A two-way membrane-integrated microfluidic device for evaluation of anti-inflammatory drug through immunohistochemical analysis and permeability assay of keratinocyte cell layers

IF 4.1 Q1 CHEMISTRY, ANALYTICAL Talanta Open Pub Date : 2024-05-16 DOI:10.1016/j.talo.2024.100330

Marika Sugimoto , Naoko Takahashi-Ando , Naoki Sasaki

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Abstract

Evaluation of anti-inflammatory drug on a two-way membrane-integrated microfluidic device (TMMD) is presented. Insertion of a porous membrane into a microfluidic device in a vertical direction and attachment of a cover glass to the lateral side of the microfluidic device enabled us to observe the device from two orthogonal directions. HaCaT, a human epidermal keratinocyte, was cultured in the TMMD. The localization of ZO-1, a tight junction protein, between the HaCaT cells was confirmed by immunohistochemical analysis. Permeability of the HaCaT cell layer increased after stimulation by potassium dichromate, whereas the pretreatment of the HaCaT by dexamethasone prior to the stimulation kept the permeability unchanged. Deoxynivalenol, an anti-inflammatory drug candidate, kept the permeability unchanged with lower concentrations compared to dexamethasone. We expect that the present TMMD is applicable to various anti-inflammatory drug candidates to evaluate their efficacy.

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通过免疫组化分析和角质细胞层渗透性测定评估抗炎药物的双向膜集成微流控装置

本文介绍了在双向膜集成微流控装置（TMMD）上对抗炎药物的评估。将多孔膜沿垂直方向插入微流控装置，并在微流控装置的侧面贴上盖玻片，这样我们就能从两个正交方向观察该装置。我们在 TMMD 中培养了人表皮角质细胞 HaCaT。免疫组化分析证实了紧连结蛋白 ZO-1 在 HaCaT 细胞之间的定位。重铬酸钾刺激后，HaCaT 细胞层的通透性增加，而在刺激前用地塞米松对 HaCaT 进行预处理可保持通透性不变。与地塞米松相比，抗炎候选药物脱氧雪腐镰刀菌醇能以较低的浓度保持渗透性不变。我们希望本 TMMD 适用于各种抗炎候选药物，以评估其疗效。

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