Native state imaging and characterisation of tissue derived extracellular matrix hydrogels for regenerative medicine applications

用于再生医学应用的组织来源的细胞外基质水凝胶的天然状态成像和表征

• 批准号: 2739790
• 金额: --
• 依托单位: University of Nottingham
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2739790
• 关键词: Native; state; imaging; characterisation; tissue

The extracellular matrix (ECM) in native mammalian tissues provides a structural and functional framework to support tissue resident cells and promote cellular function. Mammalian tissues can be decellularised, to remove cellular antigens, that would otherwise incite an adverse immune response. These ECM derived biomaterials can be ground into small particulates, solubilised and polymerised to form hydrogels which reflect the biochemical complexity and bioinductive properties of the native matrix. ECM hydrogels contain highly complex structures, including proteins, growth factors, cryptic peptides and vesicles, within a hydrated environment. ECM hydrogels have considerable utility as vehicles for cell delivery in regenerative medicine and as 3D substrates for in vitro cell culture, but the full potential has yet to be reached. To date, characterisation of ECM hydrogel structure has been limited to freeze-drying, removing water from the hydrogels, which is not representative of the native state. There is a need for new imaging modalities to inform both understanding of the ultrastructure of ECM hydrogels and interactions of cells seeded within these materials. An alternative solution to freeze-drying is the rapid freezing of samples using cryogenic liquids to lock in the water, thereby preserving the structure of the hydrogel and its contents. By embracing cryogenic techniques such as cryo-Scanning Electron Microscopy, samples can be maintained once correctly preserved in their near native state. By using a Focused Ion Beam (FIB), samples can be cross-sectioned; sequential slicing and imaging in three dimensions will lead to better understanding of hydrogel ultrastructure and the impact this has on interactions with cells seeded within the hydrogel.

天然哺乳动物组织中的细胞外基质（ECM）提供结构和功能框架以支持组织驻留细胞并促进细胞功能。哺乳动物组织可以去细胞化，以去除细胞抗原，否则会引起不利的免疫反应。这些ECM衍生的生物材料可以研磨成小颗粒，溶解和聚合以形成水凝胶，其反映了天然基质的生物化学复杂性和生物诱导性质。ECM水凝胶在水合环境中含有高度复杂的结构，包括蛋白质、生长因子、隐蔽肽和囊泡。ECM水凝胶作为再生医学中细胞递送的载体和体外细胞培养的3D基质具有相当大的实用性，但尚未达到全部潜力。迄今为止，ECM水凝胶结构的表征仅限于冷冻干燥，从水凝胶中除去水，这并不代表天然状态。需要新的成像方式来了解ECM水凝胶的超微结构和接种在这些材料内的细胞的相互作用。冷冻干燥的替代解决方案是使用低温液体快速冷冻样品以锁定水，从而保留水凝胶及其内容物的结构。通过采用低温技术，如低温扫描电子显微镜，样品一旦正确保存在其接近天然状态，就可以保持。通过使用聚焦离子束（FIB），可以对样品进行横切;连续切片和三维成像将有助于更好地了解水凝胶超微结构及其对水凝胶内接种细胞相互作用的影响。