Article(id=1320176633932263961, tenantId=1146029695717560320, journalId=1301849854269554754, issueId=1320176147946648091, articleNumber=null, orderNo=null, doi=10.11729/syltlx20250121, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1766419200000, receivedDateStr=2025-12-23, revisedDate=1769443200000, revisedDateStr=2026-01-27, acceptedDate=1770566400000, acceptedDateStr=2026-02-09, onlineDate=1792684237224, onlineDateStr=2026-10-22, pubDate=1787587200000, pubDateStr=2026-08-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1792684237224, onlineIssueDateStr=2026-10-22, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1792684237224, creator=13701087609, updateTime=1792684237224, updator=13701087609, issue=Issue{id=1320176147946648091, tenantId=1146029695717560320, journalId=1301849854269554754, year='2026', volume='40', issue='4', pageStart='1', pageEnd='137', issueExtLink='null', onlineDate='null', pubDate='1787587200000', pubDateStr='2026-08-25', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1792684121357, creator='13701087609', updateTime=1792720571222, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1320329029853495708, tenantId=1146029695717560320, journalId=1301849854269554754, issueId=1320176147946648091, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1320329029853495709, tenantId=1146029695717560320, journalId=1301849854269554754, issueId=1320176147946648091, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=97, endPage=104, ext={EN=ArticleExt(id=1320176634129396250, articleId=1320176633932263961, tenantId=1146029695717560320, journalId=1301849854269554754, language=EN, title=Mechanobiological analysis of cellular compression in narrow microfluidic channels, columnId=1320176148672262684, journalTitle=Journal of Experiments in Fluid Mechanics, columnName=Special Issue on Microscale and Nanoscale Flows, runingTitle=null, highlight=null, articleAbstract=

The compressive stress experienced by Circulating Tumor Cells (CTCs) in the microcirculation is a key mechanical factor affecting their metastatic potential. This study aims to investigate the independent regulatory effects of compressive stress intensity and duration on cell viability. Referring to the typical diameter of the MCF−7 cell line, microfluidic channels with different widths (9, 12 and 15 µm) and lengths (100 and 500 µm) were designed to construct an in vitro mechanical stimulation platform, simulating the capillary environment and decoupling the two mechanical parameters. The dynamic process and transit time of MCF−7 cells passing through the microchannels were recorded using high-speed microimaging technology. Fluorescence staining and image analysis were employed to quantitatively evaluate the proliferation and adhesion capabilities of cells after different mechanical stimulations. Results show that the cell transit time is primarily determined by the channel length, while a decrease in channel width increases the dispersion of transit velocities. Regarding cell viability, under short-duration compressive stimulation (approx. 3 ms), cell proliferation ability decreased with increasing stress intensity; whereas under long-duration stimulation (approx. 15 ms), it showed a trend of first enhancing and then weakening. Furthermore, compressive stress significantly inhibited cell adhesion ability, and the effect was more pronounced with prolonged stimulation. This study provides support for deciphering the regulation of cell fate by stress stimulation intensity and time, offering experimental evidence for a deeper understanding of the metastasis mechanism of circulating tumor cells.

, authors=Yiyang Chen1, 2, Jiaxin Rong2, Xiaobo Gong1, *, authorsList=Yiyang Chen, Jiaxin Rong, Xiaobo Gong, authorCompany=null, correspAuthors=Xiaobo Gong, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright ©2026 Journal of Experiments in Fluid Mechanics. All rights reserved., copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1320176641037414957, articleId=1320176633932263961, tenantId=1146029695717560320, journalId=1301849854269554754, language=CN, title=狭窄微流控通道中细胞挤压力生物学分析, columnId=1320176148844229150, journalTitle=实验流体力学, columnName=微纳尺度流动专刊, runingTitle=null, highlight=null, articleAbstract=

