Tissue Engineering for the Treatment of Delayed Healing

组织工程治疗延迟愈合

• 批准号: 6725749
• 负责人: Vincent Falanga
• 金额: $ 40.39万
• 依托单位: ROGER WILLIAMS HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-15 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6725749
• 关键词: blood vessels; fibrin; fibroblasts; flow cytometry; fluorescent in situ hybridization; genetically modified animals; green fluorescent proteins; hematopoietic stem cells; human tissue; immunocytochemistry; keratinocyte; laboratory mouse; mesenchyme; mitogen activated protein kinase; northern blottings; phenotype; tissue /cell culture; tissue engineering; western blottings; wound healing

DESCRIPTION (provided by applicant): A more effective tissue engineering approach to chronic wounds may require the replacement of phenotypically altered wound cells, possibly through the use of stem ceils. Using green fluorescent protein (GFP) transgenic and wild type mice, we have shown that infusion of GFP + marrow cells can result in the production of blood vessels, keratinocytes, and other structures in wounded skin. A new model of delayed healing we have developed using excisional wounds of routine tails will allow us to extend these observations. In parallel, we have found that non-healing human chronic wounds heal dramatically with the topical application of autologous bone marrow-derived cells. We propose the following specific aims: 1) to characterize the engraftment of unseparated marrow cells in a murine model of delayed healing using GFP - marrow cells and different methods of marrow cell delivery to wounds; 2) to determine the marrow cell subsets that accelerate healing in murine wounds. We will evaluate unseparated marrow, as well as Lin- and positive Lin+ marrow cells, mesenchymal stem cells, and hematopoietic stems cells, including Lin-Sca+ and Lin- Ho/l0Rho/10cells. 3) to assess the topical delivery of marrow-derived cells. In vitro and in vivo studies will determine the feasibility of this approach using specific marrow-derived cells in a Factor XIII cross-linked fibrin gel; 4) Determine whether autologous marrow-derived cells can correct the abnormal cellular phenotype of chronic wounds. We will apply specific subsets of marrow cells to non-healing human wounds and will determine whether healing and normal phenotype are restored in wound fibroblasts. These studies will advance our understanding of how to reconstitute the wound bed and whether it is possible to correct the abnormal cellular phenotype in non-healing wounds.

描述（由申请人提供）：一种更有效的组织工程方法可以用于慢性伤口，可能需要通过使用茎天花板来替换表型改变的伤口细胞。使用绿色荧光蛋白（GFP）转基因和野生型小鼠，我们已经表明，GFP +骨髓细胞的输注可以导致血管，角质形成细胞和受伤皮肤中其他结构的产生。我们使用常规尾巴的散布伤口开发了一种延迟愈合的新模型，将使我们能够扩展这些观察结果。同时，我们发现非愈合的人类慢性伤口通过自体骨髓衍生细胞的局部应用而急剧治愈。我们提出以下特定目的：1）在使用GFP延迟愈合的鼠模型中表征未分离的骨髓细胞的植入 - 骨髓细胞以及不同的骨髓细胞递送到伤口的方法； 2）确定在鼠伤口中加速愈合的骨髓细胞子集。我们将评估未分离的骨髓以及Lin+骨髓细胞，间充质干细胞以及造血干细胞，包括Lin-SCA+和Lin- HO/L0RHO/10CELLS。 3）评估骨髓衍生细胞的局部传递。体外和体内研究将使用特定的骨髓衍生的细胞在XIII XIII交联纤维蛋白凝胶中确定这种方法的可行性； 4）确定自体骨髓衍生的细胞是否可以纠正慢性伤口的异常细胞表型。我们将将特定的骨髓细胞子集应用于非愈合的人伤口，并确定在伤口成纤维细胞中是否恢复愈合和正常表型。 这些研究将促进我们对如何重建伤口床以及是否有可能纠正非愈合伤口中异常的细胞表型的理解。