Regulating the Quality and Potency of Stem Cells with Biophysical Cues from Dynamic Nanofibrous Hydrogels for Therapeutic Purposes

利用动态纳米纤维水凝胶的生物物理线索调节干细胞的质量和效力用于治疗目的

• 批准号: 10724060
• 负责人: David Castilla-Casadiego
• 金额: $ 12.48万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10724060
• 关键词: 3-Dimensional; Address; Adhesives; Adipocytes; Affect; Allogenic; Behavior; Biocompatible Materials; Biophysics; Bioreactors; Cell Count; Cell Culture System; Cell Proliferation; Cell Survival; Cell physiology; Cell surface; Cells; Characteristics; Chemicals; Chondroblast; Clinical effectiveness; Coculture Techniques; Collagen; Collagen Type I; Cues; Diameter; Disease; Electrospinning; Encapsulated; Engineering; Ensure; Extracellular Matrix; Fiber; Future; Goals; Human; Human body; Hyaluronic Acid; Hydrazones; Hydrogels; In Vitro; Individual; Integrin Binding; Length; Link; Lymphocyte Suppression; Measures; Mechanics; Memory; Mentors; Mesenchymal Stem Cells; Methods; Molecular; Morphology; Nanotopography; Osteoblasts; Phase; Phenotype; Play; Production; Proliferating; Property; Proteins; Relaxation; Reporting; Reproducibility; Research; Research Proposals; Role; Rosales; Secretory Cell; Signal Pathway; Source; Stress; Structure; Supervision; Surface; System; Testing; Therapeutic; Tissues; Transcriptional Activation; angiogenesis; biophysical properties; crosslink; cytokine; density; design; immunoregulation; improved; in vivo; insight; manufacture; manufacturing scale-up; mechanical properties; mechanical signal; nanofiber; patient population; scaffold; sensor; stem cell expansion; stem cells; three-dimensional modeling; transcription factor; viscoelasticity; 3-Dimensional; Address; Adhesives; Adipocytes; Affect; Allogenic; Behavior; Biocompatible Materials; Biophysics; Bioreactors; Cell Count; Cell Culture System; Cell Proliferation; Cell Survival; Cell physiology; Cell surface; Cells; Characteristics; Chemicals; Chondroblast; Clinical effectiveness; Coculture Techniques; Collagen; Collagen Type I; Cues; Diameter; Disease; Electrospinning; Encapsulated; Engineering; Ensure; Extracellular Matrix; Fiber; Future; Goals; Human; Human body; Hyaluronic Acid; Hydrazones; Hydrogels; In Vitro; Individual; Integrin Binding; Length; Link; Lymphocyte Suppression; Measures; Mechanics; Memory; Mentors; Mesenchymal Stem Cells; Methods; Molecular; Morphology; Nanotopography; Osteoblasts; Phase; Phenotype; Play; Production; Proliferating; Property; Proteins; Relaxation; Reporting; Reproducibility; Research; Research Proposals; Role; Rosales; Secretory Cell; Signal Pathway; Source; Stress; Structure; Supervision; Surface; System; Testing; Therapeutic; Tissues; Transcriptional Activation; angiogenesis; biophysical properties; crosslink; cytokine; density; design; immunoregulation; improved; in vivo; insight; manufacture; manufacturing scale-up; mechanical properties; mechanical signal; nanofiber; patient population; scaffold; sensor; stem cell expansion; stem cells; three-dimensional modeling; transcription factor; viscoelasticity

