Bioprosthetic cornea: using polymeric templates for directed stem cell growth

生物假体角膜：使用聚合物模板进行干细胞定向生长

• 批准号: BB/I008187/2
• 负责人: Che Connon
• 金额: $ 10.83万
• 依托单位: Newcastle University
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2014
• 资助国家: 英国
• 起止时间: 2014 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FI008187%2F2
• 关键词: Bioprosthetic; cornea; using; polymeric; templates

The cornea is our window to the world, once compromised by wounding, disease or age, a loss of vision results. By improving our understanding of corneal structure and providing new methods of corneal transplantation the sight of many more patients can be restored. Currently, corneal transplantation requires a continuous supply of healthy donor corneas. However worldwide demand has grown and taken together with an aging population and the rapid rise in laser eye surgery (which can negatively affect the donor tissue suitability for transplantation) the search for an effective engineered replacement is essential if current levels of corneal transplantation are to be maintained. This investigation stems from our previous work in understanding the molecular structure underpinning corneal transparency, the development of novel corneal biomaterials and the limitations of the current corneal stem cell transplantation techniques, specifically the materials used to grow and convey the stem cells to the patient. Previously, we have quantified the nanostructure of the cornea and related this structure to the preservation of corneal transparency; furthermore we have applied these measurements to the design of new corneal biomaterials capable of supporting corneal epithelial stem cell differentiation and growth. Therefore, we will draw on our knowledge of corneal structure, corneal stem cell isolation and cultivation and novel biomaterials to engineer a tissue suitable for corneal transplantation. To do this we will develop a template made from tractable regularly spaced aligned polymers (reflecting the natural state of corneal nanostructure). The template will contain protein fragments recognisable by the human corneal stem cells enabling them to attach in a highly ordered and controlled manner. Once attached the cells will be chemically induced to differentiate and produce collagen fibres. The alignment of these fibres will be guided by the cells orientation. The template will then slowly lose its integrity and detach from the cells by way of enzymes released by the corneal cells, thus releasing the aligned collagen as a tissue engineered collagen mat. These mats will then be stacked and compressed to produce a robust biomaterial made solely from human proteins (mostly collagen), the polymer template having been removed during processing. The biomaterials mechanical strength and ability to support corneal epithelial growth upon its surface will then be tested. We have already shown that compressed mats of rat tail collagen are both mechanically robust as well as excellent substrates for corneal epithelial cell growth. This work represents a significant step forward in the development of biomaterials. Instead of designing and using bio-compatible polymers to represent tissue for transplantation our approach is to use the polymers merely as a template allowing the cells to produce the actual biomaterial. Furthermore, since the template is easily discarded the cell based biomaterial represents the ultimate in biocompatibility as it is comprised of human proteins possibly even derived from cells taken from the patient's own body. The beneficiaries of this work would be those working in the fields of polymer chemistry as the development of aligned polymers containing protein fragments that are both recognised and degraded by cells is not trivial. Scientists in the blossoming field of biomaterials would be given a new direction in the development of truly bio-compatible materials (i.e. ones derived from stem cells). Tissue engineers would be given a new tool with which they could create similar stem cell based materials for bone, skin etc. repair and finally ocular regenerative medicine would benefit from the development of a replacement human donor corneal tissue.

