Statistical optimization of self-assembled biosynthetic cornea implants

拟议研究的总体愿景是创建一个有效的，可扩展的，负担得起的 适合临床应用的仿生角膜替代物。角膜最外层的损伤 眼睛组织的损伤导致视力丧失，这深刻地影响生活质量。角膜移植 是治疗的黄金标准，但排斥反应进展缓慢，供体组织供应短缺， 少数失败导致毁灭性后果，这仍然是关键的挑战。人工 迄今为止，角膜替代物未能再现天然组织结构和功能。一个 模仿天然角膜的有效生物合成替代物将提供一种变革性的新方法， 角膜损伤患者的选择。我们先前的工作开辟了一条新的路线， 角膜我们以前开发了一种胶原玻璃化过程， 胶原蛋白凝胶的强度。为了实现全厚度植入物的必要透明度，我们需要 以进一步控制胶原间隔和层间正交性。我们概括了角膜的发育 和组织在体外使用合成的环状蛋白聚糖分子，环糊精（CD）。初步 CD和胶原玻璃化的数据产生了具有对齐的纤维状结构的材料， 与天然角膜相似的正交组织的角膜。此外，细胞能够在 材料和植入物在兔角膜缺损中是可缝合的和生物相容的。我们的目标是 为了复制天然角膜的超微结构，包括结构（胶原纤维大小、排列和 薄片）和功能（透明度和机械强度）。我们假设CD分子 与胶原蛋白特异性相互作用，以控制玻璃化过程中的纤维间距和组装， 全功能角膜替代材料。为实现这些目标，我们提出以下具体建议： 目的： 具体目标1：使用统计优化开发CD-胶原生物材料库。A CD轴 材料库将使用多相统计方法合成不同形式的α、β和γ CD 将使用优化实验设计（DOE）来定义和优化角膜仿生 结构 具体目标2：表征CD-胶原蛋白生物材料的物理特性。Aim的配方 1，其实现了用于角膜植入的最低清晰度和机械性能， 与天然角膜和标准胶原植入物相比。 具体目标3：将仿生CD-胶原材料转化为角膜缺损模型。

The overall vision of the proposed research is to create an effective, scalable, and affordable biomimetic corneal substitute suitable for clinical translation. Damage to the cornea, the outermost tissue of the eye, causes loss of vision, which profoundly impacts quality of life. Cornea transplantation is the gold standard for treatment, but slowly progressing rejection, short supply of donor tissue, and the small number of failures that lead to devastating consequences, remain key challenges. Artificial corneal substitutes have thus far failed to recapitulate the native tissue structure and function. An effective biosynthetic replacement that mimics the native cornea would offer a transformational new option for corneal injury patients. Our prior work has opened a novel route to fully functional synthetic corneas. We previously developed a collagen vitrification process that increases the mechanical strength of collagen gels. To achieve the necessary transparency in full thickness implants, we needed to further control collagen spacing and lamellae orthogonality. We recapitulated cornea development and organization in vitro using synthetic cyclic proteoglycan molecules, cyclodextrins (CD). Preliminary data with CD and collagen vitrification produced materials with aligned fibrillar architecture and orthogonally organized lamellae similar to the native cornea. Furthermore, cells were able to grow on the materials and implants were suturable and biocompatible in a rabbit corneal defect. Our objective is to replicate the native cornea ultrastructure, including structure (collagen fibril size, alignment and lamella) and function (transparency and mechanical strength). We hypothesize that the CD molecules interact specifically with collagen to control fiber spacing and assembly during vitrification, enabling a fully functional corneal replacement material. To achieve these goals, we propose the following specific aims: Specific Aim 1: Develop a library of CD-Collagen biomaterials using statistical optimization. A CD-Col material library will be synthesized with varying forms of α, β, and γ CDs using multiphase statistical optimization Design of Experiments (DOE) will be used to define and optimize corneal biomimetic structure. Specific Aim 2: Characterize physical properties of CD-Collagen biomaterials. Formulations from Aim 1 that achieve minimum clarity and mechanics for corneal implantation will be characterized and compared to the native cornea and standard collagen implants. Specific Aim 3: Translate the biomimetic CD-Collagen material to a corneal defect model.