循环肿瘤细胞在微循环中经历的挤压应力是影响其转移潜能的关键力学因素。本文旨在探究挤应压力强度与持续时间对细胞活性的独立调控作用,参考MCF−7细胞系的典型直径,设计具有不同宽度(9、12和15 µm)和长度(100和500 µm)的微流控通道,构建了体外力学刺激平台,在模拟毛细血管环境的同时,实现对上述2个力学参数的解耦。利用高速显微成像技术记录了MCF−7细胞通过微通道的动态过程和时间,并采用荧光染色与图像分析法定量评估了细胞经受不同力学刺激后的增殖能力与黏附能力。结果表明,细胞通过微通道的时间主要由通道长度决定,而通道宽度减小会增大细胞通行速度的离散性。在细胞活性方面,短时间(约3 ms)挤压刺激下,细胞增殖能力随应力强度增大而减弱;而长时间(约15 ms)挤压刺激下,其增殖能力则呈先增强后减弱的趋势。此外,挤压应力显著抑制了细胞黏附能力,且长时间刺激的抑制效应更为明显。本文为解析应力刺激的强度和时间对细胞命运的调控作用提供了支撑,也为深入理解循环肿瘤细胞的转移机制提供了实验依据。

, authors=陈亦扬1, 2, 戎佳欣2, 龚晓波1, *, authorsList=陈亦扬, 戎佳欣, 龚晓波, authorCompany=null, correspAuthors=龚晓波, authorNote=

陈亦扬(2002—),男,福建泉州人,博士研究生。研究方向:生物力学,微流控技术。E-mail:

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*represents compared to the control group, p < 0.05), figureFileSmall=yBCpFTmgftr+GBSkU8pIsw==, figureFileBig=doQ9esI6m3urKXPzrz6Lxg==, tableContent=null), ArticleFig(id=1320177140587409778, tenantId=1146029695717560320, journalId=1301849854269554754, articleId=1320176633932263961, language=CN, label=图7, caption=接种后48 h细胞数量(数值为均值 ± 标准差;*表示与Control比较,p < 0.05), figureFileSmall=yBCpFTmgftr+GBSkU8pIsw==, figureFileBig=doQ9esI6m3urKXPzrz6Lxg==, tableContent=null), ArticleFig(id=1320177140662907251, tenantId=1146029695717560320, journalId=1301849854269554754, articleId=1320176633932263961, language=EN, label=Fig.8, caption=Cell spreading area at 6 hours post-seeding (the box represents the 25th–75th percentile, IQR; the central line is the median; the whiskers extend to the furthest data points within 1.5 IQR; * and * * * * respectively indicate that compared with control group, p < 0.05 and p < 0.0001), figureFileSmall=NjD4p7/gV/q4QzkjJj4zOA==, figureFileBig=tqPyr7YBnfUdDZF/iY1sAQ==, tableContent=null), ArticleFig(id=1320177140734210420, tenantId=1146029695717560320, journalId=1301849854269554754, articleId=1320176633932263961, language=CN, label=图8, caption=接种后6 h细胞铺展面积(箱体表示0.25分位数~0.75分位数,IQR;中线为中位数;须线延伸至不超过1.5倍IQR范围内的最远数据点;*和****分别表示与Control比较,p < 0.05和p < 0.0001), figureFileSmall=NjD4p7/gV/q4QzkjJj4zOA==, figureFileBig=tqPyr7YBnfUdDZF/iY1sAQ==, tableContent=null), ArticleFig(id=1320177140813902197, tenantId=1146029695717560320, journalId=1301849854269554754, articleId=1320176633932263961, language=EN, label=Table 1, caption=

Statistical significance of cell spreading area

, figureFileSmall=null, figureFileBig=null, tableContent=
通道类型平均值标准差p值
Control561.76325.03
Shear538.46316.290.37376
9−100520.81259.360.08854
12−100529.51255.520.17712
15−100513.58268.220.04811(*)
9−500434.97211.21< 0.0001(****)
12−500461.35233.18< 0.0001(****)
15−500500.43260.110.01096(*)
), ArticleFig(id=1320177141166223734, tenantId=1146029695717560320, journalId=1301849854269554754, articleId=1320176633932263961, language=CN, label=表1, caption=

细胞铺展面积的统计学显著性检验结果

, figureFileSmall=null, figureFileBig=null, tableContent=
通道类型平均值标准差p值
Control561.76325.03
Shear538.46316.290.37376
9−100520.81259.360.08854
12−100529.51255.520.17712
15−100513.58268.220.04811(*)
9−500434.97211.21< 0.0001(****)
12−500461.35233.18< 0.0001(****)
15−500500.43260.110.01096(*)
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狭窄微流控通道中细胞挤压力生物学分析
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陈亦扬 1, 2 , 戎佳欣 2 , 龚晓波 1, *
实验流体力学 | 微纳尺度流动专刊 2026,40(4): 97-104
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实验流体力学 |微纳尺度流动专刊 2026 , 40 (4) : 97 -104
狭窄微流控通道中细胞挤压力生物学分析
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陈亦扬1, 2 , 戎佳欣2, 龚晓波1, *
作者信息
  • 1上海交通大学 船舶海洋与建筑工程学院,上海 200240
  • 2南方科技大学 海洋高等研究院,深圳 518055
通讯作者:
作者简介:

