Design of injectable hydrogels to improve cell transplantation for peripheral arterial disease

设计可注射水凝胶以改善外周动脉疾病的细胞移植

基本信息

批准号：
8909701
负责人：
Lei Cai
金额：
$ 5.42万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8909701
关键词：
Address Adipose tissue Affect Age American Amputation Angiogenic Factor Antibodies Apoptotic Biological Assay Blood Blood Vessels Cell Death Cell Proliferation Cell Survival Cell Therapy Cell Transplantation Cell Transplants Cell membrane Cell physiology Cells Clinical Collagen Conditioned Culture Media Cytoprotection DNA Degenerative Disorder Dermal Drug Formulations Encapsulated Endothelial Cells Enzyme-Linked Immunosorbent Assay Gastrocnemius Muscle Gel Gene Expression Gene Expression Regulation Growth Growth Factor Human Hydrogels Hypoxia Image In Situ In Vitro Inflammation Injectable Injection of therapeutic agent Ischemia Label Laboratories Lasers Limb structure Location Maintenance Mechanics Medical Membrane Metabolic Modeling Monitor Motivation Multiple Trauma Mus Necrosis Needles Operative Surgical Procedures Patients Pattern Peptides Peripheral arterial disease Phase Transition PicoGreen Property Protocols documentation Recovery Recovery of Function Regenerative Medicine Reperfusion Therapy Reporting Reverse Transcriptase Polymerase Chain Reaction Saline Sampling Series Signal Transduction Site Stromal Cells Supporting Cell Syringes Therapeutic procedure Time Transplantation United States Vascular Endothelial Growth Factors Western Blotting angiogenesis base bioluminescence imaging blood perfusion caspase-3 cell type clinical application crosslink design digital experience fluorophore improved in vitro Model in vivo luciferin matrigel minimally invasive molecular recognition neovascularization novel paracrine public health relevance success tissue regeneration two-dimensional

项目摘要

DESCRIPTION (provided by applicant): Ineffective delivery of transplanted cells to the desired site is a major hurdle hampering the clinical application of cell-based, regenerative medicine therapies. Dismally low transplanted cell viability (~1-32%) is caused both by the mechanical forces during injection, which damage the cell membrane, and the lack of a three- dimensional matrix to support cell survival in situ. Peripheral arterial disease (PAD), occurs to one in 20 Americans over the age of 50. Current treatment options are often ineffective, commonly leading to amputation in cases of critical limb ischemia. A promising therapy is the transplantation of patient-derived human adipose- derived stromal cells (hASCs) into the ischemic site to promote reperfusion through secretion of pro-angiogenic paracrine factors. Here we propose a physically-crosslinked, double-network hydrogel with a wide range of stiffness from 10 to 3000 Pa to address the problem of post-transplantation cell death, to regulate cell functionality to optimize the angiogenic potential of hASCs, and to improve blood reperfusion in a murine model for PAD. In Specific Aim 1, I will synthesize and characterize an injectable hydrogel that undergoes two different physical crosslinking mechanisms. The first crosslinking step occurs ex vivo through peptide-based molecular recognition to encapsulate cells within a physical hydrogel, while the second crosslinking step occurs in situ through a thermal phase transition to form a reinforcing network. This hydrogel will be formulated with a wide range of stiffness from 10 to 3000 Pa by varying the reinforcing network densityto provide hASCs protection during syringe injection and to improve the cell viability post-transplantation. In Specific Aim 2, I will explore the effect of stiffness on the secretion of pro-angiogenic growth factors by hASCs in vitro and maximize the angiogenic potential of hASCs encapsulated within this series of novel hydrogels. In Specific Aim 3, I will choose the best-performing hydrogel formulation and deliver hASCs to a murine hind limb ischemia model of PAD. Recovery of blood perfusion and neovascularization will be assessed to validate the hASC-based therapy using our novel hydrogels, seeking to accelerate the clinical realization of cell therapy for PAD.

描述（由申请人提供）：移植细胞无效递送至所需部位是阻碍基于细胞的再生医学疗法临床应用的主要障碍。移植细胞活力极低（~1-32%）是由以下原因造成的。注射过程中的机械力会损伤细胞膜，并且缺乏支持细胞原位存活的三维基质，每 20 岁以上的美国人中就有一个发生外周动脉疾病 (PAD)。 50. 目前的治疗方案往往无效，通常会导致严重肢体缺血的病例放大。一种有前途的疗法是将患者来源的人脂肪源性基质细胞（hASC）移植到缺血部位，通过分泌原细胞促进再灌注。 -血管生成旁分泌因子。在这里，我们提出了一种物理交联的双网络水凝胶，其硬度范围为 10 至 3000 Pa，以解决血管生成旁分泌因子的问题。移植后细胞死亡，调节细胞功能以优化 hASC 的血管生成潜力，并改善 PAD 小鼠模型的血液再灌注。在具体目标 1 中，我将合成并表征一种可注射水凝胶，该水凝胶经历两种不同的物理交联机制。第一个交联步骤通过基于肽的分子识别在体外发生，将细胞封装在物理水凝胶内，而第二个交联步骤通过热相变在原位发生以形成增强。通过改变增强网络密度，该水凝胶将配制为具有 10 至 3000 Pa 的各种刚度，以在注射器注射期间提供 hASC 保护并提高移植后的细胞活力，我将在具体目标 2 中探讨其效果。体外研究了 hASC 分泌促血管生成生长因子的硬度，并最大限度地提高了封装在该系列新型水凝胶中的 hASC 的血管生成潜力。 3、我将选择性能最佳的水凝胶配方，并将 hASC 递送至 PAD 小鼠后肢缺血模型，将评估血液灌注和新生血管的恢复，以验证使用我们的新型水凝胶的基于 hASC 的治疗，寻求加速临床。实现 PAD 细胞治疗。