Substrate stiffness induces neutrophil extracellular trap (NET) formation through focal adhesion kinase activation.
Substrate stiffness induces neutrophil extracellular trap (NET) formation through focal adhesion kinase activation.
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DOI：
10.1016/j.biomaterials.2021.120715
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发表时间：
2021-04
期刊：
影响因子：
14
通讯作者：
Olivares-Navarrete R
中科院分区：
文献类型：
--
作者：
Abaricia JO;Shah AH;Olivares-Navarrete R
中性粒细胞在组织损伤和生物材料植入的早期炎症反应中占主导地位。最近的研究表明，中性粒细胞活化可以通过机械线索，如刚度或表面润湿性进行调节;然而，尚不清楚中性粒细胞如何感知和响应物理线索，特别是它们如何形成中性粒细胞胞外陷阱（NET形成）。为了检验这一点，我们使用不同的生理相关刚度（0.2-32 kPa）的聚二甲基硅氧烷（PDMS）基板，并检查小鼠中性粒细胞的反应，未经处理的表面或表面包被各种细胞外基质蛋白识别的整合素异二聚体（胶原蛋白，纤连蛋白，层粘连蛋白，玻连蛋白，合成的RGD）。较高硬度PDMS基质上的中性粒细胞增加了NET形成，并分泌了更多的促炎细胞因子和趋化因子。细胞外基质蛋白涂层表明，纤连蛋白诱导最NET的形成，这种效果是刚度依赖性。合成的RGD肽诱导的NET形成和促炎细胞因子释放水平与全长纤连蛋白相似。为了确定所观察到的响应于底物硬度的NET形成是否需要粘着斑激酶（FAK）活性，其是整联蛋白活化的下游，使用FAK抑制剂PF-573228。使用PF-573228抑制FAK消除了NET形成和促炎分子分泌的硬度依赖性增加。这些发现表明，中性粒细胞调节NET的形成响应于物理和机械生物材料的线索，这一过程是通过整联蛋白/FAK信号调节。
Neutrophils predominate the early inflammatory response to tissue injury and implantation of biomaterials. Recent studies have shown that neutrophil activation can be regulated by mechanical cues such as stiffness or surface wettability; however, it is not known how neutrophils sense and respond to physical cues, particularly how they form neutrophil extracellular traps (NET formation). To examine this, we used polydimethylsiloxane (PDMS) substrates of varying physiologically relevant stiffness (0.2-32 kPa) and examined the response of murine neutrophils to untreated surfaces or to surfaces coated with various extracellular matrix proteins recognized by integrin heterodimers (collagen, fibronectin, laminin, vitronectin, synthetic RGD). Neutrophils on higher stiffness PDMS substrates had increased NET formation and higher secretion of pro-inflammatory cytokines and chemokines. Extracellular matrix protein coatings showed that fibronectin induced the most NET formation and this effect was stiffness dependent. Synthetic RGD peptides induced similar levels of NET formation and pro-inflammatory cytokine release than the full-length fibronectin protein. To determine if the observed NET formation in response to substrate stiffness required focal adhesion kinase (FAK) activity, which is down stream of integrin activation, FAK inhibitor PF-573228 was used. Inhibition of FAK using PF-573228 ablated the stiffness-dependent increase in NET formation and pro-inflammatory molecule secretion. These findings demonstrate that neutrophils regulate NET formation in response to physical and mechanical biomaterial cues and this process is regulated through integrin/FAK signaling.
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影响因子：
14
作者：
Abaricia, Jefferson O.;Shah, Arth H.;Olivares-Navarrete, Rene
通讯作者：
Olivares-Navarrete, Rene
DOI：
10.1186/s12952-015-0042-8
发表时间：
2015-01-01
期刊：
JOURNAL OF NEGATIVE RESULTS IN BIOMEDICINE
影响因子：
--
作者：
Acevedo-Acevedo, Suehelay;Crone, Wendy C.
通讯作者：
Crone, Wendy C.
影响因子：
4.3
作者：
Cox TR;Erler JT
通讯作者：
Erler JT
影响因子：
7.3
作者：
Erpenbeck, Luise;Gruhn, Antonia Luise;Kruss, Sebastian
通讯作者：
Kruss, Sebastian
影响因子：
9.7
作者：
Hotchkiss KM;Reddy GB;Hyzy SL;Schwartz Z;Boyan BD;Olivares-Navarrete R
通讯作者：
Olivares-Navarrete R