陈亦扬(2002—),男,福建泉州人,博士研究生。研究方向:生物力学,微流控技术。E-mail:

Mechanobiological analysis of cellular compression in narrow microfluidic channels
Yiyang Chen1, 2 , Jiaxin Rong2, Xiaobo Gong1, *
Affiliations
  • 1School of Ocean and Civil Engineering, Shanghai Jiao Tong University, Shanghai 200240, China
  • 2Advanced Institute for Ocean Research, Southern University of Science and Technology, Shenzhen 518055, China
出版时间: 2026-08-25 doi: 10.11729/syltlx20250121
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循环肿瘤细胞在微循环中经历的挤压应力是影响其转移潜能的关键力学因素。本文旨在探究挤应压力强度与持续时间对细胞活性的独立调控作用,参考MCF−7细胞系的典型直径,设计具有不同宽度(9、12和15 µm)和长度(100和500 µm)的微流控通道,构建了体外力学刺激平台,在模拟毛细血管环境的同时,实现对上述2个力学参数的解耦。利用高速显微成像技术记录了MCF−7细胞通过微通道的动态过程和时间,并采用荧光染色与图像分析法定量评估了细胞经受不同力学刺激后的增殖能力与黏附能力。结果表明,细胞通过微通道的时间主要由通道长度决定,而通道宽度减小会增大细胞通行速度的离散性。在细胞活性方面,短时间(约3 ms)挤压刺激下,细胞增殖能力随应力强度增大而减弱;而长时间(约15 ms)挤压刺激下,其增殖能力则呈先增强后减弱的趋势。此外,挤压应力显著抑制了细胞黏附能力,且长时间刺激的抑制效应更为明显。本文为解析应力刺激的强度和时间对细胞命运的调控作用提供了支撑,也为深入理解循环肿瘤细胞的转移机制提供了实验依据。

微流控  /  循环肿瘤细胞  /  挤压应力  /  力学生物学  /  细胞活性  /  细胞黏附

The compressive stress experienced by Circulating Tumor Cells (CTCs) in the microcirculation is a key mechanical factor affecting their metastatic potential. This study aims to investigate the independent regulatory effects of compressive stress intensity and duration on cell viability. Referring to the typical diameter of the MCF−7 cell line, microfluidic channels with different widths (9, 12 and 15 µm) and lengths (100 and 500 µm) were designed to construct an in vitro mechanical stimulation platform, simulating the capillary environment and decoupling the two mechanical parameters. The dynamic process and transit time of MCF−7 cells passing through the microchannels were recorded using high-speed microimaging technology. Fluorescence staining and image analysis were employed to quantitatively evaluate the proliferation and adhesion capabilities of cells after different mechanical stimulations. Results show that the cell transit time is primarily determined by the channel length, while a decrease in channel width increases the dispersion of transit velocities. Regarding cell viability, under short-duration compressive stimulation (approx. 3 ms), cell proliferation ability decreased with increasing stress intensity; whereas under long-duration stimulation (approx. 15 ms), it showed a trend of first enhancing and then weakening. Furthermore, compressive stress significantly inhibited cell adhesion ability, and the effect was more pronounced with prolonged stimulation. This study provides support for deciphering the regulation of cell fate by stress stimulation intensity and time, offering experimental evidence for a deeper understanding of the metastasis mechanism of circulating tumor cells.

