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.

项目概要 尽管生物工程人角膜内皮细胞（hCEC）单层移植物已显示出视力 在动物模型恢复中，hCEC 单层由于次佳和不与宿主角膜整合。 前房中 hCEC 载体生物材料的不可调节降解，提供机械性能 因此，移植的 hCEC 单层是否会与宿主角膜融合。 hCEC 载体生物材料完全降解并保持功能后 为了验证生物工程 hCEC 单层，显然需要开发一种生物材料。 具有可调节的体内降解率来评估 hCEC 的植入，并且机械强度高，因此 生物材料膜/hCEC单层结构在移植过程中不会破裂。 该团队的工作已经确定细胞外拓扑线索可以显着调节 hCEC 该团队的前期工作开发了可光降解水凝胶（pdGel），它可以 移植后，使用组织穿透光，在数小时至数周内以可调节的方式降解。 因此，本提案的目标是开发纳米形貌 pdGel-hCEC 单层移植物， 通过调节 pdGel 的体内降解率来评估单层与宿主角膜的整合，以及 体内验证 hCEC 单层功能 人们再次认识到纳米图案的 pdGel 将能够实现 hCEC 作为汇合单层生长，通过诱导 hCEC 单层功能和稳定性 类似天然细胞外基质（ECM）的沉积，以及载体的可调光降解作用将 改善 hCEC 单层的植入 该项目的基本原理是评估 hCEC 单层植入角膜及其后的功能将验证 生物工程 hCEC 移植物可用于用一名捐赠者治疗多个角膜患者。 总体目标，第一个目标是 pdGel 上的 hCEC 单层生长、光降解动力学、 降解产物的生物相容性和单层的植入将在体外进行评估 在第二个目标中，使用高通量形貌平台，研究 253 独特的 pdGel 的效果。 将根据 hCEC 单层功能评估拓扑结构，以确定最佳移植物设计。 第三个目标，将使用光照射在体内调整光降解率，以评估其对 hCEC 植入预计将具有重要意义，因为它将验证 使用新型光降解生物材料的生物工程hCEC单层移植技术，将开发 新的生物材料和纳米形貌平台将在眼组织之外具有重要的应用 所提出的研究具有创新性，因为（1）使用双光子光刻方法 开发创新的高通量纳米形貌平台，并且（2）利用光分解 花青染料有责任开发一种用于 hCEC 移植的创新光降解水凝胶。