Statistical optimization of self-assembled biosynthetic cornea implants

拟议研究的总体愿景是创建一个高效、可扩展且经济实惠的 适用于临床移植的仿生角膜替代物。角膜受损，最外面的 眼睛的组织，会导致视力丧失，从而深刻影响生活质量。角膜移植 是治疗的黄金标准，但缓慢进展的排斥反应，供体组织短缺，以及 导致毁灭性后果的少数失败仍然是关键挑战。人造的 到目前为止，角膜替代物未能概括出天然的组织结构和功能。一个 模仿天然角膜的有效生物合成替代品将提供一种变革性的新 角膜损伤患者的选择。我们之前的工作开辟了一条全功能合成的新途径 眼角膜。我们之前开发了一种胶原玻璃化过程，它增加了机械 胶原蛋白凝胶的强度。为了在全厚度植入物中实现必要的透明度，我们需要 以进一步控制胶原蛋白间距和片层正交性。我们回顾了角膜的发育。 并在体外利用合成的环状蛋白多糖分子环糊精(CD)进行组织。初步 CD和胶原玻璃化冷冻的数据产生了具有排列的纤维结构和 平行排列的板层，类似于天然的角膜。此外，细胞能够继续生长 材料和植入物在兔角膜缺损处是可缝合和生物相容的。我们的目标是 复制天然角膜的超微结构，包括结构(胶原纤维大小、排列和 薄层)和功能(透明度和机械强度)。我们假设CD分子 专门与胶原蛋白相互作用，以控制玻璃化过程中的纤维间距和组装，从而实现 全功能角膜置换材料。为了实现这些目标，我们提出了以下具体建议 目标： 具体目标1：利用统计优化方法建立CD-胶原生物材料库。一张CD-COOL 材料库将用不同形式的α，β和γCD合成，并使用多相统计 实验优化设计(DOE)将用于定义和优化角膜仿生 结构。 具体目标2：表征CD-胶原生物材料的物理性质。来自AIM的配方 1角膜植入物达到最低透明度和机械性能的特性和 与天然角膜和标准的胶原植入物相比。 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.