Novel 3-D Macrophage Spheroid Model Reveals Reciprocal Regulation of Immunomechanical Stress and Mechano-Immunological Response.

IF 2.3 4区医学 Q3 BIOPHYSICS

Cellular and molecular bioengineering Pub Date : 2024-10-14 eCollection Date: 2024-10-01 DOI:10.1007/s12195-024-00824-z

Alice Burchett, Saeed Siri, Jun Li, Xin Lu, Meenal Datta

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引用次数: 0

Abstract

Purpose: In many diseases, an overabundance of macrophages contributes to adverse outcomes. While numerous studies have compared macrophage phenotype after mechanical stimulation or with varying local stiffness, it is unclear if and how macrophages directly contribute to mechanical forces in their microenvironment.

Methods: Raw 264.7 murine macrophages were embedded in a confining agarose gel, and proliferated to form spheroids over days/weeks. Gels were synthesized at various concentrations to tune stiffness and were shown to support cell viability and spheroid growth. These cell-agarose constructs were treated with media supplements to promote macrophage polarization. Spheroid geometries were used to computationally model the strain generated in the agarose by macrophage spheroid growth. Agarose-embedded macrophages were analyzed for viability, spheroid size, stress generation, and gene expression.

Results: Macrophages form spheroids and generate growth-induced mechanical forces (i.e., solid stress) within confining agarose gels, which can be maintained for at least 16 days in culture. Increasing agarose concentration increases gel stiffness, restricts spheroid expansion, limits gel deformation, and causes a decrease in Ki67 expression. Lipopolysaccharide (LPS) stimulation increases spheroid growth, though this effect is reversed with the addition of IFNγ. The mechanosensitive ion channels Piezo1 and TRPV4 have reduced expression with increased stiffness, externally applied compression, LPS stimulation, and M1-like polarization.

Conclusions: Macrophages alone both respond to and generate solid stress. Understanding how macrophage generation of growth-induced solid stress responds to different environmental conditions will help to inform treatment strategies for the plethora of diseases that involve macrophage accumulation and inflammation.

Supplementary information: The online version contains supplementary material available at 10.1007/s12195-024-00824-z.

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新型三维巨噬细胞球体模型揭示免疫机械应力与机械免疫反应的相互调控关系

目的：在许多疾病中，巨噬细胞过多会导致不良后果。虽然许多研究比较了巨噬细胞在机械刺激或不同局部硬度下的表型，但还不清楚巨噬细胞是否以及如何直接作用于其微环境中的机械力：方法：将原始 264.7 小鼠巨噬细胞嵌入封闭的琼脂糖凝胶中，经过数天/数周的增殖形成球体。凝胶以不同浓度合成，以调节硬度，结果表明凝胶支持细胞存活和球体生长。用培养基补充剂处理这些细胞-琼脂糖构建体，以促进巨噬细胞极化。球形体的几何形状被用于对巨噬细胞球形体生长在琼脂糖中产生的应变进行计算建模。对琼脂糖包埋的巨噬细胞的活力、球形体大小、应力产生和基因表达进行了分析：结果：巨噬细胞在封闭的琼脂糖凝胶中形成球体并产生生长诱导的机械力（即固体应力），在培养过程中至少可维持 16 天。增加琼脂糖浓度会增加凝胶硬度、限制球体扩张、限制凝胶变形并导致 Ki67 表达下降。脂多糖（LPS）刺激会增加球形体的生长，但加入 IFNγ 后这种效应会逆转。机械敏感性离子通道 Piezo1 和 TRPV4 的表达随着硬度增加、外部施加的压力、LPS 刺激和 M1 样极化而减少：结论：巨噬细胞本身既能对固体应力做出反应，也能产生固体应力。了解巨噬细胞生成生长诱导的固体应力如何应对不同的环境条件，将有助于为涉及巨噬细胞聚集和炎症的多种疾病的治疗策略提供信息：在线版本包含补充材料，可查阅 10.1007/s12195-024-00824-z。

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来源期刊

Cellular and molecular bioengineering 生物-生物物理

CiteScore

5.60

自引率

3.60%

发文量

审稿时长

>12 weeks

期刊介绍： The field of cellular and molecular bioengineering seeks to understand, so that we may ultimately control, the mechanical, chemical, and electrical processes of the cell. A key challenge in improving human health is to understand how cellular behavior arises from molecular-level interactions. CMBE, an official journal of the Biomedical Engineering Society, publishes original research and review papers in the following seven general areas: Molecular: DNA-protein/RNA-protein interactions, protein folding and function, protein-protein and receptor-ligand interactions, lipids, polysaccharides, molecular motors, and the biophysics of macromolecules that function as therapeutics or engineered matrices, for example. Cellular: Studies of how cells sense physicochemical events surrounding and within cells, and how cells transduce these events into biological responses. Specific cell processes of interest include cell growth, differentiation, migration, signal transduction, protein secretion and transport, gene expression and regulation, and cell-matrix interactions. Mechanobiology: The mechanical properties of cells and biomolecules, cellular/molecular force generation and adhesion, the response of cells to their mechanical microenvironment, and mechanotransduction in response to various physical forces such as fluid shear stress. Nanomedicine: The engineering of nanoparticles for advanced drug delivery and molecular imaging applications, with particular focus on the interaction of such particles with living cells. Also, the application of nanostructured materials to control the behavior of cells and biomolecules.