MECHANISMS OF WOUND HEALING WITH CULTURED SKIN

培养皮肤的伤口愈合机制

• 批准号: 6385853
• 负责人: Steven T Boyce
• 金额: $ 29.48万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-01-01 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6385853
• 关键词: angiogenesis; artificial skin; athymic mouse; bioengineering /biomedical engineering; biomaterial compatibility; biomaterial development /preparation; biomaterial evaluation; burn therapy; cell proliferation; clinical research; collagen; cytokine; enzyme linked immunosorbent assay; fibroblasts; human subject; human therapy evaluation; keratinocyte; medical rehabilitation related tag; melanocyte; northern blottings; pigmentation; polymers; skin transplantation; tissue /cell culture; tissue engineering; wound healing

DESCRIPTION: 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 functional barrier properties, and from absence of a vascular plexus in the dermal analog. The current model of cultured skin in this laboratory is a collagen-based sponge populated with cultured human keratinocytes and fibroblasts. In vitro studies will regulate physiology by: a) media (e.g., nutrient, mitogen) and biophysical conditions that affect cellular metabolism of keratinocytes, fibroblasts and melanocytes; b) composition (structure and biochemistry) of implantable biopolymer substrates for cell delivery; and assess c) expression of barrier lipids, cytokines and matrix molecules by skin analogues before grafting. Evaluation of these factors will be performed by : a) measurements of cell growth (DNA synthesis) and epidermal differentiation (barrier ultrastructure, lipids, surface hydration); b) molecular and ultrastructural analysis of skin substitutes; and, c) ELISA and northern blot analyses of expressed xenograft, autograft, no graft) for efficacy (wound contraction, epidermal barrier, HLA-ABC), pigmentation (hue & chromaticity by chromameter) and expression of cytokines and adhesion proteins by grafting to full-thickness wounds in athymic mice. Clinical studies will focus on stimulation of angiogenesis, and maximum survival of transplanted cells during engraftment of skin substitutes. Excised burns treated with cultured skin substitutes will be studied in the clinic by paired-site comparison to treatment with meshed, split-thickness skin autograft. Comparative parameters will include: a) quantitative wound closure with skin substitutes (incidence of engraftment, ratio of healed area to biopsy area, time to wound closure, frequency of regrafting); and, b) qualitative outcome (scarring, contraction, function and cosmesis). The investigators claim to possess all of the required expertise in cell biology, skin biochemistry and biophysics, wound physiology, and clinical burn care to perform these studies successfully. Accomplishment of these objectives may 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：用培养的烧伤伤口快速有效地治愈 人类皮肤的类似物是该提案的核心目标。 医疗的 改善愈合的好处可能包括但不限于：减少 分裂自体移植的要求，住院时间较短， 并降低了恢复后的长期发病率。 但是，解剖和 所有当前培养的皮肤模型的生理缺陷均具有 这些好处的限制。 主要缺陷来自 培养条件不会产生具有功能的表皮类似物 屏障特性，以及皮肤中缺乏血管丛 类似物。 该实验室中培养的皮肤的当前模型是 基于胶原蛋白的海绵，含有培养的人角质形成细胞和 成纤维细胞。 体外研究将通过以下方式调节生理学：a）培养基（例如， 营养，有丝分裂原）和影响细胞的生物物理条件 角质形成细胞，成纤维细胞和黑素细胞的代谢； b）组成 （结构和生物化学）细胞的可植入生物聚合物底物的（结构和生物化学） 送货;并评估c）表达屏障脂质，细胞因子和基质的表达 嫁接前皮肤类似物的分子。 评估这些因素 将通过：a）细胞生长的测量（DNA合成）和 表皮分化（屏障超微结构，脂质，表面 水合）; b）皮肤替代品的分子和超微结构分析； c）ELISA和北印迹分析的异种移植，自体移植， 无移植）提高功效（伤口收缩，表皮屏障，HLA-ABC）， 色素沉着（镀铬计的色相和色度）和细胞因子的表达 和粘附蛋白通过嫁接到胸骨小鼠的全厚性伤口。 临床研究将集中于刺激血管生成和最大 皮肤替代品植入期间，移植细胞的存活。 将研究用培养的皮肤替代品处理的切除的烧伤 通过配对地点的诊所与网状，分裂厚度的处理 皮肤自体移植。 比较参数将包括：a）定量伤口 用皮肤替代品闭合（植入的发生率，愈合的比率 活检区域的区域，闭合的时间，频率的频率）；和， b）定性结果（疤痕，收缩，功能和宇宙）。 这 调查人员声称拥有细胞中所有必需的专业知识 生物学，皮肤生物化学和生物物理学，伤口生理和临床 烧伤以成功地进行这些研究。 完成这些 目标可能导致燃烧的死亡率和发病率降低， 改进的塑料和重建手术的材料以及发育 其他组织和器官替代品。