Characterization of an Ex Vivo Bioprinted Skin Model of Sulfur Mustard Injury

硫芥损伤离体生物打印皮肤模型的表征

• 批准号: 10228424
• 负责人: ADAM JORGENSEN
• 金额: $ 5万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-17 至 2021-11-17
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10228424
• 关键词: 3-Dimensional; Address; Adherence; Adipocytes; Affect; Anatomy; Animal Model; Animals; Area; Basement membrane; Biochemical; Biological; Biological Markers; Biomedical Engineering; Biomedical Research; Biomimetics; Bulla; Burn injury; CD14 gene; Cell Culture Techniques; Cell Death; Cells; Cessation of life; Chemical Models; Chemical Warfare Agents; Chemical Weapons; Chemicals; Cicatrix; Clinical; Collagen Type VII; Complex; Data; Dermal; Dermatologic; Development; Disease; Dose; Edema; Elements; Endothelial Cells; Epidermal Ridges; Epidermis; Erythema; Exposure to; Fibroblasts; Functional disorder; Funding; Gene Expression; Genetic Transcription; Goals; Histologic; Human; In Vitro; Inflammation; Inflammation Mediators; Injury; Investigation; Metalloproteases; Military Personnel; Modality; Modeling; Mustard Gas; Organ; Organoids; Pathologic; Pathway interactions; Physiological; Physiology; Pigmentation physiologic function; Poly(ADP-ribose) Polymerases; Population; Pre-Clinical Model; Protein Analysis; Quantitative Reverse Transcriptase PCR; RNA analysis; Research Institute; Scanning Electron Microscopy; Signaling Molecule; Skin; Skin injury; Structural Protein; Structure; System; Techniques; Testing; Time; Tissue Engineering; Tissues; Toxic effect; Treatment Efficacy; Validation; Vascularization; Vesicants; World War I; base; bioprinting; bioweapon; cell type; cytokine; cytotoxicity; extracellular; healing; improved; in vitro Model; keratinocyte; laminin-5; medical countermeasure; melanocyte; monocyte; novel; organizational structure; pre-clinical; protein expression; prototype; real time monitoring; research study; response; skin barrier; three dimensional structure; two-dimensional

PROJECT SUMMARY Sulfur Mustard (SM) remains a significant threat to civilian and military populations. Skin exposure to SM induces erythema, followed by edema and large blisters in the affected area, with prolonged healing times. The sequence and manner of cell death and detachment in this injury are still unresolved, in part due to incomplete in vitro and preclinical models. Since the goal of an ex vivo 3D skin model is to replicate the authentic anatomy and physiology of native skin, there is an immense need to develop bioengineered skin with multiple cell types and appropriate physiological potential. Assessment of bioprinted skin as an ex vivo organoid (organ tissue equivalent) has demonstrated that it maintains its layered structure for over 2 months in vitro. Assessment of skin organoids by RNA and protein analysis demonstrate a physiological response similar to those seen in normal human skin. These encouraging results suggest that this skin construct is recapitulating the human organ and we propose to use the funds available through this supplement to further expand the bioprinted human skin into a model of chemical burn injuries. We will use bioprinted skin to model exposure to the known chemical vesicant sulfur mustard (SM), with the ultimate goal of showing the full utility of the bioprinted skin in elucidating biochemical and pathophysiological pathways in an effort to discover biomarkers and medical countermeasures. Based on the data we have generated to date, our central hypothesis is that ex vivo bioprinted skin will model clinical pathological effects of exposure to the chemical vesicant sulfur mustard. The overall goal of this supplemental proposal is to generate proof of principal data showing that the bioprinted skin will respond to the chemical insult in a way that is consistent with the known effects of SM. Our study will validate the dose and time of exposure for modeling the human response to SM in vitro, and will include histological, qRT-PCR, and ELIZA analysis. These studies will focus on demonstrating that the bioprinted skin behaves in a way that is consistent with Specific Aim 1 will characterize the pathological consequences of sulfur mustard exposure. Specific Aim 2 will look to identify cellular pathways affected by SM induced injury. These studies will validate the use of the bioprinted skin as a viable ex vivo model of SM toxicity which will enable high precision analysis, including real-time monitoring and -omics to elucidate the sequence and mechanism of sulfur mustard skin injuries. Furthermore, it suggests that the bioprinted skin can be developed into a general surrogate of human skin pathophysiology useful in studying other types of insults. Ultimately, we believe that this system will identify biomarkers of exposure and therapeutic efficacy, as well as serving as a useful modality to discovery and optimize medical countermeasures for a range of chemical and biological weapons.

项目摘要 硫芥末（SM）仍然对平民和军事人口构成重大威胁。皮肤暴露于SM会诱导 红斑，随后在受影响地区的水肿和大水泡，长时间的愈合时间。序列 这种损伤中细胞死亡和脱离的方式仍未解决，部分原因是体外不完整和 临床前模型。由于离体3D皮肤模型的目的是复制真实的解剖结构和 天然皮肤的生理学，有多种细胞类型的生物工程皮肤迫切需要 适当的生理潜力。评估生物打印的皮肤作为离体器官（器官组织） 等效）表明，它在体外维持了2个月以上的分层结构。评估 通过RNA和蛋白质分析的皮肤器官表现出一种与在 正常的人皮。这些令人鼓舞的结果表明，这种皮肤构造正在概括人体器官 我们建议使用通过此补充剂可用的资金进一步扩大生物打印的人类皮肤 进入化学烧伤损伤的模型。我们将使用生物打印的皮肤模型接触已知的化学物质 囊泡硫芥末酱（SM），其最终目的是显示生物打印皮肤在阐明中的全部实用性 生化和病理生理途径，以发现生物标志物和医学对策。 基于我们迄今为止生成的数据，我们的中心假设是，离体生物打印的皮肤将会 模拟暴露于化学囊泡硫芥​​末的临床病理作用。总体目标 该补充建议的是生成主要数据证明，表明生物打印的皮肤会做出反应 以与SM的已知效应一致的方式进行化学侮辱。我们的研究将验证剂量 和暴露时间用于建模人类对SM的反应体外的反应，并将包括组织学，QRT-PCR， 和Eliza分析。这些研究将着重于证明生物打印皮肤的行为方式 与特定的目标1一致，将表征硫芥末酱暴露的病理后果。 特定的目标2将旨在识别受SM诱导损伤影响的细胞途径。这些研究将验证 将生物打印的皮肤用作可行的SM毒性的可行的离体模型，这将使​​高精度分析， 包括实时监测和 - 词，以阐明硫芥末酱的序列和机制 受伤。此外，这表明生物打印的皮肤可以发展为人类的一般替代物 皮肤病理生理学有助于研究其他类型的侮辱。最终，我们认为该系统将确定 暴露和治疗功效的生物标志物，并成为发现和发现的有用方式 优化各种化学和生物武器的医学对策。