THERAPY OF CHRONIC WOUNDS WITH LIVING SKIN EQUIVALENT

用活体皮肤等效物治疗慢性伤口

• 批准号: 7725259
• 负责人: SATORI IWAMOTO
• 金额: $ 7.55万
• 依托单位: ROGER WILLIAMS HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-05 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7725259
• 关键词: Animal Experimentation; Antibodies; Beds; Biomedical Engineering; Biopsy; Caring; Cells; Centers of Research Excellence; Chronic; Clinical; Clinical Trials; Closure; Complex; Computer Retrieval of Information on Scientific Projects Database; Condition; Data; Enrollment; Experimental Designs; Funding; Future; Gene Chips; Genes; Goals; Grant; Guanosine Monophosphate; Healed; Human; In Vitro; Injury; Institution; Integrins; Knowledge; Laboratories; Life; Measurement; Methods; Modeling; Patients; Population; Preparation; Process; Protocols documentation; Publishing; RNA; Radiation Injuries; Rate; Reporting; Research; Research Personnel; Resources; Skin; Source; Stress; Testing; Thigh structure; Tissue Engineering; United States National Institutes of Health; Vitronectin; Work; Wound Healing; healing; improved; in vivo; interest; keratinocyte; migration; new technology; novel; receptor; research study; size; success; tool; volunteer; wound

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The central hypothesis of this proposal is that living skin equivalents (LSE) consisting of living cells may require an activation process in vitro to optimize their in vivo effects and accelerate healing in difficult clinical situations. Specific Aims: 1) To determine the ideal conditions for in vitro activation of a living bilayered living skin equivalent (LSE) 2) To use the optimally stimulated construct in difficult to heal human wounds 3) To characterize novel genes that are expressed by injury to the LSE and for application to human wounds At the completion of these studies, we will have produced and validated the best way to optimize the use of a living bilayered skin equivalent for use in human wounds. This knowledge could be applicable to other types of tissue engineering constructs. The overall goal of this pilot application is to explore a shift in paradigm with regard to the use of living skin equivalents (LSE). The underlying hypothesis is that it is possible to stimulate constructs and render them more effective. Specific aim 1 will identify optimal ways to stimulate bioengineered skin constructs, using novel and established methods we have established in our laboratory. The focus of that specific aim will be an already available living skn equivalent (LSC). Data generated from specific aim 1 will be used in testing the use of stimulated LSC in a clinical trial of difficult to heal wounds, making use of our GMP facility to properly prepare the LSC. Assessment of efficacy and healing will be done by measurements of the healing rate from the edge of the wound, as well as size reduction and complete wound closure. As part of specific aim 3, we will also perform a thorough gene analysis of LSC during the process of epiboly. We regard the epiboly model as a potentially very useful tool for identifying genes of interest in the migrating keratinocyte population. Here too, the experimental design is dynamic. We can block the epiboly by various methods, including the antibodies to vitronectin and its integrin receptor, or even explore ways to render the construct unable to "heal". Radiation injury can be explored, for example. In published work, we have found ways to make the construct dormant and unable to reepithelialize. RNA isolated from the BSC will be used for affymetrix gene chip analysis, capable of detecting 33,000 genes. The COBRE has already purchaced a gene microarray scanner that will facilitate our studies. The hope is that we can determine specific genes that are either upregulated or downregulated in this dynamic keratinocyte migration (or lack thereof) model. These data have definite mechanistic significance. Ideally, one could use the information for developing a putative keratinocyte gene signature that could later be applied to and explored in human wounds. All of the proposed aims and experiments are within our capabilities, as we have developed a team that can move easily from bench work to animal experimentation, to human wounds. We believe this is a major strength of this pilot proposal. Another major strength is the underlying hypothesis. Many complex wounds either do not respond to treatment at all, or with great delay. Given the expenses associated with these new technologies, it is imperative that the wound bed be optimized. The PI described the problem of wound bed preparation in a previous report. This notion, which basically is meant to underline and stress the need to improve wound care before advanced therapies are used, has received considerable success. It is our hypothesis that we now need to explore a way to stimulate LSC to optimize their success in chronic wounds. We believe that in the future this could become an essential step in the use of tissue engineering treatment of complex wounds or other situations where the wound microenvironment is severely compromised. enroll 6 volunteers to have biopsies of their thighs as an additional control. The protocol for those biopsies will be the same as described for the patients.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 该提议的中心假设是，由活细胞组成的活皮肤等效物（LSE）可能需要在体外​​激活过程来优化其体内效应并在困难的临床情况下加速愈合。 具体目的： 1）确定活着的双层生命皮肤（LSE）体外激活的理想条件（LSE） 2）在难以治愈人伤口的最佳刺激构造中使用 3）表征通过对LSE损伤表达的新型基因，并应用于人伤口 这些研究完成后，我们将生产并验证了优化使用双层皮肤的最佳方法，以在人伤口中使用。这些知识可能适用于其他类型的组织工程结构。 该试点应用的总体目标是探索使用皮肤等效物（LSE）的范式转变。基本的假设是可以刺激结构并使它们更有效。具体目标1将使用我们在实验室中建立的新颖和既定的方法来确定刺激生物工程皮肤结构的最佳方法。该特定目标的重点将是已经可用的生活SKN等效物（LSC）。 在难以治愈伤口的临床试验中，使用特定目标1产生的数据将用于测试刺激的LSC的使用，利用我们的GMP设施来正确准备LSC。通过测量伤口边缘的愈合率以及尺寸降低和完全伤口闭合，可以评估功效和愈合。 作为特定目标3的一部分，我们还将在Epiboly过程中对LSC进行彻底的基因分析。我们将表皮模型视为一种潜在的非常有用的工具，用于识别迁移角质形成细胞种群中感兴趣的基因。在这里，实验设计也是动态的。我们可以通过各种方法来阻止Epiboly，包括对玻染蛋白及其整联蛋白受体的抗体，甚至探索使构建体无法“治愈”的方法。例如，可以探索辐射损伤。在已发表的工作中，我们找到了使构造休眠且无法重新上皮化的方法。从BSC中分离出的RNA将用于Affymetrix基因芯片分析，能​​够检测33,000个基因。该鞋底已经购买了一种基因微阵列扫描仪，该扫描仪将有助于我们的研究。希望我们可以确定在这种动态角质形成细胞迁移（或缺乏其）模型中上调或下调的特定基因。这些数据具有明确的机械意义。理想情况下，可以使用该信息来开发推定的角质形成细胞基因特征，后来可以在人类伤口中应用并探索。 所有提出的目标和实验都在我们的能力内，因为我们已经建立了一个可以轻松地从板凳工作到动物实验到人类伤口的团队。我们认为这是该试点建议的主要优势。另一个主要优势是基本假设。许多复杂的伤口要么根本不反应治疗，要么延迟延迟。鉴于与这些新技术相关的费用，必须优化伤口床。 PI在上一份报告中描述了伤口床准备的问题。这种概念基本上是为了强调和强调在使用先进疗法之前改善伤口护理的需求，并取得了很大的成功。 我们的假设是，我们现在需要探索一种刺激LSC以优化其在慢性伤口中的成功的方法。我们认为，将来这可能成为使用组织工程治疗的复杂伤口或其他情况严重损害的复杂伤口或其他情况的重要步骤。招募6名志愿者将大腿的活检作为额外的控制。这些活检的方案将与患者所描述的相同。