Abstract：
Three-dimensional (3D) cell culture systems perform better in resembling tissue or organism structures compared with traditional 2D models. Organs-on-chips (OoCs) are becoming more efficient 3D models. This study aimed to create a novel simplified dentin-on-a-chip using microfluidic chip technology and tissue engineering for screening dental materials.
A microfluidic device with three channels was designed for creating 3D dental tissue constructs using stem cells from the apical papilla (SCAP) and gelatin methacrylate (GelMA). The study investigated the effect of varying cell densities and GelMA concentrations on the layer features formed within the microfluidic chip. Cell viability and distribution were evaluated through live/dead staining and nuclei/F-actin staining. The osteo/odontogenic potential was assessed through ALP staining and Alizarin red staining. The impact of GelMA concentrations (5 %, 10 %) on the osteo/odontogenic differentiation trajectory of SCAP was also studied.
The 3D tissue constructs maintained high viability and favorable spreading within the microfluidic chip for 3-7 days. A cell seeding density of 2 × 104 cells/μL was found to be the most optimal choice, ensuring favorable cell proliferation and even distribution. GelMA concentrations of 5 % and 10 % proved to be most effective for promoting cell growth and uniform distribution. Within the 5 % GelMA group, SCAP demonstrated higher osteo/odontogenic differentiation than that in the 10 % GelMA group.
In 3D culture, GelMA concentration was found to regulate the osteo/odontogenic differentiation of SCAP. The study recommends a seeding density of 2 × 104 cells/μL of SCAP within 5 % GelMA for constructing simplified dentin-on-a-chip.
This study built up the 3D culture protocol, and induced odontogenic differentiation of SCAP, thus forming the simplified dentin-on-a-chip and paving the way to be used as a well-defined biological model for regenerative endodontics. It may serve as a potential testing platform for cell differentiation.
A microfluidic device with three channels was designed for creating 3D dental tissue constructs using stem cells from the apical papilla (SCAP) and gelatin methacrylate (GelMA). The study investigated the effect of varying cell densities and GelMA concentrations on the layer features formed within the microfluidic chip. Cell viability and distribution were evaluated through live/dead staining and nuclei/F-actin staining. The osteo/odontogenic potential was assessed through ALP staining and Alizarin red staining. The impact of GelMA concentrations (5 %, 10 %) on the osteo/odontogenic differentiation trajectory of SCAP was also studied.
The 3D tissue constructs maintained high viability and favorable spreading within the microfluidic chip for 3-7 days. A cell seeding density of 2 × 104 cells/μL was found to be the most optimal choice, ensuring favorable cell proliferation and even distribution. GelMA concentrations of 5 % and 10 % proved to be most effective for promoting cell growth and uniform distribution. Within the 5 % GelMA group, SCAP demonstrated higher osteo/odontogenic differentiation than that in the 10 % GelMA group.
In 3D culture, GelMA concentration was found to regulate the osteo/odontogenic differentiation of SCAP. The study recommends a seeding density of 2 × 104 cells/μL of SCAP within 5 % GelMA for constructing simplified dentin-on-a-chip.
This study built up the 3D culture protocol, and induced odontogenic differentiation of SCAP, thus forming the simplified dentin-on-a-chip and paving the way to be used as a well-defined biological model for regenerative endodontics. It may serve as a potential testing platform for cell differentiation.
摘要：
目的：与传统的2D模型相比，三维（3D）细胞培养系统在类似组织或生物体结构方面表现更好。芯片上器官（OoC）正在成为更高效的3D模型。本研究旨在使用微流控芯片技术和组织工程来筛选牙科材料，创建一种新颖的简化的牙本质芯片。
方法：设计了具有三个通道的微流体装置，用于使用来自根尖乳头（SCAP）和明胶甲基丙烯酸酯（GelMA）的干细胞创建3D牙齿组织构建体。该研究调查了不同细胞密度和GelMA浓度对微流控芯片内形成的层特征的影响。通过活/死染色和核/F-肌动蛋白染色评估细胞活力和分布。通过ALP染色和茜素红染色评估骨/牙源性潜能。GelMA浓度的影响(5%，10%)对SCAP的骨/牙源性分化轨迹也停止了研讨。
结果：3D组织构建体在微流控芯片内保持了3-7天的高活力和良好的扩散。发现2×104个细胞/μL的细胞接种密度是最佳选择，确保有利的细胞增殖和均匀分布。5％和10％的GelMA浓度被证明对促进细胞生长和均匀分布最有效。在5%GelMA组中，SCAP显示出比10％GelMA组更高的骨/牙源性分化。
结论：在3D培养中，发现GelMA浓度调节SCAP的骨/牙源性分化。该研究建议在5％GelMA内将2×104个细胞/μLSCAP的接种密度用于构建简化的芯片上牙本质。
结论：这项研究建立了3D培养方案，和诱导SCAP的牙源性分化，从而形成了简化的牙本质芯片，并为用作再生牙髓的定义明确的生物学模型铺平了道路。它可以作为细胞分化的潜在测试平台。
方法：设计了具有三个通道的微流体装置，用于使用来自根尖乳头（SCAP）和明胶甲基丙烯酸酯（GelMA）的干细胞创建3D牙齿组织构建体。该研究调查了不同细胞密度和GelMA浓度对微流控芯片内形成的层特征的影响。通过活/死染色和核/F-肌动蛋白染色评估细胞活力和分布。通过ALP染色和茜素红染色评估骨/牙源性潜能。GelMA浓度的影响(5%，10%)对SCAP的骨/牙源性分化轨迹也停止了研讨。
结果：3D组织构建体在微流控芯片内保持了3-7天的高活力和良好的扩散。发现2×104个细胞/μL的细胞接种密度是最佳选择，确保有利的细胞增殖和均匀分布。5％和10％的GelMA浓度被证明对促进细胞生长和均匀分布最有效。在5%GelMA组中，SCAP显示出比10％GelMA组更高的骨/牙源性分化。
结论：在3D培养中，发现GelMA浓度调节SCAP的骨/牙源性分化。该研究建议在5％GelMA内将2×104个细胞/μLSCAP的接种密度用于构建简化的芯片上牙本质。
结论：这项研究建立了3D培养方案，和诱导SCAP的牙源性分化，从而形成了简化的牙本质芯片，并为用作再生牙髓的定义明确的生物学模型铺平了道路。它可以作为细胞分化的潜在测试平台。
