Integration and Remodeling of Bioprinted Skin in Full-Thickness Wound Healing

项目摘要 本项目将为皮肤组织工程双学位医学博士/博士生提供多学科培训 和烧伤重建手术，解决了对完全再生皮肤替代品的未满足的临床需求 潜力烧伤是美国的主要临床负担，有近50万患者接受治疗 每年，死亡率为4.9%，估计每年的成本为20亿美元。烧伤的护理标准 损伤包括自体皮肤移植，但这些程序需要足够的收获点，这是稀缺的， 严重创伤的患者。组织工程皮肤替代品为皮肤移植提供了一种有前途的替代品。 然而，目前的原型只含有两种细胞类型;缺乏汗腺和皮脂腺，毛囊， 和色素沉着;并且可能不会刺激血管再生和神经支配。因为皮肤移植的最终目的 是再生天然皮肤的真实解剖学和生理学， 具有更多细胞类型和完全再生潜力的生物工程皮肤。为了满足生物工程皮肤的需求， 生物打印技术已经发展到更精确地设计具有适当3D的组织替代物， 结构组织。该提案将测试3D生物打印皮肤移植物将支持 在体内全层伤口中再生天然皮肤，类似于皮肤自体移植物。为了验证这个假设， 研究金申请人提出了三个具体目标。目标1将描述生物打印皮肤如何加速 体内表皮屏障形成。这一目标将为申请人提供数字平面测量分析方面的培训 测量伤口闭合率，NOVATM皮肤相位计分析测量再闭合程度， 随着时间的推移上皮化。目的2将探索生物打印皮肤在体内如何发生新血管形成， 与IHC染色的毛细血管相比，需要能力qrtPCR来测量内皮生长因子， mm2。目的3将研究黑素细胞迁移如何影响生物打印的皮肤色素沉着。这将需要 免疫组织化学染色以确定小鼠与人的组织形成、毛囊形成和 黑色素生成。申请人已经组建了一个由赞助商、共同赞助商、 贡献者和顾问，在再生医学，ECM成像，生物材料科学， 伤口愈合、皮肤科和烧伤重建手术。他们制定了培训计划，其中包括：（1） 导师会议，（2）课程作业，（3）研究培训，（4）临床培训，使学生 从技术和概念上发展成为一名独立的皮肤组织工程师。这本小说 在世界一流的研究所进行的全层伤口治疗方法将作为 学生的培训，并允许这个有前途的申请人发展为一个物理学家，科学家准备解决未来 通过皮肤组织工程进行烧伤重建手术的缺陷。

PROJECT SUMMARY This project will provide multidisciplinary training for a dual-degree MD/PhD student in skin tissue engineering and burn reconstructive surgery, addressing an unmet clinical need for a skin substitute with full-regeneration potential. Burn injuries are a major clinical burden in the United States, with nearly 500,000 patients treated annually, a mortality rate of 4.9%, and an estimated cost of $2 billion per year. The standard of care for burn injuries includes autologous skin grafting, but these procedures require sufficient harvest sites that are scarce in patients with severe wounds. Tissue-engineered skin substitutes offer a promising alternative to skin grafts. However, current prototypes contain only up to two cell types; lack sweat and sebaceous glands, hair follicles, and pigmentation; and may not stimulate revascularization and innervation. Since the ultimate goal of a skin graft is to regenerate authentic anatomy and physiology of native skin, there is an immense need to develop bioengineered skin with more cell types and full regeneration potential. To meet the need for bioengineered skin, bioprinting techniques have been developed to more accurately engineer tissue substitutes with appropriate 3D structural organization. This proposal will test the hypothesis that a 3D bioprinted skin graft will support regeneration of native-like skin in full-thickness wounds in vivo, similar to skin autografts. To test this hypothesis, the fellowship applicant has proposed three Specific Aims. Aim 1 will delineate how bioprinted skin accelerates epidermal barrier formation in vivo. This aim will provide the applicant with training in digital planimetry analysis to measure rates of wound closure, and NOVATM dermal phase meter analysis to measure the extent of re- epithelialization over time. Aim 2 will explore how neovascularization occurs in bioprinted skin in vivo and will require competence qrtPCR to measure endothelial growth factors compared with IHC stained capillaries per mm2. Aim 3 will investigate how melanocyte migration impacts bioprinted skin pigmentation. This will require immunohistochemical staining to determine mouse vs. human tissue formation, hair follicle formation, and melanin production. The applicant has assembled a multidisciplinary team of sponsors, co-sponsors, contributors, and consultants with expertise in regenerative medicine, ECM imaging, biomaterials science, wound healing, dermatology, and burn reconstructive surgery. They have established a training plan with (1) Mentorship Meetings, (2) Coursework, (3) Research Training, and (4) Clinical Training that will allow the student to develop both technically and conceptually towards becoming an independent skin tissue engineer. This novel approach to treatment of full-thickness wounds, conducted at a world-class institute, will serve as a basis for the student’s training, and allow this promising applicant to develop as a physician-scientist poised to address future deficits in burn reconstructive surgery through skin tissue engineering.