DEVELOPMENT OF AN IN VITRO WOUND HEALING MODEL

这个研究项目的长期目标是提高我们对 伤口愈合多学科、体外和体内研究将 描述细胞-基质相互作用及其在调节 皮肤修复研究将侧重于 再上皮化和纤维增生。在未来5年内，我们的目标是回答 以下问题： 1.表皮粘附的激活是否受基底膜和 增长因素？我们将分析表达中发生的变化 表皮细胞活化过程中的特异性整合素粘附受体 粘附和迁移。我们将确定基底膜和 转化生长因子β（TGF-β）调节表皮细胞活化 细胞粘附 2.细胞粘附蛋白和生长因子对 结缔组织生物发生的收缩介导的调节？我们将 使用长期的、补充抗坏血酸的成纤维细胞培养物作为体外 新结缔组织生物发生模型。我们会研究规管 在细胞外基质收缩的影响下， 粘附蛋白（纤连蛋白和玻连蛋白）和血小板释放 因子（TGF-β和PDGF）。 3.如何释放收缩胶原蛋白凝胶从紧张调节 细胞生长和生物合成活性胶原蛋白凝胶收缩， 成纤维细胞将通过机械操作从张力中释放。 随后的细胞形态和生物合成活性的变化将是 测定了 4.含精氨酸-甘氨酸-谷氨酸（RGE）的胶原肽抑制创伤吗 宫缩？对应于I型胶原蛋白的含RGE的肽 将测试序列抑制胶原凝胶的能力， 体外收缩。将测试最活跃的肽，以了解 它抑制体内全层伤口收缩。 5.粘附蛋白在创面中的分布和活性如何 流体？我们将使用几种类型的伤口液体：抽吸水泡，慢性 皮肤溃疡、烧伤和外科手术（乳房切除术）。粘附分布 各种伤口液体中的蛋白质和 将对伤口进行分析。

The long term goal of this research project is to enhance our understanding of wound healing. Multidisciplinary, in vitro and in vivo studies will describe cell-matrix interactions and their function in the regulation of cutaneous repair. Research will focus on the mechanisms of re-epithelization and fibroplasia. During the next 5 years we aim to answer the following questions: 1. Is activation of epidermal adhesion regulated by basement membrane and growth factors? We will analyze changes that occur in the expression of specific integrin adhesion receptors during activation of epidermal adhesion and migration. We will determine if basement membrane and transforming growth factor beta (TGF-BETA) regulate activation of epidermal cell adhesion. 2. What are the effects of cell adhesion proteins and growth factors on contraction-mediated regulation of connective tissue biogenesis? We will use long term, ascorbate-supplemented, fibroblast cultures as an in vitro model of new connective tissue biogenesis. We will study the regulation of tissue biogenesis by extracellular matrix contraction under the influence of adhesion proteins (fibronectin and vitronectin) and platelet released factors (TGF-BETA and PDGF). 3.How does the release of contracted collagen gels from tension regulate cell growth and biosynthetic activity? Collagen gels contracted by fibroblasts will be released from tension by mechanical manipulation. Subsequent changes in cell morphology and biosynthetic activity will be measured. 4. Do arg-gly-glu (RGE)-containing collagen peptides inhibit wound contraction? RGE-containing peptides that correspond to collagen type I sequences will be tested for their ability to inhibit collagen gel contraction in vitro. The most active peptide will be tested to learn if it inhibits full thickness wound contraction in vivo. 5. What is the distribution and activity of adhesion proteins in wound fluid? We will use several types of wound fluid: suction blister, chronic skin ulcer, burn, and surgical (mastectomy). The distribution of adhesion proteins in the various wound fluids and proteolysis of fibronectin in the wounds will be analyzed.