Department of Diagnostic Science, Tufts University School of Dental Medicine, Boston, Massachusetts, USA.
Microcirculation Laboratory and Joslin-Beth Israel Deaconess Foot Center, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA.
Tissue Eng Part C Methods. 2021 Feb;27(2):49-58. doi: 10.1089/ten.TEC.2020.0263. Epub 2021 Jan 20.
A major challenge in the management of patients suffering from diabetes is the risk of developing nonhealing foot ulcers. Most methods to screen drugs for wound healing therapies rely on conventional 2D cell cultures that do not closely mimic the complexity of the diabetic wound environment. In addition, while three-dimensional (3D) skin tissue models of human skin exist, they have not previously been adapted to incorporate patient-derived macrophages to model inflammation from these wounds. In this study, we present a 3D human skin equivalent (HSE) model incorporating blood-derived monocytes and primary fibroblasts isolated from patients with diabetic foot ulcers (DFUs). We demonstrate that the monocytes differentiate into macrophages when incorporated into HSEs and secrete a cytokine profile indicative of the proinflammatory M1 phenotype seen in DFUs. We also show how the interaction between fibroblasts and macrophages in the HSE can guide macrophage polarization. Our findings take us a step closer to creating a human, 3D skin-like tissue model that can be applied to evaluate the response of candidate compounds needed for potential new foot ulcer therapies in a more complex tissue environment that contributes to diabetic wounds. Impact statement This study is the first to incorporate disease-specific, diabetic macrophages into a three-dimensional (3D) model of human skin. We show how to fabricate skin that incorporates macrophages with disease-specific fibroblasts to guide macrophage polarization. We also show that monocytes from diabetic patients can differentiate into macrophages directly in this skin disease model, and that they secrete a cytokine profile mimicking the proinflammatory M1 phenotype seen in diabetic foot ulcers. The data presented here indicate that this 3D skin disease model can be used to study macrophage-related inflammation in diabetes and as a drug testing tool to evaluate new treatments for the disease.
糖尿病患者管理面临的一个主要挑战是发生难以愈合的足部溃疡的风险。大多数筛选创伤愈合治疗药物的方法都依赖于传统的 2D 细胞培养，而这种方法不能很好地模拟糖尿病创面环境的复杂性。此外，虽然存在用于模拟人类皮肤的三维 (3D) 皮肤组织模型，但以前并未对其进行适应性改造以纳入源自患者的巨噬细胞来模拟这些伤口的炎症。在这项研究中，我们提出了一种 3D 人类皮肤等效物 (HSE) 模型，该模型包含源自糖尿病足溃疡 (DFU) 患者的血液衍生单核细胞和原代成纤维细胞。我们证明，当将单核细胞纳入 HSE 中时，它们会分化为巨噬细胞，并分泌出一种细胞因子谱，表明在 DFU 中观察到的促炎 M1 表型。我们还展示了 HSE 中成纤维细胞和巨噬细胞之间的相互作用如何指导巨噬细胞极化。我们的研究结果使我们离创建一个更复杂的组织环境下的人类 3D 皮肤样组织模型又近了一步，这种模型可用于评估候选化合物在糖尿病创面中的反应，候选化合物是潜在新的足部溃疡治疗方法所必需的，而这种更复杂的组织环境有助于糖尿病创面。
本研究首次将具有疾病特异性的糖尿病巨噬细胞纳入人类皮肤的三维 (3D) 模型中。我们展示了如何制造包含具有疾病特异性成纤维细胞的巨噬细胞的皮肤，以指导巨噬细胞极化。我们还表明，来自糖尿病患者的单核细胞可以直接在这种皮肤疾病模型中分化为巨噬细胞，并且它们分泌的细胞因子谱模拟了在糖尿病足溃疡中观察到的促炎 M1 表型。本文提供的数据表明，这种 3D 皮肤疾病模型可用于研究糖尿病中与巨噬细胞相关的炎症，并可作为一种药物测试工具，以评估该疾病的新治疗方法。