microfluidics  /  Circulating Tumor Cells (CTCs)  /  compressive stress  /  mechanobiology  /  cell viability  /  cell adhesion
陈亦扬, 戎佳欣, 龚晓波. 狭窄微流控通道中细胞挤压力生物学分析. 实验流体力学, 2026 , 40 (4) : 97 -104 . DOI: 10.11729/syltlx20250121
Yiyang Chen, Jiaxin Rong, Xiaobo Gong. Mechanobiological analysis of cellular compression in narrow microfluidic channels[J]. Journal of Experiments in Fluid Mechanics, 2026 , 40 (4) : 97 -104 . DOI: 10.11729/syltlx20250121
循环肿瘤细胞(Circulating Tumor Cells, CTCs)是由原发癌组织脱落并进入血液循环系统的肿瘤细胞,是癌症经血液循环系统发生转移的关键载体[1-4]。CTCs在循环过程中必须适应不同血管区段的血流动力学环境,暴露于不同程度的流动剪切应力和血管壁挤压应力之下[5-7],这些力学刺激是CTCs诱发转移的关键生物力学基础之一。
由于肿瘤细胞的典型直径(15~20 μm)通常大于毛细血管的管径(3~12 μm),当CTCs进入毛细血管网时,会因物理尺寸不匹配而发生滞留和形变,并承受显著的血管壁挤压应力[8]。从时间尺度来看,CTCs在主流血管中通常仅短暂暴露于较高的剪切应力下(数秒量级),而在微循环中则可能长时间滞留并持续受到挤压应力的作用[9]。这一时空差异提示,微循环中的挤压应力刺激可能对CTCs的力学生物学响应产生更为深远的影响。国内外研究普遍证实,CTCs穿过狭窄的毛细血管网时会经历剧烈的机械挤压[10],细胞通过机械转导感知外界力学微环境的变化[11-13],进而引发一系列适应性机制,包括重塑细胞骨架[14-15]、改变核纤层蛋白表达[16],从而增强其变形能力和迁移能力。此外,挤压应力还能诱导细胞出泡[17-18]和上皮−间充质转化(Epithelial-mesenchymal transition, EMT)[19-20]等过程,进一步提高CTCs的存活率与侵袭转移潜能。
为在体外深入研究CTCs所经历的力学微环境,微流控芯片提供了一种理想的研究平台。微流控芯片是能够在微米尺度上精确操控流体的“芯片实验室”(lab on a chip)系统,其优势在于能够仿生构建关键尺寸与真实毛细血管相似的微通道,从而高度模拟体内的生理环境。近年来,利用微流控技术研究细胞在受限环境中的挤压响应已取得显著进展。例如,Hou等[21]设计了用于区分良性和恶性乳腺肿瘤细胞刚度差异的狭窄微通道;Nath等[22]构建了毛细血管微流控模型,评估了HeLa细胞受挤压时的流动行为及挤压后活力;Yuan等[23]探究了单个血管内皮细胞通过狭窄微通道时的钙离子动态信号变化;Cognart等[20]设计了带有等间隔收缩−舒张段的通道,以研究流动剪切应力与通道挤压应力的耦合影响;Song等[24]利用高通量微流控装置,通过挤压诱导神经元细胞发生重编程。然而,这些研究大多聚焦于细胞自身力学或生理特性的变化,虽部分涉及通道狭窄程度(即应力强度)的影响,但目前仍缺乏对挤压应力强度与持续时间进行双参数解耦分析的研究,难以全面揭示力学刺激的时空特性对细胞命运的联合调控机制。
综上所述,本文从细胞力学生物学角度出发,围绕CTCs在血液系统微循环中的受力过程,采用微流控芯片体外模型实验研究方法,通过设计具有不同宽度和长度的微通道,实现对上述2个关键力学参数的独立调控与解耦分析,旨在揭示其对CTCs活性(包括增殖能力和黏附能力)的影响。
本文基于实验测得的MCF−7细胞系平均直径(17 µm,见图1),设计了7种不同的微流控芯片(结构示意见图2),以模拟微循环中不同强度和持续时间的挤压应力刺激。芯片主要分为2类:无收缩微通道的微流控芯片(剪切组)和有收缩微通道的微流控芯片(挤压组)。剪切组芯片由2个对称的出入口和1条直通道组成,直通道的宽度和高度均大于细胞直径,用于模拟流动中细胞受剪切应力刺激的过程。挤压组芯片包含2个对称的出入口、2条平行的直通道以及位于直通道之间的50个微通道,微通道高度均为15 μm;其中,微通道宽度反映挤压应力刺激的强度,设置了9、12和15 μm 3种类型;微通道长度反映挤压应力刺激的持续时间,设置了100和500 μm2种类型。通过不同的宽度与长度组合,形成6种不同的应力加载条件,从而在体外复现CTCs在微循环中的力学微环境。
