Development of a multi cellular in vitro model to study protective and regenerative therapy strategies ofchronically cutaneous wounds

辐射可导致皮肤创面愈合的妥协，具有重要的临床后果，从病理性瘢痕到危及生命的主要血管的扭曲需要广泛的重建努力。参与病理性创面愈合的机制复杂且不完全了解。促进创面愈合过程的治疗干预措施很少，研究较少，尚未最终证明能改善临床结果。这些措施包括各种药物干预以及干细胞移植。目前还没有能充分反映受损伤口愈合的病理生理学的体外模型。已建立的模型的一个重要缺陷是它们没有整合影响伤口愈合过程的多种细胞类型及其细胞间的相互作用。因此，开发一种体外伤口愈合模型，模拟外部辐射对伤口愈合过程的影响和不良反应，并允许研究治疗干预的效果是本项目的目标。计划中的项目旨在开发一种多细胞体外模型，该模型比先前描述的单细胞模型更好地反映皮肤创伤愈合的病理生理过程。慢性伤口愈合损害的关键方面包括萎缩和溃疡。这两个方面与两种关键的细胞类型相关：成纤维细胞和内皮细胞，因此将被整合到目前的模型中。人类成脂干细胞移植是最有希望的治疗干预措施，也将被整合到计划的模型中。它们的治疗效果已被描述为通过药物调节而优化。我们的初步数据表明，促炎细胞因子，如白亮氨酸-6，以及微循环标志物，如可溶性黏附分子ICAM-1和VCAM-1，与辐射诱导的伤口愈合损害的产生具有特殊的相关性。这些标记物和其他标记物将用于在多细胞体外模型中量化辐射和药物调节对辐射诱导的伤口愈合妥协的影响。

Irradiation may lead to a compromise in cutaneous wound healing with important clinical consequences, ranging from pathological scarring to life threatening arrosion of major vessels requiring extensive reconstructive efforts.The mechansims involved in pathological wound healing are complex and incompletely understood.Therapeutic interventions to enhance the wound healing process are scarce, poorly researched and have not been conclusively shown to improve the clinical outcome. These include various pharmacological interventions as well as stem cell transplantation. No in vitro model exists that adequately reflects the pathophysiology of compromised wound healing. An important limitation inherent to established models is that they do not integrate the multiple cell types and their inter-cellular interactions affecting the wound healing process.The development of an in vitro wound healing model, which simulates the effects and adverse responses to external radiation on the wound healing process and allows to investigate the effects of therapeutic intervention is hence the goal of the present project.The planned project is aimed to develop a multi cellular in vitro model, which mirrors the pathophysiological processes of cutaneous wound healing better than previously described mono-cellular models. Key aspects of chronic wound healing compromise include atrophy and ulceration. These two aspects correlate with two key cell types: fibroblasts and endothelial cells, which will hence be integrated in the present model.Human adipogenic stem cell transplantation is the most promising therapeutic intervention available and will be integrated in the planned model as well. Their therapeutic effect has been described to be optimized with pharmacological modulation. Hence the effects of pharmacological modulation are to be investigated in the present model as well.Our preliminary data suggest that pro-inflammatory cytokines, such as interleucine-6 and markers of microcirculation, like soluble adhesion molecules ICAM-1 and VCAM-1, are of special relevance for the generation of radiation induced wound healing compromise. These and other markers will serve to quantify the effects of irradiation and pharmacological modulation on radiation-induced wound healing compromise in a multi-cellular in vitro model.