角膜是我们通向世界的窗口，一旦因受伤、疾病或年龄而受损，就会导致视力丧失。通过提高我们对角膜结构的了解并提供新的角膜移植方法，可以恢复更多患者的视力。目前，角膜移植需要持续供应健康的供体角膜。然而，随着人口老龄化和激光眼科手术的迅速增加（这可能会对供体组织移植的适宜性产生负面影响），全球需求不断增长，如果要维持目前的角膜移植水平，寻找有效的工程替代品至关重要。维持。这项研究源于我们之前对支持角膜透明度的分子结构、新型角膜生物材料的开发以及当前角膜干细胞移植技术的局限性的工作，特别是用于生长和将干细胞输送给患者的材料。此前，我们已经量化了角膜的纳米结构，并将这种结构与角膜透明度的保持联系起来；此外，我们还将这些测量结果应用于能够支持角膜上皮干细胞分化和生长的新型角膜生物材料的设计。因此，我们将利用我们在角膜结构、角膜干细胞分离和培养以及新型生物材料方面的知识来设计适合角膜移植的组织。为此，我们将开发一种由易处理的规则间隔排列的聚合物制成的模板（反映角膜纳米结构的自然状态）。该模板将包含人类角膜干细胞可识别的蛋白质片段，使它们能够以高度有序和受控的方式附着。一旦附着，细胞将被化学诱导分化并产生胶原纤维。这些纤维的排列将由细胞方向引导。然后，模板将慢慢失去其完整性，并通过角膜细胞释放的酶与细胞分离，从而释放对齐的胶原蛋白作为组织工程胶原蛋白垫。然后，这些垫子将被堆叠和压缩，以生产出仅由人类蛋白质（主要是胶原蛋白）制成的坚固的生物材料，聚合物模板在加工过程中已被去除。然后将测试生物材料的机械强度和支持其表面上角膜上皮生长的能力。我们已经证明，鼠尾胶原蛋白压缩垫不仅机械坚固，而且是角膜上皮细胞生长的优良基质。这项工作代表了生物材料开发的重要一步。我们的方法不是设计和使用生物相容性聚合物来代表移植组织，而是仅使用聚合物作为模板，允许细胞产生实际的生物材料。此外，由于模板很容易被丢弃，因此基于细胞的生物材料代表了最终的生物相容性，因为它由人类蛋白质组成，甚至可能源自取自患者自己身体的细胞。这项工作的受益者将是那些在聚合物化学领域工作的人，因为开发含有可被细胞识别和降解的蛋白质片段的排列聚合物并非易事。蓬勃发展的生物材料领域的科学家将在开发真正的生物相容性材料（即源自干细胞的材料）方面获得新的方向。组织工程师将获得一种新工具，他们可以用它来创建类似的基于干细胞的材料，用于骨骼、皮肤等修复，最终眼再生医学将受益于替代人类供体角膜组织的开发。

项目成果

Self-assembly of a dual functional bioactive peptide amphiphile incorporating both matrix metalloprotease substrate and cell adhesion motifs.

• DOI: 10.1039/c5sm00459d
• 发表时间: 2015-04
• 期刊: Soft matter
• 影响因子: 3.4
• 作者: A. Dehsorkhi;R. Gouveia;Andrew M. Smith;I. Hamley;V. Castelletto;C. Connon;M. Reza;J. Ruokolainen

Hybrid membrane biomaterials from self-assembly in polysaccharide and peptide amphiphile mixtures: controllable structural and mechanical properties and antimicrobial activity

• DOI: 10.1039/c6ra27244d
• 发表时间: 2017-01-01
• 期刊: RSC ADVANCES
• 影响因子: 3.9
• 作者: Castelletto, V.;Kaur, A.;Ruokolainen, J.
• 通讯作者: Ruokolainen, J.

Application of retinoic acid improves form and function of tissue engineered corneal construct.

• DOI: 10.1080/15476278.2015.1093267
• 发表时间: 2015
• 期刊: Organogenesis
• 影响因子: 2.3
• 作者: Abidin FZ;Gouveia RM;Connon CJ
• 通讯作者: Connon CJ

Autogenous Biofabrication of Nativelike, Scaffold-Free Human Skin Equivalents Using a Smart, Enzyme-Degradable Tissue Templating Coating.

使用智能、酶可降解的组织模板涂层自体生物制造类似天然、无支架的人类皮肤等效物。

• DOI: 10.1021/acsabm.8b00685
• 发表时间: 2019
• 期刊: ACS applied bio materials
• 影响因子: 4.7
• 作者: Connon CJ

Tuning chelation by the surfactant-like peptide A6H using predetermined pH values.

• DOI: 10.1021/bm401640j
• 发表时间: 2014-02-10
• 期刊: BIOMACROMOLECULES
• 影响因子: 6.2
• 作者: Castelletto, V.;Hamley, I. W.;Segarra-Maset, M. D.;Berdugo Gumbau, C.;Miravet, J. F.;Escuder, B.;Seitsonen, J.;Ruokolainen, J.
• 通讯作者: Ruokolainen, J.