基于软光刻方法制作聚二甲基硅氧烷(Polydimethylsiloxane, PDMS, Sylgard)微流控芯片,微通道结构使用AutoCAD软件设计。首先使用SU−8光刻胶在硅板上制作模板,然后按照配胶、匀胶、脱气、浇筑、固化、剥离、打孔、清洗、干燥等标准微加工流程处理。将得到的PDMS微流控芯片和洁净载玻片进行氧气等离子体处理(PLASMA清洗机,Harrick Plasma)后键合,随后放置在110 ℃热台上加热90 min以加固键合,得到密闭性良好的微流控芯片。实验时,在微流控芯片的入口和出口处插入外径1/32 inch(1 inch = 2.54 cm)、内径0.5 mm的PEEK管,用于连接微通道与外部流体。
本文选用人乳腺癌MCF−7细胞系作为实验模型,细胞使用RPMI−1640完全培养基进行培养。完全培养基组成(以体积分数计)为88%基础培养基(Gibco)、10%胎牛血清(Gibco)、1%丙酮酸钠(Gibco)及1%青霉素−链霉素双抗(Gibco),置于37 ℃、CO2体积分数为5%的恒温培养箱中培养。每2~3天更换一次新鲜培养基,以维持细胞状态。当细胞融合度达到80%~90%时,使用质量浓度为2.5 g/L的胰蛋白酶−EDTA消化液(Gibco)进行消化传代。所有实验均使用处于对数生长期的细胞。
实验开始前,使用质量浓度为2.5 g/L的胰蛋白酶−EDTA消化液消化收集细胞,随后用10 μm细胞筛吹打过滤,获得单细胞悬液。将细胞重悬于含泊洛沙姆188(Gibco,质量浓度10 g/L)的RPMI−1640基础培养基,调整细胞密度至3 × 106 cells/mL备用。
挤压应力加载平台的组成如图3所示,主要包括可编程注射泵、1000 μL玻璃进样器、PEEK管(外径1/32 inch,内径0.5 mm)、微流控芯片和1.5 mL离心管。具体连接方式为:玻璃进样器固定于可编程注射泵上方,其出口通过PEEK管连接至微流控芯片入口;芯片出口则经另一段PEEK管连接至离心管,用以收集刺激后的细胞悬液。
实验前,所有流体接触部件均使用75%体积分数的乙醇清洗消毒,随后以磷酸盐缓冲溶液(Gibco)冲洗残留乙醇。实验时,将制备好的细胞悬液注入进样器,设置注射泵推进流量,使不同宽度收缩微通道内的平均流速均维持在20 mm/s。为保持细胞悬浮均匀,避免沉降,在进样器内放置钢珠,并利用外部磁铁进行间歇性搅拌。每次实验收集150 μL细胞悬液,整个操作流程控制在4 h内,以确保细胞活性不受长时间操作的影响。
细胞在微通道内的运动行为,采用配备高速摄像系统(Phantom)的倒置显微镜(Olympus)进行明场实时观测。实验前,将微流控芯片固定于显微镜载物台上,先以高流速通入细胞悬液以排除通道内气泡,再将流速调整至预设实验流速,待流动稳定后开始图像采集。
图像采集使用10倍物镜,通过Phantom Camera Control(PCC)软件控制相机参数,设置拍摄帧率为2000帧/s,曝光时间为0.5 ms,图像以TIFF格式保存。细胞通过微通道的持续时间定义为从细胞前端被捕获进入通道至其尾端完全离开通道的时间。为保证数据可靠性,在后续分析中进行了人工筛选,排除通道堵塞、多细胞同时进入等异常情况所对应的无效数据。
细胞经微通道挤压处理后,重悬为每 150 μL含3000个细胞的悬液,按150 μL/孔接种于96孔板中,继续于培养箱中培养。为评估细胞增殖活性,于接种后24和48 h这2个时间点,分别向各孔加入Vybrant™ DyeCycle™ Green(Invitrogen)核染色剂(终体积分数为1.25 × 10−3),置于37 ℃ 环境中避光孵育30 min。
荧光成像使用倒置荧光显微镜配合20倍物镜进行。设置激发光波长为488 nm,通过GFP滤光片组采集荧光信号。图像采集使用背照式sCMOS相机(Prime BSI),并通过VisView软件编程控制载物台移动和相机拍摄,最终拼接生成覆盖整个孔底的荧光图像,用于后续细胞计数与分析。
使用ImageJ软件对获取的全孔底荧光图像进行细胞计数分析。具体流程如下:首先进行空间标定,随后将图像转换为二值图像,并通过Watershed算法对粘连的细胞核进行分割,最后利用“Analyze Particles”功能自动识别并统计细胞数量。
