Gupta Tamaghna, Sahu Rakesh P, Dabaghi Mohammadhossein, Zhong Lily Shengjia, Shargall Yaron, Hirota Jeremy A, Richards Carl D, Puri Ishwar K
School of Biomedical Engineering, McMaster University, Hamilton, Ontario L8S 4L8, Canada.
Department of Mechanical Engineering, McMaster University, Hamilton, Ontario L8S 4L8, Canada.
ACS Omega. 2023 May 25;8(22):19976-19986. doi: 10.1021/acsomega.3c02052. eCollection 2023 Jun 6.
Soluble signaling molecules and extracellular matrix (ECM) regulate cell dynamics in various biological processes. Wound healing assays are widely used to study cell dynamics in response to physiological stimuli. However, traditional scratch-based assays can damage the underlying ECM-coated substrates. Here, we use a rapid, non-destructive, label-free magnetic exclusion technique to form annular aggregates of bronchial epithelial cells on tissue-culture treated (TCT) and ECM-coated surfaces within 3 h. The cell-free areas enclosed by the annular aggregates are measured at different times to assess cell dynamics. The effects of various signaling molecules, including epidermal growth factor (EGF), oncostatin M, and interleukin 6, on cell-free area closures are investigated for each surface condition. Surface characterization techniques are used to measure the topography and wettability of the surfaces. Further, we demonstrate the formation of annular aggregates on human lung fibroblast-laden collagen hydrogel surfaces, which mimic the native tissue architecture. The cell-free area closures on hydrogels indicate that the substrate properties modulate EGF-mediated cell dynamics. The magnetic exclusion-based assay is a rapid and versatile alternative to traditional wound healing assays.
可溶性信号分子和细胞外基质(ECM)在各种生物学过程中调节细胞动态。伤口愈合分析被广泛用于研究细胞对生理刺激的动态反应。然而,传统的划痕分析可能会损坏底层的ECM包被底物。在这里,我们使用一种快速、无损、无标记的磁排斥技术,在3小时内在组织培养处理(TCT)和ECM包被的表面上形成支气管上皮细胞的环形聚集体。在不同时间测量环形聚集体包围的无细胞区域,以评估细胞动态。针对每种表面条件,研究了包括表皮生长因子(EGF)、制瘤素M和白细胞介素6在内的各种信号分子对无细胞区域封闭的影响。使用表面表征技术测量表面的形貌和润湿性。此外,我们证明了在负载人肺成纤维细胞的胶原水凝胶表面上形成环形聚集体,该表面模拟了天然组织结构。水凝胶上的无细胞区域封闭表明底物特性调节EGF介导的细胞动态。基于磁排斥的分析是传统伤口愈合分析的一种快速且通用的替代方法。