Corneal Scar Repair through SPAACKL: Sutureless, Pro-regenerative Anterior Additive Collagen gel KeratopLasty

通过 SPAACKL 修复角膜疤痕：无缝线、促再生前部添加剂胶原蛋白凝胶 KeratopLasty

• 批准号: 10343517
• 负责人: David Myung
• 金额: $ 51.31万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10343517
• 关键词: Actins; Address; Affect; Alkynes; Anterior; Autologous; Azides; Behavior; Biological Assay; Biomedical Engineering; Blindness; Burr hole procedure; Cadaver; Cell Survival; Cells; Central Scar; Chemistry; Cicatrix; Clinical; Coculture Techniques; Collagen; Copper; Cornea; Corneal Diseases; Corneal Injury; Custom; Data; Defect; Dimensions; Disease; Encapsulated; Engineering; Enzyme-Linked Immunosorbent Assay; Epithelial; Epithelial Cells; Evaluation; Excision; Exhibits; Eye; FDA approved; Fluorescamine; Fluorescein; Fostering; Foundations; Gel; Growth; Growth Factor; HGF gene; Harvest; Hydrogels; Immunohistochemistry; In Situ; In Vitro; Inflammation; Keratoplasty; Light; Liquid substance; Mechanics; Mediating; Modality; Modeling; Morphology; Natural regeneration; Operative Surgical Procedures; Optics; Organ Culture Techniques; Oryctolagus cuniculus; Paracrine Communication; Patients; Phenotype; Photochemistry; PicoGreen; Procedures; Property; Proteins; Regenerative capacity; Research; Rheology; Riboflavin; Side; Site; Source; Spectrophotometry; Stains; Stromal Cells; Succinimides; Surface; Surgical sutures; Suspensions; Techniques; Technology; Testing; Therapeutic; Therapeutic Effect; Thick; Time; Tissue Donors; Tissue Grafts; Tissues; Transplantation; Transplantation Surgery; Vision; Visual Pathways; Visual impairment; Western Blotting; Work; analog; base; beneficiary; biological systems; catalyst; clinical translation; corneal epithelial wound healing; corneal epithelium; corneal scar; corneal surgery; cross reactivity; crosslink; curative treatments; cycloaddition; design; graft failure; healing; improved; in vivo; mesenchymal stromal cell; novel therapeutics; oxidative damage; paracrine; phenotypic biomarker; preservation; prevent; regenerative; regenerative growth; repaired; severe injury; sight restoration; wound; wound healing

PROJECT SUMMARY/ABSTRACT As the dome-shaped, transparent outermost part of the eye, the cornea provides the majority of the focusing power for the visual pathway. When it is damaged due to severe injury or disease, scarring often ensues, resulting in reduced vision and, in many cases, blindness. In spite of the various types of corneal transplants that are available, there remains a major clinical need for new modalities to restore transparency to scarred corneas without donor tissue, which is in short supply worldwide. Corneal mesenchymal stromal cells (c- MSCs) have known therapeutic effects on corneal scarring and wound healing, but the optimal way to deliver their benefits to the eye have yet to be determined. We are developing Sutureless, Pro-regenerative, Anterior Additive Collagen gel KeratopLasty (SPAACKL), a procedure that removes and replaces blinding corneal scars with a transparent, stroma-like gel matrix containing c-MSCs. After removal of corneal scar tissue, the material is applied to the defect as a viscous liquid suspension of c-MSCs, forming a crosslinked, transparent cellularized stromal substitute within minutes that not only recreates the smooth surface necessary for clear vision but also promotes rapid re-epithelialization. This technology leverages a crosslinking technology known as copper-free click chemistry that is bio-orthogonal: it does not react with proteins, cells, or biologic systems of any kind. As such, it can be safely applied to a corneal wound and around c-MSCs without producing toxic side products, and without the need for light energy, catalysts or accelerators. Our central hypothesis is that bio-orthogonal crosslinking can improve the regenerative benefits of c-MSCs by preserving the bioactivity of its encapsulated cargo compared to less specific crosslinking chemistries that are used currently in corneal surgery. In preliminary work, we have demonstrated that bio-orthogonally crosslinked gels support the growth of encapsulated stromal cells and have demonstrated the regenerative capacity of these cell-matrix composites to support rapid, multi-layered epithelialization both ex vivo and in vivo. Motivated by this data, our first aim is to test the hypothesis that matrix stiffness, composition, and crosslinking chemistry influence c-MSCs’ viability and secretion of pro- regenerative factors. Our second aim is to test the hypothesis that encapsulated c-MSCs exert their pro- regenerative influence on the corneal epithelium primarily through paracrine signaling. Our third aim is to evaluate the ability of bio-orthogonally crosslinked hydrogels to deliver therapeutic cargo that enhances epithelial and stromal regeneration through an in vivo keratectomy model. This research will build the foundational data for eventual clinical translation of a new way to treat corneal blindness without the need for sutures, light energy, or cadaveric donor tissue, and has the potential to one day help patients avoid the need for a traditional corneal transplant.

项目概要/摘要 作为眼睛的圆顶状、透明的最外层部分，角膜提供了大部分的聚焦功能 视觉通路的能量。当它因严重受伤或疾病而受损时，往往会留下疤痕， 导致视力下降，并在许多情况下导致失明。尽管角膜移植有多种类型 尽管已有可用的方法，但临床仍然迫切需要新的方法来恢复伤痕累累的透明度 没有供体组织的角膜，全世界都供不应求。角膜间充质基质细胞（c- 间充质干细胞（MSC）对角膜疤痕和伤口愈合具有已知的治疗作用，但最佳方法是 它们对眼睛的好处尚未确定。我们正在开发无缝线、促再生、 前路添加剂胶原凝胶 KeratopLasty (SPAACKL)，一种去除和替代致盲的手术 角膜疤痕具有含有 c-MSC 的透明基质样凝胶基质。角膜疤痕去除后 组织中，该材料以 c-MSC 的粘性液体悬浮液形式应用于缺损处，形成交联的、 几分钟内透明的细胞化基质替代品不仅可以重建光滑的表面 清晰视力所必需的，但也促进快速上皮化。该技术利用了 被称为无铜点击化学的交联技术是生物正交的：它不会与 蛋白质、细胞或任何类型的生物系统。因此，它可以安全地应用于角膜伤口 围绕c-MSCs，不产生有毒副产物，并且不需要光能、催化剂或 加速器。我们的中心假设是生物正交交联可以改善再生能力 与特异性较低的相比，c-MSC 的优势在于保留其封装货物的生物活性 目前用于角膜手术的交联化学物质。在前期工作中，我们有 证明生物正交交联凝胶支持封装基质细胞的生长 已经证明了这些细胞基质复合材料的再生能力，以支持快速、多层 离体和体内上皮化。受这些数据的启发，我们的首要目标是检验以下假设： 基质硬度、成分和交联化学会影响 c-MSC 的活力和亲细胞的分泌。 再生因素。我们的第二个目标是检验封装的 c-MSC 发挥其亲和力的假设。 主要通过旁分泌信号传导对角膜上皮的再生影响。我们的第三个目标是 评估生物正交交联水凝胶传递治疗物质的能力，从而增强 通过体内角膜切除模型进行上皮和基质再生。这项研究将建立 为最终临床转化治疗角膜失明的新方法提供基础数据，而无需 用于缝合、光能或尸体供体组织，并有可能有一天帮助患者避免 需要传统的角膜移植。