ERI: Biological Effects of Low-Frequency, Low-Intensity Ultrasound on Endothelial Cell and Macrophage Co-Culture

该工程研究启动（ERI）奖将支持研究超声对内皮细胞（血管细胞）和巨噬细胞（免疫细胞）的影响。内皮细胞和巨噬细胞在许多生物过程中共同作用，其中之一是血管生成，即血管的产生。血管生成对于受损组织的愈合至关重要，如在慢性伤口的情况下。慢性伤口平均持续12个月，可能导致下肢截肢甚至死亡。人体临床试验表明，低频、低强度的超声波可以促进慢性伤口愈合。然而，生物学机制仍然不清楚。从了解超声如何单独和共同影响内皮细胞和巨噬细胞中获得的知识可以增强超声治疗的高级愈合，并最终提高患者的生活质量。该奖项还将促进本科生和高中生在组织工程方面的接触和培训，特别是通过动手实验室经验为代表性不足的学生和妇女提供工程服务。还将为工程和非工程专业开设组织工程方面的正式课程，重点是少数族裔学生和工程领域的妇女。本研究的目的是评估促进血管生成作为治疗性超声的一种可能的生物学机制。这将通过两个目标来实现：（1）通过巨噬细胞和内皮直接共培养、间接共培养（transwell）和单独（单独内皮细胞和单独巨噬细胞）表征低频、低强度超声对巨噬细胞和内皮细胞促血管生成因子的单向和双向作用;（2）使用批量RNAseq和全局生物信息方法分析基因表达变化。这些目标将在体外使用3D胶原支架进行，以更接近地模拟生理条件。了解治疗性超声对血管生成的影响将为超声可能的生物学机制提供见解。初步数据和已发表的文献强烈表明，刺激血管生成可能是超声波介导愈合的机制。如果收集的数据证实了这一点，这可能表明超声可能是恢复功能性内皮行为的有效疗法，其中病理性血管生成是包括动脉粥样硬化、类风湿性关节炎和自身免疫性疾病在内的疾病的标志。这些数据将为超声的新型治疗应用提供见解。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的知识价值和更广泛的影响审查标准进行评估来支持。

This Engineering Research Initiation (ERI) award will support research to investigate the effects of ultrasound on endothelial cells (blood vessel cells) and macrophages (immune cells). Endothelial cells and macrophages work together in many biological processes, one of which is angiogenesis, the creation of blood vessels. Angiogenesis is critical for the healing of damaged tissues as in the case of chronic wounds. Chronic wounds last on average twelve months and may lead to amputation of lower limbs or possibly even death. Human clinical trials have shown that low-frequency, low-intensity ultrasound advances chronic wound healing. However, the biological mechanisms are still not understood. The knowledge gained from understanding how ultrasound affects endothelial cells and macrophages individually and together may enhance ultrasound therapies for advanced healing and ultimately increase the quality of life for patients. This award will also promote undergraduate and high school student exposure and training in tissue engineering, particularly for underrepresented students and women in engineering through hands-on laboratory experiences. Formal course offerings in tissue engineering will also be developed for both engineering and non-engineering majors with a focus on minoritized students and women in engineering. The research goal is to evaluate the promotion of angiogenesis as a possible biological mechanism of therapeutic ultrasound. This will be achieved through two objectives: (1) characterize uni- and bi-directional effects of low-frequency, low-intensity ultrasound on macrophage and endothelial cell pro-angiogenic factors via macrophage and endothelial direct co-culture, indirect co-culture (transwell), and individually (endothelial alone and macrophage alone) and (2) analyze gene expression changes using bulk RNAseq and global bioinformation methods. These objectives will be conducted in vitro using 3D collagen scaffolds to more closely mimic physiological conditions. Understanding the effects of therapeutic ultrasound on angiogenesis will provide insight to the possible biological mechanisms of ultrasound. Preliminary data and published literature strongly suggest that stimulation of angiogenesis may be the mechanism through which ultrasound mediates healing. If this is confirmed by the data collected, this may indicate that ultrasound could be an effective therapy to recover functional endothelial behavior where pathological angiogenesis is a hallmark of diseases including atherosclerosis, rheumatoid arthritis, and autoimmune diseases. These data would provide insights into novel therapeutic applications of ultrasound.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.