemistry; crosslink; dosage forms; enzyme activity; gel; guinea pigs; high performance liquid chromatography; injection /infusion; lipid bilayer membrane; liposomes; mass spectrometry; peptide chemical synthesis; peptide library; polymers; protein glutamine gamma glutamyltransferase; tissue engineering

DESCRIPTION (provided by applicant): Transglutaminase enzymes are ubiquitous Ca2+-dependent enzymes that catalyze the formation of crosslinks between glutamine and lysine residues of proteins. Extensive transglutaminase-mediated crosslinking of soluble proteins is believed to be responsible for rapid physical gelation of certain biological fluids. A common biological strategy for regulating the activity of transglutaminase enzymes is control of intracellular and extracellular Ca 2+ concentration, mediated by lipid bilayer membranes. Stimuli-responsive synthetic lipid vesicles offer a unique opportunity to regulate transglutaminase-mediated gelation by sequestering and then releasing enzyme-activating ions such as Ca 2+. We hypothesize that Ca 2+ release from temperature or light sensitive liposomes can be used to trigger TG-mediated crosslinking of peptide-modified polymers to form hydrogels suitable for use as tissue adhesives and for injectable tissue engineering. In this study, combinatorial chemistry will be employed to synthesize large peptide libraries from which short peptide substrates of transglutaminase enzymes will be identified. The peptide substrates will be covalently linked to biocompatible polymers, and the TG-catalyzed crosslinking of the polymers into hydrogels will be studied in an effort to formulate injectable solutions that undergo rapid gelation in situ. Stimuli-responsive liposomes will be utilized to trigger calcium activation of enzyme-catalyzed gelation with the goal of developing thermal and light triggered gelation for clinical use. The tissue adhesive potential of these hydrogels will be assessed by measuring the force required to separate articular cartilage surfaces bonded together by in-situ formed hydrogels, and in vitro and in vivo studies will be performed to evaluate the potential of chondrocyte-containing injectable polymer hydrogels to support the formation of cartilage tissue.

描述（由申请人提供）： 转谷氨酰胺酶是普遍存在的Ca2+依赖性酶，可催化谷氨酰胺和蛋白质的赖氨酸残基之间的交联。据信，广泛的转谷氨酰胺酶介导的可溶性蛋白的交联是导致某些生物液的快速物理凝胶。调节转谷氨酰胺酶活性的常见生物学策略是控制细胞内和细胞外Ca 2+浓度，由脂质双层膜介导。刺激反应性合成脂质囊泡提供了一个独特的机会，可以通过隔离，然后释放酶激活离子，例如Ca 2+来调节转谷氨酰胺酶介导的凝胶化。我们假设Ca 2+从温度或光敏敏感脂质体中释放可用于触发TG介导的肽改性聚合物的TG介导的交联，形成适合用作组织胶粘剂和可注射组织工程的水凝胶。在这项研究中，将采用组合化学来合成大型肽文库，从中可以从中鉴定出短肽酶的短肽底物。肽底物将与生物相容性聚合物共价连接，并将研究聚合物在水凝胶中的TG催化交联，以制定可注射的溶液，从而在原位进行快速凝胶。刺激性反应性脂质体将用于触发酶催化凝胶化的钙激活，目的是开发热和光触发凝胶化以供临床使用。这些水凝胶的组织粘合剂将通过测量分离通过原位形成的水凝胶粘合在一起的关节软骨表面所需的力量，并将进行体外和体内研究，以评估含有软骨细胞的可注射的聚合物水凝胶的潜力，以支撑骨质组织的形成。