细胞经微通道挤压处理后,以150 μL(3000个细胞)/孔的密度重悬并接种于96孔板中,继续于培养箱中培养6 h。随后,使用配备20倍物镜的荧光显微镜,在微分干涉对比(DIC)模式下观察细胞。图像采集通过VisView软件控制,曝光时间设为自动,亮度预设为50%,以记录细胞铺展形态。
细胞图像分析流程如下:首先,使用Cellpose 3.0软件[25]预估细胞直径并选择cyto3模型进行自动分割,生成细胞蒙版并经人工校正后保存;随后,将蒙版文件导入ImageJ软件,设定空间标尺后,利用MorphoLibJ插件中的“Analyze Regions”功能分析每个细胞的面积。
微通道挤压实验记录30个细胞通过不同长度与宽度组合微通道的所需时间。用微通道长度除以细胞通过时间得到细胞通过微通道的平均速度,数据以箱线图表示。由于数据可能不满足正态分布,采用非参数的Kruskal−Wallis检验分析6种不同挤压条件下细胞通过微通道平均速度的差异性。
增殖能力检测实验设置3个复孔,独立重复3次。每次实验的结果取3个复孔的均值。所有数据以均值 ± 标准差(Mean ± SD)表示。采用未配对双样本t检验比较应力刺激组与空白对照组的差异,显著性水平α设定为0.05。
黏附能力检测实验中,对每组应力刺激条件在单次实验中测量100个细胞的铺展面积,将3次独立重复实验的数据合并(每组共计300个细胞),以箱线图展示数据分布,并叠加正态分布曲线。采用未配对双样本t检验分析应力刺激组与空白对照组之间细胞铺展面积的差异,显著性水平α设定为0.05。
细胞通过微通道的过程如图4所示,整个过程可分为3个连续阶段:①细胞被微通道捕获并发生形变进入通道;②细胞在受限空间内的迁移运动;③细胞从微通道出口脱离。由影像可知,细胞为适应狭窄的微通道结构而发生显著形变,在通道内以挤压−拉伸的形态完成迁移,且在脱离微通道时仍维持其拉长的子弹状形态。这一结果表明,本文所构建的微通道系统能够实现对细胞的挤压应力加载,为研究细胞受挤压应力后的力学生物学变化提供了可靠的技术平台。
细胞通过不同类型微通道的平均速度如图5所示,其中箱体上、下分别代表第25和75百分位数,内部横线代表中位数;须线上、下端分别代表从箱体边缘向外延伸1.5倍四分位距内的最远数据点。由于注射泵设定的平均流速相同,细胞在不同宽度−长度组合的微通道内运动的平均速度基本一致。正态性检验结果表明,在α = 0.05下,细胞在4种宽度−长度组合的微通道(9−100、12−100、15−100、15−500)内的平均速度不服从正态分布,其余2种组合(9−500、12−500)服从正态分布,因此采用非参数的Kruskal−Wallis检验进行组间比较。检验结果表明,细胞通过6种不同组合微通道的平均速度中位数之间无显著性差异(p = 0.65885),说明微通道宽度与长度的变化未对细胞平均速度中位数产生显著影响,验证了通过改变微通道长度以控制细胞受挤压应力作用时间的可行性与有效性。实验中,由于部分通道存在堵塞情况,细胞通过微通道的平均速度中位数高于设定的液体流速,约为33 mm/s,对应通过100和500 μm微通道的时间分别约为3和15 ms。
图5显示,对于相同长度、不同宽度的微通道,随着通道宽度减小,细胞通过微通道的平均速度数据的离散程度增大。原因在于:直径小于通道宽度的小细胞通常不发生明显变形即可快速通过,能以较大速度完成穿行;而直径大于微通道宽度的大细胞需要发生形变后才能通过,其通过时间受细胞自身力学特性(直径、杨氏模量等)的影响更为显著,从而导致运动速度降低,增大了整体数据的变异性。
细胞在受挤压应力刺激后接种到96孔板,分别于接种后24和48 h检测细胞数量,结果如图6和7所示。统计结果显示,仅有通过9−500微通道的细胞在接种后24和48 h的数量与空白对照组(不施加任何力学刺激)相比呈现统计学显著性差异。在整体趋势方面,细胞在接种后24和48 h的数量变化趋势保持一致,表明实验具有较好的时间稳定性。剪切应力刺激下的细胞增殖能力低于未受应力刺激的空白对照组,但未达到统计学显著性水平。对于100 μm通道的短时间力学刺激,细胞增殖能力随挤压应力的增大(通道宽度减小)而减小;对于500 μm通道的较长时间力学刺激,细胞增殖能力则随挤压应力的增大(通道宽度减小)呈先增大后减小的趋势。这一结果表明,挤压应力刺激的时间效应会对细胞增殖能力产生差异性影响,长时间的力学刺激可能激活了更为复杂的细胞信号转导通路。
