Bioengineered corneal endothelial graft using photodegradable device to induce graft-host integration

使用光降解装置诱导移植物-宿主整合的生物工程角膜内皮移植物

• 批准号: 10719330
• 负责人: Muhammad Rizwan
• 金额: $ 38.64万
• 依托单位: MICHIGAN TECHNOLOGICAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10719330
• 关键词: Adhesions; Animal Model; Anterior; Basement membrane; Biocompatible Materials; Biological Assay; Biomedical Engineering; Biomimetics; Cell Maturation; Cell Transplantation; Cell physiology; Cell-Cell Adhesion; Cells; Cornea; Corneal Diseases; Corneal Endothelium; Cues; Deposition; Development; Devices; Disease model; Donor person; Endothelial Cells; Engineering; Engraftment; Evaluation; Exposure to; Extracellular Matrix; Eye; Failure; Film; Gelatin; Growth; Histologic; Hour; Human; Hydrogels; Image; Image Analysis; In Vitro; K ATPase; Keratoplasty; Kinetics; Label; Light; Measurement; Mechanics; Metabolic; Methacrylates; Mission; Modeling; Modulus; Morphology; Nanotopography; Organ Culture Techniques; Oryctolagus cuniculus; Patients; Persons; Physiologic Intraocular Pressure; Pilot Projects; Procedures; Proliferating; Public Health; Quality of life; Recovery; Research; Surgeon; Technology; Tissue Engineering; Tissues; Transplantation; United States National Institutes of Health; Vision; Waiting Lists; Work; Yam - dietary; anterior chamber; biomaterial compatibility; cyanine dye; design; extracellular; graft function; high throughput analysis; improved; in vivo; innovation; lithography; monolayer; nanopattern; progenitor; reconstruction; response; technology development; tool; two-photon

PROJECT SUMMARY Although the bioengineered human corneal endothelial cell (hCEC) monolayer graft have shown vision recovery in animal models, the hCEC monolayer do not integrate with the host cornea due to suboptimal and non-tunable degradation of the hCEC-carrier biomaterial in the anterior chamber, which provide mechanical support to the monolayer. Thus, whether the transplanted hCEC monolayer will integrate with the host cornea following the complete degradation of the hCEC-carrier biomaterial and remain functional thereafter is unknown. To validate the bioengineered hCEC monolayers, there is a clear need to develop a biomaterial that has tunable-degradation rate in-vivo to evaluate the engraftment of the hCECs, and is mechanically strong so that the biomaterial-film/hCEC-monolayer construct does not break during the transplantation. The previous work of the team has established that the extracellular-topography cues can significantly modulate hCEC responses. The preliminary work of the team has developed photodegradable hydrogel (pdGel), which can be degraded in a tunable manner after transplantation, within hours to weeks, using tissue-penetrative light. Accordingly, the objective of this proposal is to develop a nano-topography pdGel-hCEC monolayer graft, evaluate monolayer integration with host cornea by tuning the in-vivo degradation rate of the pdGel, and validate the hCEC-monolayer function in-vivo. It is hypothesized that the nano-patterned pdGel will enable the growth of hCECs as a confluent monolayer, improve the hCEC monolayer function and stability by inducing the deposition of native-like extracellular matrix (ECM), and the tunable photodegradation of the carrier will improve the engraftment of the hCEC monolayer. The rationale for this project is the evaluation of the engraftment of hCEC monolayer with the cornea and the function thereafter will validate the use of bioengineered hCEC grafts for potential treatment of multiple corneal patients with one donor. Towards the overall objective, in the first aim, the hCEC monolayer growth on the pdGel, photodegradation kinetics, the biocompatibility of the degradation products, and the engraftment of the monolayer will be evaluated in-vitro and ex–vivo. In the second aim, using a high-throughput topography platform, the effect of 253 unique pdGel topographies will be evaluated on the hCEC monolayer functions to identify the optimum graft design. In the third aim, the photodegradation rate will be tuned in-vivo using light exposure to evaluate its effect on the hCEC engraftment. The proposed research is expected to be significant because it will validate the bioengineered hCEC-monolayer graft technology using a new photodegradable biomaterial, and it will develop new biomaterial and nano-topography platform that will have significant applications beyond ocular tissue engineering. The proposed research is innovative because it, (1) uses two-photon lithography approach to develop an innovative, high throughput nano-topography platform, and (2) leverages the photo-decomposition liability of the cyanine dye to develop an innovative photodegradable hydrogel for hCEC engraftment.

项目摘要 尽管生物工程的人角膜内皮细胞（HCEC）单层移植物显示了视力 在动物模型中恢复，HCEC单层由于次优和 HCEC-CARRIER生物材料在前室中的非降低降解，该室提供机械 支持单层。那是，移植的HCEC单层是否将与宿主角膜集成 按照HCEC-Carrier生物材料的完整定义，此后保持功能是 未知。为了验证生物工程的HCEC单层，显然需要开发生物材料 具有可调的降解速率，用于评估HCEC的植入，并且在机械上很强，因此 在移植过程中，生物材料 - 胶片/HCEC单层构建体不会破裂。上一个 团队的工作已经确定，细胞外部图提示可以显着调节HCEC 回答。团队的初步工作已经开发了可降解水凝胶（PDGEL），这可以是 移植后，在数小时至几周内，使用组织渗透光在数小时内以可调节的方式降解。 彼此之间，该提案的目的是开发纳米谱pdgel-hcec单层移植物， 通过调整PDGEL的体内降解率，评估单层与宿主角膜的整合，并 在体内验证HCEC-Monolayer函数。假设纳米图案的PDGEL将使 HCEC作为汇合单层的生长，通过诱导的HCEC单层函数和稳定性提高 本地样细胞外基质（ECM）的沉积和载体的可调光降解 改善HCEC单层的植入。该项目的理由是评估 HCEC单层与角膜及其功能的植入将验证使用 生物工程的HCEC移植物可潜在用一个供体的多个角膜患者治疗。走向 总体目标，在第一个目标中，PDGEL上的HCEC单层生长，光降解动力学， 降解产物的生物相容性以及单层的植入将进行评估 和前–Vivo。在第二个目标中，使用高通量地形平台，253个独特的PDGEL的效果 地形将在HCEC单层功能上进行评估，以识别最佳的移植设计。在 第三目的是，将使用光照曝光来调整光降解率，以评估其对 HCEC植入。拟议的研究预计将是重要的，因为它将验证 生物工程的HCEC-MONOLAYER移植技术使用新的光降级生物材料，它将发展 新的生物材料和纳米读型平台将在眼组织以外的重要应用 工程。拟议的研究具有创新性，因为（1）使用两光岩岩性方法来 开发一个创新的高吞吐量纳米图平台，（2）利用照片分解 氰染料的责任，开发用于HCEC植入的创新光降解水凝胶。