3D生物打印晚期糖基化胶原构建糖尿病炎性微环境体外研究模型的研究

Diabetic lower limb chronic wound is one of the most lethal and disabling chronic complication of diabetes. Ischemic arteriosclerosis of lower limb arteries, loss of protective sensory induced by peripheral neuropathy and damaged skin barrier are the well recognized major mechanisms. Our previous study indicated that the deposition of advanced glycation endproducts (AGEs) in these key tissues is the direct risk factor of diabetic unhealing wounds. Suck risk might related with its stimulation of macrophages and hence trigger the chronic inflammation chain. Advanced glycated collagen I (CI-AGEs) is the most common form of tissue deposited AGEs. However, the mechanism how macrophages were attracted and stimulated by the microenvironment consisted with CI-AGEs remains unclear. Moreover, suitable in vitro 3 dimensional CI-AGEs microenvironment model is still absent. We intend to fabricate the 3 dimensional bio printed scaffold using CI-AGEs and peripheral blood derived macrophages, and simulate the diabetic microenvironment with such in vitro model. The mechanism how macrophages were attracted and stimulated by such microenvironment will be investigated; the result may provide new therapeutic target and risk screening tools for diabetic foot.

糖尿病下肢慢性创面是糖尿病严重致残致死性并发症。下肢动脉缺血性粥样硬化、保护性感觉神经病损和皮肤屏障功能破坏是目前公认的糖尿病足慢性创面诱发机制。其形成原因与局部毒性微环境和炎症破坏有关。我们前期研究提示晚期糖基化I型胶原（CI-AGEs）等细胞外基质在引导局部炎症活化和毒性微环境形成方面扮演重要作用，但目前缺乏对其有效模拟的立体研究模型。本研究拟以3D生物打印的CI-AGEs三维支架为平台，接种人巨噬细胞为首的炎症细胞，通过体外培养研究糖尿病足有害微环境与炎症破坏机制。具体内容包括（1）人体组织微环境结构成分的研究建模；（2）3D生物打印制作CI-AGEs立体支架模型，接种巨噬细胞并观察其迁移穿越的细胞生物学行为和分子机制；（3）激活的巨噬细胞诱导下游炎症反应能力评价。本项目研究成果对促进临床糖尿病足风险预警，建立新治疗措施的评价体系等具有重要的学术价值与应用价值。

糖尿病患病率在全球范围内快速攀升，其慢性并发症形成原因与组织结构慢性炎症破坏有关。然而，传统的药物开发却陷入了高风险、高成本的困境。我们前期研究提示晚期糖基化I型胶原（CI-AGEs）等细胞外基质在引导局部炎症活化和毒性微环境形成方面扮演重要作用，并以3D生物打印的CI-AGEs三维支架为平台，接种人巨噬细胞为首的炎症细胞，通过体外培养模拟了AGEs诱导的炎症破坏机制。