细胞接种6 h后的铺展面积统计如图8所示,统计学显著性分析结果如表1所示。实验结果显示,与未受应力刺激的空白对照组相比,经挤压应力与剪切应力刺激的细胞铺展面积均出现了一定程度的下降。其中,在500 μm长通道中受挤压应力刺激的细胞铺展面积下降最为显著(9−500组:p < 0.0001;12−500组:p < 0.0001)。这一结果说明,挤压应力刺激能够显著抑制细胞的黏附能力,从而减小细胞在6 h时的铺展面积。从时间效应上看,长时间的挤压应力刺激能够更显著地降低细胞黏附能力。
针对上述挤压应力显著抑制细胞铺展能力的现象,本文认为其核心机制与挤压诱导的细胞骨架重组密切相关。根据Huang等[15]的研究,循环肿瘤细胞在毛细血管内受到的机械挤压会激活细胞内RhoA信号通路,进而触发F−肌动蛋白的密集组装,产生强烈的向心收缩力,促使细胞由挤压状态下的拉伸形态回缩为高皮层张力的球形状态。挤压应力(尤其是500 μm长通道所对应的较长刺激时间)使细胞维持在一种高张力状态的“收缩记忆”中,从而限制了其在重新接种后与基质形成有效接触面积的能力,在宏观上表现为细胞黏附能力的暂时性下降。
本文通过设计不同宽度与长度组合的微通道,搭建了体外微流控挤压应力加载平台,实现了对挤压应力强度与时间效应的解耦研究。实验结果表明:
1)随着应力强度增大,短时间(约3 ms)挤压刺激使细胞增殖能力减弱,而长时间(约15 ms)挤压刺激则使增殖能力呈现先增强后减弱的趋势,说明长时间的挤压可能激活了更为复杂的细胞信号转导通路。
2)挤压应力显著降低细胞黏附能力,且长时间挤压的抑制效果更为显著,本文认为这与细胞受挤压刺激后的形态转变有关。
以上结果表明,应力刺激的强度和时间对细胞增殖和黏附具有不同的调控趋势,为深入理解二者对细胞命运的联合调控提供了支撑。
本文的研究仍存在一定局限性,未来工作将在以下方面深化:
1)实验条件的拓展:为兼顾实验通量与细胞活性,当前实验的微通道宽度设定为9~15 μm,对应中等程度的挤压应力刺激。若通道宽度进一步缩窄以模拟更极端的毛细血管挤压,细胞活力将发生进一步下降[22],并可能加剧细胞DNA损伤[19],进而影响上皮−间充质转化(EMT)相关基因的表达。后续将对其中的复杂机制进行深入研究。
2)研究对象的拓展:本文以MCF−7细胞系作为实验对象,未来可以探索不同类型肿瘤细胞或患者血液中分离出的CTCs在挤压应力下的响应差异,深入挖掘个体化差异背后的力学机制,为精准医疗提供更具针对性的理论依据。
3)长期力学刺激的考察:当前研究聚焦于瞬时挤压应力刺激的影响,未来可开展周期性、长时间挤压应力刺激实验,以更好地模拟体内微循环中持续存在的力学作用。

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2026年第40卷第4期
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doi: 10.11729/syltlx20250121
  • 接收时间:2025-12-23
  • 首发时间:2026-10-22
  • 出版时间:2026-08-25
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  • 收稿日期:2025-12-23
  • 修回日期:2026-01-27
  • 录用日期:2026-02-09
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    1上海交通大学 船舶海洋与建筑工程学院,上海 200240
    2南方科技大学 海洋高等研究院,深圳 518055

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2种不同金属材料的力学参数
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属数
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genus
种数
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species
占总种数比例
Percentage of
total species (%)
属
Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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