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.

项目摘要/摘要 作为圆顶状、透明的眼睛最外面的部分，角膜提供了大部分聚焦。 为视觉通路提供能量。当它因重伤或疾病而损坏时，往往会留下疤痕， 导致视力下降，在许多情况下还会失明。尽管有各种类型的角膜移植 临床上仍然需要新的方式来恢复疤痕的透明度 没有供体组织的角膜，在全球范围内供不应求。角膜间充质基质细胞 MSCs)对角膜疤痕形成和伤口愈合有已知的治疗效果，但最佳的治疗方式 它们对眼睛的好处还有待确定。我们正在开发无缝合的、支持再生的 前部胶原凝胶角膜移植(SPAACKL)--一种去除和取代失明的手术 含有c-MSCs的透明基质状凝胶基质的角膜瘢痕。角膜瘢痕摘除术后 组织，该材料作为c-MSCs的粘性液体悬浮液应用于缺损处，形成交联的， 几分钟内即可替代透明的细胞化基质，不仅可以重建光滑的表面 对于清晰的视力是必要的，但也能促进快速的再上皮化。这项技术利用了 被称为无铜点击化学的交联化技术是生物正交的：它不与 蛋白质、细胞或任何种类的生物系统。因此，它可以安全地应用于角膜伤口和 C-MSCs周围，不产生有毒副产物，也不需要光能、催化剂或 加速器。我们的中心假设是生物正交交联剂可以改善再生能力。 与非特异性相比，c-MSCs通过保留其包裹的货物的生物活性而获得的好处 目前在角膜手术中使用的交联剂。在前期工作中，我们有 证明生物正交联凝胶支持包裹的基质细胞的生长和 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 体外和体内的上皮化。在这些数据的激励下，我们的第一个目标是检验以下假设 基质硬度、成分和交联剂化学影响c-MSCs的活性和分泌前体 再生因素。我们的第二个目标是检验这一假设，即包裹的c-MSCs发挥其促进- 再生对角膜上皮细胞的影响主要通过旁分泌信号。我们的第三个目标是 评估生物正交联水凝胶输送治疗货物的能力 通过活体角膜切削术模型进行上皮和基质再生。这项研究将建立 为最终临床翻译一种治疗角膜失明的新方法提供基础数据 用于缝合、光能或身体供体组织，并有可能有朝一日帮助患者避免 需要进行传统的角膜移植。