Nanovibrational control of chondrogenic differentiation

学生奖学金战略优先领域：医疗保健技术关键词：骨关节炎（OA），组织工程（TE）纳米振动，软骨形成，PEG水凝胶软骨是一种无血管和神经组织，因此愈合不良，无人看管的软骨损伤导致骨关节炎（OA），一种退行性关节疾病。目前，世界卫生组织（WHO）已将OA列为第二大致残原因和增长速度最快的主要健康疾病。因此，组织工程（TE）是一种新兴的软骨再生技术。在这个新项目中，我们将使用一种新型的生物反应器，它可以向MSC提供微小的纳米级（30 nm，1000 Hz）振动，以使它们扩增并分化为产生正确透明软骨表型的软骨细胞。细胞的这种纳米振动控制的结果是形成关节的光滑的关节软骨并防止纤维软骨形成。将进行干涉测量和加速度测量，以表征迫使细胞在生物反应器中分化的纳米机械线索。在这些步骤之后，我们将通过使用超分辨率共聚焦显微镜、qPCR、蛋白质分析、RNA测序和代谢组学来表征和鉴定细胞对纳米振动的反应表型。然后我们将转移到纳米振动软骨细胞到我们的新型水凝胶家族的封装。这些合成的、生物工程化的粘弹性水凝胶属于基于聚乙二醇（PEG）的水凝胶家族，我们将寻求操纵生物物理性质以匹配天然软骨的刚度。这些水凝胶将由互穿聚合物网络（IPNS）制成，所述互穿聚合物网络包含弹性第一网络和提供粘性组分的高分子量聚合物的第二网络。第二个网络将呈现作为钙粘蛋白模拟物的肽基序（如HA）;对软骨形成很重要。将进行流变学测量以确定材料的弹性（G '）和损耗（G“）剪切模量。该项目旨在建立一个3D输送平台（定制水凝胶），以支持纳米振动软骨细胞培养，并防止纤维软骨形成，就像当前载体（如藻酸盐凝胶或胶原蛋白膜）的情况一样。在这里，我们提出了一个高度跨学科的项目，承诺制造一个新的技术系统，专注于生物工程细胞治疗。这些生物材料很有前途，因为它们帮助我们探索替代动物实验的新方法，最重要的是，创造一种可交付的可注射系统，可以调整以最大限度地支持软骨形成并保持有需要的人的流动性。

Studentship strategic priority area: Healthcare TechnologiesKeywords: Osteoarthritis (OA), Tissue Engineering (TE) Nanovibration, Chondrogenesis, PEG-hydrogelsCartilage is an avascular and aneural tissue and so suffers poor healing and unattended cartilage damage leads to osteoarthritis (OA), a degenerative joint disease. At present, the World Health Organization (WHO) has ranked OA as the second greatest cause of disability and the primary health disease with the fastest rate of growth. As a result, tissue engineering (TE) is an emerging technique for cartilage regeneration. In this new project, we will use a novel bioreactor that delivers tiny, nanoscale (30 nm, 1000 Hz) vibrations to MSCs to expand and differentiate them into chondrocytes that produce the correct hyaline cartilage phenotype. This nanovibrational control of cells has as a result to form the smooth, articulating cartilage of the joint and prevents fibrocartilage formation. Interferometry and accelerometry measurements will be done to characterize the nano-mechanical cues that force the cells into differentiation in the bioreactor. Following these steps, we are going to characterize and identify the cells' phenotype in response to nanovibrations by using super-resolution confocal microscopy, qPCR, western analysis, RNA seq, and metabolomics. Then we are going to move to the encapsulation of the nanovibrated-chondrocytes into our novel hydrogel family. These synthetic, bioengineered, viscoelastic hydrogels belong to a family of polyethylene glycol (PEG)-based hydrogels where we will seek to manipulate the biophysical properties to match the stiffness of native cartilage. These hydrogels will be made of interpenetrated polymer networks (IPNS) that contain an elastic first network and the second network of a high molecular weight polymer that provides the viscous component. The second network will present peptide motifs (such as HAVDI) that are cadherin mimics; important for cartilage formation. Rheological measurements will be carried out to determine the elastic (G') and loss (G'') shear modulus of the material. The project aims to build a 3D delivery platform (tailored hydrogels) to support the nanovibrated-chondrocytes culture and prevent fibrocartilage formation as is the case with current carriers such as alginate gels or collagen membranes. Here we are presenting a highly interdisciplinary project that promises to manufacture a new technology system that focuses on bioengineered cell therapy. These biomaterials are promising as they are helping us to explore new methods to replace animal experimentation, and most importantly, create a deliverable injectable system that can be tuned to maximally support chondrogenesis and maintain mobility for people in need.