Knowledge of human gingival cell responses to dental monomers is critical for the development of new dental materials. Testing standards have been developed to provide guidelines to evaluate biological functionality of dental materials and devices. However, one shortcoming of the traditional testing platforms is that they do not recapitulate the multi-layered configuration of gingiva, and thus cannot evaluate the layer-specific cellular responses. An oral mucosa-chip with two cell layers was previously developed as an alternative platform to assess the oral mucosa responses to dental biomaterials. The mucosa-chip consists of an apical keratinocyte layer attached to a fibroblast-embedded collagen hydrogel through interconnecting pores in a three-microchannel network. Here, cell responses in the mucosa-chip were evaluated against 2-hydroxyethyl methacrylate (HEMA), a common monomer used in restorative and aesthetic dentistry. The response of mucosal cell viability was evaluated by exposing the chip to HEMA of concentrations ranging from 1.56 to 25 mM and compared to cells in conventional well-plate monoculture. The co-cultured cells were then stained and imaged with epifluorescence and confocal microscopy to determine the layer-specific responses to the treatment. Mucosa-chips were demonstrated to be more sensitive to assess HEMA-altered cell viability than well-plate cultures, especially at lower doses (1.56 and 6.25 mM). The findings suggest that the mucosa-chip is a promising alternative to traditional platforms or assays to test a variety of biomaterials by offering a multi-layered tissue geometry, accessible layer-specific information, and higher sensitivity in detecting cellular responses.

了解人类牙龈细胞对牙科单体的反应对于开发新的牙科材料至关重要。已经制定了测试标准以提供评估牙科材料和设备的生物功能的指南。然而，传统测试平台的一个缺点是它们没有概括牙龈的多层配置，因此无法评估特定层的细胞反应。先前开发了具有两个细胞层的口腔粘膜芯片作为评估口腔粘膜对牙科生物材料的反应的替代平台。粘膜芯片由通过三微通道网络中的互连孔连接到成纤维细胞嵌入的胶原水凝胶的顶端角质形成细胞层组成。这里，针对 2-羟乙基甲基丙烯酸酯 (HEMA) 评估了粘膜芯片中的细胞反应，HEMA 是一种用于修复和美容牙科的常见单体。通过将芯片暴露于浓度范围为 1.56 至 25 mM 的 HEMA 并与常规孔板单培养中的细胞进行比较，评估粘膜细胞活力的反应。然后用落射荧光和共聚焦显微镜对共培养的细胞进行染色和成像，以确定对治疗的层特异性反应。与孔板培养物相比，粘膜芯片被证明对评估 HEMA 改变的细胞活力更敏感，尤其是在较低剂量（1.56 和 6.25 mM）下。研究结果表明，通过提供多层组织几何形状，粘膜芯片是传统平台或分析的有前途的替代品，可以测试各种生物材料，