新型体外延迟划痕闭合试验的开发和功能测试。

IF 2.1 4区 生物学 Q4 CELL BIOLOGY Histochemistry and Cell Biology Pub Date : 2024-09-01 Epub Date: 2024-05-07 DOI:10.1007/s00418-024-02292-y
Yi Bing Aw, Sixun Chen, Aimin Yeo, John A Dangerfield, Pamela Mok
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引用次数: 0

摘要

随着慢性伤口疗法的不断发展,对模拟体内炎症伤口微环境的先进检测系统的需求也在增加。目前,这种实验是在动物模型或体外细胞模型中进行的,如更接近急性伤口的细胞培养划痕实验。在这里,我们首次描述了一种延迟划痕闭合模型,它模拟了体外慢性伤口的某些特征。慢性伤口(如晚期糖尿病患者的伤口)的特点是愈合缓慢,这是由伤口局部持续的物理创伤和促炎信号共同造成的。为了在基于细胞的试验中重现这种情况,我们制造了一个确定的物理划痕,并通过干扰素、光稳定剂 PMA 和脂多糖等促炎因子的组合刺激来延迟划痕的闭合。这些因子的浓度是根据常用的人类角质细胞(HaCaT)和真皮成纤维细胞(HDF)细胞系来确定的。然后,对这些模型进行了划痕闭合反应性测试,测试对象是一种专有的愈合分泌物,这种分泌物来自人的沃顿果冻间充质干细胞(MSCs),此前已通过验证,并证明在体外和体内对急性伤口模型的闭合非常有效。来自 HaCaT 细胞的慢性开放性划痕在接触间充质干细胞分泌物后出现闭合。我们建议学术界和工业界的研究人员在研究慢性伤口时采用这种延迟划痕闭合模型,以便在对病人的外敷材料或体内进行测试之前,寻找对药物或生物疗法的反应性。
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Development and functional testing of a novel in vitro delayed scratch closure assay.

As the development of chronic wound therapeutics continues to expand, the demand for advanced assay systems mimicking the inflammatory wound microenvironment in vivo increases. Currently, this is performed in animal models or in in vitro cell-based models such as cell culture scratch assays that more closely resemble acute wounds. Here, we describe for the first time a delayed scratch closure model that mimics some features of a chronic wound in vitro. Chronic wounds such as those suffered by later stage diabetic patients are characterised by degrees of slowness to heal caused by a combination of continued localised physical trauma and pro-inflammatory signalling at the wound. To recreate this in a cell-based assay, a defined physical scratch was created and stimulated by combinations of pro-inflammatory factors, namely interferon, the phorbol ester PMA, and lipopolysaccharide, to delay scratch closure. The concentrations of these factors were characterised for commonly used human keratinocyte (HaCaT) and dermal fibroblast (HDF) cell lines. These models were then tested for scratch closure responsiveness to a proprietary healing secretome derived from human Wharton's jelly mesenchymal stem cells (MSCs) previously validated and shown to be highly effective on closure of acute wound models both in vitro and in vivo. The chronically open scratches from HaCaT cells showed closure after exposure to the MSC secretome product. We propose this delayed scratch closure model for academic and industrial researchers studying chronic wounds looking for responsiveness to drugs or biological treatments prior to testing on explanted patient material or in vivo.

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来源期刊
Histochemistry and Cell Biology
Histochemistry and Cell Biology 生物-细胞生物学
CiteScore
4.90
自引率
8.70%
发文量
112
审稿时长
1 months
期刊介绍: Histochemistry and Cell Biology is devoted to the field of molecular histology and cell biology, publishing original articles dealing with the localization and identification of molecular components, metabolic activities and cell biological aspects of cells and tissues. Coverage extends to the development, application, and/or evaluation of methods and probes that can be used in the entire area of histochemistry and cell biology.
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