Biotextiles for ligament repair using biofunctionalisation to aid regeneration

该项目将被整合到UKRMP治疗输送中心的无细胞方法中，该中心是曼彻斯特、诺丁汉、帝国、南安普顿、基尔和others.http://www.ukrmp.org.uk/hubs/Anterior之间的合作。交叉韧带（ACL）断裂是一个常见问题，主要是由于运动损伤而发生的。在美国，每年发生大约200，000例ACL断裂，据估计，每年进行超过100，000例ACL修复。此类断裂随后可能导致其他严重的退行性关节疾病，因此，有效治疗和处理ACL损伤非常重要。目前ACL修复的金标准涉及使用髌腱自体移植物;这种治疗有许多局限性，包括：供体部位发病率，更重要的是移植物无法与骨隧道部位（通常称为附着点的区域）完全整合。高失败率经常发生，特别是在该附着点部位，随后导致患者再次就诊以进一步改进手术。因此，需要一种干预措施，它可以与目前的金标准方法相结合，作为一种双重技术，这可以提供更强大的移植整合。一些研究已经在开发替代移植物制造的合成FDA批准的材料，包括聚己内酯（PCL）和PET，希望制造组织工程移植物，将提供所需的机械性能。然而，与这种构建体相关的限制包括它们缺乏生物相互作用，缺乏功能性新基质沉积和重塑，导致在附着点处失败。因此，在附着点处的移植物失效（自体移植物或合成物）的问题尚未完全解决。因此，本研究旨在通过提高愈合率来改善附着点处的移植物整合。在这种情况下，我们的目标是开发一种合成移植物，该移植物能够精确模拟天然组织ACL细胞外基质的纳米和微观结构沿着具有适当的机械性能和降解速率。与Eithne Comerford教授（利物浦大学）合作，我们的目标是确定ACL不同区域（特别是附着点）内的关键细胞外基质（ECM）蛋白片段，并将其功能化到合成移植物上。我们已经确定了作为起始点的Escherin-1，并且初步结果已经显示，在Escherin存在的情况下，沿着电纺纤维的沿着和对齐显著更快。进一步的分子已被确定，并将探讨沿着与信号转导途径。细胞行为将评估方面的细胞活力，形态，增殖，分化，内和细胞外信号转导途径和新基质分泌。这些将通过免疫细胞化学，ELISA，RT-PCR和组织学进行验证和鉴定。此外，静电纺丝系统可用作其他韧带和肌腱修复的平台技术。

The project will be integrated into the UKRMP Acellular approaches for therapeutic delivery Hub, a collaboration between Manchester, Nottingham, Imperial, Southampton, Keele and others.http://www.ukrmp.org.uk/hubs/Anterior cruciate ligament (ACL) ruptures are a common problem and occur mostly as a result of sports injuries. Around 200,000 ACL ruptures per year occur in the United States and it is estimated that more than 100,000 ACL repairs are performed annually in the United States.Such ruptures can subsequently lead to other, serious degenerative joint diseases and for this reason it is important to treat and tackle ACL injuries efficiently and effectively. The current gold standard for ACL repair involves the use of patellar tendon autografts; this treatment has many limitations including: donor site morbidity and more importantly the inability of the graft to fully integrate with the bone tunnel site (area often referred to as the enthesis). High failure rates often occur particularly at this enthesis site subsequently leading to patient revisits for further improved surgeries. Hence, an intervention is required which could be coupled with the current gold standard method to act as a dual technology, which could provide more robust graft integration.Some research has been performed in developing alternative grafts fabricated from synthetic FDA approved materials including Poly-e-Caprolactone (PCL) and PET with the hope to manufacture tissue-engineered grafts that would provide the required mechanical properties. However, limitations associated with such constructs include their lack of bio-interaction, lack of functional neo-matrix lay down and remodeling, leading to failure at the enthesis site. Therefore, the problem of graft failure (autograft or synthetic) at the enthesis site is yet to be fully resolved. For this reason this study aims to improve graft integration at the enthesis site through improved healing rate.In this case, we aim to develop a synthetic graft, which is able to precisely mimic the nano-and micro- architecture of native tissue ACL extracellular matrix along with the appropriate mechanical properties and degradation rate. Working in collaboration with Professor Eithne Comerford (University of Liverpool), we aim to identify key extracellular matrix (ECM) protein fragments within the different regions of the ACL (particularly the enthesis) and functionalise these onto the synthetic graft. We have identified fibrillin-1 as a starting point and preliminary results have shown significantly more rapid attachment and alignment along the electrospun fibres with fibrillin present. Further molecules have been identified and will be explored along with signalling pathways.Cell behavior will be assessed in terms of cell viability, morphology, proliferation, differentiation, intra- and extracellular signalling pathways and neo-matrix secretion. These will be validated and identified using via immunocytochemistry, ELISA, RT-PCR and histology.Additionally the electrospun system could be used as a platform technology for other ligaments and also tendon repair.