MULTILAYER VASCULAR GRAFTS BASED ON COLLAGEN-MIMETIC HYDROGELS

描述（由申请人提供）：开发现成的小口径血管移植物的主要障碍是实现支架的快速内皮化，同时最大限度地降低血栓形成、内膜增生和机械故障的风险。鉴于血小板聚集和平滑肌细胞增殖可能通过控制内皮细胞（EC）生长和表型介导，指导适当EC行为的材料的开发将对小血管修复和置换产生重大影响。然而，促进移植物内皮化的基质性质可能与维持成人血管系统相关负荷的基质性质不一致。为了解决这一限制，我们建议制造多层水凝胶-电纺网状支架，其中水凝胶层提供诱导快速内皮化的局部环境，并且电纺网状套管提供体积强度、顺应性匹配和缝线保留。因此，每个组件都可以单独调整，以实现更好的结果，而不会损害其他设计目标。我们建议通过使用胶原蛋白模拟蛋白Scl2.28（Scl 2）生成新的生物活性水凝胶来规避与天然生物聚合物凝胶相关的限制。Scl 2是最近发现的蛋白质，其具有天然胶原的三螺旋结构特征，但缺乏胶原的细胞粘附、细胞因子结合和酶切割位点的阵列。对于目前的工作，我们已经将<$1 <$1和<$2 <$1粘附位点引入到“亲本”Scl 2中，以提供EC相互作用的机制，同时保持与Scl 2相关的低血小板聚集。诱导所需细胞行为的基于Scl 2的水凝胶制剂将用于制造用不可降解的静电纺丝网“套管”增强的多层血管移植物，所述不可降解的静电纺丝网“套管”设计成具有与天然冠状动脉相似的机械性能。目标1。鉴定促进血管移植物的快速内皮化（粘附、迁移、静止表型）同时保持Scl 2蛋白的非血栓形成性质的PEGDA-Scl 2组合物。目标2.通过静电纺聚氨酯套管增强水凝胶，制造具有临床相关机械性能（破裂压力、缝线保留强度、顺应性）的多层血管移植物。目标3：评估复合移植物各组件的生物相容性和生物稳定性。目标4。在尤卡坦小型猪模型中评价植入后作为颈动脉移植物的体内多层移植物。 在5年期结束时，我们将在临床前动物研究中评价这些新管道，并证明其作为现成小口径血管移植物的潜在效用。从基本的角度来看，这一系列的混合材料将提供工具，阐明内皮化过程的关键，许多心血管设备的临床成功。此外，PEGDA-Scl 2凝胶提供的对生物活性和模量的控制，结合通过将不同的整合素结合基序掺入Scl 2中靶向一系列不同细胞类型的能力，将在创造用于广泛的生物医学应用的新生物活性材料中形成强大的平台。

DESCRIPTION (provided by applicant): A major roadblock in the development of off-the-shelf, small-caliber vascular grafts is achieving rapid endothelialization of the scaffold while minimizin the risk of thrombosis, intimal hyperplasia, and mechanical failure. Given that platelet aggregation and smooth muscle cell proliferation may be mediated by controlling endothelial cell (EC) growth and phenotype, the development of materials that direct appropriate EC behavior would have a significant impact on small vessel repair and replacement. However, matrix properties which promote graft endothelialization may not be consistent with those appropriate to sustain the loads associated with adult vasculature. To address this limitation, we propose to fabricate multilayered hydrogel-electrospun mesh scaffolds in which a hydrogel layer provides a local environment inductive of rapid endothelialization and an electrospun mesh sleeve provides bulk strength, compliance matching, and suture retention. Thus, each component can be individually tuned to achieve improved outcomes without detriment to other design goals. We propose to circumvent the limitations associated with native biopolymer gels by generating novel bioactive hydrogels using the collagen-mimetic protein Scl2.28 (Scl2). Scl2 is a recently discovered protein which has the triple helical structure characteristic of native collagen but lacks collagen's array of cell adhesion, cytokine binding, and enzyme-cleavage sites. For the present work, we have introduced ¿1¿1 and ¿2¿1 adhesion sites into the "parent" Scl2 to provide a mechanism for EC interactions while maintaining the low platelet aggregation associated with Scl2. Scl2-based hydrogel formulations that induce desired cell behaviors will be utilized in the fabrication of the multilayer vascular graft reinforced with non-degradable electrospun mesh "sleeves" designed to have mechanical properties similar to native coronary arteries. Aim 1. Identify PEGDA-Scl2 compositions that promote rapid endothelialization of the vascular graft (adhesion, migration, quiescent phenotype) while maintaining the non-thrombogenic nature of Scl2 proteins. Aim 2. Fabricate a multilayer vascular graft with clinically-relevant mechanical properties (burst pressure, suture retention strength, compliance) by reinforcing hydrogels with electrospun polyurethane sleeves. Aim 3. Assess biocompatibility and biostability of each component of the composite graft. Aim 4. Evaluate multilayer grafts in vivo after implantation as carotid grafts in a Yucatan miniature pig model. At the end of the 5 year period, we will have evaluated these new conduits in preclinical animal studies and demonstrated their potential utility as off-the-shelf, small-caliber vascular grafts. From a fundamental perspective, this family of hybrid materials will provide the tools to elucidate endothelialization processes critical to the clinical success of numerous cardiovascular devices. Furthermore, the control over both bioactivity and modulus afforded by PEGDA-Scl2 gels, combined with the ability to target a range of different cell types by incorporating different integrin binding motifs into Scl2, will form a powerful platform in the creation of new bioactive materials for a wide range of biomedical applications.