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The tooth on-a-chip: a microphysiologic model system mimicking the biologic interface of the tooth with biomaterials.
Lab on a Chip ( IF 6.1 ) Pub Date : 2019-12-19 , DOI: 10.1039/c9lc00915a Cristiane Miranda França 1 , Anthony Tahayeri 1 , Nara Sousa Rodrigues 2 , Shirin Ferdosian 1 , Regina Maria Puppin Rontani 3 , Grigoriy Sereda 4 , Jack L Ferracane 1 , Luiz E Bertassoni 5
Lab on a Chip ( IF 6.1 ) Pub Date : 2019-12-19 , DOI: 10.1039/c9lc00915a Cristiane Miranda França 1 , Anthony Tahayeri 1 , Nara Sousa Rodrigues 2 , Shirin Ferdosian 1 , Regina Maria Puppin Rontani 3 , Grigoriy Sereda 4 , Jack L Ferracane 1 , Luiz E Bertassoni 5
Affiliation
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The tooth has a unique configuration with respect to biomaterials that are used for its treatment. Cells inside of the dental pulp interface indirectly with biomaterials via a calcified permeable membrane, formed by the dentin matrix and several thousands of dentinal tubules (∼2 μm in diameter). Although the cytotoxic response of the dental pulp to biomaterials has been extensively studied, there is a shortage of in vitro model systems that mimic the dentin-pulp interface and enable an improved understanding of the morphologic, metabolic and functional influence of biomaterials on live dental pulp cells. To address this shortage, here we developed an organ-on-a-chip model system which integrates cells cultured directly on a dentin wall within a microfluidic device that replicates some of the architecture and dynamics of the dentin-pulp interface. The tooth-on-a-chip is made out of molded polydimethylsiloxane (PDMS) with a design consisting of two chambers separated by a dentin fragment. To characterize pulp cell responses to dental materials on-chip, stem cells from the apical papilla (SCAPs) were cultured in odontogenic medium and seeded onto the dentin surface, and observed using live-cell microscopy. Next, to evaluate the tooth-on-a-chip as a platform for materials testing, standard dental materials used clinically (2-hydroxyethylmethacrylate - HEMA, phosphoric acid - PA, and Adper-Scotchbond - SB) were tested for cytotoxicity, cell morphology, and metabolic activity on-chip, and compared against standardized off-chip controls. All dental materials had cytotoxic effects in both on-chip and off-chip systems in the following order: HEMA > SB > PA (p < 0.05), and cells presented consistently higher metabolic activity on-chip than off-chip (p < 0.05). Furthermore, the tooth-on-a-chip enabled real-time tracking of gelatinolytic activity in a model hybrid layer (HL) formed in the microdevice, which suggests that dental pulp cells may contribute to the proteolytic activity in the HL more than endogenous proteases. In conclusion, the tooth-on-a-chip is a novel platform that replicates near-physiologic conditions of the pulp-dentin interface and enables live-cell imaging to study dental pulp cell response to biomaterials.
中文翻译：
牙齿芯片：模拟牙齿与生物材料的生物界面的微生理模型系统。
就用于治疗的生物材料而言，牙齿具有独特的构造。牙髓内部的细胞通过钙化渗透膜与生物材料间接接触，该渗透膜由牙本质基质和数千个牙本质小管（直径约 2 μm）形成。尽管牙髓对生物材料的细胞毒性反应已被广泛研究，但仍缺乏模拟牙本质-牙髓界面并能够更好地了解生物材料对活牙髓的形态、代谢和功能影响的体外模型系统细胞。为了解决这一短缺问题，我们开发了一种芯片上器官模型系统，该系统将直接在牙本质壁上培养的细胞集成到微流体装置中，该装置复制了牙本质-牙髓界面的一些结构和动力学。片上牙齿由模制聚二甲基硅氧烷（PDMS）制成，其设计由两个由牙本质碎片隔开的腔室组成。为了表征牙髓细胞对芯片上牙科材料的反应，在成牙培养基中培养来自根尖乳头 (SCAP) 的干细胞，并将其接种到牙本质表面，并使用活细胞显微镜进行观察。接下来，为了评估片上牙齿作为材料测试的平台，对临床使用的标准牙科材料（甲基丙烯酸 2-羟乙酯 - HEMA、磷酸 - PA 和 Adper-Scotchbond - SB）进行了细胞毒性、细胞形态学测试和片内代谢活动，并与标准化片外对照进行比较。所有牙科材料在片上和片外系统中均具有细胞毒性作用，顺序如下：HEMA > SB > PA (p < 0.05)，并且细胞在片上表现出始终高于片外的代谢活性（ p< 0.05）。 此外，片上牙齿能够实时跟踪微型设备中形成的模型混合层（HL）中的明胶分解活性，这表明牙髓细胞对 HL 中蛋白水解活性的贡献可能大于内源性蛋白酶。总之，牙齿芯片是一个新颖的平台，可以复制牙髓-牙本质界面的近生理条件，并能够通过活细胞成像来研究牙髓细胞对生物材料的反应。
更新日期：2020-02-13
中文翻译：
牙齿芯片：模拟牙齿与生物材料的生物界面的微生理模型系统。
就用于治疗的生物材料而言，牙齿具有独特的构造。牙髓内部的细胞通过钙化渗透膜与生物材料间接接触，该渗透膜由牙本质基质和数千个牙本质小管（直径约 2 μm）形成。尽管牙髓对生物材料的细胞毒性反应已被广泛研究，但仍缺乏模拟牙本质-牙髓界面并能够更好地了解生物材料对活牙髓的形态、代谢和功能影响的体外模型系统细胞。为了解决这一短缺问题，我们开发了一种芯片上器官模型系统，该系统将直接在牙本质壁上培养的细胞集成到微流体装置中，该装置复制了牙本质-牙髓界面的一些结构和动力学。片上牙齿由模制聚二甲基硅氧烷（PDMS）制成，其设计由两个由牙本质碎片隔开的腔室组成。为了表征牙髓细胞对芯片上牙科材料的反应，在成牙培养基中培养来自根尖乳头 (SCAP) 的干细胞，并将其接种到牙本质表面，并使用活细胞显微镜进行观察。接下来，为了评估片上牙齿作为材料测试的平台，对临床使用的标准牙科材料（甲基丙烯酸 2-羟乙酯 - HEMA、磷酸 - PA 和 Adper-Scotchbond - SB）进行了细胞毒性、细胞形态学测试和片内代谢活动，并与标准化片外对照进行比较。所有牙科材料在片上和片外系统中均具有细胞毒性作用，顺序如下：HEMA > SB > PA (p < 0.05)，并且细胞在片上表现出始终高于片外的代谢活性（ p< 0.05）。 此外，片上牙齿能够实时跟踪微型设备中形成的模型混合层（HL）中的明胶分解活性，这表明牙髓细胞对 HL 中蛋白水解活性的贡献可能大于内源性蛋白酶。总之，牙齿芯片是一个新颖的平台，可以复制牙髓-牙本质界面的近生理条件，并能够通过活细胞成像来研究牙髓细胞对生物材料的反应。
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