Project Summary/Abstract Human mesenchymal stem cells (hMSCs) are considered a source for allogeneic therapies to treat diverse diseases. Due to the exponential increase in demand, there is a need for new strategies to produce potent hMSCs to serve diverse patient populations. Currently, conventional planar culture and bioreactors are used as scale-up manufacturing methods. However, these are not specifically tailored for hMSCs expansion. They may alter the cell phenotype and secretome, affecting clinical effectiveness. Further studies to understand the role of substrate mechanics on hMSC expansion are required to achieve reproducible production. Numerous scaffolding alternatives replicate several characteristics of the native extracellular matrix (ECM). However, its dynamic mechanics, which plays a fundamental role in regulating crucial cellular processes, has not been amply studied yet. Furthermore, most in-vitro substrates are static and supraphysiologically stiff. Static substrates have offered a substantial benefit for generating high cell numbers; however, hMSCs have been shown to retain mechanical information, limiting therapeutic capabilities. To address this problem, this proposed research seeks to investigate the role of dynamic cell-matrix interactions and nano-topographical cues on the immunomodulatory potential of hMSCs using a composite of electrospun-fibers encapsulated in a dynamic hydrogel, with the hypothesis that this composite biomaterial will promote high hMSCs production with relevant therapeutic value, while eliminating the limitations reported for the conventional cell culture systems. The K99 period will focus on engineering and characterizing the dynamic nanofibrous hydrogel composites to propel me toward establishing the mechanisms by which they modulate cell quality and potency attributes with relevant therapeutic value (during the R00 phase). In Aim 1, we will develop the dynamic nanofibrous system using a hyaluronic acid hydrogel network crosslinked via dynamic covalent hydrazone bonds that capture the viscoelasticity of ECM in tissues. Four variables, including the encapsulation of the electrospun collagen nanofibers at various densities, fiber diameter, fiber length, and the stress relaxation timescale of the hydrogel will be characterized in this aim to promote hMSC viability and proliferation. In Aim 2, hMSCs cell quality and potency will be assessed by measuring the effect of hydrogel parameters on cellular secretory activity. Immunomodulatory properties will be evaluated by quantifying lymphocyte suppression in co-culture, as well as expression of hMSC surface markers. The capacity of the hMSCs to differentiate will also be assessed. In aim 3, the mechanism linking the biophysical parameters of the nanofibrous hydrogel to hMSC secretory activity will be probed by examining cell adhesive proteins and the activation of transcription factors or sensors of mechanical cues. In sum, the proposed research will lead to new insights to produce hMSCs with high therapeutic value, which will enable new culture substrates that achieve control in reproducibility and cell quality to serve diverse patient populations.

项目摘要/摘要 人间充质干细胞（HMSC）被认为是同种异体治疗的来源 多样化的疾病。由于需求的指数增加，需要新的策略生产 有效的HMSC为多样化的患者人群提供服务。目前，传统的平面文化和生物反应器是 用作扩大制造方法。但是，这些不是专门针对HMSC扩展而定制的。 它们可能会改变细胞表型和分泌组，从而影响临床有效性。进一步了解 需要底物力学在HMSC扩展中的作用才能实现可再现的生产。很多的 脚手架替代品复制天然细胞外基质（ECM）的几个特征。但是，它 动态力学在调节关键细胞过程中起着基本作用，但尚未充分 研究了。此外，大多数体外底物都是静态的，并且在生理上僵硬。静态基材具有 为产生高细胞数量提供了可观的好处；但是，HMSC已显示保留 机械信息，限制治疗能力。为了解决这个问题，这项拟议的研究寻求 研究动态细胞矩阵相互作用和纳米型线索在免疫调节中的作用 使用封装在动态水凝胶中的电纺纤维复合物的HMSC的潜力 假设这种复合生物材料将促进具有相关治疗价值的高HMSC生产， 同时消除了传统细胞培养系统报告的局限性。 K99时期将重点放在 工程和表征动态纳米纤维水凝胶复合材料，以推动我建立 它们以相关的治疗值调节细胞质量和效力属性的机制 （在R00阶段）。在AIM 1中，我们将使用透明质酸开发动态纳米纤维系统 水凝胶网络通过动态共价桥接键进行交联，该桥梁捕获ECM在中的粘弹性 组织。四个变量，包括在各种密度下封装电纺胶原蛋白纳米纤维， 纤维直径，纤维长度和水凝胶的应力松弛时间尺度将在此目标中进行表征 促进HMSC的生存能力和增殖。在AIM 2中，HMSCS细胞质量和效力将由 测量水凝胶参数对细胞分泌活性的影响。免疫调节特性将是 通过量化共培养中的淋巴细胞抑制以及HMSC表面标记的表达来评估。 也将评估HMSC分化的能力。在AIM 3中，连接生物物理的机制 将通过检查细胞粘合剂来探测纳米纤维水凝胶与HMSC分泌活性的参数 蛋白质和机械提示的转录因子或传感器的激活。总而言之 将导致新见解，以产生具有高治疗价值的HMSC，这将使新的文化底物 这可以控制可重现性和细胞质量，以服务于多样化的患者人群。