In Situ Bioconjugation as a Therapeutic Delivery Modality to Enhance Ocular Wound Healing

摘要 这是David Myung博士提交的K08奖项申请，他被聘为 斯坦福大学拜尔斯眼科研究所眼科学助理教授。明博士正在建立 他在专注于角膜伤口愈合的转译生物工程研究中担任研究员。 K08奖项将为明博士提供必要的支持，以进一步发展特定的专业知识 在角膜伤口愈合和再生医学方面，并获得成为一名 独立调查员。为了帮助他实现这些目标，明博士组建了一个 跨学科指导团队由各自领域的以下主要研究人员组成：Ali博士 贾利安是角膜伤口愈合方面的专家，尤其是眼表疾病和干细胞。 移植；萨拉·海尔肖恩博士是生物材料专家；杰弗里·戈德堡博士是 神经再生。此外，他还将咨询斯坦福大学的临床医生爱德华·曼奇。 加州大学克里斯托弗·墨菲博士，拥有角膜交联化学专业知识的角膜专家 戴维斯是在角膜伤口愈合的动物模型中测试生长因子的专家，他也是戴维斯博士。 杰弗里·托克是斯坦福大学软材料和混合材料测试中心的负责人 工程学。 在治疗眼部创伤性损伤、溃疡和烧伤时，组织愈合不充分仍然是一种 重大挑战。角膜特别容易受到延迟愈合的致盲后果的影响。 例如结疤、熔化和穿孔。因此，临床上需要一种新的方法来治疗 促进眼部伤口愈合。这项研究调查了使用高度细胞友好型的 生物偶联化学将治疗因子直接输送到三种损伤的角膜组织 方法：(1)通过覆盖伤口并释放生长因子的可注射凝胶，(2)通过化学方法 将生长因子直接结合到伤口床上，以及(3)通过包裹人间充质干细胞 细胞(HMSCs)在眼表的凝胶中。第一个目标是表征和控制角膜复发率。 通过生长因子洗脱原位伤口上形成的胶原凝胶实现上皮化。第二个目标 目的是评价和优化生长因子偶联的上皮和神经再生效果 直接进入神经消融的角膜。第三个目标是阐明和最大化hMSCs的生物学活性。 包裹在化学损伤的眼表上的胶原胶中。这项研究可能会导致 克服局部注射生长因子限制的新的角膜伤口愈合疗法 和干细胞，并将在K奖结束前形成R01申请的基础。

ABSTRACT This is a K08 award application submitted by David Myung, MD, PhD, who is being hired as an Assistant Professor of Ophthalmology at the Byers Eye Institute at Stanford. Dr. Myung is establishing himself as an investigator in translational bioengineering research focused on corneal wound healing. The K08 award will provide Dr. Myung with the support necessary to further develop specific expertise in corneal wound healing and regenerative medicine, and to acquire the skills necessary to become an independent investigator. To help him achieve these goals, Dr. Myung has assembled an interdisciplinary mentorship team comprised of the following leading investigators in their fields: Dr. Ali Djalilian is an expert in corneal wound healing, and in particular ocular surface disease and stem cell transplantation; Dr. Sarah Heilshorn is an expert in biomaterials; Dr. Jeffrey Goldberg is an expert in nerve regeneration. In addition, he will consult with Dr. Edward Manche from Stanford, a clinical cornea specialist with expertise in corneal crosslinking chemistry, Dr. Christopher Murphy from UC Davis, an expert in testing growth factors in animal models of corneal wound healing, as well as Dr. Jeffrey Tok, who directs the soft and hybrid materials testing facility at Stanford's School of Engineering. In the treatment of traumatic injuries, ulcers, and burns of the eye, inadequate tissue healing remains a major challenge. The cornea is particularly vulnerable to the blinding consequences of delayed healing such as scarring, melts, and perforations. There is, therefore, a major clinical need for new ways to enhance wound healing in the eye. This research investigates the benefits of using highly cell-friendly bioconjugation chemistries to deliver therapeutic factors directly to wounded corneal tissue in three ways: (1) through an injectable gel that covers wounds and releases growth factors, (2) by chemically binding growth factors directly to the wound bed, and (3) by encapsulating human mesenchymal stem cells (hMSCs) within a gel at the ocular surface. The first aim is to characterize and control corneal re- epithelialization through growth-factor eluting collagen gels formed on wounds in situ. The second aim is to evaluate and optimize the epithelial and nerve regeneration effects of growth factors coupled directly to a nerve-ablated cornea. The third aim is to elucidate and maximize the bioactivity of hMSCs encapsulated within collagen gels on a chemically-injured ocular surface. This research may lead to new corneal wound healing therapies that overcome the limitations of topically delivered growth factors and stem cells, and will form the basis of an R01 application before the end of the K award.