Mechanisms of Wound Healing with Cultured Human Skin

培养人类皮肤的伤口愈合机制

基本信息

批准号：
6764053
负责人：
Steven T Boyce
金额：
$ 36.04万
依托单位：
UNIVERSITY OF CINCINNATI
依托单位国家：
美国
项目类别：
财政年份：
1994
资助国家：
美国
起止时间：
1994-01-01 至 2006-06-30
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant) Rapid and effective healing of burn wounds with cultured analogs of human skin is the central objective of this proposal. Medical benefits from improved healing may include, but not be limited to; reduced requirements for split- thickness autograft, shorter hospitalization time; and reduced long-term morbidity after recovery. However, anatomic and physiologic deficiencies of all current models of cultured skin have restricted realization of these benefits. Major deficiencies result from culture conditions that do not generate an epidermal analog with fully functional barrier properties, from irregular pigmentation after healing, and from absence of a vascular plexus in the dermal analog which delays vascularization. The current model of cultured skin substitute (CSS) in this laboratory is a collagen- based sponge populated with cultured human keratinocytes and fibroblasts. An investigative cycle has been established to: A) design experiments to address clinical deficiencies of CSS, B) perform mechanistic studies in vitro to generate new prototypes of CSS; C) transplant experimental CSS to athymic mice to measure efficacy and tissue phenotypes, and, D) study the improved prototype of CSS for reduction of mortality and morbidity in critically injured burn patients. Six specific aims will be pursued: 1 ) Regulation of cellular viability (DNA synthesis, mitochondrial metabolism) and phenotypes (epidermal barrier, basement membrane) by culture conditions (media, biophysical environment); 2) Identification of molecular mediators (cytokines, extracellular matrix) of wound healing processes (angiogenesis, matrix structure); 3) Regulation of melanocyte distribution (cell density) and pigment expression (melanin content) to restore normal skin color; 4) Stimulation of angiogenesis by addition of human dermal microvascular endothelial cells and morphogenesis of vascular analogs; 5) Regulation and automation of keratinocyte growth rates and metabolism (reduction of lactic acid and ammonia) in the Kerator bioreactor; and, 6) Treatment of extensive burns with cultured skin substitutes in the clinic by paired-site comparison to meshed, split-thickness skin autograft. Comparative parameters will include: a) quantitative wound closure with skin substitutes (rates of engraftment, ratio of healed area to biopsy area, frequency of regraftng); and, b) qualitative outcome (scarring, contraction, function and cosmesis). The investigators possess all of the required expertise in cell biology, skin biochemistry and biophysics, wound physiology, chemical engineering and clinical burn care to perform these studies successfully. Accomplishment of these objectives will contribute to reduced mortality and morbidity from burns, improved materials for plastic and reconstructive surgery, and development of other tissue and organ substitutes.

描述（由申请人提供）用人皮肤的培养类似物对烧伤伤口的快速有效愈合是该提案的核心目的。改善康复的医疗益处可能包括但不限于；减少对厚度自体移植的需求减少，住院时间较短；并降低了恢复后的长期发病率。但是，所有当前培养的皮肤模型的解剖学和生理缺陷都限制了这些益处的实现。主要缺陷是由于培养条件不会产生具有功能性屏障特性的表皮类似物，愈合后的不规则色素沉着以及皮肤类似物中缺乏血管丛的缺乏，从而延迟了血管化。该实验室中培养的皮肤替代品（CSS）的当前模型是胶原蛋白的海绵，该海绵中有培养的人角质形成细胞和成纤维细胞。已经建立了一个调查周期：a）设计实验以解决CSS的临床缺陷，b）在体外进行机械研究以生成CSS的新原型； c）移植实验性CSS对无胸腺小鼠进行测量功效和组织表型，d）研究CSS的原型改善，以降低严重受伤的烧伤患者的死亡率和发病率。将追求六个具体目标：1）根据培养条件（媒体，生物物理环境）调节细胞活力（DNA合成，线粒体代谢）和表型（表皮屏障，地下室膜）的调节； 2）鉴定伤口愈合过程（血管生成，基质结构）的分子介质（细胞因子，细胞外基质）； 3）调节黑素细胞分布（细胞密度）和色素表达（黑色素含量）以恢复正常的肤色； 4）通过添加人真皮微血管内皮细胞和血管类似物的形态发生来刺激血管生成； 5）调节角质形成细胞生长速率和代谢（乳酸和氨的降低）的调节和自动化； 6）与网状的，分裂的厚度皮肤自体移植，通过对诊所中培养的皮肤替代品进行培养的皮肤替代品处理广泛的烧伤。比较参数将包括：a）与皮肤替代品的定量伤口闭合（植入率，愈合面积与活检区域的比率，恢复频率）； b）定性结果（疤痕，收缩，功能和宇宙）。研究人员拥有细胞生物学，皮肤生物化学和生物物理学，伤口生理学，化学工程和临床烧伤护理方面的所有必需专业知识，以成功地进行这些研究。这些目标的实现将导致烧伤的死亡率和发病率降低，改善塑料和重建手术的材料以及其他组织和器官替代品的发展。